Porcine decellular endplate cartilage matrix: biocompatibility assessment and establishment of a cytotoxicity evaluation system

doi:10.3969/j.issn.2095-4344.0920

Abstract

Abstract:

BACKGROUND: Compared to other high-molecular compounds used for scaffold materials, the decellular endplate cartilage matrix (DEPM) can simulate the extracellular environment closer to the original physiological state, and the elastic modulus of the intervertebral disc scaffold made of DEPM therefore is closer to that of the cartilage tissue.

OBJECTIVE: To prepare DEPM and explore its biocompatibility at cellular level and in vivo.

METHODS: Porcine endplate cartilage samples were crushed and digested with Trypsin combined with ribozyme and TritonX-100 to obtain DEPM. (1) Cytotoxicity test: After dissolved in acetic acid, the DEPM was prepared into the acellular matrix membrane. Cartilage stem cells from the lumbar endplate of Sprague-Dawley rats were seeded onto the acellular matrix membrane, and cell proliferation was detected using cell counting kit-8 after 1-5 days of culture. Then cytotoxicity of the cells was graded. (2) Toxicity test in vivo: Acetic acid containing DEPM (experimental) or not (control) was subcutaneously injected into the back of Sprague-Dawley rats to form a bump. Rat body temperature and bump’s size were observed within 36 hours after injection.

RESULTS AND CONCLUSION: (1) Cytotoxicity test: Under the inverted microscope, cartilage stem cells from the lumbar endplate were adherent within 2 days after implantation, and grew in a fusiform shape. The relative cell growth rate was over 80% within 5 days after implantation, and the cell toxicity grade was level 0 to 1. (2) Toxicity test in vivo: A bump was formed on the rat back in the two groups immediately after injection, and completely disappeared within 36 hours after injection. Moreover, the absorption rate in the control group was faster than that in the experimental group. No red and swelling bump occurred in the two groups. the body temperature of the rats in the two groups did not exceed 37.5 ℃ within 36 hours after injection. To conclude, the DEPM has good biocompatibility, which is in line with the selection criteria of biological scaffolds.

中国组织工程研究杂志出版内容重点：生物材料；骨生物材料; 口腔生物材料; 纳米材料; 缓释材料; 材料相容性；组织工程

Key words: Materials Testing, Extracellular Matrix, Intervertebral , Cytotoxicity Tests, Immunologic, Tissue Engineering

CLC Number:

R318

Wang Xiao, Xu Hong-guang, Xiao Liang, Liu Chen, Jin Zhong-xing, Shen Yang. Porcine decellular endplate cartilage matrix: biocompatibility assessment and establishment of a cytotoxicity evaluation system[J]. Chinese Journal of Tissue Engineering Research, 2018, 22(22): 3539-3544.

Figures/Tables 6

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