A Biomimetic Basement Membrane Substitute Based on Tri-Layered Nanofibrous Scaffold for Skin Reconstruction
A Biomimetic Basement Membrane Substitute Based on Tri-Layered Nanofibrous Scaffold for Skin Reconstruction
复制标题
基于三层纳米纤维支架的仿生基底膜替代品用于皮肤重建
DOI：
10.1166/jbn.2019.2865
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发表时间：
2019-12-01
影响因子：
2.9
通讯作者：
Li, Xueyong
中科院分区：
文献类型：
--
作者：
Huang, Rong;He, Zhi;Li, Xueyong
由于基底膜结构复杂，具有调节细胞行为的多功能性，因此其替代物的开发一直是构建功能性组织工程皮肤的主要难题。本文采用稳定的静电纺丝方法，以聚己内酯（PCL）和醋酸纤维素（CA）与壳聚糖（CS）复合的新型支架为基础，制备了天然骨基质的仿生模型。对制备的单层和三层支架的形貌、结构、表面亲水性、粗糙度和机械拉伸强度进行了综合比较。此外，通过在三层支架的相对侧上接种角质形成细胞（Kcs）和成纤维细胞（Fbs）的共培养系统显示了中心纳米纤维屏障内的两种细胞类型的更有效分离以及增强的细胞附着和增殖，而不是单层支架。此外，VII型胶原和层粘连蛋白-5的沉积进行了检查，与正常皮肤BM相比。组织学观察显示，植入2周后，真皮-表皮交界处BM区重建，并与内源性细胞相互作用，伤口愈合。此外，活性整合素β 1和磷酸化粘着斑激酶（FAK）的表达与三层支架的处理促进。这项研究强调，这种三层支架可能会引起Kcs和Fbs的仿生反应，从而适用于未来发展的BM含有组织。
Developing basement membranes (BMs) substitute remains major problem for constructing functional tissue engineered skin because of its complex structure and multifunction of regulating cellular behavior. Herein, a stable electrospinning method was employed to generate a biomimetic model of natural BMs based on novel scaffold electrospun from Poly(epsilon-caprolactone) (PCL) and cellulose acetate (CA) incorporated with chitosan (CS). The morphology, structure, surface hydrophilicity, roughness and mechanical tensile strength of prepared monolayer and tri-layered scaffold were comprehensively compared. Besides, co-culture system via seeding keratinocytes (Kcs) and fibroblasts (Fbs) on opposite side of tri-layered scaffold revealed more effective segregation of both cell types within the central nanofibrous barrier together with enhanced cell attachment and proliferation than that on the monolayer scaffold. Moreover, the deposition of type VII collagen and laminin-5 was examined in comparison with normal skin BMs. Furthermore, the histological studies revealed characteristics of reconstructing BM zone at the junction of dermis-epidermis after in vivo implantation for 2 weeks, and wound healing while the seeded cells interacted with the endogenous cells. Additionally, the expression of active integrin beta 1 and phosphorylated focal adhesion kinase (FAK) was promoted with treatment of tri-layered scaffold. This study stressed that this tri-layer scaffold might provoke biomimetic responses of Kcs and Fbs and thus be applied for future development of BMs containing tissues.