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猪食管成纤维细胞在细胞外基质蛋白功能化聚乳酸上的黏附动力学

Adhesion dynamics of porcine esophageal fibroblasts on extracellular matrix protein-functionalized poly(lactic acid).

作者信息

Cai Ning, Gong Yingxue, Chian Kerm Sin, Chan Vincent, Liao Kin

机构信息

School of Chemical and Biomedical Engineering, Nanyang Technological University, Singapore 639798, Singapore.

出版信息

Biomed Mater. 2008 Mar;3(1):015014. doi: 10.1088/1748-6041/3/1/015014. Epub 2008 Mar 3.

Abstract

Effective attachment of esophageal cells on biomaterials is one important requirement in designing engineered esophagus substitute for esophageal cancer treatment. In this study, poly(lactic acid) (PLA) was subjected to surface modification by coupling extracellular matrix (ECM) proteins on its surface to promote cell adhesion. Two typical ECM proteins, collagen type I (COL) and fibronectin (FN), were immobilized on the PLA surface with the aid of glutaraldehyde as a cross linker between aminolyzed PLA and ECM proteins. By using confocal reflectance interference contrast microscopy (C-RICM) integrating with phase contrast microscopy, the long-term adhesion dynamics of porcine esophageal fibroblasts (PEFs) on four types of surfaces (unmodified PLA, PLA-COOH, PLA-COL and PLA-FN) was investigated during 24 h of culture. It is demonstrated by C-RICM results that PEFs form strong adhesion contact on all four types of surfaces at different stages of cell seeding. Among the four surfaces, PEFs on the PLA-FN surface reach the maximum adhesion energy (9.5 x 10(-7) J m(-2)) in the shortest time (20 min) during the initial stage of cell seeding. After adhesion energy reaches the maximum value, PEFs maintain their highly deformed geometries till they reached a steady state after 20 h of culture. F-actin immunostaining results show that the evolvement of spatial organization of F-actin is tightly correlated with the formation of adhesion contact and cell spreading. Furthermore, the cell attachment ratio of PEFs on PLA in 2 h is only 26% compared with 88% on PLA-FN, 73% on PLA-COL and 36% on PLA-COOH. All the results demonstrate the effect of surface functionalization on the biophysical responses of PEFs in cell adhesion. Fibronectin-immobilized PLA demonstrates promising potential for application as an engineered esophagus substitute.

摘要

食管细胞在生物材料上的有效附着是设计用于食管癌治疗的工程化食管替代物的一项重要要求。在本研究中,通过在聚乳酸(PLA)表面偶联细胞外基质(ECM)蛋白对其进行表面改性,以促进细胞黏附。借助戊二醛作为氨解PLA与ECM蛋白之间的交联剂,将两种典型的ECM蛋白,即I型胶原蛋白(COL)和纤连蛋白(FN)固定在PLA表面。通过结合相差显微镜的共聚焦反射干涉对比显微镜(C-RICM),研究了猪食管成纤维细胞(PEF)在四种类型表面(未改性PLA、PLA-COOH、PLA-COL和PLA-FN)上培养24小时期间的长期黏附动力学。C-RICM结果表明,在细胞接种的不同阶段,PEF在所有四种类型的表面上都形成了强黏附接触。在这四种表面中,在细胞接种初始阶段,PLA-FN表面上的PEF在最短时间(20分钟)内达到最大黏附能(9.5×10⁻⁷ J m⁻²)。黏附能达到最大值后,PEF保持其高度变形的几何形状,直到培养20小时后达到稳定状态。F-肌动蛋白免疫染色结果表明,F-肌动蛋白空间组织的演变与黏附接触的形成和细胞铺展紧密相关。此外,2小时时PEF在PLA上的细胞附着率仅为26%,而在PLA-FN上为88%,在PLA-COL上为73%,在PLA-COOH上为36%。所有结果都证明了表面功能化对PEF细胞黏附生物物理反应的影响。固定有纤连蛋白的PLA显示出作为工程化食管替代物应用的潜在前景。

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