钛种植体表面新型DNA水凝胶递送地塞米松诱导巨噬细胞M2极化的研究  

A novel DNA hydrogel as dexamethasone delivery platform on titanium implant surface induces M2 polarization of macrophages

作　　者：陈芳浩 何奕德 李哲 闫宁 宋文 张玉梅 CHEN Fanghao;HE Yide;LI Zhe;YAN Ning;SONG Wen;ZHANG Yumei(State Key Laboratory of Military Stomatology&National Clinical Research Center for Oral Diseases&Shaanxi Key Laboratory of Stomatology,Department of Prosthodontics,School of Stomatology,The Fourth Military Medical University,Xi'an 710032 China)

机构地区：[1]军事口腔医学国家重点实验室,口腔疾病国家临床医学研究中心,陕西省口腔医学重点实验室,第四军医大学口腔医院修复科,西安710032

出　　处：《实用口腔医学杂志》2021年第2期192-196,共5页Journal of Practical Stomatology

基　　金：国家自然科学基金(编号:81530051,31800790);陕西省重点研发计划(编号:2019SF-031);陕西省高校科协青年人才托举(编号:20190304)。

摘　　要：目的:构建壳聚糖DNA水凝胶负载递送地塞米松(Dex),探索其在钛种植体表面对巨噬细胞M2极化的作用。方法:阳极氧化法(20 V)在钛表面制备纳米管(NT);将鲑鱼精双链DNA变性-退火-交叉配对形成预凝胶,与壳聚糖交联形成水凝胶,采用真空抽吸将凝胶加载到纳米管的管腔中;通过SEM、FTIR和体外释放试验检测其结构特征及载药释放性能;将RAW264.7细胞培养在各组钛样本表面,采用CCK-8、CLSM、RT-qPCR检测凝胶对细胞增殖、极化的影响。结果:成功制备出负载Dex的DNA-壳聚糖水凝胶,延长了Dex的半数释放量天数,NT+Dex@Gel组的细胞增殖活性强于NT、NT+Gel组,CD86表达水平显著降低、CD163表达水平显著升高(P<0.05)。结论:壳聚糖交联DNA骨架形成水凝胶负载Dex在钛种植体表面能促进巨噬细胞增殖并诱导其M2型极化。Objective:To fabricate a novel DNA hydrogel with chitosan as the platform for dexamethasone(Dex)delivery from titanium implant,and to explore its effect on the M2 polarization of macrophages.Methods:The titanium implant was prepared by anodization(20 V)for the nanotube structure(NT)formation.The double-stranded DNA from salmon sperm was denatured and annealed to form a pregel through random base pairing.The final hydrogel was formed by electrostatic interaction between chitosan and the pregel.Then the hydrogel was vacuumly penetrated into the lumen of the NT.The charicteristic and Dex release performance were tested by SEM,FTIR and in vitro release test.RAW264.7 cells were cultured on different surfaces and the effects of hydrogel on cell proliferation and polarization were detected by CCK-8 assay,CLSM and RT-qPCR.Results:The DNA-chitosan hydrogel was successfully prepared and loaded both on the surface and into the tubes of NT.The half-life time of Dex release profile was extended.Compared with the NT and NT+Gel groups,the cell proliferation was promoted on NT+Dex@Gel surface.The CD86 expression were inhibited and CD163 expression was increased by NT+Dex@Gel(P<0.05).Conclusion:The novel DNA hydrogel of Dex delivery can promote the proliferation and M2 polarization of macrophages on titanium surface.

关 键 词：巨噬细胞极化 地塞米松 钛 纳米管 DNA水凝胶 壳聚糖 

分 类 号：R783.6[医药卫生—口腔医学]