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Showing posts with label Liposomal. Show all posts
Showing posts with label Liposomal. Show all posts

Thursday, 14 August 2025

Home-made Liposomal EGCG — a cost effective therapy for Autism, Parkinson’s, and Alzheimer’s? Plus alternative antioxidants — Whey protein and Liposomal vitamin C


A $30 ultrasonic jewellery cleaner can be repurposed to make inexpensive liposomal supplements

 

Today’s post is really one for those who prefer not to use prescription drugs to treat autism, or those that are just unable to access them. It is also one our longtime reader Ling might regard as MacGyver-esque (from the TV series following the adventures of Angus MacGyver, a secret agent armed with remarkable scientific resourcefulness to solve any problem out in the field using any materials at hand).

It is about increasing the bioavailability of OTC supplements (EGCG in today’s case, but applicable to many others) to get closer to achieving their often elusive health benefits in autism.

There are some effective OTC autism therapies, but most are not. This is why repurposing existing prescription drugs is likely necessary.

 

Liposomal

One of the big things in the supplement world at the moment is to call products “liposomal” and triple the price. The theory is that a preparation contains the active drug/supplement inside very tiny, fat-like particles. This form is easier for the body to absorb and allows more drug/supplement to get to the target area of the body, such as the brain. Liposomal drugs may have fewer side effects and should, in theory, work better than other forms of the drug.

This fatty encapsulation helps protect the active compound from degradation in the digestive system and improves its absorption through the gut. It can also enhance delivery to target tissues (like the brain) because liposomes can sometimes cross biological barriers more easily.

This should mean higher effectiveness with lower doses and potentially fewer side effects compared to non-encapsulated forms.

 

If you are interested in the details:

https://en.wikipedia.org/wiki/Liposome

“A liposome is a small artificial vesicle, spherical in shape, having at least one lipid bilayer. Due to their hydrophobicity and/or hydrophilicity, biocompatibility, particle size and many other properties, liposomes can be used as drug delivery vehicles for administration of pharmaceutical drugs and nutrients, such as lipid nanoparticles in mRNA vaccines, and DNA vaccines. Liposomes can be prepared by disrupting biological membranes (such as by sonication).

Liposomes are most often composed of phospholipids, especially phosphatidylcholine, and cholesterol, but may also include other lipids, such as those found in egg and phosphatidylethanolamine, as long as they are compatible with lipid bilayer structure. A liposome design may employ surface ligands for attaching to desired cells or tissues.”

 

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By making your own liposomal supplements you will save a lot of money, compared to commercial ones and have access to an undegraded product. If you customize the recipe/ingredients thoughtfully, and carefully control the processing, the result might replicate some of the benefits seen in university studies. You might wonder why compounding pharmacies are not already doing this - maybe some are.

You can pretty much buy everything you need on Amazon. Once you have figured out your ingredients and decided how big a batch to make, it is no more complex than baking a cake.

 

Liposomal vitamin C and whey protein as therapies for oxidative stress

Oxidative stress is a core feature of most autism, particularly in the early years, and a feature of aging for everyone. Vitamin C is a natural antioxidant, but it is a water soluble vitamin that your body automatically regulates and excretes via urine. If you take mega-doses of a standard supplement it just goes down the toilet, it does not reach the bloodstream.

Intravenous vitamin C causes a large increase in levels in the blood. This can be used to treat sepsis and even mast cell activation syndrome (MCAS). It has potential in oncology (cancer treatment) because at high concentrations, vitamin C can act as a pro-oxidant, generating hydrogen peroxide that is selectively toxic to tumor cells.  

It has also been used for Ehlers-Danlos syndrome, fibromyalgia and other conditions

Some practitioners consider IV vitamin C for autism because of its: 

  • Antioxidant effects – reducing oxidative stress, which is elevated in many children with autism.
  • Anti-inflammatory properties – calming neuroinflammation and microglial activation.
  • Support for neurotransmitter synthesis – vitamin C is a cofactor in dopamine and norepinephrine production.
  • Possible mast cell stabilization – relevant in children with autism and comorbid mast cell activation syndrome (MCAS).
  • Histamine degradation support – helps recycle tetrahydrobiopterin (BH4), indirectly involved in histamine metabolism.

 

It has been found that liposomal vitamin C can achieve levels in the blood somewhere in between IV-vitamin C and regular vitamin C by food or supplements. 

High levels of vitamin C can cause side effects such as kidney stones.

Liposomal vitamin C is better tolerated than very high doses of standard vitamin C. It looks like things are likely to start going wrong above 3,000mg a day of liposomal.

Healthy people just need a good diet. If they have a poor diet then take a multivitamin.

Liposomal or IV therapy is only for people with real health issues.

People with MCAS plus autism certainly do have health issues.

Ehlers–Danlos syndrome (and milder subclinical versions) is linked to MCAS, ADHD, autism and Tourette’s. So that is another group to consider.

Fibromyalgia was put forward (by me) as a step towards autism in some females, in subsequent levels of their family tree.

So overall the idea of liposomal vitamin C has much more merit than a natural sceptic would have first thought. (There are loads of YouTube videos of people doing this, and likely many did not really need it.)

 

Whey protein as an antioxidant 

This topic was recently highlighted by our reader Stephen and it naturally fits into this post.

Back in 2013 when I was developing my son’s therapy I had to choose between NAC and whey protein to boost glutathione (GSH), the body’s key antioxidant. I chose NAC.

Here is a great paper to support the use of whey protein.

 

Improving Antioxidant Capacity in Children With Autism: A Randomized, Double-Blind Controlled Study With Cysteine-Rich Whey Protein 

Previous studies indicate that children with autism spectrum disorder (ASD) have lower levels of glutathione. Nutritional interventions aim to increase glutathione levels suggest a positive effect on ASD behaviors, but findings are mixed or non-significant. A commercially available nutritional supplement comprising a cysteine-rich whey protein isolate (CRWP), a potent precursor of glutathione, was previously found to be safe and effective at raising glutathione in several conditions associated with low antioxidant capacity. Therefore, we investigated the effectiveness of a 90-day CRWP intervention in children with ASD and examined whether intracellular reduced and oxidized glutathione improvements correlated with behavioral changes. We enrolled 46 (of 81 screened) 3-5-year-old preschool children with confirmed ASD. Using a double-blind, randomized, placebo-controlled design, we evaluated the effectiveness of daily CRWP (powder form: 0.5 g/kg for children <20 kg or a 10-g dose for those >20 kg), compared with placebo (rice protein mimicking the protein load in the intervention group), on glutathione levels and ASD behaviors assessed using different behavioral scales such as Childhood Autism Rated Scale, Preschool Language Scale, Social Communication Questionnaire, Childhood Behavioral Checklist and the parent-rated Vineland Adaptive Behavior Scale, 2nd edition (VABS-II). Forty children (CRWP, 21; placebo, 19) completed the 90-day treatment period. Improvements observed in some behavioral scales were comparable. However, the VABS-II behavioral assessment, demonstrated significant changes only in children receiving CRWP compared to those observed in the placebo group in the composite score (effect size 0.98; 95% confidence intervals 1.42-4.02; p = 0.03). Further, several VABS-II domain scores such as adaptive behavior (p = 0.03), socialization (p = 0.03), maladaptive behavior (p = 0.04) and internalizing behavior (p = 0.02) also indicated significant changes. Children assigned to the CRWP group showed significant increases in glutathione levels (p = 0.04) compared to those in the placebo group. A subanalysis of the VABS-II scale results comparing responders (>1 SD change from baseline to follow up) and non-responders in the CRWP group identified older age and higher levels of total and reduced glutathione as factors associated with a response. CRWP nutritional intervention in children with ASD significantly improved both glutathione levels and some behaviors associated with ASD. Further studies are needed to confirm these results.

 

This study used a special commercial product called Immunocal, a cysteine-rich whey protein isolate (CRWP) that serves as a potent glutathione precursor.

There are less expensive alternatives to Immunocal that still offer high-quality, undenatured, cysteine-rich whey protein, especially if your goal is to support glutathione production without paying premium prices. These products are typically marketed as cold-processed, non-denatured whey protein concentrates or isolates, and some are even made from the same raw material sources as Immunocal.

If you want to further increase absorption you can even make a liposomal version of a cysteine-rich whey protein!! 

Regular body builders’ whey protein is great to help build muscles and to maintain muscle mass in seniors, but it is not the ideal source of cysteine. It has degraded during the production process, that why there are fancy ones available.

I think Stephen would indeed be well advised to add a scoop of cysteine-rich whey protein isolate (CRWP) to his sons’ diets. It should have a more prolonged effect than NAC. For young children with autism NAC really needs to be given 3-4 times a day.

You can have too much cysteine. You do not need high dose of both NAC and CRWP.

 

Back to liposomal EGCG

If you read the reviews many people find commercial liposomal supplements no more effective than the much cheaper, regular ones. I wonder why. Most likely they were not well formulated, or they degraded by the time they were used. These products are not heat or light stable.

Many manufactured products like fish oil supplements no longer maintain the health benefit of the genuine article (fish, in this case). This is because the product degraded and sometimes can even have a negative behavioral effect. 

 

Many healthy natural products like catechins or curcuma have very low bioavailability

There is a long list of healthy products that should be therapeutic in autism including:

·        Green tea catechins like EGCG

·        Turmeric/Curcuma

·        Resveratrol

·        Cocoa

·        Many herbs (sage, oregano, rosemary, Bacopa monnieri, ginseng, lions mane, etc)

They generally have very low bioavailability and so they work great in the lab, but much less so in humans; unless you consume very large amounts, for example turmeric in an Indian diet.

 

EGCG

I have written about EGCG in the past and have highlighted the research from Spain, more specifically from the beautiful city of Barcelona (just avoid visiting during the peak summer months). The research showed a benefit in Fragile X and Rett syndrome. As usual, no customized intervention has yet been brought to the market.

https://www.epiphanyasd.com/search/label/EGCG?max-results=20


Yet another study showing the potential benefit of EGCG, was published recently, this time in Pakistan.

 

Cross-linking catechins with neuro-regulatory model for autism spectrum disorder: A management in rats’ experiment 

We found that BDNF levels returned to normal levels within the groups who received Catechins treatment at III, IV, and V concentrations (compared to Group II), showing Catechins could potentially treat autism-like symptoms. The BDNF values measured in nano-grams per millilitre were Group I (13.1±0.3), followed by Group II (5.1±0.2) and Group III (9.8±0.3), Group IV (8.0±0.3), and then Group V (10.1±0.3). The BDNF concentration measured in Groups III, IV and V surpassed the BDNF level of Group II (PPA-induced) per results from a post-hoc Tukey's test at p 

Catechins successfully decreased neuroinflammatory markers throughout the brain and establish protective brain mechanisms that potentially improve ASD-associated behavioral symptoms. Rats given 100, 200, and 400 mg/kg of various catechins showed increases in BDNF levels of up to 75%, 61%, and 77%, respectively, as opposed to only 39% for rats that received no treatment. The findings of a study suggested a continuous and expandable neuroprotective effect based on dose strength. The experimental results demonstrated that in ASD models, catechins offer a potent and dosage-dependent defense against neuroinflammatory injuries.

  



This study confirms that epigallocatechin gallate (EGCG), among catechins, shows great promise for managing neuroinflammation in ASD patients. The results indicate that catechins deliver substantial reductions in neuroinflammatory markers, as they serve as protective element that improves behavioral and cognitive manifestations of ASD. Future investigations must explore mechanisms of effect and find best-use dosages for catechins while establishing their safety and lasting effect durations.

 

Then I came across this paper where the university made their own liposomal version of EGCG and tried it on their model of Parkinsons’ disease. It also worked very well. Autism is not Parkinsons’ but both conditions feature activated microglia, the brain’s immune cells that are also tasked with synaptic pruning housekeeping duties.

 

Epigallocatechin-3-Gallate-Loaded Liposomes Favor Anti-Inflammation of Microglia Cells and Promote Neuroprotection

Microglia-mediated neuroinflammation is recognized to mainly contribute to the progression of neurodegenerative diseases. Epigallocatechin-3-gallate (EGCG), known as a natural antioxidant in green tea, can inhibit microglia-mediated inflammation and protect neurons but has disadvantages such as high instability and low bioavailability. We developed an EGCG liposomal formulation to improve its bioavailability and evaluated the neuroprotective activity in in vitro and in vivo neuroinflammation models. EGCG-loaded liposomes have been prepared from phosphatidylcholine (PC) or phosphatidylserine (PS) coated with or without vitamin E (VE) by hydration and membrane extrusion method. The anti-inflammatory effect has been evaluated against lipopolysaccharide (LPS)-induced BV-2 microglial cells activation and the inflammation in the substantia nigra of Sprague Dawley rats. In the cellular inflammation model, murine BV-2 microglial cells changed their morphology from normal spheroid to activated spindle shape after 24 h of induction of LPS. In the in vitro free radical 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay, EGCG scavenged 80% of DPPH within 3 min. EGCG-loaded liposomes could be phagocytized by BV-2 cells after 1 h of cell culture from cell uptake experiments. EGCG-loaded liposomes improved the production of BV-2 microglia-derived nitric oxide and TNF-α following LPS. In the in vivo Parkinsonian syndrome rat model, simultaneous intra-nigral injection of EGCG-loaded liposomes attenuated LPS-induced pro-inflammatory cytokines and restored motor impairment. We demonstrated that EGCG-loaded liposomes exert a neuroprotective effect by modulating microglia activation. EGCG extracted from green tea and loaded liposomes could be a valuable candidate for disease-modifying therapy for Parkinson’s disease (PD).

 

Looks great, but you cannot buy their product. It then appeared that people are already making liposomal supplements at home.

Dig a little deeper to see what other clever ideas exist in the university research world that might make DIY versions better. 

 

Liposomal Formulations for an Efficient Encapsulation of Epigallocatechin-3-Gallate: An In-Silico/Experimental Approach

As a part of research project aimed to optimize antioxidant delivery, here we studied the influence of both salts and lipid matrix composition on the interaction of epigallocatechin-3-gallate (EGCG) with bilayer leaflets. Thus, we combined in silico and experimental methods to study the ability of neutral and anionic vesicles to encapsulate EGCG in the presence of Ca2+ and Mg2+ divalent salts. Experimental and in silico results show a very high correlation, thus confirming the efficiency of the developed methodology. In particular, we found out that the presence of calcium ions hinders the insertion of EGCG in the liposome bilayer in both neutral and anionic systems. On the contrary, the presence of MgCl2 improves the insertion degree of EGCG molecules respect to the liposomes without divalent salts. The best and most efficient salt concentration is that corresponding to a 5:1 molar ratio between Mg2+ and EGCG, in both neutral and anionic vesicles. Concerning the lipid matrix composition, the anionic one results in better promotion of the catechin insertion within the bilayer since experimentally we achieved 100% EGCG encapsulation in the lipid carrier in the presence of a 5:1 molar ratio of magnesium. Thus, the combination of this anionic liposomal formulation with magnesium chloride, avoids time-consuming separation steps of unentrapped active principle and appears particularly suitable for EGCG delivery applications.

 

The Mozafari method for Liposomal delivery

The latest methods used in universities to make liposomal products cannot be entirely replicated at home, but there is a well-known method developed by Dr Mohammad Mozafari that has been proved to increase bioavailability 2 to 8 times. The Mozafari method is used today by biohackers at home. Often they seem to skip some important steps.

We can fine tune his method, for example by noting the research showing that magnesium ions can help stabilize the liposomes and improve encapsulation of EGCG. Calcium ions have a very negative effect and so make sure no calcium (for example, from hard water) enters the process. YouTubers just use tap water. So use high-quality deionized (DI) water and add Magnesium Chloride (MgCl₂).

Anionic liposomes (negatively charged phospholipids) promote better EGCG insertion compared to neutral liposomes. With Mg²⁺, anionic liposomes reached 100% encapsulation efficiency experimentally. So it was actually perfect.

Magnesium chloride (MgCl₂) at about a 5:1 molar ratio relative to EGCG

(Example: for 500 mg EGCG ≈ 1.1 mmol, add ~5.5 mmol MgCl₂ — roughly 670 mg MgCl₂·6H₂O)

Both pH and temperature control are important and seem to get ignored by YouTubers.

Choose the right lipid. Here are the choices:

Most DIYers are using Lecithin (sunflower or soy), which contains phosphatidylcholine (PC), plus other substances you do not want. It is cheaper than pure PC.

If you are making liposomal vitamin C, glutathione, DHA or EGCG for therapeutic use (e.g., autism, MCAS, oxidative stress), pure PC gives superior performance.

Lecithin is zwitterionic, meaning it contains both positive and negative charges, but is overall electrically neutral. This dual nature is what makes lecithin perfect for encapsulating both water-soluble (like vitamin C) and fat-soluble (like curcumin) compounds in liposomes.

For closer to University-grade work we need to look at pure chemicals.

·        Phosphatidylcholine (PC) — neutral

·        Phosphatidylserine (PS) — anionic (negative charged)

·        CHEMS (Cholesteryl Hemisuccinate), a negatively charged cholesterol derivative.

·        Cholesterol

 

Component             Role  

PC                               Bilayer structure & fluidity         

PS                               Anionic charge, Mg²⁺ interaction          

Cholesterol               Stabilization (optional)    

CHEMS                      Additional anionic charge (optional)

 

Phosphatidylserine (PS) is itself therapeutic

PS naturally concentrates in the brain, especially in neuronal membranes.

It is known to support memory, attention, synaptic function, and neuroplasticity — ideal for neurodegenerative and developmental conditions.

PS is negatively charged (anionic), which helps form stable liposomes and can improve encapsulation of positively charged or hydrophilic molecules like EGCG.

PS has functional activity, beyond just being a carrier, PS itself may synergize with EGCG and other cognitive-enhancing compounds.

Adding cholesterol makes the liposome less leaky and more resistant to degradation. Without cholesterol, liposomes are more prone to oxidation, fusion, or breakdown over time

  

Example for 2 g Total Lipids:

Lipid Component

Weight (grams)

Percentage

PC

1.2 g

60%

PS

0.4 g

20%

Cholesterol

0.4 g

20%

 

  • PC provides a stable bilayer and good liposome formation.
  • PS introduces a negative charge that enhances electrostatic interaction with Mg²⁺ and EGCG.
  • Cholesterol improves membrane rigidity and stability, helping prevent leakage.
  • You can adjust cholesterol slightly depending on how rigid you want the membrane.
  • Maintain MgCl₂ at ~5:1 molar ratio to EGCG in the aqueous buffer for optimal encapsulation, as per references.

EGCG is highly oxidation-sensitive.

Both vitamin C (ascorbic acid) and vitamin E (tocopherol) protect:

·        the lipids in the liposome from peroxidation,

·        the EGCG itself from degradation.

So it is wise to add both vitamin C and E.

  • Vitamin E is lipid-soluble and embeds in the bilayer.
  • Vitamin C is water-soluble and protects the aqueous core.

 

Here is the home version.

 

 

Equipment

  • Glass beaker or jar
  • Ultrasonic cleaner (sold to clean jewellery)
  • Stirring rod
  • pH strips or meter
  • Dark glass storage bottle

 

Method

1.     Mix Vitamin E with PS

o    Combine PS powder and vitamin E oil or powder thoroughly in a small container.

2.     Prepare aqueous phase

o    Dissolve EGCG powder and magnesium chloride in ~20 mL PBS or distilled water with buffer salts.

o    Add vitamin C to this aqueous solution last and stir gently until dissolved.

3.     Hydrate lipids

o    Slowly add the aqueous phase (EGCG + MgCl₂ + vitamin C) to the PS + vitamin E mix.

o    Stir or vortex gently to disperse.

4.     Sonicate

o    Place the mixture in an ultrasonic cleaner bath for 20–30 minutes, stirring occasionally.

o    Solution should become milky/opalescent, indicating liposome formation.

 

How to Use Ultrasonic Cleaner for Liposomal EGCG

1.     Prepare your liposome suspension in a suitable sealed container—usually a small glass vial or bottle with a tight lid (e.g., amber glass bottle or glass vial).

2.     Fill the ultrasonic cleaner tank with clean water—enough so that when you place your container in it, the water level reaches just below the lid or about 2/3 up the container’s height. The water must not overflow into your liposome container

3.     Place your sealed bottle/vial into the ultrasonic bath, making sure it sits upright and stable.

4.     Turn on the ultrasonic cleaner for the recommended at medium power.

5.     During the process, keep an eye on the temperature—if the water or sample gets too warm (>40°C), pause and let it cool, since heat will degrade EGCG.

6.     After sonication, remove the bottle and store the liposomal EGCG in a dark, refrigerated place.

 

Important Tips

  • Use sealed containers to avoid contamination or water ingress.
  • Never put the liposomal suspension directly into the ultrasonic cleaner’s water bath.
  • If your ultrasonic cleaner has a temperature control or timer, use those settings to protect the sample.
  • Clean the ultrasonic tank well before and after use.


The final product will be stable for 7 days in the fridge.

You can freeze portion sized doses in a silicone ice cube tray. Later store in the freezer in a zip lock bag for 2-3 months. Defrost in the fridge, one by one, as you need it.

Keeping the temperature below 40°C is essential when sonifying delicate compounds like EGCG, vitamin C, and phospholipids (especially phosphatidylserine). They degrade or oxidize easily when exposed to excessive heat.

 

 1. Use a Cold Water Bath

  • Fill the ultrasonic cleaner with cold water (4–10°C).
  • Add ice cubes to keep it cold.
  • Replenish ice as needed during sonication.

 

2. Monitor Temperature

  • Use a probe thermometer or an infrared laser thermometer.
  • Check the temperature of your sample, not just the water bath.

 

Ultrasonic waves create cavitation — rapid formation and collapse of microbubbles — which:

  • Generates localized heat (tiny hot spots in the solution)
  • Transfers energy into the liquid, raising the overall temperature gradually
  • Can increase your solution temperature from room temp to 50–60°C in 10–15 minutes if not managed

Even if the water bath feels lukewarm, the inside of your beaker can be much hotter, especially in the center.

 

 Sensitive Ingredients at Risk

  • EGCG starts to degrade above ~40°C
  • Phospholipids can oxidize or become unstable
  • Vitamin C rapidly degrades in heat and light

 

Best Practices to Avoid Overheating

Tip

Why

Use an ice-cold bath

Keeps temp under control during sonication

Sonicate in short bursts (30s ON, 30s OFF)

Prevents buildup of heat

Stir gently before and after sonication

Helps distribute heat and emulsify

Monitor temperature regularly

Keeps you in the safe zone (under 40°C)

Work with small batches (≤100 mL)

Easier to cool and control

 

Sonication is the process of using high-frequency sound waves—usually ultrasound—to agitate particles in a liquid. In making liposomes, sonication helps break down large lipid aggregates into smaller, uniform liposomes by creating tiny bubbles that collapse and generate energy, mixing and dispersing the components more thoroughly.

In simpler terms, sonication is like using sound waves to gently shake and break up the mixture so that the fat-like particles (liposomes) form smaller, more consistent droplets, which can improve how well the active ingredient (like EGCG) gets absorbed by your body.

  

Conclusion

Immunocal, a cysteine-rich whey protein isolate (CRWP) does show a benefit in autism. It is an OTC product that is sold online.

EGCG is the most abundant catechin in green tea.  When taken orally, EGCG has poor absorption even at daily intake equivalent to 12 cups of green tea.

EGCG does show merit in autism research studies, but to be effective in humans the bioavailability needs to be improved. Using liposomes to encase the EGCG is a promising approach.

Will the method proposed in this post be effective?

I asked Gemini AI to comment on the method and it replied: 

"The EGCG method you described is a fantastic example of a DIY approach that is grounded in sound scientific principles. By using specific lipids, optimizing the formulation with magnesium, and strictly controlling the process, it goes far beyond the simplistic and often ineffective methods seen online.

While it won't produce a pharmaceutical-grade product, it is highly likely to create a solution that contains a significant number of genuine, stable, and correctly sized liposomes. This makes it far more likely to be an effective delivery system than the typical DIY liposomal vitamin C, which is often just an unverified emulsion."

Some people do grow their own broccoli sprouts to make sulforaphane, others grow wheat sprouts for spermidine. Some people grow their own probiotic bacteria. Making tumeric balls is a simple way to get the benefits of tumeric. There are many home-made options, and I think the parent almost certainly benefits. 

You would think that some enterprising pharmacist in Barcelona would start producing small batches of liposomal EGCG, using research grade equipment. I think Rett syndrome and Fragile X syndrome parents would buy it. Not to mention those who have parents diagnosed with Parkinson's or Alzheimer's.