摘要
目的:观察人骨髓基质干细胞(human bone marrow mesenchymal stem cells,hBMSCs)在经过表面纳米化处理的PET人工韧带材料表面的粘附、增殖及成骨分化。方法:根据PET膜片表面处理情况分为PET组(PET膜片未经表面处理)、LBL组(PET膜片表面经透明质酸、壳聚糖纳米自组装涂层处理)和HA组(PET膜片经透明质酸、壳聚糖纳米自组装处理后,表面再加纳米羟基磷灰石涂层)3组。在上述3种PET材料上分别培养hBMSCs后进行各项检测。扫描电镜(SEM)观察hBMSCs在材料上培养1天和7天时的细胞形态。培养7天后,Alamar blue法检测hBMSCs的增殖,pNPP法检测碱性磷酸酶(ALP)活性。RealTimePCR检测培养3天hBMSCs的整合素β1mRNA表达。结果:SEM显示HA组和LBL组hBMSCs能更好地伸展和粘附,形态明显优于PET组。培养7天,HA组和LBL组的细胞增殖、ALP活性定量均显著高于PET组(P<0.001);HA组细胞增殖显著高于LBL组(P=0.01),HA组ALP活性显著高于LBL组(P<0.001)。培养3天后,HA组和LBL组hBMSCs的整合素β1mRNA表达显著高于PET组(P<0.001),HA组显著高于LBL组(P=0.003)。结论:PET人工韧带材料表面经壳聚糖、透明质酸纳米自组装涂层和纳米羟基磷灰石涂层处理后,能明显促进人hBMSCs的粘附、增殖和分化。纳米羟基磷灰石涂层的作用最明显。
Objective To observe the effect of superficial nano-treated polyethylene terephthalate（PET）surface respectively coated with chitosan,hyaluronic acid and hydroxyapatite nanoscale, on the adhesion,proliferation and differentiation of human bone marrow mesenchymal stem cells （hBMSCs）. Methods According to the surface treatments of PET chaffs, the mono-structured materials were divided into three groups：LBL group（using layer-by-layer of chitosan and hyaluronic acid）, HA group （using layer-by-layer of chitosan and hyaluronic acid plus nano hydroxyapatite） and PET group （without any treatment）. The hBMSCs were cultured on those PET chaffs abovementioned,and the adhesion,proliferation and differentiation of hBMSCs were observed. The morphology of hBMSCs was observed by scanning electron microscope （SEM）1 day and 7 days after seeding. The proliferation of hBMSCs were detected by Alamar blue test and the alkali phosphatase （ALP）activity was checked using pNPP assay by the end of 7-day culture. Real time PCR was used to measure the mRNA expression of integrin β1 by the end of 3-day culture. Results Results showed that the spreading of hBMSCs in the HA and LBL groups was better than that in PET group. The proliferation and ALP activity of hBMSCs in HA and LBL groups were significantly higher than that in PET group（P 0.001）. The proliferation in HA group was significant than that in LBL group（P =0.01）,and the ALP activity in HA group was also higher than that in LBL group（P 0.001）. The mRNA expressions of integrin β1 3 days after seeding in HA and LBL groups were significantly higher than that in PET group （P 0.001）,and in HA group the expression of integrin β1 3 days after seeding was significantly higher that in LBL group （P =0.003）. Conclusion The PET materials coated with chitosan,hyaluronic acid and hydroxyapatite nanoscale significantly improve the adhesion,proliferation and differentiation of hBMSCs, especially the PET material coated with nano-hydroxyapatite.
出处
《中国运动医学杂志》
CAS
CSCD
北大核心
2010年第5期533-537,589,共6页
Chinese Journal of Sports Medicine
基金
国家科技部973项目(2009CB930000)
863项目(2007AA021902、2007AA021903)
