Chondrocytes behaviors within type I collagen microspheres and bulk hydrogels: an in vitro study
Chondrocytes behaviors within type I collagen microspheres and bulk hydrogels: an in vitro study
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I 型胶原微球和块状水凝胶内的软骨细胞行为：一项体外研究
DOI：
10.1039/c5ra04496k
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发表时间：
2015-01-01
期刊：
影响因子：
3.9
通讯作者：
Zhang, Xingdong
中科院分区：
文献类型：
--
作者：
Liu, Jun;Lin, Hai;Zhang, Xingdong
细胞微环境是组织工程支架材料设计和开发的重要方面，对细胞的增殖和分化起着至关重要的作用。支架的传质性能影响着其他物质的营养供应和交换。在这项研究中，我们制备了胶原蛋白水凝胶的微球（CHMs）和散装（CHB）的形式，调查质量交换的差异及其对嵌入的软骨细胞的影响。CHMs采用乳液法制备，可有效负载电池。以牛血清白蛋白（BSA）为扩散模型，研究了水凝胶在CHMs和CHB中的传输性能和释放动力学。在4周的体外培养过程中，以不同的时间间隔监测水凝胶的收缩、细胞活力和形态以及DNA和糖胺聚糖（GAG）含量。结果表明，CHMs显示出明显的优势，在转移性能的CHB，导致更好地保持软骨细胞表型的CHMs在体外培养的早期阶段。组织学分析表明，从第7天开始，CHM中出现大量的腔隙和均匀的阳性GAG染色。相比之下，在第21天后，在CHB的外部区域中仅发现少数腔隙和模糊的GAG染色。由于缺乏足够的营养，CHB内区的软骨细胞分泌基质很少。基于所提出的CHM系统，进一步开发的构建物被认为是工程软骨组织临床应用的一种有前途的替代方案。
Cell niche, which is considered to be critical to the proliferation and differentiation of cells, is one of the most important aspects for the design and development of ideal scaffolds in tissue engineering. The mass transfer property of the scaffold affects the nutrient supply and exchange of the other substances. In this study, we prepared collagen hydrogels in the form of microspheres (CHMs) and bulk (CHB) to investigate the mass exchange differences and their influence on embedded chondrocytes. CHMs were developed by the emulsion method, which was efficient to load cells. Bovine serum albumin (BSA) was used as a diffusion model in the CHMs and CHB to evaluate the transport property of the hydrogels and the release kinetics. During the 4 week in vitro culture process, the contraction of the hydrogels, the cell viability and morphology, and the DNA and glycosaminoglycan (GAG) content were monitored at different intervals. The results suggested that the CHMs showed an obvious superiority in the transfer property over the CHB, leading to better maintenance of the chondrocyte phenotype in CHMs at the early stage of the in vitro culture. Histological analyses indicated that lots of lacunae and homogeneous positive GAG staining appeared in the CHMs from day 7. In contrast, only a few lacunae and obscure GAG staining were found in the outer area of the CHB after day 21. Without enough nutrients, the chondrocytes in the inner area of the CHB had little secreted matrix. Based on the presented CHM system, a further developed construct is suggested as a promising alternative toward the clinical application of engineered cartilaginous tissue.