Preclinical evaluation of melanocyte transplantation by chitosan‐based melanocyte spheroid patch to skin prepared by controlled sunburn blistering

Abstract

Transplantation of autologous cultured melanocytes as cell suspension has been used for the treatment of vitiligo. The recipient site is often prepared by laser-mediated dermabrasion. Such procedures encounter disadvantages including prolonged transplantation duration, unsecured cell adherence to lesional skin and potential scarring. To improve this, here we propose a method by preparing recipient sites before transplantation by psoralen and ultraviolet A (PUVA)-induced sunburn followed by transplanting cells with a chitosan-based melanocyte spheroid patch. We evaluated the method in nude mice. Application of methoxsalen-soaked filter paper on skin for 30 min followed by ultraviolet A exposure induced controlled sunburn blisters in 2 days. Upon transplantation, the blister roof could be quickly peeled off by a waxing patch. The chitosan membrane on which melanocytes were precultured into multicellular spheroids was transplanted with cells facing the skin. The chitosan patch adhered well to skin and secured the contact of melanocytes with the recipient site. One day later, melanocyte spheroids already detached from the chitosan membrane and adhered to the recipient skin. Our results suggest that the combination of chitosan-based melanocyte spheroid patch with epidermal ablation by PUVA-induced sunburn reaction can be a feasible method to facilitate melanocyte transplantation. © 2018 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 106B: 2535-2543, 2018.

Research progress towards skin repigmentation in vitiligo

“…Many factors impair the viability of MCs transplanted in cell suspensions, including: 5,6 (1) Before transplantation, preparation of the cell suspension using trypsin digestion and mechanical shaking may damage MCs and cause them to undergo apoptosis or death. (2) During the transplantation process, it cannot be ensured that MCs in the suspension will adhere tightly to the recipient area, especially when the surface of that area is uneven.…”

“…It has been reported that MCs within MPs were successfully transferred to the depigmented areas of nude mice and survived well. 5 MPs were transplanted onto the epidermal wounds of nude mice and were active.…”

“…In order to improve the therapeutic efficacy, melanocyte patches (MPs) have been developed to replace the MC suspension and this article reviews recent studies on MPs. 5…”

Innovative Molecular and Cellular Therapeutics in Cleft Palate Tissue Engineering

“…Chitosan is a deacetylated form of the invertebrate exoskeleton biopolymer, chitin, which displays many promising characteristics that include its compatibility with other natural materials used for bone tissue engineering. [118][119][120][121] Chitosan-based delivery systems seeded with bone morphogenetic protein (BMP)-2 demonstrated increases in osteocalcin synthesis in cultured human embryonic palatal cells. 122,123 More recently, hydrophobically modified glycol chitosan nanocontrolled delivery of trichloroacetic acid and epidermal growth factor in vivo in critical-sized defects in palatal soft tissues in a canine model, demonstrating successful regeneration of these palatal defects.…”

Extracellular Matrix Composition and Remodeling: Current Perspectives on Secondary Palate Formation, Cleft Lip/Palate, and Palatal Reconstruction

“…Among these, cell spheroid is a technology in which cells are cultured in suspension to create a 3D structure using cell-cell interactions and "scaffold-free" strategy ( Figure 8B). This technique yields detailed information regarding ECM remodeling through "omics" analysis since the generation, growth, and fusion of cells allow accurate monitoring (Fan et al, 2018;Schnellmann et al, 2018;Wong et al, 2018). Understanding the ECM and its soluble factors are essential to comprehend embryonic development and tissue repair and will contribute to the discovery of new therapeutic tools.…”