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

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Injectable thermosensitive chitosan/gelatin-based hydrogel carried erythropoietin to effectively enhance maxillary sinus floor augmentation in vivo.
Dental Materials ( IF 4.6 ) Pub Date : 2020-05-26 , DOI: 10.1016/j.dental.2020.04.016
Daowei Li ₁ , Liang Zhao ₂ , Mingyu Cong ₃ , Lijun Liu ₄ , Guangxing Yan ₄ , Zhimin Li ₄ , Baoquan Li ₄ , Weixian Yu ₄ , Hongchen Sun ₅ , Bai Yang ₆

Affiliation

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 ± 15 s 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 4 h, 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 (81.98 mm³) compared with control group (43.11 mm³) 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 (∼121.4 μm) compared to control group (∼37 μm) resulting in enhancing intramembranous ossification.

Significance

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

中文翻译：

可注射的热敏壳聚糖/明胶基水凝胶携带促红细胞生成素，可在体内有效增强上颌窦底隆突。

目的

上颌窦底增强术（MSFA）通常用于在植入植入物之前增加后上颌的牙槽骨高度。在本研究中，我们评估了可注射的热敏壳聚糖/β-甘油磷酸二钠钠水合物/明胶（CS / GP / GA）水凝胶是否携带促红细胞生成素（EPO）可以增强体内MSFA的新骨形成。

方法

EPO-CS / GP / GA水凝胶是通过离子交联制备的。然后，通过形态，可注射性和pH值对胶凝时间（GT）的评价EPO-CS / GP / GA的特性。通过酶联免疫吸附试验（ELISA）评估EPO的释放曲线，并通过3- [4,5-二甲基噻唑-2-基]-分析EPO对骨髓基质细胞（BMSC）增殖和成骨细胞分化的影响。 2,5-二苯基溴化四氮唑（MTT）和逆转录定量实时PCR（RT-qPCR）。最后，将EPO-CS / GP / GA注射到兔子的上颌窦底，以测试MSFA的潜在应用。

结果

结果表明，GT随着pH值的升高而降低。GT在pH 7.0时为110±15 s。SEM图像表明CS / GP / GA水凝胶具有海绵网络结构。ELISA分析的结果表明，EPO从EPO-CS / GP / GA水凝胶的累积释放在4小时达到67％，在15天达到94％。MTT法检测显示，与对照组相比，EPO-CS / GP / GA水凝胶中的EPO可以显着促进BMSCs的增殖（p <0.001）。RT-qPCR分析结果表明，与对照组相比，EPO-CS / GP / GA组在第14天时Sp7，Runx2，Col I和Alp的表达显着增加（p <0.001）。重要的是，在体内植入后12周，与对照组（43.11 mm ³）相比，EPO-CS / GP / GA水凝胶可显着诱导骨骼形成（81.98 mm ³）。间充血凝结厚度的计算表明，与对照组（〜37μm）相比，EPO-CS / GP / GA组（〜121.4μm）间质凝结的厚度显着增加，导致膜内骨化增强。