Development and evaluation of á-arbutin loaded nanostructured lipid carriers for enhanced in-vitro cytotoxic activity in melanoma

Abstract

The aim of this study was to develop and optimize α-arbutin-loaded nanostructured lipid carriers (Ar- NLCs) using the QbD. Additionally, the formulation studies, in-vitro and ex-vivo performance of Ar-NLCs were assessed, along with their cytotoxic efficacy in melanoma cells. The Ar-NLCs were fabricated using the high-speed homogenization-ultrasonication method, incorporating Gelucire 48/16, Castor oil, Capryol 90, and Tween 80. To analyze the impact of factors on Ar-NLCs, the Box-Behnken design (BBD) was utilized. The Ar-NLCs were characterized by particle size, polydispersity index, morphology, zeta potential, release kinetics, permeation, flux and stability. Additionally, Ar-NLCs cytotoxicity was assessed using the A375 cells. The Ar-NLCs demonstrated a particle size of 228.7 ± 44.5 nm, a zeta potential of -14.2 ± 2.64 mV respectively. The entrapment efficiency was 67.62 ± 4.46%. The α- arbutin release from NLCs followed Weibull kinetics. Notably, Ar-NLCs demonstrated a 2.53-fold higher permeability compared to Ar-SOL. Furthermore, Ar-NLCs exhibited significantly stronger cytotoxic effects against melanoma cells than Ar-SOL. This study reports the successful development of Ar-NLCs using a QbD approach. Enhanced transdermal permeability, enhanced cytotoxicity on melanoma cells, and sustained release of α-arbutin from NLCs were achieved. These findings indicate that NLCs offer a viable alternative drug delivery system for transdermal applications.

Keywords

• Box-Behnken design
• release kinetics
• ex-vivo permeation
• cytotoxicity
• nanostructured lipid carriers

References

1. [1] Boo YC. Arbutin as a skin depigmenting agent with antimelanogenic and antioxidant properties. Antioxidants. 2021; 10(7): 1129-1140. https://doi.org/10.3390/antiox10071129
2. [2] Apalla Z, Nashan D, Weller RB, Castellsagué X. Skin cancer: epidemiology, disease burden, pathophysiology, diagnosis, and therapeutic approaches. Dermatol Ther. 2017; 7: 5-19. https://doi.org/10.1007/s13555-016-0165-y
3. [3] Huang J, Chan SC, Ko S, Lok V, Zhang L, Lin X, Wong MC. Global incidence, mortality, risk factors and trends of melanoma: a systematic analysis of registries. Am J Clin Dermatol. 2023; 24(6): 965-975. https://doi.org/10.1007/s40257-023-00795-3
4. [4] Liu JK. Natural products in cosmetics. Nat Prod Bioprospect. 2022; 12(1): 40-82. https://doi.org/10.1007/s13659-022-00363-y
5. [5] Li Z, Wang Z, Zhou Q, Wang R, Xiong Z, Wu Y, Shu P. The molecular mechanisms underlying optical isomer arbutin permeating the skin: The molecular interaction between arbutin and skin components. International Journal of Pharmaceutics. 2024; 664, 124584. https://doi.org/10.1016/j.ijpharm.2024.124584
6. [6] Tofani RP, Sumirtapura YC, Darijanto ST. Formulation, characterisation, and in vitro skin diffusion of nanostructured lipid carriers for deoxyarbutin compared to a nanoemulsion and conventional cream. Sci Pharm. 2016; 84(4): 634-645. https://doi.org/10.3390/scipharm84040634
7. [7] Raza K, Singh B, Lohan S, Sharma G, Negi P, Yachha Y, Katare OP. Nano-lipoidal carriers of tretinoin with enhanced percutaneous absorption, photostability, biocompatibility and anti-psoriatic activity. Int J Pharm. 2013; 456(1): 65-72. https://doi.org/10.1016/j.ijpharm.2013.08.019
8. [8] Bastogne T. Quality-by-design of nanopharmaceuticals–a state of the art. Nanomedicine: Nanotechnol Biol Med. 2017; 13(7): 2151-2157. https://doi.org/10.1016/j.nano.2017.05.014

9. [9] Saeedi M, Rezanejad Gatabi Z, Morteza-Semnani K, Rahimnia SM, Yazdian-Robati R, Hashemi SMH. Preparation of arbutin hydrogel formulation as green skin lightener formulation: in vitro and in vivo evaluation. J Dispers Sci Technol. 2024; 1-13. https://doi.org/10.1080/01932691.2024.2371954
10. [10] Shinde UK, Suryawanshi DG, Amin PD. Development of Gelucire® 48/16 and TPGS mixed micelles and its pellet formulation by extrusion spheronization technique for dissolution rate enhancement of curcumin. AAPS PharmSciTech. 2021; 22(5): 182-192. https://doi.org/10.1208/s12249-021-02032-8
11. [11] Alshawwa SZ, El-Masry TA, Elekhnawy E, Alotaibi HF, Sallam AS, and Abdelkader DH. Fabrication of Celecoxib PVP microparticles stabilized by Gelucire 48/16 via electrospraying for enhanced anti-inflammatory action. Pharmaceuticals 2023; 16(2): 258-275. https://doi.org/10.3390/ph16020258
12. [12] Ahmad U, Naqvi SR, Ali I, Saleem F, Mehran MT, Sikandar U, Juchelková D. Biolubricant production from castor oil using iron oxide nanoparticles as an additive: Experimental, modelling and tribological assessment. Fuel. 2022; 324, 124565. https://doi.org/10.1016/j.fuel.2022.124565
13. [13] Zhou X, Hao Y, Yuan L, Pradhan S, Shrestha K, Pradhan O, Liu H, Li W. Nano-formulations for transdermal drug delivery: A review. Chin Chem Lett. 2018; 29: 1713–1724. https://doi.org/10.1016/j.cclet.2018.10.037
14. [14] Han F, Li S, Yin R, Liu H, Xu L. Effect of surfactants on the formation and characterization of a new type of colloidal drug delivery system: Nanostructured lipid carriers. Colloid Surf A: Physicochem Eng Aspect. 2008; 315(1-3): 210-216. https://doi.org/10.1016/j.colsurfa.2007.08.005
15. [15] Bashiri S, Ghanbarzadeh B, Ayaseh A, Dehghannya J, Ehsani A, Ozyurt H. Essential oil-loaded nanostructured lipid carriers: The effects of liquid lipid type on the physicochemical properties in beverage models. Food Biosci. 2020; 35: 100526. https://doi.org/10.1016/j.fbio.2020.100526
16. [16] Tan SW, Billa N, Roberts CR, Burley JC. Surfactant effects on the physical characteristics of Amphotericin B-containing nanostructured lipid carriers. Colloid Surf A: Physicochem Eng Aspect. 2010; 372(1-3): 73-79. https://doi.org/10.1016/j.colsurfa.2010.09.030
17. [17] Witayaudom P, Klinkesorn U. Effect of surfactant concentration and solidification temperature on the characteristics and stability of nanostructured lipid carrier (NLC) prepared from rambutan (Nephelium lappaceum L.) kernel fat. J Colloid Interface Sci. 2017; 505: 1082-1092. https://doi.org/10.1016/j.jcis.2017.07.008
18. [18] Pogorzelski S, Watrobska-Swietlikowska D, Sznitowska M. Surface tensometry studies on formulations of surfactants with preservatives as a tool for antimicrobial drug protection characterization. J Biophys Chem. 2012; 3(04): 324-333. https://doi.org/10.4236/jbpc.2012.34040
19. [19] Souto EB, Baldim I, Oliveira WP, Rao R, Yadav N, Gama FM, Mahant S. SLN and NLC for topical, dermal, and transdermal drug delivery. Expert Opin Drug Deliv. 2020; 17(3): 357-377. https://doi.org/10.1080/17425247.2020.1727883
20. [20] Nagaich U, Gulati N. Nanostructured lipid carriers (NLC) based controlled release topical gel of clobetasol propionate: Design and in vivo characterization. Drug Deliv Transl Res. 2016; 6: 289-298. https://doi.org/10.1007/s13346-016-0291-1
21. [21] Bhaskar K, Krishna Mohan C, Lingam M, Prabhakar Reddy V, Venkateswarlu V, Madhusudan Rao Y. Development of nitrendipine controlled release formulations based on SLN and NLC for topical delivery: in vitro and ex vivo characterization. Drug Develop Ind Pharm. 2008; 34(7): 719-725. https://doi.org/10.1080/03639040701842485
22. [22] Toksoy MO, Aşır F, Güzel MC. Quality by design approach for development and characterization of gabapentin-loaded solid lipid nanoparticles for intranasal delivery: In vitro, ex vivo, and histopathological evaluation. Iran J Basic Med Sci. 2024; 27(7): 904-913. 10.22038/IJBMS.2024.76281.16511 [23] Agrawal M, Saraf S, Pradhan M, Patel RJ, Singhvi G, Alexander A. Design and optimization of curcumin loaded nano lipid carrier system using Box-Behnken design. Biomed Pharmacother. 2008; 141: 111919. https://doi.org/10.1016/j.biopha.2021.111919
23. [24] Araujo VHS, da Silva PB, Szlachetka IO, da Silva SW, Fonseca-Santos B, Chorilli M, Muehlmann LA. The influence of NLC composition on curcumin loading under a physicochemical perspective and in vitro evaluation. Colloid Surf A: Physicochem Eng Aspect. 2020; 602: 125070. https://doi.org/10.1016/j.colsurfa.2020.125070
24. [25] Porbaha P, Ansari R, Kiafar MR, Bashiry R, Khazaei MM, Dadbakhsh A, Azadi AA. comparative mathematical analysis of drug release from lipid-based nanoparticles. AAPS PharmSciTech. 2024; 25(7): 208. https://doi.org/10.1208/s12249-024-02922-7
25. [26] Dai W, Zhang D, Duan C, Jia L, Wang Y, Feng F, Zhang Q. Preparation and characteristics of oridonin-loaded nanostructured lipid carriers as a controlled-release delivery system. J Microencapsul. 2010; 27(3): 234-241. https://doi.org/10.3109/02652040903079526
26. [27] Nguyen CN, Nguyen TTT, Nguyen HT, Tran TH. Nanostructured lipid carriers to enhance transdermal delivery and efficacy of diclofenac. Drug Deliv Transl Res. 2017; 7: 664-673. https://doi.org/10.1007/s13346-017-0415-2
27. [28] Elgizawy HA, Ali AA, Hussein MA. Resveratrol: Isolation, and its nanostructured lipid carriers, inhibits cell proliferation, induces cell apoptosis in certain human cell lines carcinoma and exerts protective effect against paraquat-induced hepatotoxicity. J Med Food. 2021; 24(1): 89-100. https://doi.org/10.1089/jmf.2019.028
28. [29] Cheng SL, Liu RH, Sheu JN, Chen ST, Sinchaikul S, Tsay GJ. Toxicogenomics of A375 human malignant melanoma cells treated with arbutin. J Biomed Sci. 2007; 14: 87-105. https://doi.org/10.1007/s11373-006-9130-6
29. [30] Aung NN, Ngawhirunpat T, Rojanarata T, Patrojanasophon P, Opanasopit P, Pamornpathomkul B. HPMC/PVP dissolving microneedles: A promising delivery platform to promote trans-epidermal delivery of alpha-arbutin for skin lightening. AAPS PharmSciTech. 2020; 21: 25-37. https://doi.org/10.1208/s12249-019-1599-1
30. [31] Subramaniam B, Siddik ZH, Nagoor NH. Optimization of nanostructured lipid carriers: Understanding the types, designs, and parameters in the process of formulations. J Nanoparticle Res. 2020; 22: 1-29. https://doi.org/10.1007/s11051-020-04848-0
31. [32] Bajwa N, Mahal S, Naryal S, Singh PA, Baldi A. Development of novel solid nanostructured lipid carriers for bioavailability enhancement using a quality by design approach. AAPS PharmSciTech. 2022; 23(7): 253-266. https://doi.org/10.1208/s12249-022-02386-7
32. [33] Kim BS, Na YG, Choi JH, Kim I, Lee E, Kim SY, Cho CW. The improvement of skin whitening of phenylethyl resorcinol by nanostructured lipid carriers. Nanomaterials. 2017; 7(9): 241-257. https://doi.org/10.3390/nano7090241
33. [34] Negi LM, Jaggi M, Talegaonkar S. Development of protocol for screening the formulation components and the assessment of common quality problems of nano-structured lipid carriers. Int J Pharm. 2014; 461(1-2): 403-410. https://doi.org/10.1016/j.ijpharm.2013.12.006
34. [35] Subroto E, Andoyo R, Indiarto R. Solid lipid nanoparticles: Review of the current research on encapsulation and delivery systems for active and antioxidant compounds. Antioxidants. 2023; 12(3): 633-660. https://doi.org/10.3390/antiox12030633
35. [36] Saeedi M, Rezanejad Gatabi Z, Morteza-Semnani K, Rahimnia SM, Yazdian-Robati R, Hashemi SMH. Preparation of arbutin hydrogel formulation as green skin lightener formulation: in vitro and in vivo evaluation. J Dispers Sci Technol. 2024; 1-13. https://doi.org/10.1080/01932691.2024.2371954
36. [37] Gamal A, Saeed H, El-Ela FIA, Salem HF. Improving the antitumor activity and bioavailability of sonidegib for the treatment of skin cancer. Pharmaceutics. 2021; 13(10): 1560-1575. https://doi.org/10.3390/pharmaceutics13101560
37. [38] Ergin AD, Oltulu Ç, Koç B. Enhanced cytotoxic activity of 6-mercaptopurine–loaded solid lipid nanoparticles in hepatic cancer treatment. ASSAY Drug Develop Technol. 2023; 21(5): 212-221. https://doi.org/10.1089/adt.2023.00