Injectable thermosensitive chitosan/gelatin-based hydrogel carried erythropoietin to effectively enhance maxillary sinus floor augmentation in vivo

Abstract

Objective: Maxillary sinus floor augmentation (MSFA) is commonly used to increase the alveolar bone height in the posterior maxilla before implant placement. In the present study, we evaluated if the injectable thermosensitive chitosan/β-sodium glycerophosphate disodium salt hydrate/gelatin (CS/GP/GA) hydrogel carried erythropoietin (EPO) could enhance the new bone formation for MSFA in vivo.

Methods: EPO-CS/GP/GA hydrogel was prepared by ionic crosslinking. Then, characteristics of EPO-CS/GP/GA were evaluated by morphology, injectable property and pH on the gelling time (GT). The release profile of EPO was evaluated by enzyme linked immunosorbent assay (ELISA), and effects of EPO on proliferation and osteoblastic differentiation of bone marrow stromal cells (BMSC) were analyzed by 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide (MTT) and reverse transcription quantitative real-time PCR (RT-qPCR), respectively. Finally, EPO-CS/GP/GA was injected into the maxillary sinus floor of the rabbit to test the potential application for MSFA.

Results: Results showed that GT was decreased with the increase of pH value. The GT was 110±15s at pH 7.0. SEM images showed that the CS/GP/GA hydrogel had a sponge network structure. Results from ELISA assay revealed that the cumulative release of EPO from the EPO-CS/GP/GA hydrogel reached 67% at 4h, and 94% at 15 days. MTT assay showed that EPO within EPO-CS/GP/GA hydrogel could significantly promote proliferation of BMSCs compared to control group (p<0.001) . Results of RT-qPCR assays demonstrated that the expression of Sp7, Runx2, Col I and Alp were significantly increased from EPO-CS/GP/GA group compared to control group on day 14 (p<0.001). Importantly, EPO-CS/GP/GA hydrogel could significantly induce bone formation (81.98mm³) compared with control group (43.11mm³) after 12 weeks post-implantation in vivo. The calculation of thickness of mesenchymal condensation indicated that thickness of mesenchymal condensation was significantly increased from EPO-CS/GP/GA group (∼121.4μm) compared to control group (∼37μm) resulting in enhancing intramembranous ossification.

Significance: The EPO-CS/GP/GA hydrogel provides a novel strategy for MSFA with a minimally invasive way.

Keywords: Chitosan; Erythropoietin; Gelatin; Maxillary sinus augmentation; Thermosensitive hydrogel.