Beneficial effect on rapid skin wound healing through carboxylic acid-treated chicken eggshell membrane

At the initial stage of wound healing, growth factors stimulate tissue regeneration by interacting with the extracellular matrix (ECM), leading to rapid wound repair and structural support. Chicken eggshell membrane (ESM) is a low-cost and highly functional ECM biomaterial for tissue regeneration. However, natural ESM has limitations for tissue engineering purposes because it is difficult to control the size, shape, and biocompatibility of the surfaces. To overcome this, blends of synthetic materials and natural ESMs, such as soluble eggshell membrane protein, are combined for biomaterial applications. Unfortunately, it is difficult to pattern fibrous structure. Here, we modified the natural chicken ESM through weak acid treatment to promote wound healing and skin regeneration without loss of fibrous structure. Treatment of citric acid and acetic acid reacted the amine or amide group with carboxyl groups (R-COOH) and achieved hydrophilicity for adherence of proliferating regenerative cells. Our in vitro study revealed that the modified ESM scaffolds significantly promoted human dermal fibroblasts adhesion, viability, proliferation, and cytokine secretion, compared with natural ESM. In addition, the modified ESM accelerated skin regeneration and enhanced the wound healing process even at early stages in an in vivo rat wound model. Collectively, the modified ESM performed best for promoting skin regeneration, cytokine secretion, epidermal cell proliferation, and controlling the inflammatory response both in vitro and in vivo.