The aim of this study is to evaluate the injectable cross-linked chitosan (CS) microparticles (MPs) to apply for biomedical applications specifically for bone regeneration. The CS MPs were fabricated by emulsification method and formed the cross-links between the amide groups in the CS and phosphate groups in the sodium tripolyphosphate (TPP) ionic cross-linking agent. The MPS were analyzed for morphology by Scanning Electron Microscope (SEM). The fabricated CS MPs were in the spherical shape with the size range of 20-100 m. These CS MPs were analyzed for biodegradation by immersing in phosphate buffered saline (PBS, pH = 7.4) at 37°C for 30 weeks. The biodegradation of CS MPs in PBS was initiated at week 25. Mesenchymal stem cells (MSCs) were harvested from the bone marrow of mice tibia and femurs. The MSC attachment on CS MPs was tested using LIVE/DEAD cell sassy with a Fluorescence Microscope. The murine MSCs attachment onto CS MPs at day 2 was confirmed by visualizing fluorescence images. The CS MPs were also analyzed for the injectability and retainability at the site using a subcutaneous injection in a rat model. The fabricated CS MPs possess injectability, biodegradability and biocompatibility. Therefore, these CS MPs have a great potential to apply for various biomedical applications including bone regeneration by injection.