Preliminary Study On The Feasibility Of Injectable Gelatin Hydrogel As A Delivery Vehicle For Demineralized Bone Matrix Powder
Keywords
Injectable, In Situ Hydrogel, Gelatin, Demineralized Bone Matrix
Abstract
Objective To introduce an injectable gelatin hydrogel and explore its feasibility as a delivery carrier for demineralized bone matrix (DBM) powder. Methods: First, gelatin was taken to prepare thiolized gelatin, and its thiol content was detected by the Ellman method; then it was cross-linked with polyethylene glycol diacrylate to prepare injectable gelatin hydrogel, and the gelation time was detected by the inversion method; finally, Injectable gelatin hydrogel is mixed with DBM powder to prepare a complex (hereinafter referred to as DBM-Gel). C2C12 cells were encapsulated in DBM-Gel, and live/dead staining and Alamar blue methods were used to study the cytotoxicity of the material. C2C12 cells were adhered to the surface of DBM, and DBM-Gel that wrapped the cells was prepared (DBM-Gel group). The ALP activity of the cells was detected after culturing for 1, 3, 5, and 7 days; DBM that simply adhered cells was used as a control (DBM group). A nude mouse muscle ectopic osteogenesis model was used to observe in vivo osteoinductivity. DBM-Gel (DBM-Gel group) and a mixture of DBM and PBS (DBM group) were implanted into the abdominal muscle pockets of nude mice, and the samples were collected 4 weeks later. Histological observation and ALP activity detection. Results The thiol content of the prepared thiolated gelatin tested by Ellman method was (0.51¡À0.03) mmol/g, and the gelation time of the injectable gelatin hydrogel was (6¡À1) min. After mixing DBM with thiolated gelatin and PEGDA solution, it can be injected to the implantation site within the gel time. As the culture time prolongs, the cells in DBM-Gel take on a spreading form, and some cells are connected; the cell survival rates at 1, 3, and 7 days of culture are 95.4%¡À1.9%, 97.3%¡À1.3%, and 96.1%¡À respectively. 1.6%; the relative proliferation rates of cells after culture for 1, 3, 5, and 7 days were 1.0¡À0.0, 1.1¡À0.1, 1.5¡À0.1, and 1.6¡À0.1 respectively. After in vitro induction and culture for 1, 3, 5, and 7 days, the cells in the DBM-Gel group and the DBM group showed similar ALP activity, and there was no statistically significant difference between the groups (P>0.05); as the culture time increased, the ALP activity in both groups gradually increased. The ALP activity on days 5 and 7 was significantly higher than that on days 1 and 3 (P<0.05). There was no significant difference between days 1 and 3 and between days 5 and 7 (P>0.05). Four weeks after implantation in vivo, bone and cartilage formation were seen in the DBM-Gel group, but no bone marrow formation was observed; bone marrow, bone, and cartilage formation were seen in the DBM group. The osteoinductive histological scores of the DBM-Gel group and DBM group were 4.0 and 4.5 points respectively; the ALP activities were (119.4¡À22.7) and (146.7¡À13.0) ¦Ìmol/mg protein/min respectively, and there was no statistically significant difference between the groups. (t=¨C2.085, P=0.082). Conclusion Injectable gelatin hydrogel is feasible as a delivery carrier for DBM powder
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Original research was done by Tian Meng
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