Chinese Journal of Tissue Engineering Research ›› 2011, Vol. 15 ›› Issue (3): 399-403.doi: 10.3969/j.issn.1673-8225.2011.03.005

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Fabrication and properties of a novel tissue engineered scaffold for annulus fibrosus disci intervertebralis

Yan Zhong-sheng1, Xu Bao-shan1,2, Yang Qiang2, Xia Qun2, Zhang Chun-qiu3, He Jian1   

  1. 1Graduate School of Tianjin Medical University, Tianjin  300211, China
    2Department of Spine Surgery, Tianjin Hospital, Tianjin  300211, China
    3Tianjin University of Technology, Tianjin  300211, China
  • Received:2010-11-05 Revised:2010-12-21 Online:2011-01-15 Published:2011-01-15
  • Contact: Xu Bao-shan, Doctor, Chief physician, Master’s supervisor, Graduate School of Tianjin Medical University, Tianjin 300211, China; Department of Spine Surgery, Tianjin Hospital, Tianjin 300211, China xubshn@yahoo.com.cn Yang Qiang, Doctor, Attending physician, Department of Spine Surgery, Tianjin Hospital, Tianjin 300211, China yangqiang1980@126.com
  • About author:Yan Zhong-sheng★, Studying for master’s degree, Graduate School of Tianjin Medical University, Tianjin 300211, China sdyanzhongsheng@163.com
  • Supported by:

    Foundation for the Returned Overseas Chinese Scholars, State Ministry of Human Resources and Social Security*; Key Projects of Science and Technology Foundation of Tianjin Health Department, No. 2010KR08*; Science and Technology Foundation of Tianjin Health Department, No. 09KZ67*, 07KG1*

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Abstract:

BACKGROUND: Tissue engineering strategies to repair degenerated intervertebral disc are being developed to replace or regenerate the herniated nucleus pulposus. However, the tissue engineering of nucleus pulposus alone can not restore intact structure and function of intervertebral disc. Tissue engineering of annulus fibrosus is deemed as a major limiting factor for tissue engineered intervertebral disc treatment strategy.
OBJECTIVE: To fabricate a natural acellular demineralized bone matrix gelatin and to verify the feasibility of it as a scaffold for annulus fibrosus tissue engineering.
METHODS: Pig proximal femoral cancellous bone rings (external diameter 10 mm, internal diameter 5 mm, thickness 3 mm) were fabricated, then dealed with high-pressure water washing, degreasing, decalcification, decellularization and other related treatments. The ring scaffolds were investigated by gross, histological, light microscopy and scanning electron microscopy observations, as well as porosity measurement, water absorption rate and biomechanical analysis. Dog bone marrow mesenchymal stem cells were isolated and cultured, MTT test was also done to assess cytotoxicity of the scaffolds. 
RESULTS AND CONCLUSION: The scaffold showed white, porous cancellous ring. The hematoxylin-eosin staining revealed that there were no cell fragments in the scaffolds. The light microscopy and scanning electron microscopy observations demonstrated that the novel porous scaffold had a good pore interconnectivity with (401.4±13.1) μm pore diameter, (62.12±1.52%) porosity and (409.77±11.34)% water absorption rate. The biomechanical test showed that the longitudinal elastic modulus was (47.75 ±6.32) kPa. The intrinsic cytotoxicity assessment of novel scaffolds using MTT test showed that there were no significant differences in the absorbance value between scaffold leaching liquid ad control DMEM culture solution, thus the scaffolds do not exert any cytotoxic effect on cells. Novel acellular demineralized bone matrix gelatin scaffold had good pore diameter and porosity, appropriate biomechanical character, non-toxicity and good biocompatibility, which make it a suitable candidate as an alternative scaffold for annulus fibrosus tissue engineering.

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