中国组织工程研究

中国组织工程研究 ›› 2010, Vol. 14 ›› Issue (34): 6333-6338.doi: 10.3969/j.issn.1673-8225.2010.34.015

• 材料生物相容性 material biocompatibility • 上一篇    下一篇

超声脱细胞异种松质骨材料的抗原性与生物相容性

赵自平1，蔡道章2，那晓东3，欧宣成1，吴冬冬1   

  1. 1广州医学院荔湾医院骨科，广东省广州市   510170；2中山大学附属第三医院骨科，广东省广州市 510630；3中山大学中山医学院病理生理教研室，广东省广州市   510008
  • 出版日期:2010-08-20 发布日期:2010-08-20
  • 作者简介:赵自平☆，男，1968年生，湖南省平江县人，2003年中南大学湘雅医学院毕业，博士，主要从事骨组织工程与骨科微创研究。 zhaozip6007@sina.com
  • 基金资助:

    国家自然科学基金项目(30200064)；广东省医学科研基金项目(A2008593)；广东市荔湾区科技计划项目(200712088048)。

Antigenicity and biocompatibility of acellular xenogeneic cancellous bone prepared by ultrasonic biotechnology

Zhao Zi-ping1, Cai Dao-zhang2, Na Xiao-dong3, Ou Xuan-cheng1, Wu Dong-dong1   

  1. 1 Department of Orthopaedics, Liwan Hospital of Guangzhou Medical College, Guangzhou   510170, Guangdong Province, China; 2 Department of Orthopaedics, Third Hospital of Sun Yat-sen University, Guangzhou   510630, Guangdong Province, China; 3 Department of Pathophysiology, Zhongshan School of Medicine, Sun Yat-sen University, Guangzhou   510008, Guangdong Province, China
  • Online:2010-08-20 Published:2010-08-20
  • About author:Zhao Zi-ping☆, Doctor, Department of Orthopaedics, Liwan Hospital of Guangzhou Medical College, Guangzhou 510170, Guangdong Province, China zhaozip6007@sina.com
  • Supported by:

    the National Natural Science Foundation of China, No. 30200064*; Medical Scientific Research Foundation of Guangdong Province, No. A2008593*; Science and Technology Plan in Liwan District of Guangzhou City, No. 200712088048*

PDF

434

被引次数

15

摘要：

背景：异种骨是一种复合组织，来源广泛，一直是骨修复研究领域热衷于制备、改良、开发的植骨材料，其抗原主要分布于骨组织的有核细胞，充分脱细胞是去除异种骨抗原的有效手段。
目的：观察超声脱细胞异种松质骨材料的抗原性与生物相容性。
方法：提取新鲜松质骨、常规材料和超声材料上清，检测抗原特异性淋巴细胞刺激增殖指数；将常规材料和超声材料分别植入SD大鼠体内，以假手术组做对照。流式细胞分析仪检测其外周血CD4+ 、CD8+淋巴细胞亚群的百分率及CD4+ /CD8+淋巴细胞比值的变化；与骨髓基质细胞的接触培养，观察两种材料对基质细胞增殖指数的影响，扫描电镜观察骨髓基质细胞在超声材料和常规材料表面的黏附和增殖情况。
结果与结论：新鲜松质骨刺激指数值最高，超声材料组刺激指数值接近于1，与常规材料组比较有显著性差异(P < 0.05)。术后第2周，常规材料组CD4+T细胞亚群百分率为32.89%，与假手术组(28.72%)比较，差异有显著性意义 (P < 0.05)，超声材料组与假手术组比较无显著差异(P > 0.05)；常规材料组和超声材料组CD8+T细胞亚群百分率分别为16.28%和13.49%，两组差异有显著性意义(P < 0.05)，均高于假手术组(10.07%) (P < 0.01)，常规材料组和超声材料组CD4+ /CD8+T细胞比值无显著性差异；2周后各组动物CD4+  、CD8+ T细胞亚群百分率逐渐回落，至术后6周3组之间差异无显著性意义。常规材料和超声材料对骨髓基质细胞的增殖均产生接触抑制效应，常规材料的抑制作用稍强，但差异无显著性意义(P > 0.05)。扫描电镜观测骨髓基质细胞在超声材料表面黏附生长，培养至5 d呈亚融合状态，细胞密度明显高于常规材料组。
结果提示，实验所制备的超声材料，其抗原性较常规材料弱，有利于骨髓基质细胞的黏附生长，表现出良好的生物相容性。

关键词: 异种骨, 超声生物技术, 生物材料, 抗原性, 生物相容性

Abstract:

BACKGROUND: Xenogeneic bone is a kind of combined tissues with abundant sources, it is always considered as the graft bone for the preparation, modification and development in field of bone repair. The antigenicity mainly locates in karyotes of bone tissue, full acellularization is an effective way to remove the heterogenous antigenicity.
OBJECTIVE: To study the antigenicity and biocompatibility of acellular xenogeneic cancellous bone (ACCB) prepared by ultrasonic biotechnology.
METHODS: The supernatant of fresh cancellous bone, ACCB and ultrasonic ACCB was extracted to detect the antigen-specific stimulated index on lymphocytes. The ACCB and ultrasonic ACCB were implanted into SD rats separately, while sham operated rats served as controls. The percentage of CD4+ and CD8+ lymphocyte subpopulations, and the ratio of CD4+ and CD8+ lymphocyte in peripheral blood from recipient were detected by flow cytometry. The ACCB and ultrasonic ACCB were co-cultured with bone marrow stroma cells to investigate the effects of these two kinds of materials on the proliferation of cells. The adhesion and proliferation of bone marrow stroma cells on ACCB and ultrasonic ACCB were observed under scanning electron microscopy.
RESULTS AND CONCLUSION: The stimulation index was the highest in fresh cancellous bone group, and close to 1 in ultrasonic material group, the difference between ACCB group and ultrasonic ACCB group was significant (P < 0.05). The percentage of CD4+ T lymphocytes of ACCB group was 32.89%, the difference between ACCB and sham operated group (28.72%) was significant (P < 0.05), the difference between ultrasonic ACCB and sham operated group was insignificant (P > 0.05). The percentage of CD8+ T lymphocytes of ACCB and ultrasonic ACCB groups was 16.28% and 13.49%, with significant differences between two groups (P < 0.05), which were both higher than that in sham operated group (10.07%, P < 0.01). In ACCB and ultrasonic ACCB groups, the ratio of CD4+ /CD8+ T cells was not significantly different; at 2 weeks, the percentage of CD4+ and CD8+ T cell subpopulations gradually decreased and 3 groups was not significantly different at 6 weeks. The depressive effects of ACCB was a littler stronger than ultrasonic ACCB on the proliferation of bone marrow stroma cells, without significant difference (P > 0.05). Adhesion of bone marrow stroma cells on ultrasonic ACCB could be observed on day 2 by scanning electron microscopy, the intensive growth of bone marrow stroma cells to subconfluent state could be observed on day 5, the density of bone marrow stroma cells was higher than that of ACCB group. The ultrasonic ACCB prepared in the study showed less antigenicity and better biocompatibiliby for adhesion and proliferation of bone marrow stroma cells.

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