Contribution of Fibroblasts to the Mechanical Stability of In Vitro Engineered Dermal-Like Tissue Through Extracellular Matrix Deposition

摘要

Tissue-engineered skin with mechanical and biological properties that match the native tissue could be a valuable graft to treat non-healing chronic wounds. Fibroblasts grown on a suitable biodegradable scaffold are a feasible strategy for the development of a dermal substitute above which epithelialization may occur naturally. Cell growth and phenotype maintenance are crucial to ensure the functional status of engineered tissue. In this study, an electrospun biodegradable polymer scaffold composed of a terpolymer PLGC [poly(lactide-glycolide-caprolactone)] with appropriate mechanical strength was used as a scaffold so that undesirable contraction of the wound could be prevented when it was implanted. To enhance cell growth, synthetic PLGC was incorporated with a fibrin-based biomimetic composite. The efficacy of the hybrid scaffold was evaluated by comparing it with bare PLGC in terms of fibroblast growth potential, extracellular matrix (ECM) deposition, polymer degradation, and mechanical strength. A significant increase was observed in fibroblast attachment, proliferation, and deposition of ECM proteins such as collagen and elastin in the hybrid scaffold. After growing fibroblasts for 20 d and 40 d, immunochemical staining of the decellularized scaffolds showed deposition of insoluble collagen and elastin on the hybrid scaffold but not on the bare scaffold. The loss of mechanical strength consequent to in vitro polymer degradation seemed to be balanced owing to the ECM deposition. Thus, tensile strength and elongation were better when cells were grown on the hybrid scaffold rather than the bare samples immersed in culture medium. Similar patterns of in vivo and in vitro degradation were observed during subcutaneous implantation and fibroblast culture, respectively. We therefore postulate that a hybrid scaffold comprising PLGC and fibrin is a potential candidate for the engineering of dermal tissue to be used in the regeneration of chronic wounds.