Chen Shangwu, Zhang Qin, Nakamoto Tomoko, Kawazoe Naoki, Chen Guoping
1 Tissue Regeneration Materials Unit, International Center for Materials Nanoarchitectonics, National Institute for Materials Science , Tsukuba, Japan .
2 Department of Materials Science and Engineering, Graduate School of Pure and Applied Sciences, University of Tsukuba , Tsukuba, Japan .
Tissue Eng Part C Methods. 2016 Mar;22(3):189-98. doi: 10.1089/ten.TEC.2015.0281. Epub 2016 Jan 21.
Engineering of cartilage tissue in vitro using porous scaffolds and chondrocytes provides a promising approach for cartilage repair. However, nonuniform cell distribution and heterogeneous tissue formation together with weak mechanical property of in vitro engineered cartilage limit their clinical application. In this study, gelatin porous scaffolds with homogeneous and open pores were prepared using ice particulates and freeze-drying. The scaffolds were used to culture bovine articular chondrocytes to engineer cartilage tissue in vitro. The pore structure and mechanical property of gelatin scaffolds could be well controlled by using different ratios of ice particulates to gelatin solution and different concentrations of gelatin. Gelatin scaffolds prepared from ≥70% ice particulates enabled homogeneous seeding of bovine articular chondrocytes throughout the scaffolds and formation of homogeneous cartilage extracellular matrix. While soft scaffolds underwent cellular contraction, stiff scaffolds resisted cellular contraction and had significantly higher cell proliferation and synthesis of sulfated glycosaminoglycan. Compared with the gelatin scaffolds prepared without ice particulates, the gelatin scaffolds prepared with ice particulates facilitated formation of homogeneous cartilage tissue with significantly higher compressive modulus. The gelatin scaffolds with highly open pore structure and good mechanical property can be used to improve in vitro tissue-engineered cartilage.
使用多孔支架和软骨细胞在体外构建软骨组织为软骨修复提供了一种有前景的方法。然而，体外构建软骨的细胞分布不均匀、组织形成异质性以及力学性能较弱限制了它们的临床应用。在本研究中，利用冰颗粒和冷冻干燥制备了具有均匀且开放孔隙的明胶多孔支架。这些支架用于培养牛关节软骨细胞以在体外构建软骨组织。通过使用不同比例的冰颗粒与明胶溶液以及不同浓度的明胶，可以很好地控制明胶支架的孔隙结构和力学性能。由≥70%冰颗粒制备的明胶支架能够使牛关节软骨细胞在整个支架中均匀接种，并形成均匀的软骨细胞外基质。软支架会发生细胞收缩，而硬支架能抵抗细胞收缩，并且具有显著更高的细胞增殖和硫酸化糖胺聚糖合成。与未使用冰颗粒制备的明胶支架相比，使用冰颗粒制备的明胶支架有助于形成具有显著更高压缩模量的均匀软骨组织。具有高度开放孔隙结构和良好力学性能的明胶支架可用于改善体外组织工程软骨。
