用于颌骨缺损修复的可注射多功能脂质体-水凝胶复合体的构建及体外评价

doi:10.19438/j.cjoms.2022.06.003

摘要/Abstract

摘要： 目的利用脂质体作为促血管化小分子去铁胺（DFO）和成骨诱导小分子地塞米松（Dex）的缓释载体,同时利用海藻酸钠水凝胶作为万古霉素（Van）的缓释载体,构建一种可注射多功能脂质体-水凝胶复合体,探讨复合体体外抑菌、促血管生成及促成骨作用。方法通过薄膜分散法制备DFO/Dex脂质体,与Van海藻酸钠溶液混合,在钙离子作用下形成Van/DFO/Dex脂质体-水凝胶复合体,通过Zeta粒径仪检测脂质体的粒径、分散度（PDI）和Zeta电位;通过紫外可见光分光光度计检测复合体中Van、DFO和Dex的缓释曲线;WST-8定量实验评价复合体的体外抑菌作用;CCK-8和细胞活/死荧光染色评价复合体的细胞相容性;实时荧光定量PCR（RT-PCR）检测复合体促血管生成作用;ALP活性定量和染色评价Van/DFO/Dex脂质体-水凝胶复合体的体外促成骨分化作用。采用OriginPro软件对数据进行统计学处理。结果成功构建可注射性Van/DFO/Dex脂质体-水凝胶复合体,测定脂质体的粒径为（121.6±6.35）nm,PDI为0.2~0.3,DFO包封率约为57%,Dex包封率约为23%。药物缓释曲线表明,Van、DFO和Dex呈分阶段持续释放。CCK-8和细胞活/死荧光染色显示,Van/DFO/Dex脂质体-水凝胶复合体具有良好的细胞相容性。WST-8定量检测表明,复合体具有良好的体外抑菌作用。血管生成相关基因的表达提示,复合体具有促血管生成的能力。ALP活性定量和染色表明,Van/DFO/Dex脂质体-水凝胶复合体能够提高成骨分化活性。结论 Van/DFO/Dex脂质体-水凝胶复合体是一种具有成骨、促血管生成及抗感染的多功能骨再生材料,其可注射性有利于不规则颌骨缺损的临床应用。

关键词: 颌骨缺损, 骨替代材料, 血管生成, 成骨, 抗感染

Abstract: PURPOSE: Liposome was used as a sustained release carrier for pro-vascularization small molecule desferrioxamine (DFO) and osteogenesis-inducing small molecule dexamethasone (Dex), while sodium alginate hydrogel was used as a sustained release carrier for vancomycin (Van) to construct an injectable liposome-hydrogel complex with simultaneous antibacterial, pro-angiogenic and osteogenic effects, and to investigate the ability of the complex to promote bone tissue regeneration and repair. METHODS: DFO/Dex liposomes were prepared by thin film dispersion method, and mixed with Van sodium alginate solution to form Van/DFO/Dex liposome-hydrogel complex under the action of calcium ions. The particle size, polydispersity index(PDI) and Zeta potential of the liposomes were measured by Zeta particle size analyzer. The sustained release curves of Van, DFO and Dex in the complex were determined by UV-vis spectrophotometer. Wst-8 quantitative assay was used to evaluate the antibacterial activity of the complex in vitro. CCK-8 and cell live/dead fluorescence staining were used to evaluate the cytocompatibility of the complex. Real-time fluorescence quantitative PCR (RT-PCR) was used to detect the angiogenesis promoting effect of the complex. ALP activity quantification and staining were used to evaluate the effect of Van/DFO/Dex liposome-hydrogel complex on promoting bone differentiation in vitro. OriginPro software was used for analysis of the data. RESULTS: An injectable Van/DFO/Dex liposome-hydrogel complex was successfully constructed. The particle size of the liposome was (121.6±6.35)nm, PDI was 0.2-0.3. The encapsulation rate of DFO was about 57% and that of Dex was about 23%. Sustained release curve showed that Van DFO and Dex were released continuously in stages. CCK-8 and cell live/dead fluorescence staining showed that Van/DFO/Dex liposome-hydrogel complex had good cytocompatibility. Quantitative detection of WST-8 showed that the complex had good antibacterial activity in vitro. The expression of genes related to angiogenesis suggests that the complex had the ability to promote angiogenesis. ALP activity quantification and staining showed that Van/DFO/Dex liposome-hydrogel complex could enhance osteogenic differentiation activity. CONCLUSIONS: Van/DFO/Dex liposome-hydrogel complex is a multifunctional bone regeneration material with osteogenesis, angiogenesis promotion and anti-infection, and its injectable properties are beneficial to the clinical application of irregular jaw defects.

Key words: Jaw defect, Bone substitute, Angiogenesis, Osteogenesis, Bacteriostasis

中图分类号:

R782.4

寻兴祥, 孙伟, 王荷静, 楚晨, 赵谦, 许晓, 袁荣涛. 用于颌骨缺损修复的可注射多功能脂质体-水凝胶复合体的构建及体外评价[J]. 中国口腔颌面外科杂志, 2022, 20(6): 534-540.

XUN Xing-xiang, SUN Wei, WANG He-jing, CHU Chen, ZHAO Qian, XU Xiao, YUAN Rong-tao. Construction and in vitro evaluation of an injectable multifunctional liposome-hydrogel composite for jaw defect repair[J]. China J Oral Maxillofac Surg, 2022, 20(6): 534-540.

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