中国组织工程研究

中国组织工程研究 ›› 2015, Vol. 19 ›› Issue (34): 5512-5512.doi: 10.3969/j.issn.2095-4344.2015.34.019

• 细胞外基质材料 extracellular matrix materials • 上一篇    下一篇

动物源Ⅰ型胶原蛋白可引起BALB/c小鼠细胞免疫反应和组织免疫毒性

雷 静1，2，李奕恒3，刘旭昭2，3，汤顺清1，2   

  1. 1暨南大学生物医学工程研究所，广东省广州市  510632； 2广东省医用胶原工程技术研究开发中心，广东省广州市  510663； 3广州创尔生物技术股份有限公司，广东省广州市  510663
  • 出版日期:2015-08-20 发布日期:2015-08-20
  • 通讯作者: 汤顺清，教授，博士生导师，暨南大学生物医学工程研究所，广东省广州市 510632
  • 作者简介:雷静，女，1980年生，湖北省襄阳市人，汉族，暨南大学在读博士，主要从事生物医用材料与皮肤组织修复方面的研究。
  • 基金资助:

    广东省科技计划项目(2012B070800007)；广东省海洋经济创新发展区域示范专项(GD2012-B03-002)；广州市科技计划项目(2014JR000021)

Cellular immune response and immune toxicity to BALB/c mice for animal-based collagen

Lei Jing1, 2, Li Yi-heng3, Liu Xu-zhao2, 3, Tang Shun-qing1, 2   

  1. 1Institute of Biomedical Engineering, Jinan University, Guangzhou 510632, Guangdong Province, China; 2Guangdong Medical Collagen Engineering Technology Research and Development Center, Guangzhou 510663, Guangdong Province, China; 3Guangzhou Trauer Biotechnology Co., Ltd., Guangzhou 510663, Guangdong Province, China
  • Online:2015-08-20 Published:2015-08-20
  • Contact: Tang Shun-qing, Professor, Doctoral supervisor, Institute of Biomedical Engineering, Jinan University, Guangzhou 510632, Guangdong Province, China; Guangdong Medical Collagen Engineering Technology Research and Development Center, Guangzhou 510663, Guangdong Province, China
  • About author:Lei Jing, Studying for doctorate, Institute of Biomedical Engineering, Jinan University, Guangzhou 510632, Guangdong Province, China; Guangdong Medical Collagen Engineering Technology Research and Development Center, Guangzhou 510663, Guangdong Province, China
  • Supported by:

    the Science and Technology Plan of Guangdong Province, No. 2012B070800007; the Special Fund for the Innovative and Developmental Region of Marine Economy in Guangdong Province, No. GD2012-B03-002; the Science and Technology Plan of Guangzhou City, No. 2014JR000021

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591

被引次数

9

摘要：

背景：天然纯胶原被认为具有较低的免疫原性和较好的生物相容性。

目的：体外评估动物源Ⅰ型胶原蛋白的免疫原性。

方法：将牛跟腱经免疫原性清除后，提取制成Ⅰ型胶原蛋白，HPLC测定胶原蛋白纯度，荧光染色定量胶原蛋白中残留DNA。将50只BALB/c小鼠随机均分为5组，分别皮下注射生理盐水(阴性对照)、小牛来源Ⅰ型胶原蛋白标准品(阳性对照)、33.4，66.8，133.4 mg/kg的动物源Ⅰ型胶原蛋白，1次/d，连续注射12 d后，检测注射胶原蛋白后小鼠淋巴细胞的增殖、细胞分型及NK细胞杀伤功能；连续注射3周后，取脾脏、肝脏及肺组织进行病理组织学检查。

结果与结论：相比Ⅰ型胶原蛋白标准品，纯化后的牛源性Ⅰ型胶原蛋白纯度可达到99%以上，而残留DNA低于1 mg/L，远远低于目前常规脱细胞基质中DNA的残留水平50-100 µg/g(干质量)。注射12 d后，各组均未发生淋巴细胞增殖、NK细胞杀伤功能及淋巴细胞亚群的比例的变化。注射3周后，66.8，133.4 mg/kg动物源胶原蛋白组小鼠脾小动脉周围淋巴鞘面积明显增厚变大，有可能引发偶发性的肝损伤和肺损伤，而脾小体生发中心面积未发生明显变化。表明连续皮下注射动物源胶原蛋白可引发BALB/c小鼠发生较低水平的脾淋巴细胞免疫应答，可能引起偶发性肝、肺损伤。

中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程

关键词: 生物材料, 材料相容性, 动物源, 胶原蛋白, 免疫原性, 免疫应答, 免疫毒性, BALB/c小鼠, 残留DNA, 淋巴细胞

Abstract:

BACKGROUND: Natural collagen is considered to have low immunogenicity and good biocompatibility relatively.

OBJECTIVE: To evaluate the immunogenicity of animal-based collagen in vitro.

METHODS: Type I collagen was extracted from bovine tendon after immunogenicity removal. The collagen purity was detected by high performance liquid chromatography and residual DNA was measured quantitatively by fluorescence staining. Fifty BALB/c mice were randomly divided into five groups: subcutaneous injection of normal saline solution (negative control), bovine collagen (positive control), 33.4, 66.8, 133.4 mg/kg of bovine tendon collagen, respectively, once a day. After 12 days of continuously subcutaneous injection, lymphocyte proliferation, and cell classification and NK cell killing function of mice were detected; after 3 weeks of continuous injection, the spleen, liver, spleen and lung tissue of mice were taken for histological examination.

RESULTS AND CONCLUSION: Compared with the standard type I collagen, the purity of purified type I bovine collagen reached more than 99%, but the residual DNA was below 1 mg/L which was far less than the residue level of conventional cell-free DNA in the matrix (dry weight: 50-100 μg/g). After 12 days of continuous injection, there were no changes in lymphocyte proliferation, NK cell killing function and the proportion of lymphocyte subsets. After 3 weeks of injection, the spleen and lymph sheath of mice around the small artery became thickened in the 66.8 and 133.6 mg/kg bovine tendon collagen groups, which could cause accidental liver injury and lung injury, but the splenic corpuscle germinal center area had no change. These findings indicate that continuously subcutaneous injection of animal-based collagen can cause the lower lymphocyte immune response to the spleen of BALB/c mice, which may cause accidental liver and lung injuries.

中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程

Key words: Collagen, Immunity, Cellular, Lymphocytes

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