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三维培养中胶原蛋白/明胶海绵支架持续释放碱性成纤维细胞生长因子对声带成纤维细胞的生物相容性和疗效

Biocompatibility and efficacy of collagen/gelatin sponge scaffold with sustained release of basic fibroblast growth factor on vocal fold fibroblasts in 3-dimensional culture.

作者信息

Hiwatashi Nao, Hirano Shigeru, Mizuta Masanobu, Tateya Ichiro, Kanemaru Shin-Ichi, Nakamura Tatsuo, Ito Juichi, Kawai Katsuya, Suzuki Shigehiko

机构信息

Department of Otolaryngology-Head and Neck Surgery, Graduate School of Medicine, Kyoto University, Kyoto, Japan.

Department of Otolaryngology-Head and Neck Surgery, Graduate School of Medicine, Kyoto University, Kyoto, Japan

出版信息

Ann Otol Rhinol Laryngol. 2015 Feb;124(2):116-25. doi: 10.1177/0003489414546396. Epub 2014 Aug 12.

Abstract

OBJECTIVE

Treatment of vocal fold scarring remains challenging. We have previously reported the therapeutic effects of local injection of basic fibroblast growth factor (bFGF) in animal models and humans. A novel collagen/gelatin sponge (CGS) is capable of sustained release of bFGF, which compensates for its quick absorption in vivo, avoiding multiple injections. This study aimed to evaluate the biocompatibility and efficacy of the CGS in rat vocal fold fibroblasts prior to human trials.

METHODS

Fibroblasts extracted from Sprague-Dawley rat vocal folds were seeded onto a CGS and then cultivated with bFGF at concentrations of 0, 10, and 100 ng/mL. Vocal fold fibroblast morphology, adhesion, proliferation, and gene expression were measured under these 3-dimensional conditions.

RESULTS

Cells adhered to the CGS from day 1. Although no significant differences in cell morphology were detected, cell proliferation was accelerated by bFGF administration. Expression of endogenous bFGF and hepatocyte growth factor was significantly up-regulated at 10 ng/mL bFGF. The expression of procollagen I and procollagen III was significantly suppressed, whereas HAS-1 and HAS-2 were up-regulated at 10 and 100 ng/mL bFGF.

CONCLUSION

The collagen/gelatin sponge is biocompatible with vocal fold fibroblasts and may be useful as a bFGF drug delivery system for the treatment of scarred vocal folds.

摘要

目的

声带瘢痕的治疗仍然具有挑战性。我们之前已报道了在动物模型和人体中局部注射碱性成纤维细胞生长因子(bFGF)的治疗效果。一种新型的胶原/明胶海绵(CGS)能够持续释放bFGF,弥补其在体内的快速吸收,避免多次注射。本研究旨在在人体试验之前评估CGS在大鼠声带成纤维细胞中的生物相容性和疗效。

方法

从Sprague-Dawley大鼠声带中提取的成纤维细胞接种到CGS上,然后分别用浓度为0、10和100 ng/mL的bFGF进行培养。在这些三维条件下测量声带成纤维细胞的形态、黏附、增殖和基因表达。

结果

从第1天起细胞就黏附到CGS上。虽然未检测到细胞形态有显著差异,但给予bFGF可加速细胞增殖。在10 ng/mL bFGF时,内源性bFGF和肝细胞生长因子的表达显著上调。在10和100 ng/mL bFGF时,I型前胶原和III型前胶原的表达显著受到抑制,而HAS-1和HAS-2则上调。

结论

胶原/明胶海绵与声带成纤维细胞具有生物相容性,可能作为一种用于治疗瘢痕化声带的bFGF药物递送系统。

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