| Objective:To prepare of chitosan thermosensitive hydrogel and platelet-rich plasma (PRP) and investigate the micro-morphous of chitosan thermosensitive hydrogel combined with PRP. To investigate the effects of the leachate from chitosan thermosensitive hydrogel/PRP on bone marrow mesenchymal cells'(BMSCs) growth and proliferation. Methods:(1) Preparing chitosan thermosensitive hydrogel and PRP.(2) Combining chitosan thermosensitive hydrogel and PRP with different volume ratio and then dividing them into5groups:A. pure chitosan thermosensitive hydrogel; B, chitosan thermosensitive hydrogel:PRP=2:1; C, chitosan thermosensitive hydrogel:PRP=1:1; D, chitosan thermosensitive hydrogel:PRP=1:2; E, pure PRP. Strucure of each group was observed by scanning electron microscopy after lyophilizing.(3) BMSCs were obtained from dog bone marrow aspiration. The cells of the third generation were cultured by the leachate from each group above. The proliferation rates and ALP activity of BMSCs were examined by MTT assay and ALP kit. Results:(1) It clearly indicated that the liquid solution took eight-ten minutes to become a semi-solid gel in the vicinity of37?.(2) With the increasing of PRP volume ratio, the aperture size of Chitosan thermosensitive hydrogel grows up. The aperture size of group A to E are respectively about50um,30-60um,100-200um,200-300um and200-500um.(3) The proliferation and the ALP activity of BMSCs was significantly enhanced by B, C, D, E group (P<0.05). Conclusion:PRP can increase the aperture size of Chitosan thermosensitive hydrogel. The aperture sizes of Group C and D have already reached the ideal demand of tissue engineering scaffold. Chitosan thermosensitive hydrogel can promote proliferation rates and ALP activity of BMSCs by releasing PRP slowly. The Chitosan thermosensitive hydrogel combined with PRP is likely to be used as a new scaffold material in the periodontal regeneration. |
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