Topical green tea formulation with anti-hemorrhagic and antibacterial effects

Document Type : Original Article


1 Biotechnology Research Center, Pharmaceutical Technology Institute, Mashhad University of Medical Sciences, Mashhad, Iran

2 Department of Pharmaceutical Nanotechnology, School of Pharmacy, Mashhad University of Medical Sciences, Mashhad, Iran

3 School of Pharmacy, Mashhad University of Medical Sciences, Mashhad, Iran

4 Targeted Drug Delivery Research Center, Pharmaceutical Technology Institute, Mashhad University of Medical Sciences, Mashhad, Iran

5 Department of Pharmaceutics, School of Pharmacy, Mashhad University of Medical Sciences, Mashhad, Iran

6 Complementary Medicine Research Center, Faculty of Traditional Medicine, Mashhad University of Medical Sciences, Mashhad, lran


Objective(s): Potentially preventable death from uncontrolled hemorrhage clearly indicates the importance of simple, fast and efficient ways to achieving hemostasis. The aim of this study was to develop a topical formulation of green tea extract for reducing bleeding that can be helpful in hemorrhage control.
Materials and Methods: Hydroalcoholic extract of green tea was isolated from Camellia sinensis and formulated in polyvinyl alcohol (PVA) to achieve two concentrations of 2% and 4% v/v. Folin-Ciocalteau assay was used to determine the total amount of tannins in extract. Rheological behavior of solutions was investigated by measuring viscosity at shear rates of 0–200 sec−1. Quantitative and qualitative microbial limit tests and minimum inhibitory concentration (MIC) assay were done. The effect of formulations on bleeding time was evaluated in an animal model.
Results: The total amount of tannin in green tea extract was 3.8% w/w and addition of green tea significantly increased the viscosity of PVA. The results of MIC assay showed that PVA could not inhibit the growth of bacteria, while, 716 µg/ml of green tea and 2860 µg/ml of green tea/PVA 4% inhibited the growth of Staphylococcus aureus and Pseudomonas aeruginosa. In an animal study both 2% and 4% formulations were able to stop hemorrhage approximately at an equal time compared with tranexamic acid (TXA) 50 mg/ml as a control and the lowest bleeding time was 6.4±0.51 sec for green tea/PVA 4%.
Conclusion: Based on our results, the topical formulation of green tea extract in PVA has a great potential for anti-hemorrhage applications.


1. Holcomb JB. Methods for improved hemorrhage control. Crit Care 2004;8:S57-60.
2. Ong SY, Wu J, Moochhala SM, Tan MH, Lu J. Development of a chitosan-based wound dressing with improved hemostatic and antimicrobial properties. Biomaterials 2008;29:4323-4332.
3. Williams-Johnson J, McDonald A, Strachan GG, Williams E. Effects of tranexamic acid on death, vascular occlusive events, and blood transfusion in trauma patients with significant haemorrhage (CRASH-2): a randomised, placebo-controlled trial. West Indian Med J 2010;59:612-624.
4. Cabrera C, Artacho R, Giménez R. Beneficial effects of green tea: a review. J Am Coll Nutr 2006;25:79-99.
5. Al-Awaida W, Akash M, Aburubaiha Z, Talib WH, Shehadeh H. Chinese green tea consumption reduces oxidative stress, inflammation and tissues damage in smoke exposed rats. Iran J Basic Med Sci 2014;17:740-746.
6. Graham HN. Green tea composition, consumption, and polyphenol chemistry. Prev Med 1992;21:334-350.
7. Cooper R, Morre DJ, Morre DM. Medicinal benefits of green tea: part II. review of anticancer properties. J Altern Complement Med 2005;11:639-652.
8. Saito Y, Hasebe-Takenaka Y, Ueda T, Nakanishi-Ueda T, Kosuge S, Aburada M, et al. Effects of green tea fractions on oxygen-induced retinal neovascularization in the neonatal rat. J Clin Biochem Nutr 2007;41:43-49.
9. Muramatsu K, Fukuyo M, Hara Y. Effect of green tea catechins on plasma cholesterol level in cholesterol-fed rats. J Nutr Sci Vitaminol 1986;32:613-622.
10. Chung KT, Wong TY, Wei CI, Huang YW, Lin Y. Tannins and human health: a review. Crit Rev Food Sci Nutr 1998;38:421-464.
11. Al-Safaar MA, Mohey Al-Din AA, Alwan ZH. The effect of green tea (Camilla sinensis L.) on blood clotting after tooth extractions. Int J Sci Technol 2013;143:1-12.
12. Brough C, Miller DA, Keen JM, Kucera SA, Lubda D, Williams RO. Use of polyvinyl alcohol as a solubility-enhancing polymer for poorly water soluble drug delivery (part 1). AAPS PharmSciTech 2016;17:167-179.
13. Jaiturong P, Sirithunyalug B, Eitsayeam S, Asawahame C, Tipduangta P, Sirithunyalug J. Preparation of glutinous rice starch/polyvinyl alcohol copolymer electrospun fibers for using as a drug delivery carrier. Asian J Pharm Sci 2018;13:239-247.
14. Anwar H, Ahmad M, Minhas MU, Rehmani S. Alginate-polyvinyl alcohol based interpenetrating polymer network for prolonged drug therapy, optimization and in vitro characterization. Carbohydr Polym 2017;166:183-194.
15. Schulze J, Hendrikx S, Schulz-Siegmund M, Aigner A. Microparticulate poly(vinyl alcohol) hydrogel formulations for embedding and controlled release of polyethylenimine (PEI)-based nanoparticles. Acta Biomater 2016;45:210-222.
16. Makkar HPS, Blummel M, Borowy NK, Becker K. Gravimetric determination of tannins and their correlations with chemical and protein precipitation Methods J Sci Food Agric 1993;61:161-165.
17. Blainski A, Lopes GC, de Mello JC. Application and analysis of the folin ciocalteu method for the determination of the total phenolic content from Limonium brasiliense L. Molecules 2013;18:6852-6865.
18. Singleton VL, Orthofer R, Lamuela-Raventós RM. Analysis of total phenols and other oxidation substrates and anti-oxidants by means of folin-ciocalteu reagent. Methods Enzymol 1999;299:152-178.
19.    Radji M, Agustama RA, Elya B, Tjampakasari CR. Antimicrobial activity of green tea extract against isolates of methicillin-resistant Staphylococcus aureus and multi-drug resistant Pseudomonas aeruginosa. Asian Pac J Trop Biomed 2013;3:663-667.
20. United States Pharmacopeial Convention, U.S. Pharmacopeia National Formulary 2018: USP 41 NF 36.  
21. Andrews JM. Determination of minimum inhibitory concentrations. J Antimicrob Chemother 2001;48:5-16.
22. Fazly Bazzaz BS, Iranshahi M, Naderinasab M, Hajian S, Sabeti Z, Masumi E. Evaluation of the effects of galbanic acid from Ferula szowitsiana and conferol from F. badrakema, as modulators of multi-drug resistance in clinical isolates of Escherichia coli and Staphylococcus aureus. Res Pharm Sci 2010;5:21-28.
23. Fazly Bazzaz BS, Sarabandi S, Khameneh B, Hosseinzadeh H. Effect of catechins, green tea extract and methylxanthines in combination with gentamicin against Staphylococcus aureus and Pseudomonas aeruginosa: combination therapy against resistant bacteria. J Pharmacopuncture 2016;19:312-318.
24. Ishihara M, Nakanishi K, Ono K, Sato M, Kikuchi M, Saito Y, et al. Photocrosslinkable chitosan as a dressing for wound occlusion and accelerator in healing process. Biomaterials 2002;23:833-840.
25. Chen Nx, Zhang Jh. The role of hydrogen-bonding interaction in poly(vinyl alcohol)/poly(acrylic acid) blending solutions and their films. Chin J Polymer Sci 2010;28:903-911.
26. Botten D, Fugallo G, Fraternali F, Molteni C. Structural properties of green tea catechins. J Phys Chem B 2015;119:12860-12867.
27. Soltani R, Haghighat A, Fanaei M, Asghari G. Evaluation of the effect of green tea extract on the prevention of gingival bleeding after posterior mandibular teeth extraction: a randomized controlled trial. Evid Based Complement Alternat Med 2014;2014:1-4.
28. Park SY, Lee HU, Lee Y-C, Kim GH, Park EC, Han SH, et al. Wound healing potential of antibacterial microneedles loaded with green tea extracts. Mater Sci Eng C. 2014;42:757-762.
29. Ashok PK, Upadhyaya K. Tannins are astringent. J Pharm Phytochem 2012;1:45-51.
30. Khasnabis J, Rai1 C, Roy A. Determination of tannin content by titrimetric method from different types of tea. J Chem Pharm Res 2015;7:238-241.
31. Kumar TMP, Umesh HM, Shivakumar HG, Ravi V, Siddaramaiah. Feasibility of polyvinyl alcohol as a transdermal drug delivery system for terbutaline sulphate. J Macromol Sci Part A 2007;44:583-589.
32. Reygaert WC. The antimicrobial possibilities of green tea. Front Microbiol 2014;5:434.
33. Forouzanfar A, Arab HR, Shafaee H, Mokhtari MR, Golestani S. The effect of green tea mouthwash (Camellia sinensis) on wound healing following periodontal crown lengthening surgery; a double blind randomized controlled trial. Open J Stomatol 2010;2:369-372.