The Collaborative Research Of C-G-Ha-HS Scaffolds And PCA On NS/PCs Differentiation

At present, there are some achievements in neural stem/progenitor cells (NS/PCs) transplantation treating Parkinson’s disease (PD), but this method exist defects. Though NS/PCs can differentiate into dopaminergic (DA) neuron, but the short survival time and limited differentiation conditions of NS/PCs made it lose contact with the surrounding tissue of damaged parts of PD, this method need to be further perfected. In order to improve NS/PCs’vitality and differentiation ability, it is essential to establish3-D scaffold for NS/PCs expansion and differentiation.Chitosan-gelatin-hyaluronate-heparan sulfate (C-G-Ha-HS) scaffolds were fabricated by freeze-drying. After physical detection of FITC, pore diameter, porosity, water absorbing ratio, adhering ratio and degradation rate, screened out the ideal scaffolds, modified with laminin (LN), constructed3-D culture system of NS/PCs. Meanwhile, the effect of modified scaffolds on biocompatibility and the synergistic effect of protocatechuic acid (PCA) and scaffolds on inducing NS/PCs differentiation were determined. By FTIR, all scaffolds were cross linked. The amide group, sulfo group, ester bond, acid anhydride of scaffolds can promote NS/PCs adhesion, migration and differentiation. By physical characterization, C-G-Ha-HS scaffolds of120～130μm pore size and92%porosity possess the ability of water absorbing and degradation ability, they are suitable for NS/PCs culture. By CCK-8assay, the adhesive capacity and viability of NS/PCs in LN modified C-G-Ha-HS scaffolds were improved remarkably, compared with LN-unmodified C-G-Ha-HS scaffold, LN-modified scaffolds enhanced NS/PCs adhesion rate by113.53±4.32%after4hours, LN-modified scaffolds increased the cell viability by120.30±6.65%; By immunocytochemistry assay, the synergistic effect of1%fetal bovine serum (FBS) and0.06mM PCA enhance the differentiation rate of NS/PCs into neurons. Compared with scaffold group, the differentiate rate of TH and Nurrl positive cell were enhanced by172.72±6.79%and181.23±1.04%in LN modified C-G-Ha-HS (5:5) scaffolds. In conclusion, LN modified C-G-Ha-HS (5:5) scaffolds promote NS/PCs adhesion and proliferation, meanwhile, the synergistic effect of scaffold, LN, PCA and FBS can improve NS/PCs’ ability of differentiating into DA neuron and the viability of differentiation phenotype. Therefore, it provides experimental basis for the application of PCA to treat PD by NS/PCs transplantation.