Mohan Beena Gopalan, Suresh Babu Sivadasan, Varma Hari Krishna, John Annie
Transmission Electron Microscopy Laboratory, Biomedical Technology Wing, Sree Chitra Tirunal Institute for Medical Sciences & Technology, Thiruvananthapuram, 695012, India.
J Mater Sci Mater Med. 2013 Dec;24(12):2831-44. doi: 10.1007/s10856-013-5018-y. Epub 2013 Aug 29.
The development of bone replacement materials is an important objective in the field of orthopaedic surgery. Due to the drawbacks of treating bone defects with autografts, synthetic bone graft materials have become optional. So in this work, a bone tissue engineering approach with radiopaque bioactive strontium incorporated calcium phosphate was proposed for the preliminary cytocompatibility studies for bone substitutes. Accumulating evidence indicates that strontium containing biomaterials promote enhanced bone repair and radiopacity for easy imaging. Hence, strontium calcium phosphate (SrCaPO4) and hydroxyapatite scaffolds have been investigated for its ability to support and sustain the growth of rabbit adipose-derived mesenchymal stem cells (RADMSCs) in vitro. They were characterized via Micro-CT for pore size distribution. Cells used were isolated from New Zealand White rabbit adipose tissue, characterized by FACS and via differentiation into the osteogenic lineage by alkaline phosphatase, Masson's trichome, Alizarin Red and von Kossa staining on day 28. Material-cell interaction was observed by SEM imaging of cell morphology on contact with material. Live-Dead analysis was done by confocal laser scanning microscopy and cell cluster analysis via μCT. The in vitro biodegradation, elution and nucleation of apatite formation of the material was evaluated using simulated body fluid and phosphate buffered saline in static regime up to 28 days at 37 °C. These results demonstrated that SrCaPO4 is a good candidate for bone tissue engineering applications and with osteogenically-induced RADMSCs, they may serve as potential implants for the repair of critical-sized bone defects.
骨替代材料的研发是骨外科领域的一个重要目标。由于自体骨治疗骨缺损存在弊端,合成骨移植材料成为了一种选择。因此,在本研究中,提出了一种将具有射线不透性的生物活性锶掺入磷酸钙的骨组织工程方法,用于骨替代物的初步细胞相容性研究。越来越多的证据表明,含锶生物材料可促进骨修复增强,并具有射线不透性以便于成像。因此,已对磷酸锶钙(SrCaPO4)和羟基磷灰石支架在体外支持和维持兔脂肪来源间充质干细胞(RADMSCs)生长的能力进行了研究。通过显微CT对其孔径分布进行了表征。所用细胞取自新西兰白兔的脂肪组织,通过流式细胞术进行表征,并在第28天通过碱性磷酸酶、马松三色染色、茜素红染色和冯·科萨染色分化为成骨谱系。通过扫描电子显微镜对细胞与材料接触时的形态进行成像,观察材料与细胞的相互作用。通过共聚焦激光扫描显微镜进行活死分析,并通过显微CT进行细胞簇分析。在37℃下,使用模拟体液和磷酸盐缓冲盐水在静态条件下对材料的体外生物降解、洗脱和成磷灰石形成的成核作用进行了长达28天的评估。这些结果表明,SrCaPO4是骨组织工程应用的良好候选材料,与经成骨诱导的RADMSCs一起,它们可能作为修复临界尺寸骨缺损的潜在植入物。