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200~500nm。将制作好的材料切制成8mm×8mm×2mm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3d,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7d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,7d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。
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 intopieces 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. RESULTS: 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). ③ Observation 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 misdjvision were seen in both NPAB and control groups. ④ T
出处
《中国组织工程研究与临床康复》
CAS
CSCD
北大核心
2007年第1期37-40,共4页
Journal of Clinical Rehabilitative Tissue Engineering Research
基金
国家自然科学基金资助项目(30271321)~~