Green Arbutin-Chitosan Nanoparticles gel as an Eco-Friendly and Promising Product for Skin Lightening: In Vitro and In Vivo Assessment

Document Type : Research Paper

Authors

1 Food Health Research Center, Hormozgan University of Medical Sciences, Bandar Abbas, Iran

2 Department of Pharmacology and Toxicology, Faculty of Pharmacy, Hormozgan University of Medical Sciences, Bandar Abbas, Iran

3 Department of Pharmaceutics, Faculty of Pharmacy, Mazandaran University of Medical Sciences, Sari, Iran

4 Student Research Committee, Faculty of Pharmacy, Hormozgan University of Medical Sciences, Bandar Abbas, Iran

5 Department of Clinical Pharmacy, Faculty of Pharmacy, Hormozgan University of Medical Sciences, Bandar Abbas, Iran

6 Cellular and Molecular Research Center, Research Institute for Health Development, Kurdistan University of Medical Sciences, Sanandaj, Iran

7 Department of Medicinal Chemistry, School of Pharmacy and Pharmaceutical Sciences, Hormozgan University of Medical Sciences, Bandar Abbas, Iran

8 Department of Pharmaceutics, Faculty of Pharmacy, Hormozgan University of Medical Sciences, Bandar Abbas, Iran

10.22038/nmj.2025.83053.2075

Abstract

Objective(s): In the present study, a cross-linking gelation method combined with ultrasound was employed to create arbutin (ARB)-incorporated chitosan (CHT) nanoparticles (NPs). This approach aims to enhance cutaneous absorption and improve anti-melanogenesis effects.
Material and Methods: Environmentally-friendly preparation of NP, monitoring NP features, checking structure, animal safety application, cellular viability, and inhibitory assessment on melanin creation were performed.
Results: The results showed that increasing the volume ratio of chitosan (CHT) to tripolyphosphate (TPP) from 10:1.25 to 10:5 resulted in a reduction of particle size from 1097.133 ± 28.655 nm to 215.666 ± 5.976 nm. Moreover, this CHT/TPP volume ratio increase from 10:1.25 to 10:5 enhanced the encapsulation efficiency, from 55.084 ± 4.283% to 97.151 ± 0.066%. Assessment of cutaneous absorption revealed that the ARB-CHT-NP gel delivered significantly more arbutin (ARB) to both the cutaneous layers (46.168 ± 3.313% or 810.094 ± 58.147 μg/cm²) and the receiver compartment (34.155 ± 2.699% or 599.314 ± 47.371 μg/cm²) compared to the ARB plain gel. In vitro cytotoxicity testing demonstrated that, in the presence of the optimal formulation, a higher percentage of cell survival was observed in the HFF cell line compared to kojic acid and ARB. Additionally, the ARB-CHT-NP gel exhibited greater cytotoxicity in the B16F10 cell line compared to the other groups. A cutaneous itching assay on Wistar rats showed no signs of sensitivity to the ARB-CHT-NP gel. Furthermore, ARB-CHT-NP inhibited melanogenesis more effectively than kojic acid and ARB. L-dopa auto-oxidation was also significantly inhibited by ARB-CHT-NP (56.971 ± 1.265%) compared to kojic acid (46.141 ± 1.169%) and ARB (41.308 ± 1.967%).
Conclusion: Based on the results, the ARB-CHT-NP could serve as a prospective nanocarrier for ARB cutaneous application. Therefore, it is recommended that its use for treating melasma be considered.

Keywords


  1. Picardi A, Abeni D, Melchi C, Puddu P, Pasquini P. Psychiatric morbidity in dermatological outpatients: an issue to be recognized. Br J Dermatol. 2000;143(5):983-991.
  2. Deshpande SS, Khatu SS, Pardeshi GS, Gokhale NR. Cross-sectional study of psychiatric morbidity in patients with melasma. Indian J Psychiatry. 2018;60(3):324-328.
  3. Rendon M, Berneburg M, Arellano I, Picardo M. Treatment of melasma. J Am Acad Dermatol. 2006;54(5):S272-S281.
  4. Levy LL, Emer JJ. Emotional benefit of cosmetic camouflage in the treatment of facial skin conditions: personal experience and review. Clin Cosmet Investig Dermatol. 2012:173-182.
  5. Sheth VM, Pandya AG. Melasma: a comprehensive update: part II. J Am Acad Dermatol. 2011;65(4):699-714.
  6. Clardy J, Walsh C. Lessons from natural molecules. Nature. 2004;432(7019):829-837.
  7. Zhu W, Gao J, editors. The use of botanical extracts as topical skin-lightening agents for the improvement of skin pigmentation disorders. J Investig Dermatol Symp Proc; 2008: Elsevier.
  8. Inoue Y, Hasegawa S, Yamada T, Date Y, Mizutani H, Nakata S, et al. Analysis of the effects of hydroquinone and arbutin on the differentiation of melanocytes. Biol Pharm Bull. 2013;36(11):1722-30.
  9. Tada M, Kohno M, Niwano Y. Alleviation effect of arbutin on oxidative stress generated through tyrosinase reaction with L-tyrosine and L-DOPA. BMC Biochem. 2014;15:1-7.
  10. Ertam I, Mutlu B, Unal I, Alper S, Kivcak B, Ozer O. Efficiency of ellagic acid and arbutin in melasma: A randomized, prospective, open‐label study. J Dermatol. 2008;35(9):570-574.
  11. Polnikorn N. Treatment of refractory melasma with the MedLite C6 Q-switched Nd: YAG laser and alpha arbutin: a prospective study. J Cosmet Laser Ther. 2010;12(3):126-131.
  12. Davis EC, Callender VD. Postinflammatory hyperpigmentation: a review of the epidemiology, clinical features, and treatment options in skin of color. J Clin Aesthet Dermato. 2010;3(7):20.
  13. Fong P, Tong HH. In silico prediction of the cosmetic whitening effects of naturally occurring lead compounds. Nat Prod Commun. 2012;7(10):1934578X1200701010.
  14. Khezri K, Saeedi M, Morteza-Semnani K, Akbari J, Rostamkalaei SS. An emerging technology in lipid research for targeting hydrophilic drugs to the skin in the treatment of hyperpigmentation disorders: kojic acid-solid lipid nanoparticles. Artif Cells Nanomed Biotechnol. 2020;48(1):841-853.
  15. Sharma V, Anandhakumar S, Sasidharan M. Self-degrading niosomes for encapsulation of hydrophilic and hydrophobic drugs: an efficient carrier for cancer multi-drug delivery. Mater Sci Eng C. 2015;56:393-400.
  16. Mohammadian E, Rahimpour E, Alizadeh-Sani M, Foroumadi A, Jouyban A. An overview on terbium sensitized based-optical sensors/nanosensors for determination of pharmaceuticals. Appl Spectrosc Rev. 2022;57(1):39-76.
  17. Ta Q, Ting J, Harwood S, Browning N, Simm A, Ross K, et al. Chitosan nanoparticles for enhancing drugs and cosmetic components penetration through the skin. Eur J Pharm Sci. 2021;160:105765.
  18. Shah S, Pal A, Kaushik V, Devi S. Preparation and characterization of venlafaxine hydrochloride‐loaded chitosan nanoparticles and in vitro release of drug. J Appl Polym Sci. 2009;112(5):2876-2887.
  19. Saeedi M, Morteza-Semnani K, Siahposht-Khachaki A, Akbari J, Valizadeh M, Sanaee A, et al. Passive targeted drug delivery of venlafaxine hcl to the brain by modified chitosan nanoparticles: Characterization, cellular safety assessment, and in vivo evaluation. J Pharm Innov. 2023;18(3):1441-1453.
  20. Nawaz A, Wong TW. Chitosan-carboxymethyl-5-fluorouracil-folate conjugate particles: microwave modulated uptake by skin and melanoma cells. J Invest Dermatol. 2018;138(11):2412-2422.
  21. Haque S, Md S, Fazil M, Kumar M, Sahni JK, Ali J, et al. Venlafaxine loaded chitosan NPs for brain targeting: pharmacokinetic and pharmacodynamic evaluation. Carbohydr Polym. 2012;89(1):72-79.
  22. Calvo P, Remunan‐Lopez C, Vila‐Jato JL, Alonso M. Novel hydrophilic chitosan‐polyethylene oxide nanoparticles as protein carriers. J Appl Polym Sci. 1997;63(1):125-132.
  23. Hashemi SMH, Enayatifard R, Akbari J, Saeedi M, Seyedabadi M, Morteza-Semnani K, et al. Venlafaxine HCl encapsulated in niosome: green and eco-friendly formulation for the management of pain. AAPS Pharm Sci Tech. 2022;23(5):149.
  24. Saeedi M, Morteza-Semnani K, Akbari J, Rahimnia SM, Babaei A, Eghbali M, et al. Eco-friendly preparation, characterization, evaluation of anti-melanogenesis/antioxidant effect and in vitro/in vivo safety profile of kojic acid loaded niosome as skin lightener preparation. J Biomater Sci Polym Ed. 2023;34(14):1952-1980.
  25. Gatabi ZR, Saeedi M, Morteza-Semnani K, Rahimnia SM, Yazdian-Robati R, Hashemi SMH. Green preparation, characterization, evaluation of anti-melanogenesis effect and in vitro/in vivo safety profile of kojic acid hydrogel as skin lightener formulation. J Biomater Sci Polym Ed. 2022;33(17):2270-2291.
  26. Masarudin MJ, Cutts SM, Evison BJ, Phillips DR, Pigram PJ. Factors determining the stability, size distribution, and cellular accumulation of small, monodisperse chitosan nanoparticles as candidate vectors for anticancer drug delivery: application to the passive encapsulation of [14C]-doxorubicin. Nanotechnol Sci Appl. 2015:67-80.
  27. Hejjaji EM, Smith AM, Morris GA. Evaluation of the mucoadhesive properties of chitosan nanoparticles prepared using different chitosan to tripolyphosphate (CS: TPP) ratios. Int J Biol Macromol. 2018;120:1610-1617.
  28. Abd El-Aziz BA. Improvement of kojic acid production by a mutant strain of Aspergillus flavus. J Nat Sci Res. 2013;3(4):31-41.
  29. Pant A, Negi JS. Novel controlled ionic gelation strategy for chitosan nanoparticles preparation using TPP-β-CD inclusion complex. Eur J Pharm Sci. 2018;112:180-185.
  30. Pan C, Qian J, Zhao C, Yang H, Zhao X, Guo H. Study on the relationship between crosslinking degree and properties of TPP crosslinked chitosan nanoparticles. Carbohydr Polym. 2020;241:116349.
  31. Al-Nemrawi N, Alsharif S, Dave R. Preparation of chitosan-TPP nanoparticles: the influence of chitosan polymeric properties and formulation variables. Int J Appl Pharm. 2018;10(5):60-65.
  32. Akbari J, Saeedi M, Morteza-Semnani K, Hashemi SMH, Babaei A, Eghbali M, et al. Innovative topical niosomal gel formulation containing diclofenac sodium (niofenac). J Drug Target. 2022;30(1):108-117.
  33. Maji R, Ray S, Das B, Nayak AK. Ethyl cellulose microparticles containing metformin HCl by emulsification-solvent evaporation technique: effect of formulation variables. Int Sch Res Notices 2012;2012.
  34. Raju G, Mas Haris MRH, Azura A, Ahmed Mohamed Eid AM. Chitosan epoxidized natural rubber biocomposites for sorption and biodegradability studies. ACS Omega. 2020;5(44):28760-28766.
  35. Saeedi M, Morteza-Semnani K, Akbari J, Rahimnia SM, Ahmadi F, Choubdari H, et al. Development of kojic acid loaded collagen-chitosan nanoparticle as skin lightener product: in vitro and in vivo assessment. J Biomater Sci Polym Ed. 2024;35(1):63-84.
  36. Sahudin S, Sahrum Ayumi N, Kaharudin N. Enhancement of skin permeation and penetration of β-arbutin fabricated in chitosan nanoparticles as the delivery system. Cosmetics. 2022;9(6):114.
  37. Akbari J, Saeedi M, Enayatifard R, Morteza-Semnani K, Hashemi SMH, Babaei A, et al. Curcumin Niosomes (curcusomes) as an alternative to conventional vehicles: A potential for efficient dermal delivery. J Drug Deliv Technol. 2020;60:102035.
  38. Liu P, Boyle AJ, Lu Y, Adams J, Chi Y, Reilly RM, et al. Metal-chelating polymers (MCPs) with zwitterionic pendant groups complexed to trastuzumab exhibit decreased liver accumulation compared to polyanionic MCP immunoconjugates. Biomacromolecules. 2015;16(11):3613-3623.
  39. Miyayama T, Matsuoka M. Involvement of lysosomal dysfunction in silver nanoparticle-induced cellular damage in A549 human lung alveolar epithelial cells. J occup med toxicol. 2016;11:1-6.
  40. Zubareva A, Svirshchevskaya E. Mechanisms of the interaction of chitosan and its derivatives with the cell. Prikl Biokhim Mikrobiol. 2016;52(5):448-454.
  41. Mohammadian E, Foroumadi A, Hasanvand Z, Rahimpour E, Zhao H, Jouyban A. Simulation of mesalazine solubility in the binary solvents at various temperatures. J Mol Liq. 2022;357:119160.
  42. Rudeekulthamrong P, Kaulpiboon J. Optimization of amylomaltase for the synthesis of α-arbutin derivatives as tyrosinase inhibitors. Carbohydr Res. 2020;494:108078.
  43. Rigopoulos D, Gregoriou S, Katsambas A. Hyperpigmentation and melasma. Cosmet Dermatol. 2007;6(3):195-202.
  44. Dastan Z, Pouramir M, Ghasemi-Kasman M, Ghasemzadeh Z, Dadgar M, Gol M, et al. Arbutin reduces cognitive deficit and oxidative stress in animal model of Alzheimer's disease. Int J Neurosci. 2019;129(11):1145-1153.
  45. Fytianos G, Rahdar A, Kyzas GZ. Nanomaterials in cosmetics: Recent updates. Nanomaterials. 2020;10(5):979.
  46. Nakamura M, Haarmann‐Stemmann T, Krutmann J, Morita A. Alternative test models for skin ageing research. Exp Dermatol. 2018;27(5):495-500.