Du Ping, Casavitri Cininta, Suhaeri Muhammad, Wang Peng-Yuan, Lee Jong Ho, Koh Won-Gun, Park Kwideok
Center for Biomaterials, Korea Institute of Science and Technology (KIST), Seoul 02792, Republic of Korea.
Center for Human Tissues & Organs Degeneration, Shenzhen Institute of Advanced Technology, Chinese Academy of Sciences, Shenzhen, Guangdong 518055, China.
ACS Biomater Sci Eng. 2019 Feb 11;5(2):900-910. doi: 10.1021/acsbiomaterials.8b01118. Epub 2018 Dec 27.
Skin wound healing is an intricate orchestration that involves different cell types, an extracellular matrix (ECM), cytokines, and growth factors. On the basis of the great benefits of cell-derived ECM in regenerative applications, here we propose an electrospun fibrous membrane that combines poly(l-lactide--caprolactone) (PLCL) and human fibroblast-derived ECM (hFDM). hFDM-deposited PLCL (hFDM-PLCL) was obtained via decellularization of a confluent layer of fibroblasts cultivated on PLCL. An organized assembly of fibrillar structure on hFDM-PLCL was notable via immunostaining. As human umbilical vein endothelial cells (HUVECs) were seeded on the hFDM-PLCL, they proliferated faster and exhibited more elongated, capillary-like morphology than those on PLCL or fibronectin-coated PLCL (Fn-PLCL). HUVECs have a relatively large aspect ratio, spreading area, and vinculin-positive area per cell on the fibrillary structure of hFDM-PLCL. In addition, transwell cell migration assay showed the chemoattractant effect of hFDM for HUVECs and human dermal fibroblasts. Furthermore, HUVECs-loaded hFDM-PLCL membranes showed the most promising therapeutic effects on a mouse skin wound model as proved via the wound closure rate, neovascularization effect, regenerated epidermis, and skin appendage. This study shows that biodegradable PLCL fibers not only support the weak mechanical properties of hFDM but also allow hFDM to reserve ECM macromolecules and to maintain structural integrity. Current results also demonstrate the critical role of hFDM with biochemical and biophysical cues on HUVECs adhesion, proliferation, and vascular morphogenesis and even on the wound healing process . Taken together, hFDM-functionalized PLCL patch should be a promising platform for cell delivery and regenerative applications.
皮肤伤口愈合是一个复杂的过程，涉及不同的细胞类型、细胞外基质（ECM）、细胞因子和生长因子。基于细胞衍生的ECM在再生应用中的巨大益处，我们在此提出一种将聚（L-丙交酯-ε-己内酯）（PLCL）与人成纤维细胞衍生的ECM（hFDM）相结合的电纺纤维膜。通过对在PLCL上培养的成纤维细胞汇合层进行脱细胞处理，获得了沉积有hFDM的PLCL（hFDM-PLCL）。通过免疫染色可以明显看到hFDM-PLCL上有纤维状结构的有序组装。当人脐静脉内皮细胞（HUVECs）接种在hFDM-PLCL上时，它们的增殖速度更快，并且与接种在PLCL或纤连蛋白包被的PLCL（Fn-PLCL）上的细胞相比，呈现出更细长的毛细血管样形态。在hFDM-PLCL的纤维结构上，HUVECs具有相对较大的纵横比、铺展面积和每个细胞的粘着斑蛋白阳性面积。此外，Transwell细胞迁移试验显示hFDM对HUVECs和人皮肤成纤维细胞具有趋化作用。此外，在小鼠皮肤伤口模型中，负载HUVECs的hFDM-PLCL膜通过伤口闭合率、新血管形成效应、再生表皮和皮肤附属器等指标显示出最有前景的治疗效果。这项研究表明，可生物降解的PLCL纤维不仅支持hFDM较弱的机械性能，还能使hFDM保留ECM大分子并维持结构完整性。目前的结果还证明了hFDM在生化和生物物理线索方面对HUVECs粘附、增殖和血管形态发生甚至对伤口愈合过程的关键作用。综上所述，hFDM功能化的PLCL贴片应该是一个有前景的细胞递送和再生应用平台。
