Department of Orthopedics, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China.
College of Life Science and Technology, Huazhong University of Science and Technology, Wuhan 430074, China.
J Biomed Mater Res A. 2019 Jun;107(6):1294-1302. doi: 10.1002/jbm.a.36642. Epub 2019 Feb 21.
In this current study, a novel multilayer porous composite scaffold was fabricated with chitosan (CS), silk fibrin (SF) and nano-hydroxyapatite (n-HA). Scanning electron microscope was utilized to detect the characteristics of the composed scaffold. Rat bone marrow stromal cells (rBMSC) were loaded onto the CS/SF/n-HA scaffold and cultured in a bioreactor under an on-off dynamic compression (10% compressive strain, 0.5 Hz, [2 h action + 4 h pause]/cycle, 4 cycles/day). Metabolism of the loaded rBMSC was assessed through CCK-8 test. Qualitative polymerase chain reaction and western blot were applied to assess the chondrogenic differentiation of the seeded cells. Compressive modulus of the cell/scaffold constructs was analyzed. Additionally, a pig model was employed to evaluate the effect of the tissue-engineered cartilage on repairing of cartilage defect. Results showed that the four layers within the scaffold were tightly connected without gaps between porous interfaces of the layers. Scaffold porosity was 92.20% ± 1.30%. The cyclic compression upregulated chondrogenesis markers (Aggrecan, Sox-9, and collagen II). Increased compressive modulus of the cell/scaffold complex was detected after dynamic compression. The pig bone marrow stromal cells/scaffold complex exposed to cyclic compression presented most favorable reparative effect on the mini pig femoral condyle cartilage defects. Our study suggested that the on-off dynamic compression might be a promising approach to fabricate tissue-engineered cartilage in vitro. © 2019 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 107A: 1294-1302, 2019.
在本研究中,采用壳聚糖(CS)、丝素纤维(SF)和纳米羟基磷灰石(n-HA)制备了一种新型多层多孔复合支架。利用扫描电子显微镜检测了复合支架的特性。将大鼠骨髓基质细胞(rBMSC)加载到 CS/SF/n-HA 支架上,并在生物反应器中进行开/关动态压缩(10%压缩应变,0.5 Hz,[2 h 作用+4 h 暂停]/循环,每天 4 个循环)下培养。通过 CCK-8 试验评估加载 rBMSC 的代谢情况。应用定性聚合酶链反应和 Western blot 评估接种细胞的软骨分化情况。分析细胞/支架构建体的压缩模量。此外,还采用猪模型评估组织工程软骨修复软骨缺损的效果。结果表明,支架内的 4 层紧密相连,层间多孔界面无间隙。支架的孔隙率为 92.20%±1.30%。周期性压缩上调软骨形成标志物(聚集蛋白聚糖、Sox-9 和胶原 II)。动态压缩后检测到细胞/支架复合物的压缩模量增加。暴露于周期性压缩的猪骨髓基质细胞/支架复合物对小型猪股骨髁软骨缺损具有最佳的修复效果。本研究表明,开/关动态压缩可能是体外构建组织工程软骨的一种很有前途的方法。© 2019 威立出版社