Nano Phosphorylated Pullulan-Integrated Carboxymethyl Chitosan Scaffold for Regeneration of the Pulp-Dentin Complex: An

Background: To optimize a novel composite scaffold comprising of nano phosphorylated pullulan incorporated carboxymethyl chitosan (nPP-CMC) and to evaluate its cell viability and proliferation in human dental pulp stem cells (hDPSCs). Methodology: Nano phosphorylated pullulan (nPP) in concentrations of 200 mg, 400 mg, 600 mg, 800 mg, and 1 g was vortexed in 20 mL of distilled water, followed by 30 mins of ultrasonication for even dispersion. 1 g of CMC was added to each nPP mixture, blended for 1 hour to form a hydrogel, and freeze-dried for 18 hours at -20°C to yield nPP-CMC scaffolds in ratios of 1:5, 2:5, 3:5, 4:5, and 1:1. These scaffolds were stored at room temperature. The scaffolds were characterized using SEM-EDX, FT-IR, and XRD. The optimized nPP-CMC (group 1), CMC (group 2), and osteogenic medium (group 3) were tested for cell viability with hDPSCs using the MTT assay at 0, 7, 14, and 21 days. Results: Based on the results of MTT assay, the cell viability and proliferation of nPP-CMC scaffold was comparable to that of CMC in the tested time intervals of 7 and 14 days. The 4:5 nPP-CMC revealed a favourable pore size (60-180 μm) that was suitable for cell seeding and was chosen for further experiments. Cell viability/proliferation rates of hDPSCs loaded 4:5 HAnp- CMC at 21 Conclusion: Hence, within the limitations of this