Lahiji A, Sohrabi A, Hungerford D S, Frondoza C G
Department of Orthopaedic Surgery, Good Samaritan Hospital, Johns Hopkins University, Professional Office Building, 5601 Loch Raven Boulevard, Baltimore, Maryland 21239, USA.
J Biomed Mater Res. 2000 Sep 15;51(4):586-95. doi: 10.1002/1097-4636(20000915)51:4<586::aid-jbm6>3.0.co;2-s.
The search for biocompatible materials that can support the growth and phenotypic expression of osteoblasts and chondrocytes is a major challenge in the application of tissue engineering techniques for the repair of bone and cartilage defects. Chitosan, a copolymer of glucosamine and N-acetylglucosamine, may provide an answer to this search. Chitosan is the deacetylated product of chitin, a ubiquitous biopolymer found in the exoskeleton of insects and marine invertebrates. Little is known about the utility of chitosan in propagating human osteoblasts and chondrocytes. In this study, we test the hypothesis that chitosan promotes the survival and function of osteoblasts and chondrocytes. Chitosan (4%, w/v in 2% HAc) was coated onto plastic coverslips that had been fitted into 24-well plates. Human osteoblasts and articular chondrocytes were seeded on either uncoated or chitosan-coated coverslips at 1 x 10(5)/cells per well. Cultures were incubated at 37 degrees C, 5% CO(2) for a period of 7 days. Cell viability was assessed at that time using a fluorescent molecular probe. The phenotypic expression of osteoblasts and chondrocytes was analyzed by reverse transcriptase-polymerase chain reaction and immunocytochemistry. Osteoblasts and chondrocytes appeared spherical and refractile on chitosan-coated coverslips. In contrast, greater than 90% of cells on plastic coverslips were elongated and spindle shaped after 7 days of culture. Similar to cells propagated on uncoated control wells, greater than 90% of human osteoblasts and chondrocytes propagated on chitosan remained viable. Human osteoblasts propagated on chitosan films continued to express collagen type I whereas chondrocytes expressed collagen type II and aggrecan, as shown by reverse transcriptase-polymerase chain reaction analysis and immunostaining. The present in vitro work demonstrates the biocompatibility of chitosan as a substrate for the growth and continued function of human osteoblasts and chondrocytes. Chitosan may have potential use as a tissue engineering tool for the repair of osseous and chondral defects.
寻找能够支持成骨细胞和软骨细胞生长及表型表达的生物相容性材料，是组织工程技术应用于修复骨和软骨缺损时面临的一项重大挑战。壳聚糖是葡糖胺和N - 乙酰葡糖胺的共聚物，可能为这一探索提供答案。壳聚糖是几丁质的脱乙酰产物，几丁质是一种广泛存在于昆虫外骨骼和海洋无脊椎动物中的生物聚合物。关于壳聚糖在增殖人成骨细胞和软骨细胞方面的效用，人们了解甚少。在本研究中，我们检验了壳聚糖促进成骨细胞和软骨细胞存活及功能的假说。将壳聚糖（4%，溶于2%醋酸中，w/v）包被在已装入24孔板的塑料盖玻片上。将人成骨细胞和关节软骨细胞以每孔1×10⁵个细胞的密度接种在未包被或壳聚糖包被的盖玻片上。培养物在37℃、5%二氧化碳条件下孵育7天。此时使用荧光分子探针评估细胞活力。通过逆转录 - 聚合酶链反应和免疫细胞化学分析成骨细胞和软骨细胞的表型表达。在壳聚糖包被的盖玻片上，成骨细胞和软骨细胞呈球形且有折光性。相比之下，培养7天后，塑料盖玻片上超过90%的细胞呈细长纺锤形。与在未包被的对照孔中增殖的细胞类似，在壳聚糖上增殖的超过90%的人成骨细胞和软骨细胞仍保持存活。逆转录 - 聚合酶链反应分析和免疫染色显示，在壳聚糖膜上增殖的人成骨细胞继续表达I型胶原蛋白，而软骨细胞表达II型胶原蛋白和聚集蛋白聚糖。目前的体外研究证明了壳聚糖作为人成骨细胞和软骨细胞生长及持续功能的基质具有生物相容性。壳聚糖可能有潜力作为一种组织工程工具用于修复骨和软骨缺损。
