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三维鸭脚胶原蛋白/PLGA 支架软骨化:孔径和孔隙率的作用。

Three-dimensional duck's feet collagen/PLGA scaffold for chondrification: role of pore size and porosity.

机构信息

a Department of BIN Convergence Technology, Department of Polymer Nano Science & Technology and Polymer Fusion Research Center , Chonbuk National University , Jeonju , Republic of Korea.

b Department of Orthopedic Surgery , Yeouido St. Mary's Hospital, Catholic University of Korea , Seoul , Korea.

出版信息

J Biomater Sci Polym Ed. 2018 May-Jun;29(7-9):932-941. doi: 10.1080/09205063.2017.1394712. Epub 2017 Oct 31.

Abstract

An ideal tissue-engineered scaffold must provide sufficient porosity to allow free movement of cells, nutrients, and oxygen for proper cell growth and further maintenance. Owing to variation in pore sizes and shapes of as-fabricated scaffold, the amount of oxygen available for the cells attached to the scaffold and transfer of by-products and excrement will be different, which ultimately results in cell activity. Thus, optimizing pore size and porosity of a scaffold for a specific tissue regeneration are one of the key highlights, which should be considered while designing a scaffold as well as choosing a specific cell type. In this study, three-dimensional (3D) scaffolds based on blends of duck's feet collagen (DC) and poly (lactic-co-glycolic acid) (PLGA) with different pore sizes i.e. 90-180, 180-250, 250-355 and 355-425 μm were prepared using solvent casting/salt leaching approach and examined its effects on chondrification. The morphological analysis of the as-fabricated scaffolds was performed using SEM for studying porosity and pore size. The cell proliferation and gene expression were investigated after culturing costal chondrocytes on each scaffolds using 3-(4, 5-dimethylthiazol-2-yl)-2,5 diphenyltetrazolium bromide (MTT) assay and qRT-PCR. Histological staining of in vivo implants was performed in nude mice as models. The biological evaluation showed a pore-size dependent chondrification at different time points. Especially, the 355-425 μm DC/PLGA scaffold showed a highest positive impact on maintenance of cell proliferation, costal chondrocyte phenotype and increased glycosaminoglycan accumulation than the other groups. These results indicated that DC/PLGA scaffolds with pore size ranging from 250 to 425 μm can be considered as highly-suitable constructs for enhanced chondrification.

摘要

理想的组织工程支架必须提供足够的孔隙率,以允许细胞、营养物质和氧气自由移动,从而促进细胞的正常生长和进一步的维持。由于制造的支架的孔径和形状的变化,附着在支架上的细胞可用的氧气量以及代谢产物和排泄物的转移将不同,这最终会影响细胞活性。因此,优化特定组织再生用支架的孔径和孔隙率是关键要点之一,在设计支架以及选择特定细胞类型时都应考虑这一点。在这项研究中,使用溶剂浇铸/盐析法制备了基于鸭脚胶原蛋白(DC)和聚(乳酸-共-乙醇酸)(PLGA)混合物的具有不同孔径(即 90-180、180-250、250-355 和 355-425 μm)的三维(3D)支架,并研究了其对软骨化的影响。使用 SEM 对制造的支架进行形态分析,以研究其孔隙率和孔径。使用 3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐(MTT)测定法和 qRT-PCR 研究了在每个支架上培养肋软骨细胞后的细胞增殖和基因表达。在裸鼠模型中进行了体内植入物的组织学染色。生物评估显示,不同时间点的软骨化具有孔径依赖性。特别是,355-425 μm 的 DC/PLGA 支架对维持细胞增殖、肋软骨细胞表型和增加糖胺聚糖积累的影响最大。这些结果表明,孔径在 250 到 425 μm 之间的 DC/PLGA 支架可以被认为是增强软骨化的高度合适的构建体。

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