珍珠层人工骨与人骨髓基质细胞的生物相容性-中国组织工程与临床康复2007年1期目次

珍珠层人工骨与人骨髓基质细胞的生物相容性*☆

杨春露，陈建庭，王建钧，赵勇，金大地

Biocompatibility between NacrePDLLA artificial bone
and human marrow stroma cells*☆

Abstract

AIM：To investigate the biocompatibility between human marrow stroma cells (MSCs) and Nacre-PDLLA artificial bone (NPAB), so as to offer experimental foundation for the choice of better bone tissue engineering materials.

METHODS：The experiment was conducted from May to October 2005 in the spinal and orthopedic laboratory of Nanfang Hospital, Southern Medical University. The nacre powder and despinning polylactic acid were made into three diamensions and porous material named NPAB with 200－500 nm in diameter by means of press-shaping patent technology. The material was cut into pieces of 8 mm×8 mm×2 mm, and air dried after humidification by alcohol and deionized water following by sterillization by megatemperature and moist heat before using. MSCs were obtained from normal adult male (volunteers). MSCs were cultured in vitro, and co-cultured with NPAM. Two groups were divided: NPAB group added with lump NPAB, and control group with nothing added. Morphology was assayed by means of inverted microscope and scanning electron microscope, and cell proliferation and protein content were detected using MTT and coomassie brilliant blue micro-estimation.

RESULT:①MSCs population was increased gradually, but no significant difference was found between the two groups (P > 0.05). ②There was no significant difference in the protein content of MSCs between the NPAM group and control group after co-cultured for 3 days (0.39±0.02, 0.37±0.03, P > 0.05); but after co-cultured for 7 days, the protein content of the NPAM group was higher than the control group, which had significant differences (0.97±0.05, 0.83±0.03, P < 0.05). ③Observation result by inversion phase contrast microscope showed that the cells grew intensively around the NPAB after cultured for 7 days, and compared with the control group, no significant difference in the growth condition was found in the NPAB group. No great quantity cell apoptosis, ageing or misdivision were seen in both NPAB and control groups. ④The MSCs co-cultured with NPAB were observed by scanning electron microscope. Fusiform shape MSCs were seen on the surface of NPAB, which aligned to clusters, and some parapodiums were found in the marginal MSCs.

CONCLUSION: NPAB displays a good biocompatibility, which can be applied into bone tissue engineering as a superior MSC carrier.

Department of Orthopedics and Spinal Surgery, Nanfang Hospital, Southern Medical University, Guangzhou 510515, Guangdong Province, China

Yang Chun-lu☆, Studying for doctorate, Physician, Department of Orthopedics and Spinal Surgery, Nanfang Hospital, Southern Medical University, Guangzhou 510515, Guangdong Province, China
ycl_2002@163.com

Correspondence to: Chen Jian-ting, Master, Professor, Chief physician, Department of Orthopedics and Spinal Surgery, Nanfang Hospital, Southern Medical University, Guangzhou 510515, Guangdong Province, China
chenjt99@sina.com

Supported by: the National Natural Science Foundation of China, No. 30271321*

Received: 2006-05-23
Accepted: 2006-07-11

摘要
目的：探讨人骨髓基质细胞与珍珠层人工骨的生物相容性，为选择更好的骨组织工程材料提供实验依据。
方法：实验于2005－05/10在南方医科大学南方医院脊柱骨科实验室完成。采用模压成型专利技术，将珍珠层粉与消旋聚乳酸制成三维多孔材料，孔径200～500 nm。将制作好的材料切制成8 mm×8 mm×2 mm大小的块状，乙醇及去离子水湿化，自然干燥，高温湿热灭菌后备用。骨髓基质细胞取自健康成年男性（自愿捐献）。体外培养人骨髓基质细胞，与珍珠层－聚乳酸复合人工骨共同培养。实验分两组：珍珠层人工骨组加入块状的珍珠层人工骨，对照组不加处理因素。采用倒置相差显微镜、扫描电镜进行形态学观察，噻唑蓝法进行细胞增殖等指标测定，考马斯亮蓝法微量测定细胞蛋白含量。
结果：①随着培养时间的延长，各组细胞数量逐渐增加，各时间点珍珠层人工骨组与对照组相比，差异无显著性意义（P > 0.05）。②培养3 d，珍珠层人工骨组和对照组蛋白含量相比较差异无显著性意义(分别为0.39±0.02，0.37±0.03，P > 0.05)；培养7 d后，珍珠层人工骨组蛋白含量高于对照组，差异有显著性意义（分别为0.97±0.05，0.83±0.03，P < 0.05）。③倒置相差显微镜观察，7 d时可见细胞在珍珠层人工骨周围密集生长。珍珠层人工骨组细胞生长状态与对照组相比差异不明显，未见到细胞大量凋亡、衰老及异常分裂现象。④扫描电镜观察，共同培养第7天，可见梭形骨髓基质细胞大量紧密生长在珍珠层人工骨表面，呈集束状排列，边缘部分细胞可见到伪足。
结论：珍珠层人工骨具有良好的生物相容性，可作为骨髓基质细胞的载体而应用于骨组织工程的研究。
关键词：生物相容性材料；骨髓细胞；珍珠层

杨春露，陈建庭，王建钧，赵勇，金大地.珍珠层人工骨与人骨髓基质细胞的生物相容性[J].中国组织工程研究与临床康复，2007，11(1):37-40 [www.zglckf.com/zglckf/ejournal/upfiles/07-1/1k-37(ps).pdf]

南方医科大学南方医院脊柱骨病外科，广东省广州市 510515

杨春露☆，男，1971年生，辽宁省鞍山市人，汉族，南方医科大学在读博士，医师，主要从事骨生物材料、骨修复研究。 ycl_2002@163.com

通讯作者：陈建庭，硕士，教授，主任医师，南方医科大学南方医院脊柱骨病外科，广东省广州市 510515 chenjt99@sina.com

国家自然科学基金资助项目（30271321）*

中图分类号:R318.08 文献标识码:A
文章编号:1673-8225
(2007)01-00037-04

收稿日期:2006-05-23 修回日期:2006-07-11 (06-50-5-4337/Y·LL)

课题背景：研究获得国家自然科学基金资助（30271321）。骨生物替代材料的研究是矫形外科及骨科学研究热点，获得具有良好生物相容性、适宜降解速度及良好的成骨特性的材料一直是研究者努力的重要方向之一。本研究即以此为出发点，探索珍珠层与骨髓基质细胞的生物相容性，明确将珍珠层作为一种新的生物骨替代材料的可能性及其对于临床、科研的重要意义。

创新要点：珍珠层是一种新型的骨生物替代材料，来源于海洋软体动物珍珠贝科或蚌科动物的贝壳内层部分。来源广泛，成本低廉，加工简易，具有较好的成骨特性。珍珠层作为生物替代材料具有其他无机材料无法比拟的优势，是一种理想的天然成骨材料。本实验对于难治性骨缺损的基础及临床研究具有十分重要的借鉴意义，为最终解决临床大面积或大段性骨缺损的治疗难题具有积极意义。

同行评价：该文以珍珠层作为骨替代材料，探讨人骨髓基质细胞与珍珠层人工骨的生细胞相容性，论文的创新点是提出通过将体外培养的人骨髓基质细胞与珍珠层聚乳酸人工骨共同培养，观察细胞增殖、生长状态等的变化，从而考察珍珠层人工骨的生物相容性，研究结果表明：珍珠层人工骨具有良好的生物相容
性，可作为骨髓基质细胞的载体而应用于骨组织工程的研究。论文选题新颖，具有较高的学术水平。