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热致相分离(TIPS)法制备的 PLLA 支架可根据孔径大小,促进人软骨细胞生长和细胞外基质形成。

PLLA scaffolds produced by thermally induced phase separation (TIPS) allow human chondrocyte growth and extracellular matrix formation dependent on pore size.

机构信息

Department of Civil, Environmental, Aerospace, Materials Engineering, Bio and Tissue Engineering Lab, Universita' di Palermo, Palermo, Italia.

Institute of Anatomy, Paracelsus Medical University, Salzburg and Nuremberg, Nuremberg, Germany; Department of Orthopaedic, Trauma and Reconstructive Surgery, Charité-Universitätsmedizin Berlin, Campus Benjamin Franklin, Germany.

出版信息

Mater Sci Eng C Mater Biol Appl. 2017 Nov 1;80:449-459. doi: 10.1016/j.msec.2017.06.011. Epub 2017 Jun 17.

Abstract

Damage of hyaline cartilage species such as nasoseptal or joint cartilage requires proper reconstruction, which remains challenging due to the low intrinsic repair capacity of this tissue. Implantation of autologous chondrocytes in combination with a biomimetic biomaterial represents a promising strategy to support cartilage repair. The aim of this work was to assess the viability, attachment, morphology, extracellular matrix (ECM) production of human articular and nasoseptal chondrocytes cultured in vitro in porous poly(l-lactic) (PLLA) scaffolds of two selected pore sizes (100 and 200μm). The PLLA scaffolds with 100 and 200μm pore sizes were prepared via ternary thermally induced phase separation (TIPS) technique and analyzed using scanning electron microscopy (SEM). Articular and nasoseptal chondrocytes were seeded on the scaffold and cultures maintained for 7 and 14days. Live/dead staining, (immuno-)histology and gene expression analysis of type II, type I collagen, aggrecan and SOX9 were performed to assess scaffold cytocompatibility and chondrocyte phenotype. The majority of both chondrocyte types survived on both scaffolds for the whole culture period. Hematoxylin-eosin (HE), alcian blue (visualizing glycosaminoglycans) stainings, immunoreactivity and gene expression of ECM proteins and cartilage marker (type II, I collagen, aggrecan, SOX9) of the chondrocyte scaffold constructs indicated that the smaller pore dimensions promoted the differentiation of the chondrocytes compared with the larger pore size. The present work revealed that the scaffold pore size is an important factor influencing chondrocyte differentiation and indicated that the scaffolds with 100μm pores serve as a cytocompatible basis for further future modifications.

摘要

透明软骨物种(如鼻中隔或关节软骨)的损伤需要适当的重建,由于这种组织的内在修复能力较低,因此仍然具有挑战性。将自体软骨细胞植入与仿生生物材料相结合,代表了一种支持软骨修复的有前途的策略。本工作的目的是评估体外培养在两种选定孔径(100 和 200μm)的多孔聚(L-丙交酯)(PLLA)支架中的人关节和鼻中隔软骨细胞的活力、附着、形态、细胞外基质(ECM)产生。使用扫描电子显微镜(SEM)分析通过三元热致相分离(TIPS)技术制备的 100 和 200μm 孔径的 PLLA 支架。将关节和鼻中隔软骨细胞接种在支架上,并培养 7 和 14 天。通过活/死染色、(免疫)组织化学和 II 型、I 型胶原、聚集蛋白聚糖和 SOX9 的基因表达分析来评估支架的细胞相容性和软骨细胞表型。两种类型的软骨细胞在整个培养期间都在两种支架上大部分存活。苏木精-伊红(HE)、阿利新蓝(显示糖胺聚糖)染色、ECM 蛋白的免疫反应性和基因表达以及软骨标志物(II 型、I 型胶原、聚集蛋白聚糖、SOX9)的表达表明,较小的孔径尺寸促进了软骨细胞的分化与较大的孔径相比。本工作表明支架孔径是影响软骨细胞分化的重要因素,并表明具有 100μm 孔径的支架可作为进一步未来修饰的细胞相容性基础。

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