Porous thermoresponsive-co-biodegradable hydrogels as tissue-engineering scaffolds for 3-dimensional in vitro culture of chondrocytes.

Xiao Huang, Yue Zhang, Henry J Donahue, Tao L Lowe
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引用次数: 39

Abstract

A new porous, thermoresponsive, partially biodegradable, chemically crosslinked hydrogel system was developed, characterized, and tested as a cartilage tissue-engineering scaffold for in vitro chondrocyte culture over a 4-week period. The hydrogel system was composed of poly(N-isopropylacrylamide), poly(D,L-lactic acid), and dextran segments. Pores in the hydrogels were generated using a salt leaching technique. The hydrogels showed thermoresponsive properties, with a lower critical solution temperature at approximately 32 degrees C. They continuously swelled at physiological temperature in phosphate buffered saline (pH 7.4) for at least 1 month. Chondrocytes isolated from embryonic chick sterna were seeded into the hydrogel scaffolds at room temperature and cultured at 37 degrees C for 4 weeks. Real-time reverse-transcriptase polymerase chain reaction quantification was conducted every week to study messenger ribonucleic acid levels of 3 chondrocyte phenotypic markers: type II collagen, type X collagen, and Indian hedgehog. Results suggested that chondrocytes maintained their phenotype during the 4-week in vitro culture and could mimic in vivo development. Chondrocytes were non-enzymatically harvested from the hydrogel scaffold at the end of the fourth week by simply lowering the temperature from 37 degrees C to room temperature. The harvested chondrocytes kept a round morphology, confirming the maintenance of the chondrocyte phenotype in the hydrogel scaffolds.

多孔热响应-共生物可降解水凝胶作为软骨细胞体外三维培养的组织工程支架。
一种新型多孔、热响应、部分可生物降解、化学交联的水凝胶体系被开发、表征,并作为软骨组织工程支架进行了为期4周的体外软骨细胞培养测试。该水凝胶体系由聚(n -异丙基丙烯酰胺)、聚(D, l -乳酸)和葡聚糖段组成。水凝胶中的孔隙是用盐浸技术生成的。水凝胶表现出热响应特性,其临界溶液温度较低,约为32℃。它们在磷酸盐缓冲盐水(pH 7.4)中在生理温度下持续膨胀至少1个月。从鸡胚胸骨分离软骨细胞,在室温下植入水凝胶支架,37℃培养4周。每周进行实时逆转录酶聚合酶链反应定量,研究3种软骨细胞表型标记物:II型胶原、X型胶原、印度刺猬的信使核糖核酸水平。结果表明,软骨细胞在体外培养的4周内保持其表型,并能模拟体内发育。在第四周结束时,通过简单地将温度从37摄氏度降至室温,从水凝胶支架上非酶地收获软骨细胞。收获的软骨细胞保持圆形形态,证实了水凝胶支架中软骨细胞表型的维持。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
Tissue engineering
Tissue engineering CELL & TISSUE ENGINEERING-BIOTECHNOLOGY & APPLIED MICROBIOLOGY
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