Ways to Inhibit Elastase for More Elastin?
Nate-2004 15 Dec 2018
Large quantities of oxidants and proteases released by leukocytes that are recruited to the site of inflammation can inactivate these endogenous inhibitors. Moreover, tight adhesion of neutrophils to the ECM leads to the compartmentalization of the released proteases between the neutrophil and the ECM, thereby excluding the large, circulating protease inhibitors. Tight binding of extracellular HNE to the cell membrane can render it inaccessible to circulating endogenous inhibitors. Altogether, the imbalance between HNE and its inhibitors caused by these events provokes severe tissue injuries resulting in a variety of diseases.
It has become clear that serine proteases, such as HNE, have an important regulatory role in the local inflammatory response. Thus, its dysregulation resulting in its accumulation can be involved in the development of chronic inflammatory diseases, such as rheumatoid arthritis, pulmonary emphysema, adult respiratory distress syndrome (ARDS), cystic fibrosis, COPD, asthma, and delayed wound healing.
However, it has to be kept in mind that flavonoids are highly metabolized during oral application, and that some possible metabolites (such as 4-methylcatechol, 4-hydroxyphenylacetic acid and 3,4-dihydroxyphenylacetic acid ) exhibited a very low activity in the assay (IC50 range from 135 qM to > 400 juM, see Fig. 2A and Fig. 2B). Therefore, the in vitro studies mentioned above may be of limited therapeutic relevance and it is questionable that flavonoids may be orally active principles in a Drosera extract used to treat cough.
Based on these results a formulation of oleic acid with albumin was developed for the treatment of chronic wounds. Albumin was used as a carrier for the hydrophobic oleic acid. Oleic acid/albumin formulations with mole ratios of 100:1, 50:1, and 25:1 showed a strong inhibition of HNE with IC50 values at 0.029-0.049 /iM. Albumin alone increased to a small extent the substrate conversion by HNE, which could be equalized by a higher concentration of inhibitor. The authors suppose that an increase of the albumin concentration may even have positive effects, since albumin level is decreased in chronic wounds. The formulation was still active (IC50 = 0.26-0.42 /iM), even after being bound to derivatized cotton.
Edited by Nate-2004, 15 December 2018 - 06:06 PM.
Nate-2004 15 Dec 2018
I dunno about well written, already seeing two errors (break down elastin not elastase and also, "this is largely" not "this is part largely") but thanks.
Wondering if I can do anything topical with mango butter.
Nate-2004 18 Dec 2018
What references are missing??
Working on a formulation for a serum based on the above info. Here's what I'm going to try:
EGCG is soluble at 100g per liter of distilled water. That's a nice 1/10th. I wouldn't go that far with it though because I'd want to add other ingredients for ease of application.
Note: I haven't tried this yet so I don't know how well it will go.
Part 1:
350ml distilled water heated to boiling
Mix in around 20 to 25g EGCG and let it dissolve thoroughly as it cools
Optional: Add 3g niacinamide and stir
Add in 3.5g hyaluronic acid powder, stir and then refrigerate 12 to 24 hours
The result should be a gel or a slightly thickened brown liquid.
Part 2: Here's where I'm not so sure this is how to go about it.
Melt 50g mango butter in a double boiler
Pour the mango butter into the gel.
Mix thoroughly.
If it's too thick or not the consistency of thick liquid at room temperature it might need to be reheated in the double boiler and mixed with more of the hot distilled water and EGCG made in part 1
Edited by Nate-2004, 18 December 2018 - 09:18 PM.
Nate-2004 19 Dec 2018
Interesting article here: https://www.ncbi.nlm...les/PMC3189735/
EGCG apparently has a much, much better bioavailability if taken in a fasted state with ascorbic acid and fish oil.
Unfortunately for anyone taking this while fasting, EGCG also happens to suppress hepatic gluconeogenesis, so look forward to feeling pretty low energy.
Edited by Nate-2004, 19 December 2018 - 05:44 PM.
Nate-2004 19 Dec 2018
I'm still debating about how to create a transdermal gel for EGCG. There are companies that sell Lipoderm which has been tested with EGCG but I have no access to this.
I am also unsure about whether adding any amount of DMSO is a good idea, because sulfation is an issue with EGCG. If anyone knows an ideal way to improve transdermal absorption that would be great. Some transdermal drug delivery systems use ethanol, or "nanoethosomes", which is a bit vague. I worry that ethanol would dry out the skin though.
Coincidentally I learned that oleic acid is a permeation enhancer, which means the mango butter may help more, plus I have soy lecithin already mixed in with the mango butter I have. Adding a small amount of ethanol probably wouldn't hurt much. I cannot find any information on how nanoethosomes are produced but it definitely involves ethanol, phospholipids and water. Possibly requires a scanning and tunneling microscope so that's out.
Edited by Nate-2004, 19 December 2018 - 06:26 PM.
Nate-2004 24 Dec 2018
Here's something else I can't shake as what should be a really solid question. If an overrun of HSE is one of the major causes of loss in elastin production, and oleic acid is one of the strongest inhibitors, then why on earth aren't oleic acid rich foods like walnuts reversing atherosclerosis and wrinkles?
Well according to this study walnuts do actually improve endothelial function... https://www.ahajourn...124477.91474.FF
Maybe this is why, considering that the unsaturated fatty acids are said to be the active ingredient. However this study highlights a big point that elastin loss isn't the ONLY issue affecting the stiffening of tissues. If it was, wouldn't we see reversal? Or maybe it's because it's just not as effective as it needs to be. Perhaps our endogenous elastase inhibitors trapped by the strong adhesion of neutrophil to the ECM is what needs to be addressed. Not only that, I'm sure it all ties into glycation and AGEs being the other big issue.
Nate-2004 24 Dec 2018
So in my experimentation with the first batch of topical EGCG and Oleic, I have the same problem as I had trying to just mix it into ready made CeraVe lotion. It doesn't dissolve in anything but water and the moment you put it in anything with lipids it begins to ball up into little balls of wet extract powder. I'm not sure if there's a word for that. It seems ok with just the hyaluronic acid serum so long as I mix it with glycerin to keep it from just making a film on my skin. I'll have to test this more but I don't think it's going to carry through well. You're also missing out on the oleic acid.
Maybe I need a better EGCG extract than 50%
Also,
I don't know if anyone knows about sulfation, but wouldn't DMSO destabilize the EGCG?
Edited by Nate-2004, 24 December 2018 - 01:52 AM.
Nate-2004 27 Dec 2018
Know what else destabilizes EGCG? HOT WATER! WTF?? My first question was how the hell did they come to the conclusion that EGCG polyphenols are why green tea, which is brewed in HOT WATER, is good for you?
Well turns out it just needs to be cooled fast within 30 mins, or put on ice. Luckily I think I got it into the cold fridge pretty quick after mixing in the hyaluronic acid. So gotta remember to do that, or better yet put it on ice as they did here. Or perhaps heating it isn't even needed to mix it in thoroughly.
http://www.vup.sk/en...d.php?bulID=786
I wonder if I'd solve the clumping issue if I drained it through a cheesecloth beforehand.
Edited by Nate-2004, 27 December 2018 - 04:35 AM.
Nate-2004 03 Jan 2019
Got some help with the formulation from someone who's obviously better at this:
There are a few issues or concerns. I will state the normal practices that the formulators will have.
Normally when we write out the formulation for troubleshooting, we will write it out in percentages. I've converted it for the sake of the process, which you can double check if required. I've also written the ingredients in different phases for easy preparation. We will also state the required ending pH as well.
Formulation:
Oil Phase
- Lecithin - 14.78g | 4%
- Mango Butter - 59.15g | 15%
Water Phase
- Distilled Water - 300g | 74%
- EGCG - 15g | 4%
- HA - 3g | 1%
- Glycerin - 14.78g | 4%
Preservative Phase
- Potassium Sorbate
Few things to take note:
- Lecithin may not be a strong emulsifier as it's normally used as a co-emulsifier in skin care. Some expert formulator will advice not using it at all as it increases the chances of bacteria and mould growth. You can choose a proper emulsifier (if you require it to be natural), such as Olivem 1000, Montanov 68, etc
- For the EGCG, you can make an extract out of it, depending on the concentration that i had mentioned earlier. You can take a look at this video here. If you decide to make it with just Distilled Water, you will need to continue straining with a finer filter paper.
- As you didn't measure the pH, Potassium Sorbate has a preservative will not be effective as it's most effective if the pH is below pH 5. Any pH above pH 5, the Potassium Sorbate does not have any anti-mould activity.
- If you insist on using all natural or natural derived ingredients, then i will take it that you will use a natural type preservative.
Formulation that i will suggest based on your ingredients + natural derived ingredients only:
Oil Phase
- Mango Butter - 15% (Emollient)
- Olivem 1000 - 5% (Emulsifier)
- Glyceryl Stearate - 1% (Co-emulsifier)
- Lecithin - 2% (Co-emulsifier)
Water Phase
- Distilled Water - 64.3%
- HA - 1%
- 1,3-Propanediol 3% (Humectant / Preservative Booster)
- Sodium Phytate - 0.2% (Chelating Agent)
Cool Down Phase
- EGCG Extract - 4%
Preservative
- Leucidal Liquid - 4% (Anti bacterial preservative)
- Potassium Sorbate - 0.25% (Anti mould preservative)
- Sodium Benzoate - 0.25% (Anti mould preservative)
Targeted pH: pH 5 (For the anti mould preservatives to work)
Steps:
- Sift the HA into the Distilled Water in a heat proof container and let it hydrate for 1 hour
- Mix the oil phase into a heat proof container
- Mix the remaining water phase ingredients into the premixed HA liquid
- Use the heat and hold method, heat both phases till 65 - 75 degrees (until the emulsifier melts)
- Pour the water phase into the oil phase and blend using a electric blender (preferably a stick blender) till emulsified
- When below 40 degrees, add the Cool Down Phase & Preservatives, blend till well mixed
- Adjust using a citric acid (50%) solution or lactic acid solution to pH 5 using a pH strip or pH meter for measurement
Read more about formulation here: