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纯化海藻酸盐海绵对软骨细胞再生的影响:体外和体内研究

Effects of purified alginate sponge on the regeneration of chondrocytes: in vitro and in vivo.

作者信息

Song Jeong Eun, Kim A Ram, Lee Cheon Jung, Tripathy Nirmalya, Yoon Kun Ho, Lee Dongwon, Khang Gilson

机构信息

a Department of BIN Fusion Technology , Chonbuk National University , 567, Beackje-daero, Deokjin, Jeonju 561-756 , Republic of Korea.

出版信息

J Biomater Sci Polym Ed. 2015;26(3):181-95. doi: 10.1080/09205063.2014.987570. Epub 2014 Dec 11.

Abstract

Regeneration science has been studied using tissue engineering techniques due to the self-renewal difficulties of damaged or degenerated cartilage. A scaffold with biodegradability and biocompatibility features plays a key role in developing cartilage tissue similar to human biological materials. Herein, we have fabricated three-dimensional sponge using purified alginate for the regeneration of chondrocytes cells and formation of cartilage. We demonstrated that the alginate purification can effectively minimize inflammatory reaction through reducing the content of mannuronic acid causing immune rejection. Cartilage regeneration research was performed using three-dimensional non-purified and purified alginate sponges synthesized by modified Korbutt method. In vitro cell viability and specific gene expression in the cartilage cells were investigated using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and reverse transcriptase-polymerase chain reaction (RT-PCR) after seeding chondrocytes on the as-fabricated sponges. Specific extracellular matrix (ECM) of chondrocytes, sGAG, and the content of collagen were also measured. Histological staining was carried out after purified alginate sponge seeded with chondrocytes and was implanted in subcutaneous nude mouse followed by extraction. Compared to the non-purified ones, the purified alginate sponges showed positive effects on maintaining affinities and phenotype of chondrocytes. From these results, it can be suggested that the purified alginate sponges provide a promising platform for cartilage regeneration.

摘要

由于受损或退变软骨的自我更新困难,再生科学一直通过组织工程技术进行研究。具有生物可降解性和生物相容性特征的支架在开发类似于人类生物材料的软骨组织中起着关键作用。在此,我们使用纯化的藻酸盐制备了三维海绵,用于软骨细胞的再生和软骨的形成。我们证明,藻酸盐纯化可以通过降低引起免疫排斥的甘露糖醛酸含量,有效减少炎症反应。使用改良的Korbutt方法合成的三维未纯化和纯化藻酸盐海绵进行软骨再生研究。将软骨细胞接种到制备好的海绵上后,使用3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐(MTT) 法和逆转录聚合酶链反应 (RT-PCR) 研究软骨细胞的体外细胞活力和特定基因表达。还测量了软骨细胞的特定细胞外基质 (ECM)、硫酸糖胺聚糖 (sGAG) 和胶原蛋白含量。将接种软骨细胞的纯化藻酸盐海绵植入皮下裸鼠后进行提取,然后进行组织学染色。与未纯化的藻酸盐海绵相比,纯化的藻酸盐海绵在维持软骨细胞的亲和力和表型方面显示出积极作用。从这些结果可以看出,纯化的藻酸盐海绵为软骨再生提供了一个有前景的平台。

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