Chinese Journal of Tissue Engineering Research ›› 2011, Vol. 15 ›› Issue (8): 1355-1359.doi: 10.3969/j.issn.1673-8225.2011.08.006

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Preparation of acellular natural bone matrix to remove immunological rejection antigen

Li Kang-jie1, Sun Shu2   

  1. 1Second Department of Orthopaedics, Affiliated Hospital of Yanbian University, Yanji  133000, Jilin Province, China
    2Department of Pathology, School of Medicine, Yanbian University, Yanji   133000, Jilin Province, China
  • Received:2010-08-28 Revised:2010-10-10 Online:2011-02-19 Published:2011-02-19
  • About author:Li Kang-jie☆, Studying for doctorate, Associate professor, Second Department of Orthopaedics, Affiliated Hospital of Yanbian University, Yanji 133000, Jilin Province, China likangjie2010@sina.cn
  • Supported by:

    the National Natural Science Foundation of China, No. 81000791*

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Abstract:

BACKGROUND: Allograft implants have a variety of sources and are easy to be processed and stored. If the immunological rejection is able to be removed, they will be applied as bone scaffolds to repair bone defect, which will resolve human bone supply ultimately. But there is no perfect approach to avoid immunological rejection at present.
OBJECTIVE: To remove antigen leading to immunological rejection in acellular natural bone matrix with hypotension-decellularization combined with hydrogen peroxide.
METHODS: SD rat femurs were processed by hydrogen peroxide handling following hypotonic-decellularized procedures. Then, the manufactured acellular natural bone matrix were evaluated by hematoxylin-eosin staining, toluidine blue staining, scanning electron microscope, transmission electron microscope, organic component analysis, eluant residue measurement and transplantation immunity analysis. The femoral specimens were compared with fresh rat femur.
RESULTS AND CONCLUSION: Histological observation showed that collagen fibers of acellular natural bone matrix arranged regularly and bone lacuna were lack of osteocytes, the fibronectin and laminin were present mainly in space around lacuna. Scanning electron microscope displayed that different layers of acellular natural bone matrix connected loosely and presented amounts of pore space between layers. Transmission electron microscope showed that no high density image appeared in lacuna area. There was no TritonX-100 in acellular natural bone matrix with the measurement of high-performance liquid chromatography. Light microscopy and scanning electron microscopy observations showed a large amount of cellular components were seen in fresh bone. Lymphocyte stimulation assay showed that immunological rejection of flesh bone was much greater than that of acellular natural bone matrix (P < 0.01). Above results indicated that antigen leading to immunological rejection could be removed mostly with procedures of hypotonic-decellularized processing and hydrogen peroxide handling.

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