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Having difficulty making the data in the below charts to apply to daily life. What is the right interpretation and implementation? How are the “in vitro” results of the below charts relevant to levels that cells would experience "in vivo" after the consumption of caffeinated beverages?

Fig 1

Fig 2

Link to the medical research article which the two above charts were taken from

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  • $\begingroup$ Good - hope you get some useful answers. I could add that in the lab I work in, we often do in vitro experiments with cells or tissue in a dish, and it's a normal part of our daily business to ask questions exactly like this - it's a great question, though can also be difficult to answer sometimes. The science that figures out answers to these types of questions is called "pharmacology", by the way. $\endgroup$ – Bryan Krause Jan 24 at 3:35
  • $\begingroup$ You're asking how skin growth rate is affected in vivo "confluent fibroblasts" by caffeine and it looks like that without the factors and matrix growth environment, that the caffeine metabolically stimulates the fibroblasts away from confluence in graph 1. $\endgroup$ – W4t3randWind Jan 28 at 9:07
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In the mentioned study, they tried to support the evidence from earlier studies in which caffeine consumption was associated with skin aging and slow wound healing:

Our research confirms our earlier observations that caffeine may have an adverse effect on the wound healing process, as well as on the aging process, of the human skin.

...and by "earlier observations" they mean another in vitro study.

In the study, they used 1, 2 and 5 mM caffeine solutions. 1 cup (~250 mL) of brewed tea can contain 95-165 mg caffeine (source), which makes it 2-3.4 mM, so much like the solutions used in the study. When you drink 1 cup of coffee (let's say, 2 mM caffeine solution), the caffeine, which is water- and lipid-soluble (StatPearls), will be dissolved in the body water and fat, which can together make ~50 kg in a 70 kg person. So, you would need to drink ~200 cups of coffee (~20 g caffeine) in ~2 hours (caffeine half-life is ~5 hours - source) to achieve 2 mM caffeine solution in the skin cells.

I haven't found any actual human clinical trial in which caffeine consumption would be associated with adverse effects on the skin. In a recent review: Coffee consumption and health: umbrella review of meta-analyses of multiple health outcomes (BMJ, 2017), they don't even mention skin or wound healing and regular drinking of 3-4 cups of coffee per day was not associated with an increased risk of various health conditions (except in pregnancy).

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    $\begingroup$ Since caffeine is water soluble, wouldn't you have to consume (approximately) 194 mg per liter of water to reach that concentration in skin? That is above the LD50 for caffeine. $\endgroup$ – user1850479 Jan 24 at 14:55
  • $\begingroup$ I edited the bold paragraph about the amounts. The concentrations of caffeine solutions used in the study and in coffee seem to be about the same, but when you drink a cup of coffee, caffeine would be dissolved in the whole body and would not be so concentrated in the skin cells any more. I think you would need unrealistic amounts of coffee to reach such concentrations. $\endgroup$ – Jan Jan 24 at 14:59
  • $\begingroup$ The LD50 of caffeine is 150-200 mg per kilogram of body weight. So, the lethal dose for a 70 kg man would need to be at least 10 grams of caffeine (~100 cups of coffee) consumed in a short time. $\endgroup$ – Jan Jan 24 at 15:19
  • $\begingroup$ Ah yes, weight not volume, so the 2 molar solution is about the LD50. $\endgroup$ – user1850479 Jan 24 at 15:22
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How can one relate in vitro studies of caffeine (dose expressed as concentration) to dietary intake of caffeine (dose in mass)?

Caffeine rapidly diffuses throughout the body's water, both intra and extracellular.

Therefore, since the caffeine will be relatively evenly distributed, you can calculate the amount of caffeine needed to reach a given molar concentration within any particular cell by knowing the total water in a person and the molar mass of caffeine (194.19 g/mol ). A 72 kg person has about 40 liters of water in their body.

Combining those numbers, reaching 1mM requires 7.7676 grams of caffeine. Reaching 5mM requires 38.838 grams.

How are the “in vitro” results of the below charts relevant to levels that cells would experience "in vivo" after the consumption of caffeinated beverages?

Wikipedia puts the LD50 of caffeine at 150-200 mg/kg, or 10.8-14.4g for our hypothetical 72kg man. These numbers are likely very approximate as there isn't going to be a lot of well controlled clinical data on caffeine toxicity in humans, but someone reaching those concentrations stands a good chance of dying from caffeine poisoning.

For that reason I don't think these results are relevant to normal caffeine consumption. I interpreted the high concentrations used in that study as a deliberate attempt to induce cell injury, not as a means to simulate normal caffeine consumption.

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Tar86, on the chemistry site, stated that: “they work in concentrations, not total dosage”. If this is a correct judgement, which it seems to be, then a 1mM caffeine concentration containing 194mg caffeine needs to be reduced, because in the test they use a much lesser volume of solution then the basic unit of one litre.

The culture dishes they used hold only 3.5 ml each.

In each dish was poured a 1ml mixture of 100000 cells in growth medium. Leaving space for 2.5 ml caffeine solution.

2.5 ml is only 0.25% of the base unit of 1 litre. This turns out to be about 0.485 mg pure caffeine. (0.25/194; or 1/776; or 0.13 %)

There is 17% protein in a human body. In an 80 kg person that equals to about 13.6 kg protein. Collagen makes up 30% of that protein, which equals to about 4 kg collagen. So, there is in an 80 kg body about 4 kg collagen. That is 1/20; or 5%.

One cup of a 250 ml cup of coffee at 1mM concentration contains 48.5 mg caffeine. If a double doze, or 97 mg caffeine is ingested only 5%, or 4.49 mg, will reach the collagen. This 4.49 mg of caffeine will be distributed over 4 kg of collagen, becoming 0.00112 mg caffeine per mg collagen.

There are 37.2 trillion cells in the body. And in each culture dish 100000 were plated in a 1 ml growth medium. That is, each culture dish contains 1/372000 of all the cells in the body.

The answer seems to be dependent on:

    1. The number of cells in 1 mg of collagen in the human body
    1. The toxic impact levels of caffeine on human body cells.
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    $\begingroup$ You don't need to know the number of cells and amount of collagen - you only need to know the volume of the body space in which caffeine solution will be dissolved. They work with concentrations, so they relate the effects more to concentrations rather than to absolute amounts of caffeine. It's like when you spill hydrochloric acid over your clothes, the damaging effect will be more related to acid concentration than the amount. Anyway, this is more the part of the question than the answer. $\endgroup$ – Jan Jan 28 at 10:15

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