Yu Xiaohua, Mengsteab Paulos Y, Narayanan Ganesh, Nair Lakshmi S, Laurencin Cato T
Connecticut Convergence Institute for Translation in Regenerative Engineering, University of Connecticut Health, Farmington, CT 06030, USA.
Raymond and Beverly Sackler Center for Biological, Physical and Engineering Sciences, University of Connecticut Health, Farmington, CT 06030, USA.
Engineering (Beijing). 2021 Feb;7(2):153-161. Epub 2020 May 7.
We have previously developed a poly(-lactic) acid (PLLA) bioengineered anterior cruciate ligament (ACL) matrix that has demonstrated enhanced healing when seeded with primary ACL cells prior to implantation in a rabbit model, as compared with the matrix alone. This suggests that improving cell adhesion on the matrix may beneficially affect the healing response and long-term performance of the bioengineered ACL matrix. One regenerative engineering approach involves enhancing the surface properties of the matrix to support cell adhesion and growth in combination with point-of-care stem cell therapy. Herein, we studied the cell adhesion properties of PLLA braided microfiber matrices enhanced through the physical adsorption of fibronectin and air plasma treatment. We evaluated the kinetics and binding efficiency of fibronectin onto matrices at three time points and three fibronectin concentrations. Incubating the matrix for 120 min in a solution of 25 mg mL fibronectin achieved the greatest binding efficiency to the matrix and cellular adhesion. Exposing the matrices to air plasma treatment for 5 min before fibronectin adsorption significantly enhanced the cell adhesion of rabbit bone marrow-derived mesenchymal stem cells (R-BMMSCs) 24 h post cell seeding. Finally, cellular proliferation was monitored for up to 21 d, the matrices were exposed to air plasma treatment, and fibronectin adsorption was found to result in enhanced cell number. These findings suggest that exposure to air plasma treatment and fibronectin adsorption enhances the cellular adhesion of PLLA braided microfiber matrices and may improve the clinical efficacy of the matrix in combination with point-of-care stem cell therapies.
我们之前开发了一种聚(-乳酸)(PLLA)生物工程化前交叉韧带(ACL)基质,与单独的基质相比,在兔模型中植入前用原代ACL细胞接种时,该基质已显示出增强的愈合效果。这表明改善细胞在基质上的粘附可能会有益地影响生物工程化ACL基质的愈合反应和长期性能。一种再生工程方法涉及增强基质的表面特性,以支持细胞粘附和生长,并结合即时护理干细胞疗法。在此,我们研究了通过纤连蛋白的物理吸附和空气等离子体处理增强的PLLA编织微纤维基质的细胞粘附特性。我们在三个时间点和三种纤连蛋白浓度下评估了纤连蛋白在基质上的动力学和结合效率。在25 mg/mL纤连蛋白溶液中孵育基质120分钟可实现与基质的最大结合效率和细胞粘附。在纤连蛋白吸附前将基质暴露于空气等离子体处理5分钟,可显著增强接种细胞24小时后兔骨髓间充质干细胞(R-BMMSCs)的细胞粘附。最后,监测细胞增殖长达21天,对基质进行空气等离子体处理,发现纤连蛋白吸附可导致细胞数量增加。这些发现表明,暴露于空气等离子体处理和纤连蛋白吸附可增强PLLA编织微纤维基质的细胞粘附,并可能提高该基质与即时护理干细胞疗法联合使用时的临床疗效。