Electrospun nanofibrous SF/P(LLA-CL) membrane: a potential substratum for endothelial keratoplasty.
Electrospun nanofibrous SF/P(LLA-CL) membrane: a potential substratum for endothelial keratoplasty.
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电纺纳米纤维SF/P（LLA-CL）膜：内皮角化膜成形术的潜在底层。
DOI：
10.2147/ijn.s77706
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发表时间：
2015
影响因子：
8
通讯作者：
Fan X
中科院分区：
文献类型：
--
作者：
Chen J;Yan C;Zhu M;Yao Q;Shao C;Lu W;Wang J;Mo X;Gu P;Fu Y;Fan X
近几十年来，角膜移植技术取得了显着进步，使外科医生能够用薄层状结构替换患病的角膜内皮。目前，用于内皮角膜移植术的薄的、透明的、生物相容的、具有角膜内皮细胞的组织工程基质引起了人们的兴趣。静电纺丝纳米纤维结构可以模拟细胞外基质，对细胞培养产生有益的影响。丝素蛋白（SF）具有良好的生物相容性但机械性能较差，而聚（L-乳酸-co-ε-己内酯）（P（LLA-CL））具有良好的机械性能但生物相容性较差。将SF与P(LLA-CL)共混可以保持这两种材料的优点并克服它们的缺点。混合电纺纳米纤维膜可能适合角膜内皮的再生。本研究的目的是生产适合内皮角膜移植术的组织工程结构。制造了五种含有不同 SF:P(LLA-CL) 混合比例（100:0、75:25、50:50、25:75、0:100）的支架。在膜上培养人角膜内皮（B4G12）细胞系。在不同混合比例的膜上检测透光率、细胞贴壁速度、细胞活力（活死试验）、细胞增殖（Ki-67、BrdU染色）和细胞单层形成，并通过实时聚合酶链式反应检测一些功能基因的表达。不同混合比例的支架具有不同的透光性能。 25:75混合比例的膜在这些支架中具有最好的透光率。与对照组相比，所有电纺纳米纤维膜均显示出细胞粘附速度的提高，特别是当 P(LLA-CL) 比率增加时。 25:75 混合比例的膜也具有最高的细胞增殖能力。 B4G12细胞可以在所有支架上形成单层，并且大多数功能基因也在所有支架上稳定表达。只有两个基因表现出表达变化。对 SF:P(LLA-CL) 支架的所有混合比例进行了评估，并显示出良好的细胞粘附和单层形成生物相容性。其中，25:75混合比例的SF:P(LLA-CL)支架的透光率最好，细胞增殖能力最高。这些属性进一步促进了 SF:P(LLA-CL) 支架在角膜内皮移植中的潜在应用。
Cornea transplant technology has progressed markedly in recent decades, allowing surgeons to replace diseased corneal endothelium by a thin lamellar structure. A thin, transparent, biocompatible, tissue-engineered substratum with corneal endothelial cells for endothelial keratoplasty is currently of interest. Electrospinning a nanofibrous structure can simulate the extracellular matrix and have beneficial effects for cell culture. Silk fibroin (SF) has good biocompatibility but poor mechanical properties, while poly(l-lactic acid-co-ε-caprolactone) (P(LLA-CL)) has good mechanical properties but poor biocompatibility. Blending SF with P(LLA-CL) can maintain the advantages of both these materials and overcome their disadvantages. Blended electrospun nanofibrous membranes may be suitable for regeneration of the corneal endothelium. The aim of this study was to produce a tissue-engineered construct suitable for endothelial keratoplasty. Five scaffolds containing different SF:P(LLA-CL) blended ratios (100:0, 75:25, 50:50, 25:75, 0:100) were manufactured. A human corneal endothelial (B4G12) cell line was cultured on the membranes. Light transmission, speed of cell adherence, cell viability (live-dead test), cell proliferation (Ki-67, BrdU staining), and cell monolayer formation were detected on membranes with the different blended ratios, and expression of some functional genes was also detected by real-time polymerase chain reaction. Different blended ratios of scaffolds had different light transmittance properties. The 25:75 blended ratio membrane had the best transmittance among these scaffolds. All electrospun nanofibrous membranes showed improved speed of cell adherence when compared with the control group, especially when the P(LLA-CL) ratio increased. The 25:75 blended ratio membranes also had the highest cell proliferation. B4G12 cells could form a monolayer on all scaffolds, and most functional genes were also stably expressed on all scaffolds. Only two genes showed changes in expression. All blended ratios of SF:P(LLA-CL) scaffolds were evaluated and showed good biocompatibility for cell adherence and monolayer formation. Among them, the 25:75 blended ratio SF:P(LLA-CL) scaffold had the best transmittance and the highest cell proliferation. These attributes further the potential application of the SF:P(LLA-CL) scaffold for corneal endothelial transplantation.