Posted on 8 April 20268 April 2026 by Joe

What Fibromyalgia Reveals About the Endocannabinoid System and Why It Matters

When the System Breaks — What Fibromyalgia Reveals About the Endocannabinoid System | The Certified

The Endocannabinoid System · Part 3 · Clinical Deficiency

When the System Breaks — What Fibromyalgia Reveals About the Endocannabinoid System and Why It Matters

A 2025 peer-reviewed review has mapped the relationship between fibromyalgia and the endocannabinoid system in detail. The findings suggest that what millions experience as widespread chronic pain may be, at least in part, a disease of endocannabinoid deficiency.

The Grower's Connect · 2025 · 11 min read

6.4% of US adults affected by fibromyalgia

94% of patients reported pain relief with cannabis

35% Reduction in opioid use when combined with cannabis

Listen to this article When the System Breaks — What Fibromyalgia Reveals About the Endocannabinoid System

Over the past two weeks we have been building a picture of the endocannabinoid system from the inside out. We looked at anandamide — the bliss molecule — what it is, where it comes from, and what it does in the brain and body. Then we looked at what happens when you run at the right intensity, and how moderate exercise triggers your body's own endocannabinoid release — reducing anxiety, elevating mood, and producing effects that closely mirror what cannabis achieves pharmacologically.

This week we arrive at the darker side of the same story. What happens when the endocannabinoid system doesn't work properly? What does a chronically dysregulated endocannabinoid system look like from the outside — as experienced by a real person, in a real body, every day?

A 2025 review published in Current Issues in Molecular Biology by Mario García-Domínguez at the Universidad de Navarra provides one of the most comprehensive analyses to date of the endocannabinoid system's role in fibromyalgia. It connects everything we have covered in the last two weeks — the receptors, the molecules, the signalling cascades — to a clinical condition affecting hundreds of millions of people worldwide. Understanding this connection matters for anyone trying to understand what cannabis is actually doing in the human body, and why.

What Fibromyalgia Is — And Why It Has Been So Hard to Explain

Fibromyalgia is a chronic condition characterised by widespread musculoskeletal pain, persistent fatigue, sleep disturbances, and cognitive impairments — a cluster that includes difficulty with memory and concentration often called fibrofog. The pain varies in intensity and location and is linked to sensitivity at specific areas known as tender points.

Widespread Pain

Musculoskeletal pain across multiple body regions, linked to sensitivity at tender points. Varies in intensity and location, often described as burning, aching, or stabbing.

Persistent Fatigue

Chronic exhaustion that is not relieved by rest, often described as profound and disproportionate to any physical activity undertaken.

Sleep Disturbance

Non-restorative sleep, difficulty maintaining sleep, and frequent waking — creating a cycle where poor sleep worsens pain sensitivity and pain disrupts sleep.

Fibrofog

Cognitive impairments including memory loss, difficulty concentrating, and slowed mental processing — often as debilitating as the physical symptoms.

It affects 6.4% of the US population and between 2.4% and 3.3% in Europe and South America — significantly more prevalent in women. It is not rare. It is one of the most common chronic pain syndromes on the planet, affecting hundreds of millions of people globally.

What has made fibromyalgia so difficult to treat, and historically so difficult to take seriously in medical settings, is that its underlying mechanisms have resisted clear explanation. There is no obvious tissue damage visible on scans. There is no single biomarker. For decades, patients were told the pain was psychological. The condition was real and debilitating, but the biology behind it was opaque. What is now emerging from the research is a different picture. The problem may not be in the joints or muscles themselves. The problem may be in the system responsible for regulating how pain signals are processed, amplified, and dampened — and that system is the endocannabinoid system.

The Clinical Endocannabinoid Deficiency Hypothesis

The central theoretical framework the review examines is called Clinical Endocannabinoid Deficiency — CECD. The hypothesis is straightforward: in some individuals, the endocannabinoid system operates chronically below its optimal level. The system that is supposed to modulate pain, regulate sleep, stabilise mood, and dampen inflammation is not producing enough, not signalling effectively, or not maintaining adequate receptor sensitivity. The result is a body that cannot properly regulate its own experience of pain and discomfort.

This hypothesis would explain much of what makes fibromyalgia so puzzling. If the problem is systemic underfunction of the endocannabinoid system — rather than localised tissue damage — then of course there would be no obvious structural abnormality on imaging. The problem would be functional, not structural. The pain would be real, widespread, and variable because the system responsible for dampening and contextualising pain signals across the entire nervous system is impaired.

"If the problem is a systemic underfunction of the endocannabinoid system, then of course there is no structural abnormality on imaging. The problem is functional. The pain is real — the dampening system is what's failing."

The CECD hypothesis also directly connects to the synaptic signalling mechanism we described in the anandamide piece. Endocannabinoids work as retrograde messengers — released by postsynaptic neurons, travelling backwards across the synapse, and binding to presynaptic CB1 receptors to suppress the release of glutamate and other excitatory neurotransmitters. This mechanism is the brain's primary tool for preventing pain signals from being over-amplified. If that tool is impaired, pain signals propagate more freely. The threshold for what feels painful is lowered. Everything hurts more than it should.

What the Endocannabinoid System Is Actually Doing in Pain Regulation

The review provides a detailed account of the endocannabinoid system's role in pain modulation that clarifies precisely why a deficiency in this system would produce the pattern of symptoms seen in fibromyalgia.

ECS Pain Regulation — Key Mechanisms Involved in Fibromyalgia

Spinal Cord CB1 receptors are present in the dorsal horn — the primary relay station for pain signals entering the central nervous system. Endocannabinoid signalling here suppresses pain transmission before it reaches the brain.
Fascial Tissue CB1 and CB2 receptors have been identified in fascial tissue — the connective network covering and connecting muscles throughout the body. A direct mechanism by which ECS deficiency could produce the diffuse musculoskeletal pain of fibromyalgia.
Joint Protection CB1 activation blocks inflammatory degradation of connective tissues. When synovial cells are exposed to the inflammatory cytokine TNF-alpha, they secrete enzymes that degrade cartilage. Anandamide inhibits this process — ECS deficiency removes this protective mechanism.
Retrograde Brake Endocannabinoids released by postsynaptic neurons travel backwards across synapses to suppress further excitatory neurotransmitter release. This is the nervous system's volume control on pain. When this brake is compromised, pain sensitisation is amplified system-wide.

The Paradox in the Blood Data

Here is where the research becomes genuinely counterintuitive — and requires careful interpretation.

Several studies cited in the review have measured circulating endocannabinoid levels in fibromyalgia patients and found them to be elevated, not depleted. Anandamide concentrations were significantly higher in fibromyalgia patients than in healthy controls. Levels of 2-AG, OEA, PEA, and SEA — related endocannabinoid-like molecules — were also increased. At first glance this seems to contradict the deficiency hypothesis. If endocannabinoid levels are higher in fibromyalgia patients, how can the condition be caused by deficiency?

The Compensatory Mechanism

The review interprets the elevated blood levels as a probable compensatory response — the system producing more of these molecules in response to inadequate function at the receptor level. The same phenomenon is well known in other hormonal systems: when receptors become less responsive, the body increases production of the signalling molecule in an attempt to compensate. The elevated circulating levels may reflect not abundance but distress — a body working harder than it should to achieve an effect it is struggling to produce.

This interpretation is also consistent with the exercise research from last week. Long-term regular exercise was associated with decreased baseline endocannabinoid levels — the body adapting by upregulating FAAH, the enzyme that degrades anandamide, in response to repeated elevation. A body chronically producing excess endocannabinoids as a compensatory response may develop a similar pattern of accelerated degradation, creating a cycle that perpetuates the deficiency rather than correcting it.

The Menstrual Cycle Connection

One of the most striking findings in the review involves the relationship between the menstrual cycle and fibromyalgia diagnosis. It illuminates the endocannabinoid system's hormonal sensitivity in ways with real clinical implications.

Anandamide levels fluctuate across the menstrual cycle in healthy women. During the follicular phase — the first half — AEA levels are relatively high. During the luteal phase — the second half — progesterone upregulates FAAH, the anandamide-degrading enzyme, causing AEA levels to fall. A study found that this drop in anandamide during the luteal phase was associated with significantly increased sensitivity to pressure pain. And in a particularly striking finding: some participants met the diagnostic criteria for fibromyalgia during the luteal phase — the low-AEA phase — but not during the follicular phase, when AEA was higher.

This is not a peripheral observation. It suggests that the boundary between fibromyalgia and normal pain sensitivity may, for some individuals, be a matter of endocannabinoid tone — and that tone fluctuates with hormonal cycles. It may help explain the significantly higher prevalence of fibromyalgia in women. It also opens a question about whether hormonal fluctuations more broadly interact with endocannabinoid function in ways that contribute to chronic pain vulnerability across multiple conditions.

The Sleep Dimension

The review addresses the endocannabinoid system's role in sleep regulation — directly relevant to fibromyalgia because sleep disturbance is one of its most disabling features.

The pineal gland produces both melatonin and 2-AG in a circadian rhythm, partially regulated through CB2 receptor activation in the suprachiasmatic nucleus — the brain's master clock. Anandamide has also been shown to play a role in sleep onset. In a person with endocannabinoid deficiency, both the sleep regulatory function and the pain modulatory function would be impaired simultaneously.

The Cycle That Sustains the Condition

The characteristic pattern of fibromyalgia — pain that worsens with poor sleep, and sleep that is disrupted by pain — may not be two separate problems feeding each other. It may be one problem: a dysregulated endocannabinoid system failing at both pain regulation and sleep regulation at the same time, from the same underlying deficiency.

What Cannabis-Based Therapies Have Shown

The review surveys the clinical evidence for cannabis-based treatments in fibromyalgia, covering studies from 2011 to 2024. The picture is promising but not yet definitive.

Clinical Evidence — Key Study Findings

2019 study: 50% reduction in pain intensity in 81% of fibromyalgia patients after six months of medical cannabis treatment
Israeli survey: 94% reported pain relief, 93% improved sleep, 87% reduced depressive symptoms, 62% reduced anxiety
2024 study: cannabis combined with oxycodone reduced opioid consumption by 35% without affecting cannabis use frequency
2024 low-dose medical cannabis study: substantial reduction in pain intensity and improvements in physical and mental state in the majority of participants
Nabilone (synthetic cannabinoid): significant reductions in pain, anxiety, and overall fibromyalgia impact in randomised controlled trials
Systematic review of 17 studies (2021): cannabis-based medicines may be effective for pain relief and sleep improvement — moderate quality evidence

The anti-inflammatory properties of CBD combined with the analgesic and muscle-relaxant properties of THC appear to produce a synergistic effect across fibromyalgia's multiple symptom domains. This is biologically coherent — the condition involves dysregulation across pain, mood, sleep, and inflammation, and a therapy that modulates the endocannabinoid system broadly would be expected to address multiple symptoms simultaneously rather than one in isolation.

The honest limitation is that most of this evidence comes from observational studies, surveys, and small trials. Randomised controlled trials with large sample sizes and long follow-up periods are largely absent. The evidence is promising and biologically well-motivated, but it is not yet at the standard required to establish definitive clinical guidelines. Patients should consult healthcare professionals before considering cannabis as a treatment, as individual responses can vary significantly.

What This Means for the Cannabis Community

The fibromyalgia research adds a critical dimension to the understanding of cannabis that this series has been building week by week.

We established that the endocannabinoid system is the body's own regulatory network — producing anandamide and 2-AG to manage pain, mood, sleep, anxiety, and inflammation. We established that exercise activates this system. This week's paper adds: when this system chronically underperforms, the result is not just a mildly worse baseline mood. The result can be a debilitating condition — widespread pain, exhaustion, cognitive impairment, and disrupted sleep — affecting millions of people who often spend years being told nothing is physically wrong with them.

Cannabis, in this context, is not a recreational novelty or a pharmaceutical shortcut. It is a plant-derived intervention targeting a specific physiological system that, in a significant proportion of the population, is not functioning adequately. The CB1 and CB2 receptors that THC and CBD interact with are the same receptors that are failing to do their job in fibromyalgia patients. The anandamide that cannabis mimics is the same molecule that fluctuates with the menstrual cycle and drops to a level that temporarily meets the diagnostic threshold for fibromyalgia.

"For many people in genuine physiological distress, cannabis may not be making them high. It may be making them feel normal — because it is restoring a function the body is struggling to maintain on its own."

This is what understanding the endocannabinoid system means in practice. Not just a more sophisticated explanation for why cannabis makes some people feel good. A clearer picture of why, for many people in genuine physiological distress, it may be making them feel normal — because it is restoring a function the body is struggling to maintain on its own.

The Endocannabinoid System Series — The Grower's Connect

Part 01 → Anandamide — Unlocking the Bliss Molecule
Part 02 → Your Body Makes Its Own Cannabis — And Running Is the Key That Unlocks It
Part 03 → When the System Breaks — What Fibromyalgia Reveals About the Endocannabinoid System — You're reading it

Source Study: García-Domínguez M (2025) Role of the Endocannabinoid System in Fibromyalgia. Curr. Issues Mol. Biol. 47, 230. doi:10.3390/cimb47040230 — Program of Immunology and Immunotherapy, CIMA-Universidad de Navarra, Pamplona, Spain. Published March 27, 2025.

Posted on 1 April 20261 April 2026 by Joe

Your Body Makes Its Own Cannabis — And Running Is the Key That Unlocks It

Your Body Makes Its Own Cannabis — And Running Is the Key That Unlocks It | The Certified

Plant Science · The Endocannabinoid System

Your Body Makes Its Own Cannabis — And Running Is the Key That Unlocks It

Scientists have been searching for decades for what causes the runner's high. The endorphin theory turned out to be mostly wrong. What the evidence now points to is something far more interesting — your body producing its own versions of the compounds found in cannabis, triggered by exercise.

The Grower's Connect · 2025 · 11 min read

14/17 Studies confirmed AEA rise after exercise

571 Human participants across 21 trials

70–85% Max heart rate sweet spot for release

Listen to this article Your Body Makes Its Own Cannabis — And Running Is the Key That Unlocks It

Five weeks into this series and we have moved from the plant outward. We examined what sucrose does inside the stem. We looked at how the brain responds to decades of heavy use. We watched scientists coax a naked cell back to life. We mapped the microscopic architecture of the flower itself. This week we turn to something that connects the plant to the person in a way most growers have never considered.

Your body has its own endocannabinoid system. It produces its own cannabinoid-like molecules. And a growing body of peer-reviewed evidence suggests that moderate-intensity endurance exercise — running, specifically — is one of the most reliable ways to trigger their release. This isn't fringe science or wellness marketing. It's a systematic review published in The Neuroscientist, covering 21 human clinical trials and 571 participants.

The implications run in two directions simultaneously. For the cannabis consumer, it tells you something important about what the plant is actually binding to — and why it works the way it does. For the grower, it reframes what you are cultivating. You are not producing a foreign substance that overrides the brain. You are producing plant-based versions of molecules the brain already knows, already produces, and already uses to regulate mood, pain, anxiety, and motivation.

First, the Endorphin Myth

Before we get into what the evidence actually shows, it's worth clearing the ground of what it doesn't show — because most people have been told the wrong story for decades.

The runner's high was first attributed to endorphins in the 1980s. The idea was straightforward: intense exercise releases endorphins, endorphins bind to opioid receptors, opioid receptors produce euphoria. It was widely reported, widely believed, and is still repeated today.

The problem is that endorphins are hydrophilic molecules — water-soluble. The blood-brain barrier is largely impermeable to water-soluble molecules. Peripheral endorphins physically cannot cross into the brain in meaningful quantities. The machinery needed to produce the runner's high is inside the brain. The endorphins produced in the body largely cannot reach it.

The evidence backed this up directly. When researchers blocked opioid receptors entirely using naltrexone — preventing anything from binding to those receptors — the runner's high happened anyway. Euphoria was still present. Anxiety was still reduced. The opioid system wasn't the mechanism. So what was?

"When researchers blocked the opioid system entirely, the runner's high happened anyway. Euphoria was still present. Anxiety was still reduced. The endorphin theory was wrong."

The Endocannabinoid System — Your Body's Built-In Cannabis

In the early 1990s, researchers made two discoveries that changed the picture completely. In 1992, a molecule was identified in the brain that bound to the same receptors as THC. It was named anandamide — from the Sanskrit word for bliss. In 1995, a second molecule was discovered: 2-arachidonoyl glycerol, known as 2-AG.

Both are endocannabinoids — cannabinoid-like molecules produced naturally inside the body. Both are lipophilic, meaning fat-soluble. And critically, fat-soluble molecules can cross the blood-brain barrier with ease. Unlike endorphins, endocannabinoids produced in the body can actually reach the brain.

From the plant THC & CBD

Plant-derived cannabinoids that bind to CB1 and CB2 receptors in the brain. THC produces psychoactive effects via CB1. CBD modulates the system more broadly. Both are fat-soluble and cross the blood-brain barrier.

From your body AEA & 2-AG

Endogenous cannabinoids produced by the brain itself. Anandamide binds primarily to CB1 — the same receptor as THC. 2-AG is a full agonist at both CB1 and CB2. Exercise triggers their release into circulation.

The endocannabinoid system they activate regulates synaptic transmission, mood, reward, anxiety, appetite, memory, neuroprotection, and neuroinflammation. It also plays important roles in neural development. When a cannabis grower talks about what THC does in the brain, they are talking about what the brain's own endocannabinoid system does naturally. THC is a plant-based key to a lock the body built for itself. Anandamide is the body's own key to that same lock.

What the Review Found — 21 Studies, 571 People

The systematic review published in The Neuroscientist screened 278 records and included 21 human clinical trials meeting strict criteria: aerobic exercise, minimum 20 minutes, endocannabinoid blood levels measured before and after, published in peer-reviewed journals. The 571 participants included healthy adults, athletes, people with PTSD, major depression, chronic pain, and substance use disorder.

The Headline Numbers

14 of 17 studies found a significant increase in anandamide after acute exercise — an 82% replication rate
Only about half the studies found an increase in 2-AG, likely due to small sample sizes and greater biological variability
76% of studies found increases in OEA, another endocannabinoid-like molecule, after exercise
All 4 long-term exercise studies found endocannabinoid levels decreased after programs of 12 weeks or more
Opioid blockade with naltrexone did not inhibit endocannabinoid release, euphoria, or anxiety reduction after running

For a field studying a molecule that degrades quickly and is difficult to measure, an 82% replication rate across independent research groups, different participant populations, and different countries is notable.

Intensity Is Everything — The 70 to 85% Window

One of the most practically useful findings in the review is that endocannabinoid release is not simply triggered by moving your body. It is triggered by moving your body at the right intensity.

A pivotal study tested the same participants on four separate days at four different intensities: walking at under 50% of maximum heart rate, and running at roughly 70%, 80%, and 90% of maximum heart rate. The result was clean: only the two middle intensities — approximately 70% and 80% of maximum heart rate — produced a significant increase in anandamide. Walking produced nothing. High-intensity running at 90% produced nothing.

The Sweet Spot

The review's recommendation, drawn from accumulated evidence, is 70% to 85% of age-adjusted maximum heart rate for at least 30 minutes. In practical terms this is a pace where you can speak but feel genuinely challenged — sustainable for 30 to 45 minutes, but not a stroll. Duration should be at least 20 minutes. Peak mood benefits appear around 30 to 35 minutes. Endocannabinoid levels in the blood peak immediately after exercise and can be detected for up to 15 minutes post-exercise.

What the Endocannabinoids Are Actually Doing

The runner's high has four classically described components: euphoria, reduced anxiety, reduced pain sensitivity, and sedation. Here is what the evidence shows for each in relation to endocannabinoids.

Euphoria

Strong evidence

Endocannabinoid levels were roughly twice as high after running as walking. Euphoria tracked the same pattern. Blocking opioid receptors with naltrexone did not reduce either the endocannabinoid release or the euphoria — ruling out endorphins as the mechanism.

Anxiety Reduction

Strong evidence

8 out of 10 studies found reduced anxiety after acute exercise. Higher endocannabinoid increases correlated with greater anxiety reductions. This held even in PTSD, major depression, and substance use disorder populations.

Pain Reduction

Mixed evidence

Results were inconsistent across studies. One study found significant hypoalgesia after 30 minutes of running. Another found no effect in chronic pain patients. Appears to depend heavily on intensity, timing, and participant health status.

Sedation

No evidence yet

None of the 21 studies measured or detected sedation effects after exercise. A mouse study suggests post-exercise sedation may be a non-specific fatigue response that does not require endocannabinoid signalling at all.

The Long-Term Paradox

Here is the finding that surprised the researchers most — and has the most significant implications for anyone thinking about regular exercise and the endocannabinoid system.

After acute exercise, endocannabinoids go up. But after long-term regular exercise programs lasting 12 weeks or more, all four studies that measured endocannabinoid levels found they went down. Not just back to baseline. Measurably below it.

The mechanism proposed is an upregulation of FAAH — fatty acid amide hydrolase — an enzyme that breaks down anandamide. In physically active people, FAAH activity in lymphocytes was found to be higher than in sedentary controls. The body, it appears, compensates for repeated endocannabinoid elevation by becoming more efficient at clearing it. The same homeostatic intelligence that governs tolerance to cannabis in regular users appears to operate in regular exercisers — not through receptor downregulation but through accelerated degradation of the molecule itself.

What this means practically is not yet clear. It may be neutral — the body adapting its baseline without losing the capacity for acute elevation during exercise. Or it may have implications for mood regulation in long-term athletes. The review flags this as a priority for future research.

"The same homeostatic system that governs cannabis tolerance in regular users appears to operate in regular exercisers — not through receptor changes, but through faster degradation of the molecule itself."

The Stress Connection

The review identifies a relationship between the endocannabinoid system and the stress response that goes beyond exercise-induced euphoria.

Cortisol — the primary stress hormone — and anandamide levels were found to correlate positively. When exercise drove cortisol up, anandamide went up with it. In another study, a social stress test also increased anandamide. The researchers frame this through allostasis — the body's system for maintaining stability through change. Exercise is itself a stressor. The endocannabinoid release it triggers may be part of the body's mechanism for modulating how that stress is experienced, buffering the physiological stress signal with a neurobiological one that promotes calm and positive affect.

A study of cosmonauts during spaceflight adds a striking data point. In cosmonauts experiencing low stress, endocannabinoids were elevated. In those experiencing high stress and motion sickness, endocannabinoid elevation was absent and cortisol surged. The endocannabinoid system appears to function, at least in part, as a stress buffer — one that can be depleted by excessive stress rather than activated by it.

This is directly relevant to understanding what cannabis does therapeutically. When patients report using cannabis for anxiety, stress, or mood regulation, they are not introducing a foreign chemical that overrides normal function. They are supplementing a system that exists specifically to perform those regulatory functions — one that can be overwhelmed, depleted, or dysregulated by the conditions of modern life.

What This Means for Growers and Consumers

The relevance of this research to cannabis cultivation runs deeper than it first appears.

For anyone who grows and also uses cannabis, understanding that THC and anandamide bind to the same receptor — that the plant molecule is essentially mimicking a molecule your body already produces — reframes the experience of using cannabis in a meaningful way. It is not an alien substance producing an artificial state. It is a plant-derived key fitting a lock your brain evolved for its own purposes.

For medical growers and producers, the endocannabinoid research strengthens the biological rationale for cannabis as medicine in ways that go beyond anecdote. The anxiety reduction, mood elevation, and pain modulation that cannabis produces are all functions of the endocannabinoid system — documented in drug-free exercise studies, across hundreds of participants, in peer-reviewed journals.

And for the series of conversations we have been having here on The Grower's Connect — about what the plant is at a cellular level, what its flowers are under a microscope, what happens in the brain over a lifetime of use — this piece adds something important. The plant and the person share a receptor. The endocannabinoid system is the bridge between them. And running, it turns out, is one of the oldest ways the body has of activating that bridge on its own.

The Grower's Connect — Plant Science Series

Week 01 → Feeding Your Plant From the Inside Out: Sucrose Stem Infusion and 30%+ Yield Increases
Week 02 → What Heavy Cannabis Use Actually Does to Your Brain — And What the Science Really Says
Week 03 → Scientists Just Grew Cannabis From a Single Cell — And It Changes Everything
Week 04 → What's Actually Inside Your Cannabis Flower — And Why Understanding It Could Change How You Grow
Week 05 → Your Body Makes Its Own Cannabis — And Running Is the Key That Unlocks It — You're reading it

Source Study: Siebers M, Biedermann SV, and Fuss J (2022) Do Endocannabinoids Cause the Runner's High? Evidence and Open Questions. The Neuroscientist. 2023;29(3):352–369. doi:10.1177/10738584211069981 — Institute of Forensic Psychiatry and Sex Research, University of Duisburg-Essen, Germany. Published 2022.

Posted on 18 March 202618 March 2026 by Joe

Scientists Just Grew Cannabis From a Single Cell — And It Changes Everything

Scientists Just Grew Cannabis From a Single Cell — And It Changes Everything About the Future of This Plant | The Certified

Plant Science · Cannabis Biotechnology

Scientists Just Grew Cannabis From a Single Cell — And It Changes Everything About the Future of This Plant

Czech researchers have achieved only the second-ever successful cultivation of cannabis protoplasts — naked plant cells stripped of their walls — and coaxed them into dividing. Here's why that quiet laboratory milestone could reshape how cannabis is bred, cloned, and engineered at a genetic level.

The Grower's Connect · June 2025 · 10 min read

2nd Ever achieved globally

83.2% Peak cell viability

9 Cannabis cultivars tested

Listen to this article Scientists Just Grew Cannabis From a Single Cell

Three weeks on The Grower's Connect, and a clear theme has emerged. We started by looking at how feeding sucrose directly into the stem at precisely the right pressure could push yields up by over 30%. Then we examined what heavy cannabis use does to the brain, and found the science far more nuanced than headlines suggest. This week, we go deeper still, into the cellular machinery of the plant itself. Because a quietly remarkable study just published in Frontiers in Plant Science has brought us one step closer to something that could fundamentally change cannabis cultivation as we know it.

The word "protoplast" won't mean much to most growers. By the end of this piece, it will, and you'll understand why it matters.

What Is a Protoplast, and Why Does It Matter?

A protoplast is simply a plant cell with its cell wall enzymatically removed. What's left is the naked cell, just the plasma membrane and everything inside it. That sounds destructive, but it's actually the opposite. Strip away the cell wall and something remarkable happens: the cell loses its identity. It forgets, in a sense, what it was programmed to be. It becomes capable of becoming anything.

Under the right conditions, a single protoplast can divide, redifferentiate, and regenerate into a complete, genetically identical plant. This process, called somatic embryogenesis or protoplast-to-plant regeneration, is the foundation of some of the most powerful tools in modern plant biotechnology, including genetic transformation and CRISPR-based genome editing.

The reason this matters for cannabis specifically is that the plant has been notoriously difficult to work with at the cellular level. Unlike tobacco, tomato, or Arabidopsis, the laboratory workhorse of plant biology, cannabis has resisted almost every attempt to regenerate a whole plant from isolated cells. That resistance has been a major bottleneck for cannabis research and breeding for decades.

"Strip away the cell wall and the cell loses its identity. It forgets what it was programmed to be — and becomes capable of becoming anything. That is the entire point."

What the Study Actually Did

Researchers at Palacký University Olomouc in the Czech Republic, led by Daniel Král, set out to crack that bottleneck. Published in June 2025 in Frontiers in Plant Science, the study reports only the second successful establishment and partial regeneration of cannabis protoplast cultures ever documented in scientific literature.

The team worked with nine cannabis cultivars, all industrial hemp varieties with varying CBD content, including USO 31, Finola, Fédora 17, Futura 75, and others. The work involved two distinct phases: first, optimising the conditions for protoplast isolation, and second, attempting to get those isolated cells to survive, divide, and form early-stage cell clusters called microcalli.

Getting the donor material right was everything. The researchers tested protoplast isolation from plants at multiple developmental stages, from one-week-old seedlings all the way through to six-month-old in vitro cultures and greenhouse-grown plants. The results were unambiguous: only leaves from one to two-week-old seedlings grown in sterile in vitro conditions produced protoplasts at a useful yield and viability. Older material consistently failed. Greenhouse-grown plants failed entirely.

Study Design at a Glance

9 industrial hemp cultivars tested, including both high and low CBD strains
Donor material tested from 1-week-old seedlings through to 6-month-old in vitro cultures and greenhouse plants
5 enzyme formulations evaluated for cell wall digestion
Gene expression tracked at 0, 24, 48 and 72 hours post-isolation across 6 target genes
Extended 14-day cultivation experiment confirmed microcallus formation and cell viability
Palacký University Olomouc, Czech Republic — published June 2025, Frontiers in Plant Science

The best results came from the USO 31 cultivar, producing yields of up to 9.9 million cells per gram of leaf tissue, with a peak viability of 83.2%, meaning more than four in five isolated cells were alive and functional immediately after extraction.

Key Finding

Just as we saw with sucrose stem infusion — where the difference between 0.5 bar and 2 bar of pressure determined whether yields increased by 34% or fell below the control — the protoplast work shows that precise conditions at the starting point determine everything that follows. You cannot compensate downstream for poor decisions upstream.

The Isolation Process — How You Strip a Cell Wall

The enzymatic solution used to digest away the cell wall is one of the most critical variables in the entire process. The team tested five different enzyme formulations, ultimately finding that a solution containing cellulase and macerozyme in a mannitol buffer — originally developed by Matchett-Oates et al. in 2021 — performed best.

The digestion ran for 16 hours in complete darkness at 25°C without shaking. Shorter digestion periods consistently failed. Adding a washing solution both before and after filtration was critical; skipping this step caused the protoplasts to clump and aggregate, making separation impossible. Centrifugation at 1000 rpm for 10 minutes, using a sucrose density gradient, provided the cleanest separation of viable cells from cellular debris.

One important finding: adding pectolyase, an enzyme used successfully in other species and suggested by several previous cannabis studies, actually made things worse at higher concentrations, likely by causing excessive enzymatic activity that damaged the cells before they could be collected.

Getting Them to Divide — The Hard Part

Isolating protoplasts is one thing. Getting them to survive in a culture and divide is another challenge entirely. The vast majority of cannabis protoplast research has stopped at isolation. Getting cells to actually re-enter the cell cycle, to start dividing, is where the field has repeatedly hit a wall.

This study used a regeneration medium originally developed for Arabidopsis thaliana, modified slightly for cannabis, supplemented with two plant growth hormones: indole-3-acetic acid (a natural auxin) and benzylaminopurine (a cytokinin). These two hormones together are what push a dedifferentiated cell back toward division and development.

The results: cultures remained viable after three days of incubation, microscopy confirmed cells had undergone at least one division, and an extended 14-day cultivation experiment produced visible microcalli, small clusters of dividing cells, that retained viability under FDA staining. This is the proof-of-concept moment. The cells didn't just survive. They started rebuilding.

What the Gene Expression Data Revealed

This is where the study goes beyond what any grower would attempt in their facility, but it's also where the most interesting science lives.

The researchers tracked gene expression in the cultured protoplasts at 0, 24, 48, and 72 hours using RT-qPCR, a technique that quantifies how actively specific genes are being transcribed. They monitored six genes across three categories: cell proliferation, abiotic stress, and oxidative stress.

Cell Proliferation Markers

PCNA, a protein directly involved in DNA replication and used as a standard marker of cell division, was undetectable in normal leaf tissue but increased significantly after protoplast isolation, peaking at 72 hours with a threefold increase in high-viability cultures. The auxin-responsive gene IAA-2 also showed a significant increase during cultivation, peaking at 48 hours with a 3.5-fold increase, indicating that the cells were actively responding to the growth hormones in the medium.

Abiotic Stress Markers

Two genes associated with the plant stress hormone abscisic acid, PP2C-1 and LEA34, both dropped significantly after isolation and stayed low throughout cultivation. This is good news: it means the cells were not experiencing escalating abiotic stress during the culture period. They adapted to their new environment quickly rather than deteriorating under it.

Oxidative Stress — The Critical Battle

When cell walls are enzymatically digested, reactive oxygen species, essentially cellular rust, accumulate rapidly. If not neutralised, these damage the plasma membrane and kill the cell. Two antioxidant genes, APX and CAT, showed complementary activation patterns throughout cultivation, indicating the cells' own antioxidant systems were engaged and functioning. In high-viability cultures, APX expression increased 3.5-fold within 24 hours. This coordinated antioxidant response, the researchers argue, was a critical factor in enabling the cells to survive and eventually divide.

"The cells didn't just survive the isolation process. They mounted an antioxidant defence, responded to growth hormones, and started dividing. That coordinated response is the entire story."

Why Viability at Isolation Predicts Everything That Follows

One of the most practically useful findings in the study is the relationship between viability at the point of isolation and everything that happens afterwards.

High-viability cultures, those above 60% viability at isolation, showed roughly twice the PCNA expression of low-viability cultures, stronger antioxidant responses, and more robust cell division overall. Low-viability cultures, below 15%, showed reduced expression across all proliferation and stress markers, consistent with impaired capacity to divide and survive.

The implication is clear: the quality of your starting material and your isolation technique determine your ceiling. You cannot compensate downstream for poor isolation conditions upstream. In the context of cannabis biotechnology, this means the painstaking work of optimising donor plant age, cultivation conditions, enzymatic solutions, and washing protocols is not procedural box-ticking; it is the entire game.

Why This Matters for Growers — Now and in the Future

Most growers will never set foot in a protoplast laboratory. So why should any of this matter to someone running a cultivation facility, a breeding programme, or a craft grow? Because this is how the next generation of cannabis genetics gets made.

The ability to work with cannabis at the single-cell level unlocks capabilities that are simply not possible through conventional breeding. Consider what becomes possible once protoplast-to-plant regeneration is fully achieved in cannabis:

Precision Genetic Transformation

Rather than working with whole plants and hoping a genetic modification takes hold uniformly, you can transform a single cell and regenerate an entire plant from it, guaranteeing that every cell carries the intended modification. This is non-chimeric transformation — the gold standard in plant genetic engineering.

CRISPR Genome Editing

Several research groups have already demonstrated transient CRISPR delivery into cannabis protoplasts, with transformation efficiencies reaching 75% in some studies. Once stable whole-plant regeneration is achieved, targeted editing of cannabinoid pathway genes becomes a realistic breeding tool.

Somatic Hybridisation

Protoplasts from two different plant species or varieties can be fused together, creating hybrid cells that combine the genetics of both parent plants. In cannabis, where specific terpene and cannabinoid profiles are of enormous commercial value, the implications are significant.

Ploidy Manipulation

Working at the cellular level allows controlled changes to chromosome number — a technique used in other crops to create larger, more vigorous varieties. In cannabis, this pathway to polyploid breeding has barely been explored.

You should know how I feel about all this. I am simply reporting what the study found. If this is overall good for us, I will let you decide for yourself. Just be mindful that we as people can always decide the direction of our own spaces, but not the overall market's direction. Similarly with food.

The Honest Limitations

Complete plant regeneration from cannabis protoplasts has not yet been achieved. The study reports microcallus formation — early-stage cell clusters — but not organised shoot or root development from those clusters. That next step remains the critical bottleneck, and the researchers are explicit that it is unresolved.

The work was done on industrial hemp varieties, all with low THC content. Whether high-THC varieties will respond similarly to the same isolation and culture conditions is unknown. Genotype variability is a significant factor throughout the study — results that worked well for USO 31 did not always transfer to Finola or other cultivars.

Where This Is All Going

Plant biotechnology in cannabis is accelerating. The first stable genetically modified cannabis plant was created just a few years ago using Agrobacterium-mediated transformation. CRISPR constructs validated in protoplasts are already being used to edit cannabis genes. And now, the second-ever report of cannabis protoplast cultivation and division has been published, with gene expression data that confirms the cells are genuinely viable, actively dividing, and mounting appropriate stress responses.

Each of these steps is incremental. None of them is the finish line. But they are building toward a moment when cannabis breeders will have access to the same precision genetic tools that have been available for maize, tomato, and rice for decades. When that moment arrives, everything as we know it will change.

The Grower's Connect — Plant Science Series

Week 01 → Feeding Your Plant From the Inside Out: Sucrose Stem Infusion and 30%+ Yield Increases
Week 02 → What Heavy Cannabis Use Actually Does to Your Brain — And What the Science Really Says
Week 03 → Scientists Just Grew Cannabis From a Single Cell — You're reading it

Source Study: Král D, Šenkyřík JB and Ondřej V (2025) Early protoplast culture and partial regeneration in Cannabis sativa: gene expression dynamics of proliferation and stress response. Front. Plant Sci. 16:1609413. doi: 10.3389/fpls.2025.1609413 — Department of Botany, Faculty of Science, Palacký University Olomouc, Czechia. Published June 6, 2025.

Posted on 11 March 202616 March 2026 by Joe

What Heavy Cannabis Use Actually Does to Your Brain

What Heavy Cannabis Use Actually Does to Your Brain — And What the Science Really Says | The Certified

Plant Science · Cannabis & The Brain

What Heavy Cannabis Use Actually Does to Your Brain — And What the Science Really Says

A major new study scanned over 1000 brains across 7 cognitive tasks. Only one showed significant lifetime impact. Here's what it means for growers, consumers, and anyone who wants to think clearly about this plant.

The Grower's Connect · March 2025 · 9 min read

1003 Adults brain-scanned

7 Cognitive tasks tested

1 of 7 Tasks with significant lifetime impact

Listen to this article What Heavy Cannabis Use Actually Does to Your Brain

Last week we looked at the science of feeding your plant from the inside out — how sucrose stem infusion at precisely 0.5 bar can push cannabis yields up by over 30%. This week we're flipping the lens. Instead of what cannabis produces, we're looking at what cannabis consumption does — specifically, what a rigorous new brain imaging study out of JAMA Network Open has to say about how heavy and recent cannabis use affects brain function.

This is research worth understanding properly. Not because it's cause for alarm — the nuance here is actually more reassuring than most headlines suggest — but because the details are genuinely useful whether you're a grower who also consumes, a medical patient, or someone who simply wants to make informed decisions about how and when they use.

Let's get into it.

The Study — What Was Actually Done

Published in January 2025 in JAMA Network Open, this research used data from the Human Connectome Project — one of the most comprehensive brain mapping datasets ever assembled. Researchers at the University of Colorado analysed 1003 young adults aged 22 to 36, putting them through 7 different brain function tasks inside an MRI scanner while measuring real-time brain activation.

The tasks covered working memory, reward processing, emotional recognition, language, motor function, logical reasoning, and theory of mind — essentially a full sweep of the cognitive functions we use daily.

Crucially, the researchers did something most cannabis studies don't: they separated recent use from lifetime use. Participants provided urine samples on the day of scanning to determine whether THC was still active in their system, allowing the team to distinguish between the short-term effects of being recently high versus the longer-term effects of years of heavy use.

How Participants Were Grouped

Heavy lifetime users — more than 1000 lifetime uses (88 participants, 8.8%)
Moderate users — 10 to 999 lifetime uses (179 participants, 17.8%)
Non-users — fewer than 10 lifetime uses (736 participants, 73.4%)
Recent users — THC detected in urine on scan day (106 participants)
All models adjusted for age, sex, income, education, alcohol and nicotine use

What the Scans Actually Showed

Here is where the story gets more interesting — and more nuanced — than most headlines will tell you.

Out of 7 cognitive tasks, only one — working memory — showed a statistically significant association with heavy lifetime cannabis use. Heavy lifetime users showed lower brain activation during the working memory task, with a small to medium effect size. The brain regions most affected were the anterior insula, the dorsomedial prefrontal cortex, and the dorsolateral prefrontal cortex — all areas known to have a high density of CB1 receptors, the primary binding site for THC.

Emotion, language, motor function, logical reasoning, reward processing, and theory of mind all showed no statistically significant association with lifetime heavy use once properly adjusted for demographics, alcohol use, and nicotine use. That is a meaningful finding. Six out of seven tasks showed no significant long-term impact.

"Six out of seven cognitive tasks showed no statistically significant association with heavy lifetime cannabis use. The science is more nuanced than the headlines suggest."

Recent Use Is a Different Story

Where the data gets more pointed is around recent use — people who had THC actively in their system on the day of scanning.

Recent cannabis users showed lower brain activation during both the working memory and motor tasks, and also showed poorer behavioural performance on the working memory task, episodic verbal memory, and theory of mind. Being recently high measurably affected how well people performed on cognitive tests — this is the clearest and most practical finding in the study.

However, even here the picture is more nuanced. After adjusting for race and education — both independently associated with a positive THC test — the theory of mind association disappeared entirely. And after applying false discovery rate correction across all 7 tasks, the working memory association was the only one that survived statistically.

Practical Takeaway

The impairment from recent use is real and measurable, but temporary. Studies suggest residual cognitive effects may persist for 2 to 4 weeks in regular users. If you need to perform well cognitively — a complex task, important decision-making, a demanding work situation — abstaining beforehand makes empirical sense.

What This Means for You

If You Grow

Managing a cultivation environment — VPD, nutrient schedules, plant health, environmental controls — is a working memory intensive activity. The study is a practical reminder that separating consumption from the cognitive demands of running a grow has real basis in the science.

If You Consume

For moderate users the findings are genuinely reassuring — no significant cognitive differences were found compared to non-users across any task. For heavy, long-term users the working memory finding is worth taking seriously as useful information, not cause for alarm.

For medical patients, the picture is more complex. This study focused on recreational community users, not patients managing specific conditions. The therapeutic benefits of cannabis for pain, anxiety, epilepsy, and other conditions may well outweigh cognitive trade-offs — that is a conversation for a medical professional, not a conclusion to draw from a single study.

On potency — THC content in cannabis products has increased significantly over the past two decades, and higher potency means more THC binding to those CB1 receptors. The study didn't measure dose or potency, but it's a reasonable inference that frequency and potency together are more impactful than frequency alone. As growers producing increasingly potent product, that context matters.

The Honest Limitations

The researchers are admirably upfront about what the study can't tell us. It is cross-sectional — a snapshot, not a long-term follow-up. We cannot say cannabis caused these brain differences. People who become heavy users may differ in other ways the study couldn't fully account for.

The sample was young adults aged 22 to 36, so findings may not apply to older adults — and more critically, not to adolescents, for whom the developing brain presents an entirely different risk profile. The study also lacked data on typical THC dose, potency, CBD content, or route of administration. A joint, an edible, and a high-potency concentrate deliver THC very differently, and the study treats all of these the same.

The Bigger Picture

Two weeks of research coverage on The Grower's Connect, and a theme is emerging: the cannabis plant rewards those who understand it deeply.

Last week that meant understanding how sucrose functions as a signalling molecule, and what precise delivery pressure does to yield outcomes. This week it means understanding what THC does in the CB1-receptor-rich regions of the prefrontal cortex, and why that matters specifically for working memory.

The takeaway from this study is not that cannabis is dangerous. It is that heavy, long-term use appears to have a specific and measurable effect on one important cognitive function, that recent use has broader but temporary effects, and that moderate use showed no significant difference from non-use. That is useful information. Use it.

"Growers and consumers who engage with the science honestly — including the parts that aren't entirely flattering — build more credibility and make better decisions than those who don't."

Source Study: Gowin JL, Ellingson JM, Karoly HC, Manza P, Ross JM, Sloan ME, Tanabe JL, Volkow ND. Brain Function Outcomes of Recent and Lifetime Cannabis Use. JAMA Network Open. 2025;8(1):e2457069. doi:10.1001/jamanetworkopen.2024.57069. University of Colorado Anschutz Medical Campus. Published January 28, 2025.

Posted on 21 January 202621 January 2026 by Joe

HHC: The Synthetic Ghost Haunting Our Shelves

This week, I found myself wandering into a vape shop. Fully stocked with all sorts of devices and vape carts. Flavours of all kinds. Mind you all, I am not a vapour? I think this is the term. But curiosity had me and a buddy stumble in. Upon walking in, we started chatting and being the weedy I am, I asked, “Are there THC vapes here?” The guy lit up and said: “No, but I have this, something better.” And handed me an HHC cart…

Naturally, I had to ask. “What can you tell me about this?”

The conversation that followed was, frankly, disheartening. The person selling this product had no idea what hydrogenation was, let alone why a synthetic derivative might be concerning to someone who values the natural plant. They couldn’t explain why we need to chemically alter a compound that nature already perfected. It was a stark reminder that while the shelves are filling up, the knowledge gap is widening.

We’ve touched on Hexahydrocannabinol (HHC) before, warning about its semi-synthetic nature and the grey market it inhabits. But encounters like this, coupled with emerging research from Europe, make it clear: we need to revisit this topic. We need to remind ourselves what HHC is, what it isn’t, and why the “legal” label doesn’t always mean “safe.”

What is HHC? A Quick Refresher

Let’s strip away the marketing. HHC is a semi-synthetic cannabinoid. While trace amounts can theoretically be found in the cannabis plant, the HHC you see on shelves is not natural. It is created in a lab through a process called hydrogenation.

Think of it like margarine. You take a natural oil (in this case, usually CBD extracted from hemp) and bombard it with hydrogen atoms using heavy metal catalysts (like palladium or nickel) under high pressure. This breaks the double bonds in the molecule, turning it into a more stable, hydrogen-saturated compound.

Why do they do this? Two reasons:

Shelf Life: Hydrogenation makes the molecule incredibly stable. It resists oxidation, meaning it can sit on a shelf for months or years without degrading.
Legal Loopholes: By chemically altering the structure, manufacturers create a compound that mimics THC’s effects but often slips through the cracks of specific drug laws at least until regulators catch up.

The New Reality: Poisonings and Public Health Alarms

While the salesperson in the vape shop might tell you it’s “just like THC but legal,” recent data tells a different, more alarming story. A study released in 2025 by the Czech Toxicological Information Centre (TIC) paints a grim picture of what happens when these semi-synthetic products flood an unregulated market.

Following the appearance of HHC products in the Czech Republic in 2022, the poison control centre saw a sharp rise in calls. We’re not talking about feeling a bit too sleepy; we’re talking about neurological, cardiovascular, and gastrointestinal toxicity.

The study analysed nearly 200 cases of HHC poisoning. The victims weren’t just seasoned users pushing limits; many were children and teenagers who consumed HHC in the form of gummies, cookies, and vape products identical to what I saw in that shop.

The symptoms reported were serious:

Neurological: 74% of patients experienced issues ranging from severe drowsiness and confusion to hallucinations and even seizures.
Cardiovascular: Over 40% suffered from tachycardia (rapid heart rate) and hypertension.
Gastrointestinal: Severe vomiting and nausea were common.

Crucially, the study highlighted that HHC intoxication can last significantly longer than THC intoxication. One case report detailed a healthy man in his 40s who consumed HHC cookies and suffered from cognitive and physical impairment for nine days. He experienced visual disturbances, disorientation, and an inability to function normally long after the “high” should have worn off. This prolonged effect is likely due to the structural changes from hydrogenation, which may alter how our bodies metabolise and eliminate the compound.

The “Entourage” vs. The Isolate

The beauty of the cannabis plant lies in its complexity. We’ve spent this year celebrating the Entourage Effect, the synergy between hundreds of cannabinoids, terpenes, and flavonoids working together. We’ve marvelled at the discovery of flavoalkaloids in natural leaves.

HHC products are the antithesis of this. They are typically made from isolates. You are getting a single, chemically modified molecule, stripped of the natural buffers and modulators found in the whole plant.

Furthermore, because HHC is synthesised, it exists in two forms (enantiomers): 9R-HHC and 9S-HHC.

9R-HHC binds actively to your endocannabinoid receptors, mimicking THC.
9S-HHC does not bind well and is largely inactive.

Commercial products are a mix of both. You have no idea what ratio you are getting, which leads to wildly inconsistent effects. One vape cart might do nothing; the next might send you to the ER with panic attacks and heart palpitations because the batch had a higher ratio of the active 9R isomer.

Why Are We Seeing It Here?

You might ask, “If we have access to amazing, natural South African cannabis, why is this stuff here?”

The answer is simple: Economics and Opportunism.
The 2018 US Farm Bill legalised hemp cultivation, leading to a massive surplus of CBD. Chemists realised they could convert this cheap CBD into psychoactive HHC and sell it in markets where THC is restricted or where “legal” sounds safer to the uninitiated consumer.

It is a product born from a loophole, not from a love for the plant. The process was founded with the intention of creating stable medicine. things similar to Marinol. We all know how that turned out.

The Verdict: Keep It Real

The encounter at the vape shop was a wake-up call. It showed that while the culture is growing, so is the misinformation.

As a community that prides itself on understanding the plant, from the soil microbiome to the terpene profile, we need to be discerning.

HHC is not “natural weed.” It is a lab-made chemical analogue.
It carries risks. The potential for contaminants (heavy metals from the hydrogenation process) and the documented cases of severe, prolonged intoxication are real.
We have better. We live in a country with some of the best sun-grown genetics on earth. Why trade the rich, therapeutic complexity of a Durban Poison or a well-grown White Widow for a synthetic mystery fluid?

Let’s stick to what we know, what we love, and what the earth provides. Let’s keep our culture green, not grey. Stay safe, stay informed, and keep it natural.

Posted on 7 January 20267 January 2026 by Joe

New Hope for Knees? Unpacking the Latest Clinical Trial on CBD

As we embrace the fresh start of 2026, many of us are looking for new ways to enhance our health and well-being. For those battling the persistent ache of osteoarthritis, particularly in the knees, the quest for relief is often top of mind. In South Africa, where an active lifestyle is cherished, joint pain can be a significant hurdle.

Late last year, a new clinical trial titled “Effects and safety of a CBD-rich Cannabis sativa oil in knee osteoarthritis” (the CANOA trial) was published, adding a crucial piece to the puzzle of cannabis medicine. Conducted by researchers at the Universidade Federal da Integração Latino-Americana in Brazil, this double-blind, randomised, placebo-controlled study offers valuable insights into the potential and the limitations of CBD oil for pain management.

This week, we’re diving deep into this study to understand what it means for patients, the medical community, and the future of cannabis-based therapies for osteoarthritis.

The Challenge: Living with Osteoarthritis

Osteoarthritis is a degenerative joint disease that affects millions worldwide, causing chronic pain, inflammation, and reduced mobility. Conventional treatments often rely on Non-Steroidal Anti-Inflammatory Drugs (NSAIDs) or opioids. While these can provide relief, they often come with significant long-term side effects, leaving many patients searching for safer, more sustainable alternatives.

Enter cannabis. With its well-known anti-inflammatory and analgesic properties, the cannabis plant has emerged as a beacon of hope. Specifically, Cannabidiol (CBD), a non-intoxicating compound, has shown promise in preclinical studies. However, rigorous clinical evidence in humans has remained scarce and sometimes contradictory. The CANOA trial aimed to fill this gap.

The Study: Testing a Full-Spectrum Solution

The CANOA trial was meticulously designed to test the efficacy and safety of a full-spectrum CBD-rich cannabis oil. Unlike isolated CBD products, full-spectrum oils contain a range of phytocannabinoids, terpenes, and flavonoids. This composition is believed to leverage the “entourage effect,” where these compounds work synergistically to enhance therapeutic benefits.

Key Study Details:

Participants: 45 patients aged 30-70 with diagnosed knee osteoarthritis and moderate-to-severe pain.
The Treatment: Participants were randomly assigned to receive either the cannabis oil (containing 45 mg of CBD daily) or a placebo (MCT oil) for 60 days. Crucially, the oil contained virtually no THC (below detection limits), isolating the potential effects of CBD and other minor cannabinoids.
Measurement: The primary goal was to measure changes in pain intensity using the WOMAC scale, a standard tool for assessing osteoarthritis. Secondary measures included quality of life, sleep quality, and depression levels.

The Findings: Relief, But No “Magic Bullet”

The results of the CANOA trial are fascinating and nuanced.

Pain Relief was Universal:
At the end of the 60-day trial, both groups reported a significant reduction in pain. Patients taking the CBD oil experienced relief, but so did those taking the placebo. Statistically, there was no significant difference in pain reduction between the two groups.
Quality of Life Improvements:
Similarly, both groups reported improvements in sleep quality, depression symptoms, and overall quality of life. Again, the cannabis oil did not outperform the placebo in these metrics.
The Power of the Placebo:
The researchers highlighted a strong “placebo effect” and potentially the “Hawthorne effect” (where individuals modify an aspect of their behavior in response to their awareness of being observed). The care, attention, and positive expectations associated with participating in a clinical trial can themselves be powerful healers.
Safety and Tolerability:
This is a critical win. The CBD-rich oil was exceptionally well-tolerated. No serious adverse events were reported, and blood tests showed no negative impact on liver or kidney function. The most common side effects were mild, such as dry mouth or slight weight changes, but these were comparable to the placebo group. This confirms the high safety profile of CBD, a vital consideration for long-term use.

What Does This Mean for You?

The CANOA trial might seem discouraging at first glance—after all, the CBD oil didn’t beat the placebo. However, it provides essential context for managing expectations and refining treatments.

Dosage Matters:
The study used a daily dose of 45 mg of CBD. While safe, this might be too low for severe osteoarthritic pain. Cannabis medicine is highly individualised, and higher doses or different formulations might be necessary to unlock significant analgesic effects.
The “Entourage” Needs THC?
The oil used had undetectable levels of THC. Many experts argue that a small amount of THC is crucial to fully activate the entourage effect and provide potent pain relief. Future research needs to explore formulations with balanced ratios of CBD and THC.
It’s Safe to Try:
The study reinforces that high-quality CBD oil is safe. For those struggling with osteoarthritis, it remains a low-risk option to explore, potentially as an add-on to other therapies. While it might not be a cure-all on its own at this dosage, its safety profile makes it a viable candidate for personalised medicine approaches.
The Mind-Body Connection:
The strong placebo response underscores the importance of holistic care. Managing chronic pain isn’t just about molecules; it’s about patient support, mental well-being, and the therapeutic relationship.

Looking Ahead

The CANOA trial is a stepping stone, not a roadblock. It challenges the cannabis community to look deeper to investigate higher dosages, different cannabinoid combinations (like adding THC, CBG, or CBC), and longer treatment periods.

As we move through 2026, let’s use this knowledge to advocate for more research and to approach cannabis medicine with both optimism and a critical scientific eye. For now, the takeaway is clear: CBD is safe and holds potential, but finding the “sweet spot” for osteoarthritis relief is a journey we are still navigating.

Posted on 3 September 20253 September 2025 by Joe

South African Scientists Uncover Rare Compounds in Cannabis Leaves

For too long, the conversation around cannabis in South Africa has been narrowly confined, often overshadowed by debates on recreational use or the well-known psychoactive components. Yet, beneath this limited perception lies a world of intricate chemistry, brimming with therapeutic potential that is only just beginning to be unveiled. Right here, on our own soil, groundbreaking research from Stellenbosch University is challenging these outdated narratives, positioning South Africa at the forefront of global cannabis science.

This pioneering study, published in the prestigious Journal of Chromatography A, goes beyond the familiar cannabinoids like THC and CBD. It delves deep into the often-overlooked parts of the cannabis plant – particularly its leaves and inflorescence – to uncover a treasure chest of complex, health-boosting compounds. The findings are nothing short of astonishing, as local scientists have not only extensively characterized previously understudied “phenolics” but have also provided the first-ever evidence of a rare and chemically intriguing class of compounds: flavoalkaloids, in Cannabis sativa L.

This monumental work, led by Magriet Muller and Professor André de Villiers from the Department of Chemistry and Polymer Science at Stellenbosch, utilized state-of-the-art analytical techniques to dissect the chemical makeup of three commercial cannabis strains cultivated in South Africa: ‘Cape Cookie’, ‘CBG’, and ‘Blue Sky’, sourced from Cannsun Medicinals. It’s a testament to the ingenuity and scientific rigour thriving in our nation, offering a glimpse into a future where cannabis is recognized for its full spectrum of medicinal and wellness applications.

The Groundbreaking Discovery: Flavoalkaloids Make Their Debut in Cannabis

The most electrifying revelation from this Stellenbosch study is the unequivocal evidence of flavoalkaloids in the Cannabis sativa L. plant for the first time in scientific history. For many, the term “flavoalkaloid” might sound unfamiliar, and for good reason: these compounds are relatively rare in the plant kingdom, making their discovery in a species as widely studied as cannabis a true scientific breakthrough.

So, what exactly are flavoalkaloids? Imagine a powerful fusion of two distinct classes of plant compounds, each with its own significant biological activities:

Flavonoids: These are well-known plant pigments and secondary metabolites found in fruits, vegetables, and many herbs. They are celebrated for their potent antioxidant, anti-inflammatory, and neuroprotective properties.
Alkaloids: These are nitrogen-containing organic compounds, often found in plants, that can have profound physiological effects. Many pharmaceutical drugs, such as caffeine and morphine, are alkaloids.

A flavoalkaloid, therefore, is a hybrid molecule where an alkaloid moiety is directly linked to a flavonoid backbone. This unique structural combination suggests the potential for novel and diverse biological activities, potentially offering a broader range of therapeutic effects than either component on its own. The Stellenbosch research tentatively identified 16 flavone derivatives belonging to four distinct classes of flavoalkaloids, primarily in the leaf extracts of the ‘Blue Sky’ cannabis strain. This specificity is crucial, indicating that the therapeutic profile of cannabis can vary significantly not only between strains but also across different parts of the plant.

Why is this discovery so profoundly significant?

Uncharted Chemical Territory: Finding an entirely new class of compounds in cannabis opens up an uncharted frontier in phytochemistry. It implies that despite extensive global research, we are only just scratching the surface of Cannabis sativa’s true chemical complexity.
Promise for Novel Therapies: In other plant species, flavoalkaloids have been shown to exhibit a wide array of bioactivities, including anti-cancer, anti-inflammatory, antimicrobial, and neuroprotective effects. The identification of these compounds in cannabis offers exciting prospects for developing entirely new therapeutic agents, potentially leading to cannabis-derived medicines with unique mechanisms of action. This moves beyond the current focus on cannabinoids and terpenes, expanding the plant’s medicinal potential exponentially.
Reimagining Plant Utilisation: The detection of flavoalkaloids predominantly in the leaves of a specific strain suggests that cannabis leaves, often considered by-products in some cultivation practices, may hold significant untapped value. This could lead to more sustainable and economically viable cultivation models, where the entire plant biomass is utilised for its full chemical potential, benefiting local farmers and industries in South Africa.
Understanding the “Entourage Effect” Further: This discovery adds another layer of complexity to the “entourage effect,” the hypothesis that various compounds in cannabis work synergistically to enhance therapeutic outcomes. The presence of flavoalkaloids suggests a richer interplay of compounds than previously understood, potentially leading to more effective, whole-plant-based remedies.

This unprecedented finding by the Stellenbosch team is a testament to the power of advanced analytical chemistry and underscores the vast, unexplored medicinal potential within Cannabis sativa L.

Diving Deeper: The Rich Tapestry of Cannabis Phenolics

Beyond the groundbreaking discovery of flavoalkaloids, the Stellenbosch study performed an extensive characterisation of other phenolic compounds in the cannabis plant. Phenolics are a diverse group of plant metabolites widely known for their crucial roles in plant defence and their beneficial effects on human health, primarily through their antioxidant and anti-inflammatory properties. Their presence is a key component of the “entourage effect,” where these compounds interact with cannabinoids and terpenes to modulate and enhance therapeutic benefits.

The research identified a remarkable total of 79 phenolic compounds, with an impressive 25 of these being reported in Cannabis sativa L. for the very first time. This expanded inventory paints a much more detailed picture of cannabis’s non-cannabinoid chemical profile. These compounds can be broadly categorised as follows:

Flavones and Their Glycosides: The study detected key flavones such as luteolin, apigenin, chrysoeriol, and acacetin, many of which were present as O-glucuronide or O-diglycoside derivatives. These are common flavonoids, but their specific glycosylation patterns (attachment to sugar molecules) can influence their bioavailability and biological activity.
- Strain-Specific Variability: A particularly interesting finding was the significant difference in flavone profiles between the strains. ‘Blue Sky’ (Strain C) exhibited markedly higher levels of C-glycosylated flavones (e.g., orientin, vitexin, cytisoside) and their O-glycosylated derivatives, which were either absent or in much lower concentrations in ‘Cape Cookie’ (Strain A) and ‘CBG’ (Strain B). This underscores the importance of genotype in determining the unique chemical signature of each cannabis variety. Conversely, strains A and B showed higher levels of acacetin O-diglycosides.
Flavonols and Their Glycosides: The researchers also tentatively identified flavonols like quercetin and kaempferol derivatives, including a quercetin O-hexosyl O-glucuronide derivative. These compounds are potent antioxidants and contribute to the plant’s overall protective qualities.
Phenolic Amides and Hydroxycinnamic Acid Derivatives: The study found a range of these compounds, including N-trans-coumaroyltyramine and N-trans-feruloyltyramine, along with various derivatives of coumaric acid and caffeic acid. These compounds are known for their antioxidant properties and have been investigated for roles in anti-inflammatory processes.
Novel Phenolic Acid Sulfates: In another significant first, two isomeric caffeic acid sulfates were tentatively identified exclusively in the inflorescence of the ‘Blue Sky’ strain. This discovery of sulfur-containing phenolics in cannabis represents a new frontier for research into their potential bioactivities and functions within the plant.

The comprehensive nature of this phenolic characterisation highlights the extraordinary chemical diversity within Cannabis sativa L. It reinforces that different strains and even different parts of the same plant possess unique chemical profiles, suggesting that targeted cultivation and processing could yield specialized extracts for specific health conditions. The contour plots generated by the analytical method provided a rapid visual comparison of these complex profiles, making these subtle yet significant differences immediately apparent to researchers.

The Cutting-Edge Toolkit: Unravelling Complexity with Advanced Analytical Chemistry

The groundbreaking discoveries from Stellenbosch University were made possible by employing a highly sophisticated and optimised analytical methodology: Comprehensive Two-Dimensional Liquid Chromatography hyphenated to High-Resolution Mass Spectrometry (HILIC × RP-LC-HR-MS). This advanced technique is crucial for dissecting the immense chemical complexity of the cannabis plant, where hundreds of compounds, many structurally similar, coexist across a wide range of concentrations.

Here’s a detailed look at the methodology that enabled these profound insights:

Sample Preparation: Isolating the Targets
To specifically target polar phenolic compounds and avoid interference from well-known apolar compounds like cannabinoids and chlorophyll, a precise sample preparation strategy was crucial:

Freezing and Grinding: Plant samples (inflorescence and leaves) were snap-frozen with liquid nitrogen and finely ground using a mortar and pestle. This step preserves the integrity of the compounds and maximizes extraction efficiency.
Defatting with Hexane: The ground material was defatted three times with hexane through sonication and centrifugation. This process effectively removed non-polar cannabinoids, chlorophyll, and other lipids, ensuring that the subsequent analysis focused on the more polar phenolic compounds.
Extraction with Aqueous Acetone: After defatting, the polar phenolics were extracted using an aqueous acetone solution (30/70 v/v H2O/acetone), followed by sonication and centrifugation. The supernatant was then evaporated, freeze-dried, and re-dissolved in a dilute H2O/MeOH solution for analysis. This selective extraction method was designed to concentrate the target compounds and minimize interference.

Comprehensive Two-Dimensional Liquid Chromatography (LC × LC): The Ultimate Separator
Traditional one-dimensional (1D) liquid chromatography often struggles with complex plant extracts, as many compounds co-elute (come out of the column at the same time), making individual identification nearly impossible. 2D-LC overcomes this limitation by employing two different separation mechanisms in sequence:

First Dimension (¹D) – Hydrophilic Interaction Liquid Chromatography (HILIC):
- Mechanism: HILIC separates compounds based on their polarity. Polar compounds interact strongly with the stationary phase (Acquity BEH Amide column, 150 × 1.0 mm, 1.7 µm) and are retained longer, while less polar compounds elute faster.
- Purpose: This step effectively “spreads out” the highly polar phenolic compounds, providing an initial broad separation based on a property distinct from the second dimension.
- Dilution and Modulation: The effluent from the ¹D column was diluted with a weak reversed-phase solvent and then introduced into an interface with two 80 µL loops, acting as a modulator. This process collects small fractions from the ¹D separation and rapidly injects them onto the ²D column, preventing peak distortion.
Second Dimension (²D) – Reversed-Phase Liquid Chromatography (RP-LC):
- Mechanism: RP-LC separates compounds based on their hydrophobicity. Less polar compounds are retained longer on the stationary phase (Zorbax Eclipse Plus C18 column, 50 × 3.0 mm, 1.8 µm), while more polar compounds elute faster.
- Purpose: By applying a different separation mechanism, RP-LC can resolve compounds that may have co-eluted in the HILIC dimension.
- Fast Gradient: The ²D separation uses a very fast gradient (0.45 min) and a high flow rate (3 mL/min) to ensure rapid analysis of each ¹D fraction, maintaining high resolution.
Orthogonality and Peak Capacity: The combination of HILIC and RP-LC is highly “orthogonal” because these two modes separate compounds based on fundamentally different chemical properties. This means compounds that co-elute in one dimension are highly likely to be separated in the other, leading to vastly improved resolution. The Stellenbosch method achieved an “excellent separation performance” with a “practical peak capacity above 3000” and an average orthogonality of 75%. This level of separation is exponentially greater than what can be achieved with 1D methods, allowing for the detection of many more individual compounds.
Method Optimization: The team used an in-house developed predictive optimization algorithm (in Matlab R2019b) [31-33]. This software systematically explored a wide range of experimental conditions to find the optimal settings for analysis time, peak capacity, and resolution, further enhancing the method’s effectiveness.

High-Resolution Mass Spectrometry (HR-MS) – Quadrupole Time-of-Flight (Q-TOF): Identifying the Unknowns
As compounds exit the ²D column, they are immediately directed to a Q-TOF mass spectrometer. This instrument is essential for identifying the separated compounds:

Accurate Mass Measurement: Q-TOF provides highly accurate mass measurements of both precursor (intact) ions and fragment ions. This precision allows researchers to determine the exact molecular formula of an unknown compound, which is a crucial first step in identification.
MSE Fragmentation: The instrument was operated in MSE mode, which collects both low (4 eV) and high (10-30 eV ramped) collision energy data simultaneously. Low-energy data shows the intact molecular ions, while high-energy data provides characteristic fragmentation patterns. These “fingerprints” are invaluable for elucidating the structure of compounds, even those never before seen.
Tentative Identification: By combining accurate mass data, fragmentation patterns, UV spectral data (from the DAD detector), and relative retention times in both dimensions, researchers could tentatively identify 79 compounds, including the novel flavoalkaloids and phenolic acid sulfates.

This sophisticated analytical pipeline allowed the Stellenbosch team to peer into the complex chemistry of cannabis with unprecedented clarity, leading to discoveries that would have been impossible with less advanced techniques.

The Road Ahead: An Exciting Journey of Discovery

The work by Magriet Muller and Professor André de Villiers is not merely an academic exercise; it is a powerful stride into the future of cannabis. It reminds us that even in plants we think we know well, nature often holds profound secrets waiting to be uncovered. As research continues to peel back the layers of Cannabis sativa’s intricate chemistry, the potential for new health solutions and economic opportunities grows exponentially.

For South Africa, this research is a beacon of hope and innovation. It champions local scientific excellence and offers a path toward a future where cannabis is understood and utilised for its full, multifaceted potential, contributing significantly to health, wellness, and a sustainable economy. The journey to fully understand cannabis and its immense potential has just become even more fascinating, and we eagerly await the next chapter of discoveries that will undoubtedly emerge from our vibrant scientific community.

You can read the full published paper here: “Comprehensive two-dimensional liquid chromatographic analysis of Cannabis phenolics and first evidence of flavoalkaloids in Cannabis” by Magriet Muller and André de Villiers, 2 August 2025, Journal of Chromatography A.
DOI: 10.1016/j.chroma.2025.466023

Posted on 16 July 202516 July 2025 by Joe

Cannabis Tissue Culture: Unlocking Potential

In the rapidly evolving landscape of cannabis cultivation, growers are constantly seeking methods to optimise yield, quality, and consistency. While traditional cloning through cuttings remains a cornerstone, a sophisticated biotechnological approach known as tissue culture (micropropagation) is emerging as a game-changer. This method promises unprecedented control and is unlocking new frontiers for genetic improvement within Cannabis sativa.

This advanced technique moves beyond conventional cloning to address some of the most persistent challenges in cannabis production. Today, we will delve into what cannabis tissue culture is, its profound benefits, the unique hurdles it presents for Cannabis sativa, and the cutting-edge innovations that are shaping its future.

The Promise of Tissue Culture: A Leap Beyond Conventional Cloning

Cannabis tissue culture involves cultivating plants from very small pieces of plant tissue, called explants, in a sterile, nutrient-rich laboratory environment. This method offers several compelling advantages over traditional cloning:

Production of Disease-, Pest-, and Virus-Free Stock: One of the most critical benefits of tissue culture is its ability to establish and maintain clean plant programs. Traditional cloning risks transmitting pathogens, pests, and viruses from mother plants to subsequent generations. Tissue culture provides a sterile starting point, ensuring disease-free and vigorous plant material for every cultivation cycle. This is particularly vital in mitigating devastating diseases like bud rot, where even careful environmental controls might not eliminate lingering inoculum.
Rapid, Large-Scale Clonal Propagation: Once a successful tissue culture protocol is established, it allows for the exponential multiplication of genetically identical plants from a single parent. This scalability is essential for commercial operations aiming for uniform, high-quality harvests, leading to higher multiplication rates and more consistent production.
Genetic Preservation: Tissue culture enables the long-term storage of valuable cannabis genetics in a small, controlled space. This is a far more efficient method than maintaining large mother plant populations, protecting rare or desirable chemovars from loss due to disease, pests, or environmental calamities. Advanced techniques like cryopreservation, which store tissues at ultra-low temperatures, can preserve genetic material indefinitely while preventing genetic drift over time.
Foundation for Advanced Breeding: Tissue culture is the bedrock for modern plant breeding and genetic engineering. It provides a sterile and controlled environment to work with individual cells or small tissue samples, facilitating techniques like genome editing (e.g., CRISPR/Cas9) and genetic transformation. This accelerates the development of new, improved cannabis varieties with enhanced traits, such as increased cannabinoid or terpene production or greater disease resilience.

The Art and Science of Micropropagation: A Multi-Stage Journey

Cannabis micropropagation typically involves a precise, multi-stage process, each step requiring careful control over environmental factors and nutrient media:

Stage 0: Selection and Maintenance of Parent Stock: The process begins with selecting healthy, vigorous mother plants that possess the desired traits. Maintaining the health of these initial stock plants is crucial, as any latent pathogens could compromise the sterility of the subsequent cultures.
Stage 1: Culture Initiation: Very small pieces of plant tissue, or explants, are carefully sterilised and placed onto a specialised nutrient medium. Commonly used explants include nodal segments, hypocotyls, cotyledons, leaves, or even floral tissues. This initial stage aims to induce growth and shoot proliferation in a completely aseptic environment.
Stage 2: Multiplication: This is where the exponential propagation occurs. The developing shoots are repeatedly divided and subcultured onto fresh nutrient media to encourage rapid multiplication. This stage is key to producing the large numbers of genetically identical clones needed for commercial-scale cultivation.
Stage 3: Shoot Elongation and Rooting: Once a sufficient quantity of shoots has been produced, they are transferred to different media formulations designed to promote shoot elongation and the development of a robust root system. This prepares the young plantlets for life outside the sterile laboratory environment.
Stage 4: Acclimatisation (Hardening Off): In this critical final stage, the young plantlets are gradually transitioned from the high-humidity, sterile conditions of the lab to a greenhouse or indoor grow room environment. This hardening-off process is essential to prepare them for less controlled conditions and independent growth.

Throughout these stages, the nutrient media is a paramount factor. Typically, a basal salt mixture (such as Murashige and Skoog (MS) or Driver and Kuniyuki Walnut (DKW) media), is supplemented with Plant Growth Regulators (PGRs) like auxins (e.g., Indole-3-acetic acid (IAA), Indole-3-butyric acid (IBA), Naphthaleneacetic acid (NAA)) and cytokinins (e.g., 6-benzylaminopurine (BAP), Thidiazuron (TDZ), meta-Topolin), carbohydrates (sucrose), and various vitamins. The precise balance of these components is vital, as it profoundly impacts the efficiency of shoot proliferation, rooting, and the overall health and development of the explants.

Cannabis’s Unique Hurdles: Why Tissue Culture Has Been Challenging

Despite its immense potential, the application of tissue culture to Cannabis sativa has historically faced significant challenges:

Historical Prohibition: Decades of legal restrictions severely limited scientific research into cannabis plant biology and tissue culture. Unlike other agricultural crops that benefited from extensive public and private research, cannabis remained largely understudied, leading to a substantial knowledge gap in optimised protocols.
“Recalcitrance” to Regeneration: Cannabis sativa has shown a notable recalcitrance to regeneration in tissue culture. This is particularly true of non-meristematic tissues (like mature leaves or cotyledons) that could offer a larger starting material pool. Many published protocols report low multiplication rates and difficulty in achieving sustained, vigorous growth across multiple subcultures.
Genotype and Tissue Specificity: A significant hurdle is that tissue culture protocols often do not translate well between different cannabis chemovars (strains) or even between different plant parts from the same genotype. For instance, a method optimised for a high-THC Mexican strain may not work efficiently for high-CBD lines. This highlights the critical need for extensive genotype-specific research and protocol development.
Strong Apical Dominance: Cannabis naturally exhibits strong apical dominance, where the main stem grows preferentially, suppressing side branching. This trait can lead to low shoot multiplication rates from nodal explants in tissue culture, as explants tend to produce a single shoot rather than multiple branches, limiting the efficiency of mass propagation.
Reproducibility Issues: Even within published scientific literature, successful tissue culture protocols for cannabis have sometimes proven difficult for independent research groups to replicate consistently. This variability further underscores the inherent biological complexities and the genotype-dependent nature of cannabis tissue culture.

The Cutting Edge: Innovations Shaping the Future

To overcome these enduring hurdles, researchers are actively pursuing and developing innovative approaches and technologies in cannabis tissue culture:

Floral Reversion: A promising alternative involves using immature floral tissues as explants. These tissues, which contain numerous meristematic regions, can be induced to “revert” from a flowering state back to a vegetative state when cultured under specific conditions. This approach has shown potential for significantly higher multiplication rates compared to traditional nodal explants.
De Novo Regeneration: While challenging, regenerating whole plants from non-meristematic somatic tissues (such as leaves or hypocotyls) offers a theoretically almost limitless source of starting material. Advances in optimising the precise balance of PGRs and media composition are gradually improving regeneration rates in this complex area.
Advanced Cryopreservation: For truly long-term genetic preservation, cryopreservation involves storing plant tissues at ultra-low temperatures, which effectively halts metabolic processes. This method ensures exceptional genetic stability and prevents the accumulation of mutations or decline that can occur even in long-term active cultures, offering superior genetic fidelity over time.
Artificial Intelligence and Machine Learning: To address the immense complexity and multi-variable nature of tissue culture protocols, AI and machine learning algorithms are being integrated. These computational approaches can analyse vast datasets to predict and optimise ideal culture conditions and media formulations, accelerating the development of robust and efficient protocols.
Nanoparticle Technologies: Research is exploring the use of nanoparticles to enhance tissue culture processes. These tiny carriers can improve nutrient delivery, boost PGR uptake efficiency, and even provide targeted antimicrobial protection within the sterile culture environment. This precision could significantly improve regeneration success rates.
Genetic Engineering: Beyond simple micropropagation, advanced techniques like gene editing (e.g., CRISPR/Cas9) and genetic transformation are advancing rapidly. These tools allow for precise modifications to the cannabis genome, enabling the development of plants with enhanced disease resistance (e.g., to bud rot), altered cannabinoid profiles, or improved growth characteristics. Tissue culture provides the essential sterile platform for implementing and propagating these genetically modified plants efficiently.

A New Era of Precision Cultivation

Cannabis tissue culture is poised to profoundly revolutionise the way we grow and understand Cannabis sativa. While historical prohibitions and inherent biological challenges have shaped its development, the recent surge in scientific inquiry and technological innovation is rapidly transforming its potential.

By embracing this advanced approach, cultivators can achieve:

Unprecedented Health and Purity: Starting with certified disease-free material eliminates many common threats, leading to healthier, more vigorous, and reliable plants.
Scalable and Consistent Production: The ability to mass-produce genetically identical clones ensures uniformity in plant growth, cannabinoid, and terpene profiles, which is crucial for a standardised and quality-driven market.
Accelerated Genetic Improvement: Providing a sophisticated platform for advanced breeding, tissue culture significantly accelerates the development of new cannabis varieties tailored for specific purposes, from optimising extract yields to enhancing disease resistance.

The journey of cannabis tissue culture, from its early rudimentary attempts to its current cutting-edge applications, underscores a powerful shift towards a new era of precision cultivation. By leveraging these scientific advancements, growers can unlock the full, incredible potential of Cannabis sativa, ensuring a vibrant, sustainable, and high-quality future for the industry.

Posted on 2 July 20252 July 2025 by Joe

CBG, The Original Cannabinoid

We can all agree that most of the spotlight has traditionally shone on two major compounds: the psychoactive THC and the calming CBD. But as science and consumer curiosity evolve, we are beginning to appreciate the vast and complex family of over 100 cannabinoids found in the plant. Among these, one compound stands out for its foundational role and unique potential: Cannabigerol (CBG).

Often called the “stem cell of all cannabinoids,” CBG is a non-intoxicating compound that acts as the chemical precursor from which many other major cannabinoids are synthesised within the cannabis plant. While it’s typically found in smaller quantities in mature plants, its potential therapeutic benefits and unique interactions with our bodies are generating significant excitement.

This guide will provide a deep dive into what CBG is, how it works, its potential benefits as highlighted by recent research, and its unique place within the broader cannabinoid family.

What is CBG, and Why is it Called “The Stem Cell”?

CBG’s story begins with its acidic form, Cannabigerolic Acid (CBGA). Within the growing cannabis plant, CBGA is the first major cannabinoid acid to form. It serves as a crucial building block. As the plant matures, natural enzymes synthesise other cannabinoid acids from CBGA, primarily:

Tetrahydrocannabinolic Acid (THCA), which becomes THC when heated.
Cannabidiolic Acid (CBDA), which becomes CBD when heated.
Cannabichromenic Acid (CBCA), which becomes CBC when heated.

Because CBGA is the starting point for these major compounds, it is often referred to as the “mother” or “stem cell” cannabinoid. Any CBGA that is not converted into these other forms will, upon heating (decarboxylation), become CBG. This is why most finished cannabis flower contains high levels of THC or CBD but only trace amounts of CBG—most of it has already been transformed.

However, breeders are now developing CBG-dominant strains, and extractors are isolating it, allowing us to explore the unique properties of this foundational molecule on its own. And please, let me say this again. I support the isolation of compounds to study their effects. However, I don’t believe in medication through isolation. Full-spectrum medication is what I believe in.

How Does CBG Interact with Our Bodies? A Unique Mechanism

Like other cannabinoids, CBG interacts with our body’s Endocannabinoid System (ECS), the master regulatory network responsible for maintaining internal balance (homeostasis). But CBG’s method of interaction is distinctly different from that of THC or CBD.

THC primarily acts by directly binding to and activating the CB1 receptor, producing strong psychoactive effects.
CBD has a very low affinity for CB1 and CB2 receptors. As we’ve discussed, one of its key actions is inhibiting the FAAH enzyme, which increases levels of our body’s own anandamide.

CBG, on the other hand, exhibits a broader and more complex range of interactions:

It acts as a partial agonist for both CB1 and CB2 receptors, meaning it can bind to them but doesn’t produce the strong intoxicating effect of THC.
Crucially, research has shown CBG to be a potent alpha-2-adrenergic receptor (α2-AR) agonist. These receptors are involved in regulating sympathetic nerve activity, which controls processes like heart rate and blood pressure. This unique action is not seen with other major cannabinoids and is a key area of research for potential cardiovascular applications.
It also interacts with serotonin receptors (as a 5-HT1A antagonist) and various TRP channels, which are involved in mediating pain, inflammation, and temperature sensation.

This multi-target mechanism means CBG has a unique and versatile potential to influence our physiology in ways that differ from its more famous counterparts. To learn more about CBG, click here to download a Study Review.

Exploring the Potential Benefits of CBG

While human clinical research on CBG is still in its early stages, preclinical studies (in vitro and in animal models) and initial human trials have highlighted several promising areas where CBG may offer therapeutic benefits.

1. Anti-Inflammatory and Antioxidant Properties

Much like CBD, CBG has demonstrated significant anti-inflammatory and antioxidant effects in laboratory settings. It has been shown to reduce the production of inflammatory cytokines and inhibit oxidative stress by neutralising reactive oxygen species (ROS). This action is central to its potential in managing a variety of inflammatory conditions, from skin disorders to inflammatory bowel disease.

2. Potential in Pain Management and Stress Relief

CBG is gaining attention for its analgesic (pain-relieving) properties. A recent double-blind, placebo-controlled clinical trial with healthy adults found that a 20 mg dose of CBG significantly reduced subjective feelings of anxiety and stress compared to a placebo. This human trial corroborates earlier survey data where individuals reported using CBG successfully for managing anxiety.

Furthermore, a pioneering study on horses with chronic osteoarthritis provided compelling evidence. Horses given an oil containing both CBG and CBD showed a significant reduction in pain scores and a decrease in inflammatory markers in their blood, without any adverse side effects. This not only supports CBG’s potential for pain modulation but also highlights its good safety profile in veterinary applications.

3. Neuroprotective Effects

Preclinical research suggests that CBG has neuroprotective qualities, meaning it may help protect nerve cells from damage. This has led to its investigation for neurodegenerative conditions, though this research is still very early.

4. Cardiovascular Health

Because of its unique action as an α2-AR agonist, CBG is being explored for its potential to lower blood pressure. While initial studies in mice have shown hypotensive effects, more research is needed to understand how chronic administration of CBG would affect cardiovascular parameters in humans, especially those with hypertension.

The Bigger Picture: CBG, CBDA, and the Power of the “Entourage”

The recent study on horses is particularly insightful because it didn’t just test one compound. It compared two different formulations: one with CBDA (the acidic precursor to CBD) and another with a combination of CBG and CBD. Both treatments were effective in reducing pain, but the study design underscores a key concept in cannabis science: the “entourage effect.“

Just as terpenes can modulate the effects of cannabinoids, different cannabinoids can work together synergistically. Combining CBG and CBD may offer a broader spectrum of action than either compound alone. For example, CBG’s unique receptor interactions combined with CBD’s well-documented anti-inflammatory properties could create a more comprehensive therapeutic effect.

This highlights the value of full-spectrum or broad-spectrum products, which retain a range of cannabinoids and terpenes, versus isolates, which contain only a single compound.

Your Guide to Exploring CBG

As CBG becomes more available in tinctures, edibles, and even flower, here’s how you can approach it mindfully:

Start with a Reputable Source: Ensure any CBG product you purchase is from a well-respected Rasta, Budtender shop or healer. Have a conversation with the person about the CBG, and ask about lab testing. Ask about sourcing and extraction. When you get an honest answer, you will be able to make an informed decision.
Understand the Dose: As the clinical trial showed, even a relatively low dose of 20 mg can produce noticeable effects on stress and anxiety. Always start low and go slow, especially if you are new to CBG, to gauge your individual response. Trust me on this. My first time smoking CBG flower, I was blown away by how much of a clear mental state it gave me, and it was a little uneasy. Ease yourself into it.
Consider the Goal: Are you looking for stress relief, pain modulation, or general wellness? Your intent can guide your choice between a CBG-isolate product or a broad-spectrum product containing CBG alongside other cannabinoids like CBD.
Manage Expectations: While the research is exciting, it’s still emerging. CBG is not a cure-all, and its effects can be subtle and vary from person to person.

CBG, the “stem cell cannabinoid,” is finally stepping into the spotlight, revealing itself as a compound with a unique and promising profile. As science continues to unravel the complex chemistry of cannabis, we are reminded that there is so much more to this plant than just THC and CBD. Exploring compounds like CBG opens a new chapter in our understanding of how cannabis can contribute to health and well-being.

Posted on 11 June 202511 June 2025 by Joe

Anandamide: Unlocking the “Bliss Molecule”

In the vast landscape of the human body’s biochemistry, few molecules have as intriguing a name as anandamide. Derived from the Sanskrit word “ananda,” meaning “internal bliss” or “joy,” this compound is a cornerstone of a critical regulatory network known as the Endocannabinoid System (ECS). While the ECS gained fame through its connection to cannabis, understanding anandamide itself is key to unlocking the science behind our body’s sense of balance, well-being, and how it responds to cannabinoids like THC and CBD.

This post will dive into what anandamide is, how it functions within your body, and its intricate relationship with cannabis, from the plant’s psychoactive effects to its therapeutic potential.

What is Anandamide?

Anandamide (AEA) is an endocannabinoid, meaning it’s a cannabinoid-like molecule produced inside your body. Discovered in the early 1990s, it was one of the first endogenous “keys” found that fit the “locks” of the cannabinoid receptors, which had just been identified as the primary targets of THC.

Anandamide was the proof. It was the body’s own, internally produced molecule that perfectly fit into the CB1 receptor lock. This confirmed that the cannabinoid receptors weren’t just for cannabis; they were part of a vast, pre-existing communication network essential for our health – The Endocannabinoid System.

So, when we say:

“Anandamide (AEA) was one of the first endogenous ‘keys’ found that fit the ‘locks’ of the cannabinoid receptors, which had just been identified as the primary targets of THC.”

We are saying this:

Scientists first found the “lock” (the CB1 receptor) by seeing where THC from cannabis was binding in the brain.

They correctly assumed our bodies must have a natural reason for these locks.

They then discovered Anandamide – the first internally-produced “key” that our body makes to regulate itself by unlocking these same receptors.

Unlike traditional neurotransmitters that are stored in vesicles and released when needed, anandamide is synthesised on demand. When your body senses a need to restore balance, whether in response to stress, pain, or inflammation, your cells produce and release anandamide. It has a short half-life and is quickly broken down by an enzyme called Fatty Acid Amide Hydrolase (FAAH). This rapid synthesis and degradation allow for precise, localised control over various physiological processes.

The Role of Anandamide and the Endocannabinoid System

Anandamide and the broader ECS act as a master regulatory system, helping to maintain homeostasis (internal balance). Research has shown it plays a vital role in modulating a wide range of functions:

Mood and Anxiety: Studies have consistently linked higher anandamide levels with reduced anxiety. Research in both animal models and humans has shown that elevating anandamide levels, for instance through exercise or by inhibiting the FAAH enzyme, produces anxiolytic (anxiety-reducing) effects. In fact, some individuals with naturally higher anandamide levels (due to a genetic variation in the FAAH enzyme) self-report lower anxiety.
Pain and Inflammation: The ECS is deeply involved in modulating pain signals and inflammatory responses. Anandamide can help regulate these processes, which is why cannabinoid-based therapies are being explored for pain management and inflammatory conditions.
Reward and Motivation: The ECS, including anandamide, plays a modulatory role in the brain’s reward circuitry. It can influence how we experience pleasure and motivation, a factor that is central to research on addiction and substance use disorders.
Sleep: As some of the provided research highlights, anandamide is also implicated in sleep regulation. Studies have shown that administration of anandamide can induce sleep and increase slow-wave (deep) sleep, possibly by influencing adenosine levels, another key sleep-promoting molecule.
Appetite and Metabolism: Anandamide is known to stimulate appetite, a well-known effect also associated with THC.

The Connection to Cannabis: How THC and CBD Interact with Anandamide

The cannabis plant produces phytocannabinoids (plant-based cannabinoids) that interact with our ECS, often by mimicking or influencing anandamide.

THC and Anandamide: THC, the main psychoactive component of cannabis, is a partial agonist of the CB1 receptor – the same receptor that anandamide activates. Essentially, THC fits into the same “lock” as anandamide, but it does so more powerfully and for a longer duration, as it’s not broken down as quickly. This strong activation of CB1 receptors, particularly in the brain, is what produces the euphoric “high” and other effects associated with cannabis, such as increased appetite and altered perception. Chronic, heavy cannabis use can lead to the brain downregulating its CB1 receptors to compensate for this constant stimulation. In turn, this can lead to lower anandamide levels in some individuals, a finding that is particularly relevant in studies of cannabis use disorder.
CBD and Anandamide: Unlike THC, CBD does not bind strongly to CB1 receptors and is non-intoxicating. Instead, one of its primary mechanisms of action is to inhibit the FAAH enzyme, the very enzyme that breaks down anandamide. By slowing down FAAH’s activity, CBD can lead to an increase in your body’s own anandamide levels. This is a crucial distinction: instead of directly activating the receptors like THC, CBD helps boost your natural “bliss molecule.”

This FAAH-inhibiting action is a key hypothesis behind many of CBD’s potential therapeutic benefits. For example, research has explored CBD’s role in treating psychiatric disorders. A recent clinical trial investigated this very mechanism in individuals with cannabis use disorder, looking at how CBD administration affects plasma anandamide levels. The study found that an 800 mg dose of CBD appeared to protect against reductions in anandamide levels that were observed in the placebo group during a cannabis cessation attempt. By potentially increasing anandamide signalling, CBD may help alleviate symptoms of anxiety, psychosis, and withdrawal, offering a promising avenue for treatment.

The Takeaway: A Molecule of Balance

Anandamide is more than just our body’s “bliss molecule”; it is a fundamental regulator of our physiological and psychological well-being. It represents the delicate balance our system constantly strives to maintain. The cannabis plant, through compounds like THC and CBD, offers us a way to directly interact with this system. THC acts as a powerful external key, while CBD works more subtly, by helping our own natural key, anandamide, stay in the lock a little longer.

Understanding the role of anandamide deepens our appreciation for both the complexity of our own biology and the profound ways in which cannabis can influence it. Whether you’re a medical user seeking relief or a recreational consumer exploring different experiences, recognising the interplay between anandamide, THC, and CBD can empower you to make more informed and mindful choices on your cannabis journey.