Exploring the Possibility of Kombucha Mushroom as Material for Drug Delivery

Caroline Liu, Katherine Wang, Shalier Xia

BackgroundCurrently, there are many problems with targeted drug delivery. The current

methods are either a combination of the problems below or too costly.

• non-biodegradable or toxic to the body

• hard to trigger for release and to adjust release rate

• carrying mechanism reacts with drug

The biological agent we are using is Kombucha, a bacterial fungal symbiont. We chose this particular biomaterial due to its rapid growth rate, ability to form bacterial fungal mat, and beneficial effects on health. The source of its growth is a solution composed of a SCOBY (symbiotic colony of bacteria and yeast), black tea, and white sugar. The mat forms at the top of the solution. According to Dr. C. Dufresne and Dr. E. Farnworth, Kombucha naturally has positive health benefits upon ingestion such as “detoxify[ing] the body, reduc[ing] cholesterol levels, [and] promot[ing] liver functions...”

ObjectiveTo test the viability of Kombucha biomaterial as a bacterial fungal mat for drug delivery.

Goals of Testing• To test the durability of the bacterial mat under extreme

circumstances

• To embed drugs into the Kombucha mat and test the rate at which the drug is released

• To test the capabilities of Kombucha as a viable biological drug delivery system

• To give experimental evidence of benefits of having Kombucha as a drug delivery system over other conventional methods

MethodsGrowing the Kombucha Mat

• Prepare solution for Kombucha with a 4cm x 4cm scoby mother

Testing the Durability of Kombucha Mat

• Expose the bacterial mat to acid (pH of 1.5-3.5), base (pH of 7-8), drying, heat (98°F)

Testing the Semipermeability of Kombucha Mat

• Stretch each mat over volumetric flask with water and salt and one drop of food coloring and water on top of the bacterial fungal mat

Testing the Ability of Kombucha to Release Drugs

• Grow Kombucha mat with food coloring and test release.

Testing the Methods of Embedment of Medicine into Kombucha Mat

• Add caffeine in Kombucha growing solution and let the bacterial fungal mat grow, place four solutions on TLC paper and label, place into ethanol mixture and observe

Making the Kombucha Pill

• Use empty gel capsule, wrap dried mat, leave for 1-2 days to grow around capsule, and then remove gel capsule

Testing Release Rate of Kombucha

• Kombucha pill with sodium acetate into stomach solution, record pH for ~2 hours

The Kombucha bacterial fungal mat naturally grows at the top of the solution.

Supplies for the growth and nurturing of the Kombucha symbiont.

General setup.

Displays the separation of the bacterial fungal mat for pill preparation.

These results show that the mats wet (right) and dried (left) are semi-permeable. The solutions within the flasks contain ions (potassium chloride), so we could test whether or not osmosis could occur through the bacterial fungal mat.

These results show that the mats, having been exposed to both strong acid (left) and strong base (right), are both durable and semi-permeable.

Caffeine-injected Kombucha growth solution.

Indication of the solutions applied to the TLC paper. 1) Kombucha growth solution, 2) 5% caffeine solution control, 3 & 4) solution drawn from the kombucha after being placed into a new beaker of 100 mL dI water for ~48 hours

These are the results from our TLC paper chromatography. 1: Kombucha growth solution2: 5% caffeine solution3 and 4: release solution after 48 hrs.

The picture comparisons show that both the dried and wet blue food coloring embedded kombucha mats released the food coloring into the water.

Using the gelatin capsules as a mold, we made pills out of the Kombucha mat. We first wrapped a semi-dry Kombucha around the pill, sealed it using water (it became sticky) and then removed the gel capsule. The Kombucha pill was allowed to grow and dry in the shape of a capsule.

Setup for testing the release of sodium acetate in the “stomach” solution, made of pepsin, potassium chloride, sodium chloride, and 0.10 M hydrochloric acid. Both thermometers depicted were placed to ensure that the temperature remained around 37° C.

The graph shows that that the pH of the solution rises as the Kombucha pill releases sodium acetate (a weak base).

Allowing copper chloride (top left) to be released in a solution of a pH level of 7.3 (blood pH), we saw the pill (right) unravel over the course of 24 hours. The pill released the copper chloride into the solution, causing the blue tint.

In an attempt to increase the marketability of the Kombucha, we altered the natural fermented smell of the biomaterial with the addition of fragrant fruit teas.

DiscussionWe found that the Kombucha has many of the properties ideal for a drug-delivering mat. It is semi-permeable and relatively durable, yet it can be broken down.

We also discovered that the Kombucha is able to deliver the drug when the drug is both embedded in the mat (caffeine) and enclosed by a Kombucha pill (sodium acetate). When the drug is enclosed in the pill, it is not released immediately giving the pill time to reach the stomach.

Conclusion• Kombucha is edible and therefore naturally biodegradable,

solving the issue of toxicity. Our durability tests show that the Kombucha is indeed reliable as a means of drug release under various body environments and temperatures.

• The experiments with drug embedment and release rate further show the adaptability and versatility of the Kombucha. Finally, with the total budget of our research being ~$20 for tea along with other basic materials, our Kombucha drug delivery system was shown to be cost effective and dependable.

• However, Kombucha does not significantly stand out as a candidate for biological drug delivery because of the many problems with drug delivery, like toxicity to the body,the triggering for release of the drug, the release rate, delivery vehicle reacts with drug and cost, Kombucha only solves 2 of the 5 issues.

Future Work

• Test whether the thickness of the mat affects the delivery rate of the drug and use our results to improve the Kombucha’s drug delivery capability.

• Test the capacity of Kombucha to be a mode of transdermal drug delivery

• Improve Kombucha’s marketability

• Test various drugs to see whether the Kombucha will cause any alterations

References• C. Dufresne, E. Farnworth. Tea, Kombucha, and health: a review, Food

Research International, Volume 33, Issue 6, July 2000, Pages 409-421, ISSN 0963-9969.

• Department of Chemistry and Biochemistry. Thin Layer Chromatography (TLC). Organic Chemistry. CU Boulder, 13 Jan. 2013. Web. 26 Jan. 2014.

• Hanne Devle, Ellen Kathrine Ulleberg, Carl Fredrik Naess-Andresen, Elling-Olav Rukke, Gerd Vegarud, Dag Ekeberg. Reciprocal interacting effects of proteins and lipids during ex vivo digestion of bovine milk, International Dairy Journal, Volume 36, Issue 1, May 2014, Pages 6-13, ISSN 0958-6946.

• Jenway. The Quantitative Determination of Caffeine in Beverages and Soft Drinks Using UV Wavelength Spectroscopy. Staffordshire: Bibby Scientific, n.d. PDF. Web. 26 Jan. 2014.

Acknowledgements

• Dr. Myra Halpin and the Research in Chemistry program at the North Carolina School Science and Mathematics.

• Nikita Khlystov, MIT Class of 2014

• Ms. Korah Wiley, NCSSM

• Bruce Cao, Pharm. D., POZEN