Fabrication and characterization of nano-fibrous bilayer composite for skin regeneration application
Fabrication and characterization of nano-fibrous bilayer composite for skin regeneration application
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DOI：
10.1016/j.ymeth.2015.08.017
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发表时间：
2016-04-15
期刊：
影响因子：
4.8
通讯作者：
Mobini, Sahba
中科院分区：
文献类型：
--
作者：
Arasteh, Shaghayegh;Kazemnejad, Somaieh;Mobini, Sahba
全层创面愈合，最小的疤痕和完全恢复正常的皮肤属性仍然是临床上的一个挑战。在本研究中，制备了一种双层皮肤替代物，以仿生皮肤天然细胞外基质的微结构。将人羊膜(HAM)与丝素纳米纤维复合形成双层结构，并对其进行处理和表征。火腿取自健康母亲，用精细酶法去上皮化，以保持细胞外结构。从新鲜蚕茧中提取丝素蛋白，转化为均匀的纳米纤维结构，作为脱上皮膜的涂层。尝试通过氧等离子体处理对表面进行改性，以进一步诱导亲水性。随后，对支架的形态、力学性能、亲水性和细胞培养反应进行了全面的表征。组织学和免疫组织化学染色显示去上皮化后的HAM纤维连接蛋白定位、细胞剥离和结构完整性。扫描电子显微镜显示无珠状丝素纳米纤维，平均直径为250 nm。经氧等离子体处理后，双层支架的水接触角显著降低至26.34，这与表面亲水性增强有关。由于丝素纳米纤维的包覆，使火腿的力学性能显著提高。拉伸杨氏模数和拉伸强度分别从16.14兆帕和68.46兆帕提高到25.69兆帕和108.03兆帕。体外培养小鼠胚胎成纤维细胞14天后发现，双层支架能够支持细胞贴壁和增殖。等离子蚀刻支架通过允许细胞穿透毛孔下方并沿纤维方向整合，为细胞-基质串扰提供了最佳的利基环境。研究结果表明，基于HAM的纳米纤维双层复合材料在皮肤再生应用中具有潜在的替代性。(C)2015 Elsevier Inc.保留所有权利。
Full thickness wound healing with minimal scarring and complete restoration of normal skin properties still remains as a clinical challenge. In this study, a bilayer skin substitute has been fabricated to biomimic the microstructure of natural extracellular matrix of the skin. Human amniotic membrane (HAM) and silk fibroin nano-fibers were combined to produce bilayer construct, which was further treated and characterized. HAM was obtained from healthy mothers and de-epithelized by means of fine enzymatic method to preserve the extracellular structure. Fibroin protein was extracted from fresh Bombyx mori cocoons and transformed to uniform nano-fiberous structure, which was used as a coating layer on the de-epithelized membrane. Surface modification through oxygen plasma treatment was attempted to further induce hydrophilicity. Subsequently, scaffolds were fully characterized in terms of morphology, mechanical properties, hydrophilicity and cell culture response. Histological and immunohistological staining demonstrated localization of fibronectin, cell denudation and structural integrity of HAM after de-epithelization. Scanning electron microscopy images showed bead-free silk fibroin nano-fibers with the average diameter of 250 nm. Water contact angle of bilayer scaffolds reduced dramatically to 26.34 after oxygen plasma treatment, which is correlated with more hydrophilic surface. Due to fibroin nano-fiber coating, mechanical properties of HAM improved significantly. Tensile Young's modulus and tensile strength increased from 16.14 MPa and 68.46 MPa to 25.69 MPa and 108.03 MPa, respectively. 14 days in vitro cultivation of mouse embryonic fibroblasts on the scaffolds revealed that bilayer scaffolds are able to support cell attachment and proliferation. Plasma-etched scaffolds provided the best niche for cell-matrix crosstalk by allowing cells to penetrate beneath the pores and to integrate in fibers direction. The obtained results suggest that the presented nano-fibrous bilayer composite based on HAM is a potential substitute for skin regeneration application. (C) 2015 Elsevier Inc. All rights reserved.