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用于体外培养形成髓核样组织的人髓核细胞的去端肽胶原蛋白:对HNPSV-1细胞增殖和蛋白聚糖产生的影响

Atelocollagen for culture of human nucleus pulposus cells forming nucleus pulposus-like tissue in vitro: influence on the proliferation and proteoglycan production of HNPSV-1 cells.

作者信息

Sakai Daisuke, Mochida Joji, Iwashina Toru, Watanabe Takuya, Suyama Kaori, Ando Kiyoshi, Hotta Tomomitsu

机构信息

Department of Orthopaedic Surgery, Surgical Science, Tokai University School of Medicine, Bohseidai, Isehara, Kanagawa 259-1193, Japan.

出版信息

Biomaterials. 2006 Jan;27(3):346-53. doi: 10.1016/j.biomaterials.2005.06.040. Epub 2005 Aug 18.

Abstract

Nucleus pulposus (NP) is responsible for maintaining function and structure of the disc. Scaffolds to culture disc cells three-dimensionally are emphasized in recent reports on development of a new method for treating disc degeneration using cell transplantation and tissue engineering. Among artificial scaffolds and cell carrying materials, Atelocollagen is a collagen gel that has an advantage in safety issues over others. However, to date there has been no study that investigated culture of human nucleus pulposus cells in Atelocollagen. To investigate whether Atelocollagen could be used as a culture scaffold and if it has any effect on cell proliferation and proteoglycan (PG) production, as well as to find the optimal commercially available Atelocollagen for NP cell transplantation and tissue engineering, we cultured human NP cell line HNPSV-1, in three different Atelocollagen and compared with alginate. Furthermore, NP-like tissues were generated using these cells and different Atellocollagen solutions. Results showed that both DNA synthesis and content is significantly greater when cultured in Atelocollagen than in alginate. On the other hand, proteoglycan synthesis and accumulation was significantly greater in alginate compared with the 0.3% Atelocollagen scaffolds; with 3% Atelocollagen, however, results were similar. NP-like tissue generated by Atelocollagen showed good water and proteoglycan preservation. The current study demonstrates that the use of Atelocollagen as an in vitro culture scaffold for three-dimensional culture of human NP cell lines is indeed feasible and moreover, Atelocollagen possesses the potential to become a candidate scaffold for cell transplantation or tissue engineering for the treatment of intervertebral disc degeneration.

摘要

髓核(NP)负责维持椎间盘的功能和结构。在最近关于使用细胞移植和组织工程治疗椎间盘退变新方法的报道中,强调了用于三维培养椎间盘细胞的支架。在人工支架和细胞载体材料中,去端胶原是一种胶原凝胶,在安全性方面比其他材料具有优势。然而,迄今为止,尚无研究调查人类髓核细胞在去端胶原中的培养情况。为了研究去端胶原是否可用作培养支架,以及它对细胞增殖和蛋白聚糖(PG)产生是否有任何影响,同时为找到用于NP细胞移植和组织工程的最佳市售去端胶原,我们将人类NP细胞系HNPSV - 1培养在三种不同的去端胶原中,并与藻酸盐进行比较。此外,使用这些细胞和不同的去端胶原溶液生成了类NP组织。结果表明,在去端胶原中培养时,DNA合成和含量均显著高于藻酸盐。另一方面,与0.3%的去端胶原支架相比,藻酸盐中的蛋白聚糖合成和积累显著更高;然而,对于3%的去端胶原,结果相似。由去端胶原生成的类NP组织显示出良好的水分和蛋白聚糖保存情况。当前研究表明,将去端胶原用作人类NP细胞系三维培养的体外培养支架确实可行,而且,去端胶原具有成为用于治疗椎间盘退变的细胞移植或组织工程候选支架的潜力。

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