BOX-BEHNKEN DESIGN APPROACH TO DEVELOP NANO-VESICULAR HERBAL GEL FOR THE MANAGEMENT OF SKIN CANCER IN EXPERIMENTAL ANIMAL MODEL

TRINAYAN DEKA; MALAY K. DAS; SANJOY DAS; PUNAMJYOTI DAS; L. RONIBALA SINGHA

doi:10.22159/ijap.2022v14i6.45867

Authors

TRINAYAN DEKA Department of Pharmaceutical Sciences, Dibrugarh University, Dibrugarh, Assam 786004, India https://orcid.org/0000-0002-2898-8810
MALAY K. DAS Department of Pharmaceutical Sciences, Dibrugarh University, Dibrugarh, Assam 786004, India
SANJOY DAS Department of Pharmaceutical Sciences, Dibrugarh University, Dibrugarh, Assam 786004, India https://orcid.org/0000-0001-6589-9967
PUNAMJYOTI DAS Department of Pharmaceutical Sciences, Dibrugarh University, Dibrugarh, Assam 786004, India https://orcid.org/0000-0003-1840-3206
L. RONIBALA SINGHA Department of Pharmaceutical Sciences, Dibrugarh University, Dibrugarh, Assam 786004, India https://orcid.org/0000-0002-7445-2291

DOI:

https://doi.org/10.22159/ijap.2022v14i6.45867

Keywords:

Skin cancer, Green tea catechins, Phytomedicine, Transfersome, Box-Behnken design, Herbal nanogel

Abstract

Objective: To manage the increasing burden of skin cancer cases globally and to replace conventional invasive treatments and their side effects, the present study is aimed to develop a transfersomal herbal gel of Green Tea Catechins (GTC) extracted from indigenous green tea and evaluate it for in vivo management of skin cancer in an experimental animal model.

Methods: GTC-loaded transfersomes (GTCTF) were prepared by the thin-film hydration method. After optimizing the GTCTFs using the Box-Behnken design, they were characterized for zeta potential, structure, in vitro drug release, and in vitro skin permeation. Carbopol 940 gel was developed for the topical delivery of GTCTF and characterized for pH, viscosity, spreadability and in vitro skin permeation. In vitro MTT assay and in vivo chemopreventive and anticancer efficacy of the GTCTF gel were evaluated in mice.

Results: The GTCTF has shown a particle size of 151.4±1.9 nm, entrapment efficiency of 68.25±0.06 %, and drug loading of 10.41±0.02 %. The in vitro MTT assay in B16F10 melanoma cell lines showed promising anticancer efficacy of the GTCTF. GTCTF gel was found suitable for topical delivery with favorable pH, viscosity, spreadability, and permeability and effective in preventing and curing skin cancer in mice, with a significant reduction of tissue biochemical parameters like TNF-α, IL-1β, and IL-6.

Conclusion: Collectively, successful prevention and curing of the induced skin cancer in the experimental animal model by the GTCTF gel have established a novel herbal nanomedicine approach for the management of skin cancer.

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References

Jones OT, Ranmuthu CKI, Hall PN, Funston G, Walter FM. Recognising skin cancer in primary care. Adv Ther. 2020;37(1):603-16. doi: 10.1007/s12325-019-01130-1, PMID 31734824.

Bradford PT. Skin cancer in skin of color. Dermatol Nurs. 2009;21(4):170-8. PMID 19691228.

Apalla Z, Lallas A, Sotiriou E, Lazaridou E, Ioannides D. Epidemiological trends in skin cancer. Dermatol Pract Concept. 2017;7(2):1-6. doi: 10.5826/dpc.0702a01, PMID 28515985.

Linares MA, Zakaria A, Nizran P. Skin cancer. Prim Care. 2015;42(4):645-59. doi: 10.1016/j.pop.2015.07.006, PMID 26612377.

Saginala K, Barsouk A, Aluru JS, Rawla P, Barsouk A. Epidemiology of melanoma. Med Sci (Basel). 2021;9(4):63. doi: 10.3390/medsci9040063, PMID 34698235.

Perera E, Gnaneswaran N, Staines C, Win AK, Sinclair R. Incidence and prevalence of non-melanoma skin cancer in Australia: a systematic review. Australas J Dermatol. 2015;56(4):258-67. doi: 10.1111/ajd.12282, PMID 25716064.

Labani S, Asthana S, Rathore K, Sardana K. Incidence of melanoma and nonmelanoma skin cancers in Indian and the global regions. J Cancer Res Ther. 2021;17(4):906-11. doi: 10.4103/jcrt.JCRT_785_19, PMID 34528540.

India Population Fact Sheet. WHO International Agency for Cancer Research. Available from: https://gco.iarc.fr/today/ data/factsheets/populations/356-india-fact-sheets.pdf. [Last accessed on 02 Mar 2022].

Priya P, Mohan Raj RM, Vasanthakumar V, Raj V. Curcumin-loaded layer-by-layer folic acid and casein coated carboxymethyl cellulose/casein nanogels for treatment of skin cancer. Arab J Chem. 2020;13(1):694-708. doi: 10.1016/j.arabjc.2017.07.010.

Shukla T, Upmanyu N, Agrawal M, Saraf S, Saraf S, Alexander A. Biomedical applications of microemulsion through dermal and transdermal route. Biomed Pharmacother. 2018;108:1477-94. doi: 10.1016/j.biopha.2018.10.021, PMID 30372850.

Iqbal J, Abbasi BA, Ahmad R, Batool R, Mahmood T, Ali B. Potential phytochemicals in the fight against skin cancer: current landscape and future perspectives. Biomed Pharmacother. 2019;109:1381-93. doi: 10.1016/ j.biopha.2018.10.107, PMID 30551389.

Chinembiri TN, du Plessis LH, Gerber M, Hamman JH, du Plessis J. Review of natural compounds for potential skin cancer treatment. Molecules. 2014;19(8):11679-721. doi: 10.3390/ molecules190811679, PMID 25102117.

Bhutadiya VL, Mistry KN. A review on bioactive phytochemicals and it’s mechanism on cancer treatment and prevention by targeting multiple cellular signaling pathways. Int J Pharm Pharm Sci. 2021;13:15-9. doi: 10.22159/ijpps.2021v13i12.42798.

Ng CY, Yen H, Hsiao HY, Su SC. Phytochemicals in skin cancer prevention and treatment: an updated review. Int J Mol Sci. 2018;19(4):941. doi: 10.3390/ijms19040941, PMID 29565284.

Cheng Z, Zhang Z, Han Y, Wang J, Wang Y, Chen X. A review on the anti-cancer effect of green tea catechins. J Funct Foods. 2020;74:104172. doi: 10.1016/j.jff.2020.104172.

Mittal A, Piyathilake C, Hara Y, Katiyar SK. Exceptionally high protection of photocarcinogenesis by topical application of (-)-epigallocatechin-3-gallate in hydrophilic cream in SKH-1 hairless mouse model: relationship to inhibition of UVB-induced global DNA hypomethylation. Neoplasia. 2003;5(6):555-65. doi: 10.1016/s1476-5586(03)80039-8, PMID 14965448.

Magcwebeba TU, Riedel S, Swanevelder S, Swart P, De Beer D, Joubert E. The potential role of polyphenols in the modulation of skin cell viability by Aspalathus linearis and cyclopia spp. Herbal tea extracts in vitro. J Pharm Pharmacol. 2016;68(11):1440-53. doi: 10.1111/jphp.12629, PMID 27671741.

Katiyar SK, Mohan RR, Agarwal R, Mukhtar H. Protection against induction of mouse skin papillomas with low and high risk of conversion to malignancy by green tea polyphenols. Carcinogenesis. 1997;18(3):497-502. doi: 10.1093/carcin/ 18.3.497, PMID 9067548.

Przystupski D, Michel O, Rossowska J, Kwiatkowski S, Saczko J, Kulbacka J. The modulatory effect of green tea catechin on drug resistance in human ovarian cancer cells. Med Chem Res. 2019;28(5):657-67. doi: 10.1007/s00044-019-02324-6.

Manikandan R, Beulaja M, Arulvasu C, Sellamuthu S, Dinesh D, Prabhu D. Synergistic anticancer activity of curcumin and catechin: an in vitro study using human cancer cell lines. Microsc Res Tech. 2012;75(2):112-6. doi: 10.1002/jemt.21032, PMID 21780253.

Braicu C, Gherman CD, Irimie A, Berindan Neagoe I. Epigallocatechin-3-gallate (EGCG) inhibits cell proliferation and migratory behaviour of triple-negative breast cancer cells. J Nanosci Nanotechnol. 2013;13(1):632-7. doi: 10.1166/jnn.2013.6882, PMID 23646788.

Shim JH, Su ZY, Chae JI, Kim DJ, Zhu F, Ma WY. Epigallocatechin gallate suppresses lung cancer cell growth through Ras-GTPase-activating protein SH3 domain-binding protein 1. Cancer Prev Res (Phila). 2010;3(5):670-9. doi: 10.1158/1940-6207.CAPR-09-0185, PMID 20424128.

Tyagi T, Garlapati PK, Yadav P, Naika M, Mallya A, Kandangath Raghavan A. Development of nano-encapsulated green tea catechins: studies on optimization, characterization, release dynamics, and in vitro toxicity. J Food Biochem. 2021;45(11):e13951. doi: 10.1111/jfbc.13951, PMID 34569069.

PM, V BB, KN CM, NA, SB. Enrichment of in vivo efficacy of catechin-rich extract with the application of nanotechnology. Int J Appl Pharm. 2018;10(5):281-8. doi: 10.22159/ijap.2018v10i5.29569.

Krishnan V, Mitragotri S. Nanoparticles for topical drug delivery: potential for skin cancer treatment. Adv Drug Deliv Rev. 2020;153:87-108. doi: 10.1016/j.addr.2020.05.011, PMID 32497707.

Natarajan SB, Chandran SP, Vinukonda A, Rajan DS. Green tea catechin loaded nano delivery systems for the treatment of pandemic diseases. Asian J Pharm Clin Res. 2019;12(5):1-7.

Chen J, Shao R, Zhang XD, Chen C. Applications of nanotechnology for melanoma treatment, diagnosis, and theranostics. Int J Nanomedicine. 2013;8:2677-88. doi: 10.2147/IJN.S45429, PMID 23926430.

Tsai YJ, Chen BH. Preparation of catechin extracts and nanoemulsions from green tea leaf waste and their inhibition effect on prostate cancer cell PC-3. Int J Nanomedicine. 2016;11:1907-26. doi: 10.2147/IJN.S103759, PMID 27226712.

Chen CC, Hsieh DS, Huang KJ, Chan YL, Hong PD, Yeh MK. Improving anticancer efficacy of (-)-epigallocatechin-3-gallate gold nanoparticles in murine B16F10 melanoma cells. Drug Des Dev Ther. 2014;8:459-74. doi: 10.2147/DDDT.S58414, PMID 24855338.

Jiang Y, Jiang Z, Ma L, Huang Q. Advances in nanodelivery of green tea catechins to enhance the anticancer activity. Molecules. 2021;26(11):3301. doi: 10.3390/ molecules26113301, PMID 34072700.

Harwansh RK, Mukherjee PK, Kar A, Bahadur S, Al-Dhabi NA, Duraipandiyan V. Enhancement of photoprotection potential of catechin loaded nanoemulsion gel against UVA-induced oxidative stress. J Photochem Photobiol B. 2016;160:318-29. doi: 10.1016/j.jphotobiol.2016.03.026, PMID 27167597.

Abd El-Alim SH, Kassem AA, Basha M, Salama A. Comparative study of liposomes, ethosomes and transfersomes as carriers for enhancing the transdermal delivery of diflunisal: in vitro and in vivo evaluation. Int J Pharm. 2019;563:293-303. doi: 10.1016/j.ijpharm.2019.04.001, PMID 30951860.

Shaji J, Garude S. Transethosomes and ethosomes for enhanced transdermal delivery of ketorolac tromethamine: a comparative assessment. Int J Curr Pharm Res. 2014;6:88-93.

Abdel-Hafez SM, Hathout RM, Sammour OA. Curcumin-loaded ultra deformable nanovesicles as a potential delivery system for breast cancer therapy. Colloids Surf B Biointerfaces. 2018;167:63-72. doi: 10.1016/j.colsurfb.2018.03.051, PMID 29626721.

Cevc G, Blume G. New, Highly efficient formulation of diclofenac for the topical, transdermal administration in ultradeformable drug carriers, transfersomes. Biochim Biophys Acta. 2001;1514(2):191-205. doi: 10.1016/s0005-2736(01)00369-8, PMID 11557020.

Fujiki H, Sueoka E, Watanabe T, Suganuma M. Synergistic enhancement of anticancer effects on numerous human cancer cell lines treated with the combination of EGCG, other green tea catechins, and anticancer compounds. J Cancer Res Clin Oncol. 2015;141(9):1511-22. doi: 10.1007/s00432-014-1899-5, PMID 25544670.

Bharaduwaj M, Das MK. Preformulation optimization of catechin extracts from assam green tea as a candidate for topical chemoprevention. World J Pharm Res. 2018;7:1035-48.

Row KH, Jin Y. Recovery of catechin compounds from Korean tea by solvent extraction. Bioresour Technol. 2006;97(5):790-3. doi: 10.1016/j.biortech.2005.04.001, PMID 15919205.

Agarwal OP. Advanced practical organic chemistry. 26th ed. India: GOEL Publishing House; 2019.

Methods for identification of Herbals-HPTLC Association. Camellia sinensis. Available from: https://www.hptlc-association.org/methods/methods_for_identification_of_herbals.cfm. [Last accessed on 04 Oct 2018]

Cavazzuti M. Design of experiments. In: Cavazzuti M, editor. Optimization methods: from theory to design. 1st ed. Berlin: Springer; 2013. p. 13-42.

Sachan R, Parashar T, Singh V, Singh G, Tyagi S, Patel C. Drug carrier transfersomes: a novel tool for transdermal drug delivery system. Int J Res Dev Pharm L Sci. 2013;2:309-16.

Kunasekaran V, Krishnamoorthy K. Compatibility studies of rasagiline mesylate with selected excipients for an effective solid lipid nanoparticles formulation. Int J Pharm Pharm Sci. 2015;7:73-80.

Tsai MJ, Wu PC, Huang YB, Chang JS, Lin CL, Tsai YH. Baicalein loaded in tocol nanostructured lipid carriers (tocol NLCs) for enhanced stability and brain targeting. Int J Pharm. 2012;423(2):461-70. doi: 10.1016/j.ijpharm.2011.12.009, PMID 22193056.

Pardeike J, Weber S, Haber T, Wagner J, Zarfl HP, Plank H. Development of an itraconazole-loaded nanostructured lipid carrier (NLC) formulation for pulmonary application. Int J Pharm. 2011;419(1-2):329-38. doi: 10.1016/j.ijpharm. 2011.07.040, PMID 21839157.

Nagasamy VD, Kalyani K, Tulasi K, Swetha PV, Ali SKA, Kiran HC. Transfersomes: a novel technique for transdermal drug delivery. Int J Res Pharm Nano Sci. 2014;3:266-76.

Walve JR, Bakliwal SR, Rane BR, Pawar SP. Transfersomes: a surrogated carrier for transdermal drug delivery system. Int J Appl Biol PharmTechnol. 2011;2:204-13.

Sheo DM, Shweta A, Ram CD, Ghanshyam M, Girish K, Sunil KP. Transfersomes-a novel vesicular carrier for enhanced transdermal delivery of stavudine: development, characterization and performance evaluation. J Sci Specul Res. 2010;1:30-6.

M VL, Zafaruddin MD, Kuchana V. Design and characterization of transferosomal gel of repaglinide. Int Res J Pharm. 2015;6(1):38-42. doi: 10.7897/2230-8407.0619.

Darusman F, Raisya R, Priani SE. Development, characterization, and performance evaluation of transfersome gel of ibuprofen as a transdermal drug delivery system using nanovesicular carrier. Drug Invent Today. 2018;10:3750-5.

Khan MA, Pandit J, Sultana Y, Sultana S, Ali A, Aqil M. Novel carbopol-based transfersomal gel of 5-fluorouracil for skin cancer treatment: in vitro characterization and in vivo study. Drug Deliv. 2015;22(6):795-802. doi: 10.3109/10717544.2014.902146, PMID 24735246.

Mishra N, Rana K, Seelam SD, Kumar R, Pandey V, Salimath BP. Characterization and cytotoxicity of pseudomonas mediated rhamnolipids against breast cancer MDA-MB-231 cell line. Front Bioeng Biotechnol. 2021;9:761266. doi: 10.3389/fbioe.2021.761266, PMID 34950641.

OECD. Guideline 404 for Testing of Chemicals. Acute dermal irritation. Corrosion; 2015.

Shinde U, Pokharkar S, Modani S. Design and evaluation of microemulsion gel system of nadifloxacin. Indian J Pharm Sci. 2012;74(3):237-47. doi: 10.4103/0250-474X.106066, PMID 23439454.

Uttley M, Van Abbe NJ. Primary irritation of the skin: mouse ear test and human patch test procedures. J Soc Cosmet Chem. 1973;24:217-27.

Chanda S, Nagani K. In vitro and in vivo methods for anticancer activity evaluation and some Indian medicinal plants possessing anticancer properties: an overview. J Pharmacogn Phytochem. 2013;2:140-52.

Bharadwaj R, Haloi J, Medhi S. Topical delivery of methanolic root extract of Annona reticulata against skin cancer. S Afr J Bot. 2019;124:484-93. doi: 10.1016/j.sajb.2019.06.006.

Kowalczyk MC, Kowalczyk P, Tolstykh O, Hanausek M, Walaszek Z, Slaga TJ. Synergistic effects of combined phytochemicals and skin cancer prevention in SENCAR mice. Cancer Prev Res. 2010;3(2):170-8. doi: 10.1158/1940-6207.CAPR-09-0196.

Das MK, Kumar R. Development of curcumin nanoniosomes for skin cancer chemoprevention. Int J ChemTech Res. 2015;7:747-54.

Honary S, Zahir F. Effect of zeta potential on the properties of nano-drug delivery systems-a review (part 1). Trop J Pharm Res. 2013;12:255-64.

Opatha SAT, Titapiwatanakun V, Chutoprapat R. Transfersomes: A promising nanoencapsulation technique for transdermal drug delivery. Pharmaceutics. 2020;12(9):855. doi: 10.3390/pharmaceutics12090855, PMID 32916782.

Duangjit S, Opanasopit P, Rojanarata T, Ngawhi Frunpat T. Characterization and in vitro skin permeation of meloxicam-loaded liposomes versus transfersomes. J Drug Deliv. 2011;2011:418316. doi: 10.1155/2011/418316, PMID 21490750.

Patel R, Singh S, Singh S, Sheth NR, Gendle R. Development and characterization of curcumin loaded transfersome for transdermal delivery. J Pharm Sci Res. 2009;1:71-80.

Tyagi T, Garlapati PK, Yadav P, Naika M, Mallya A, Kandangath Raghavan AK. Development of nano-encapsulated green tea catechins: studies on optimization, characterization, release dynamics, and in vitro toxicity. J Food Biochem. 2021;45(11):e13951. doi: 10.1111/jfbc.13951, PMID 34569069.

Guo L, Santschi PH. Ultrafiltration and its applications to sampling and characterisation of aquatic colloids. In: Wilkinson KJ, Lead JR, editors. Environmental colloids and particles: behaviour, separation and characterisation. Chichester: John Wiley & Sons; 2006. p. 159-221.

Rangwala S, Tsai KY. Roles of the immune system in skin cancer. Br J Dermatol. 2011;165(5):953-65. doi: 10.1111/j.1365-2133.2011.10507.x, PMID 21729024.

Lippitz BE, Harris RA. Cytokine patterns in cancer patients: a review of the correlation between interleukin 6 and prognosis. Oncoimmunology. 2016;5(5):e1093722. doi: 10.1080/2162402X.2015.1093722, PMID 27467926.

Balkwill F. Tumour necrosis factor and cancer. Nat Rev Cancer. 2009;9(5):361-71. doi: 10.1038/nrc2628, PMID 19343034.