Chinese Journal of Tissue Engineering Research ›› 2015, Vol. 19 ›› Issue (34): 5512-5512.doi: 10.3969/j.issn.2095-4344.2015.34.019

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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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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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