The degradation properties of the MPs is important to the long-term benefits of the use of the chitosan (CS) based hybrid MPs in bone tissue-engineering, because the degradation kinetics could affect a multitude of processes within the cell, such as cell growth, tissue regeneration, and host response. The aim of this study was to investigate the degradation of solid, hybrid CS microparticles (MPs), CS-10% calcium phosphate (CaHPO4, w/w), and CS-10% calcium carbonate (CaCO3, w/w) MPs in phosphate buffered solution (PBS) over a 30-week period. The hybrid MPs were synthesized by emulsification technique, cross-linked with 64% sodium tripolyphosphate (TPP), purified and air dried overnight. Each sample had 30 mg of MPs was placed in a glass vial with 9 ml of PBS added and then the vial was closed to prevent evaporation. Every week 4 ml of the incubated solution was removed for sample measurement and all samples were replaced with an equivalent amount of fresh medium. The samples were maintained at 37°C under continuous shaking. The hybrid MPs were measured for pH and calcium release, every week in triplicate. At 0, 5, 10, 15, 20, 25, and 30 weeks, surface and bulk morphology were analyzed with a scanning electron microscope (SEM). The degradation data suggested that the hybrid MPs were stable at least up to 25 week and maintain the physiologically relevant pH. Therefore, we can use these hybrid MPs to apply in the bone tissue engineering applications since they do not degrade within a short period.
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 μm. 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.
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