The impact of decellularization strategy on the structural and biological integrity of human dermis: a comparative analysis of three decellularization methods.

Abstract

Introduction: Acellular dermal matrices are increasingly applied in regenerative medicine and wound healing because they mimic native dermal structure and exhibit low immunogenicity. However, decellularization methods vary, and no consensus exists on the optimal protocol for human full-thickness dermis. This study compared three decellularization approaches to identify the most effective method for generating biocompatible, structurally preserved acellular dermis suitable for clinical use.

Methods: Full-thickness human dermis was processed using: (1) alkaline NaOH treatment, (2) Triton X-100 followed by mild alkali, and (3) freeze-thaw cycling with surfactant and sonication. The resulting scaffolds were assessed macroscopically, histologically, and through cytocompatibility assays. In vivo wound healing was evaluated in a murine full-thickness wound model.

Results: Alkaline treatment effectively removed cells but damaged the extracellular matrix, reducing mechanical strength and biocompatibility. Triton X-100-based and freeze-thaw/surfactant protocols preserved dermal architecture and mechanical integrity while ensuring complete decellularization and cytocompatibility. Both supported improved cellular infiltration and tissue remodeling in vivo. Neoangiogenesis and graft orientation showed minimal influence on healing.

Conclusion: Triton X-100-based and freeze-thaw/surfactant protocols achieved optimal balance between cell removal and matrix preservation, offering promising strategies for producing clinically applicable acellular dermal scaffolds.