Date of Award
Master of Science (MS)
Biomedical and Chemical Engineering
Alizarin Red staining, Hematoxylin and Eosin, Bone Osteosarcoma Cells (SaOs-2), Gelatin Methacryloyl Hydrogel, Mineralized Microenvironment, Scanning electron microscopy, Energy Despersive X-ray Spectroscopy, Simulated Body Fluid
Musculoskeletal diseases are widespread, and they affect hundreds of millions of people all around the world. Bone defects occurs due to various reasons such as surgeries, fracture and diseases. These bone defects need a surgical intervention and are generally treated with state-of-the-art grafting techniques such as natural bone grafts and implants. However, even the best available corrective treatments have several limitations such as availability, disease transfer, donor site scarcity, and immune rejection. Bone remodeling techniques which would support natural bone regeneration upon graft implantation, can be used to maximize the efficiency of current grafting techniques. In this study, our goal was to understand how cell mediated bone mineralization takes place in-vitro and to investigate if mineralized micro-environment has any effect on natural bone mineralization. Many studies have shown that the use of Simulated Body Fluid (SBF) has been consistent for developing the mineral element when used with gelatin, collagen, or other hydrogels. These resulting mineral coated hydrogels have similar morphology and chemistry as that of the native mineralized tissue. Thus, we used Simulated Body Fluid to generate pre-mineralized gelatin methacrylate samples. On top of these pre-mineralized samples, we encapsulated Bone Osteosarcoma cells (SaOs-2) and the culture was maintained for a period of two weeks in osteogenic media. From the results of Scanning Electron Microscopy (SEM), we found that the mineral component produced by 2X modified Simulated Body Fluid (2X m-SBF) and Bone Osteosarcoma Cells was morphologically different. Alizarin red staining showed that calcium apatite was present on both Simulated Body Fluid mineralized and SaOs-2 cell-mineralized sides of the samples. It was also observed that the mineral laid by SaOs-2 cells in the mineralized (7-day 2X m-SBF mineralized) environment was denser than that seen in the unmineralized environment. H&E tests supported Alizarin red test results and detected calcified regions on cell-laden side of the pre-mineralized samples. Though the results were not clear enough to conclude that the rate of mineral deposition by SaOs-2 cells in mineralized environment was higher than that of the non-mineralized environment, it can be proved in the future with more trials by using different types of bone cell-lines and improved cutting and imaging tools.
Marathe, Vaikhari Abhay, "AN IN-VITRO MODEL TO UNDERSTAND OSTEOBLAST MEDIATED BONE MINERALIZATION" (2019). Theses - ALL. 371.