中国组织工程研究 ›› 2023, Vol. 27 ›› Issue (7): 998-1003.doi: 10.12307/2023.033

• 组织工程骨材料 tissue-engineered bone • 上一篇    下一篇

水凝胶复合载辛伐他汀蛋白微球对成骨细胞增殖分化的影响

柳晓琳1,穆新月2,马子雨3,刘树泰4,王文龙5,韩晓谦1,董志恒6   

  1. 滨州医学院附属医院,1口腔修复科,5口腔颌面外科,6儿童口腔科,山东省滨州市  256600;2济宁医学院附属医院口腔内科,山东省济宁市 272007;3滨州医学院,山东省烟台市  264000;4烟台市口腔医院牙周科,山东省烟台市  264000
  • 收稿日期:2021-11-24 接受日期:2022-01-13 出版日期:2023-03-08 发布日期:2022-07-16
  • 通讯作者: 韩晓谦,硕士,滨州医学院附属医院口腔修复科,山东省滨州市 256603 董志恒,硕士,滨州医学院附属医院儿童口腔科,山东省滨州市 256603
  • 作者简介:柳晓琳,女,1995年生,汉族,山东省烟台市人,滨州医学院在读硕士,主要从事口腔牙周病研究。
  • 基金资助:
    山东省医药卫生科技发展计划项目(ZR2019PH075),项目负责人:王文龙;滨州医学院科技计划项目(BY2019KJ15),项目负责人:韩晓谦

Effect of hydrogel-loaded simvastatin microspheres on osteoblast proliferation and differentiation

Liu Xiaolin1, Mu Xinyue2, Ma Ziyu3, Liu Shutai4, Wang Wenlong5, Han Xiaoqian1, Dong Zhiheng6   

  1. 1Department of Prosthodontics, 5Department of Oral and Maxillofacial Surgery, 6Department of Children’s Stomatology, Affiliated Hospital of Binzhou Medical College, Binzhou 256600, Shandong Province, China; 2Department of Stomatology, Affiliated Hospital of Jining Medical College, Jining 272007, Shandong Province, China; 3Binzhou Medical College, Yantai 264000, Shandong Province, China; 4Periodontal Department, Yantai Stomatological Hospital, Yantai 264000, Shandong Province, China
  • Received:2021-11-24 Accepted:2022-01-13 Online:2023-03-08 Published:2022-07-16
  • Contact: Han Xiaoqian, Master, Department of Prosthodontics, Affiliated Hospital of Binzhou Medical College, Binzhou 256600, Shandong Province, China Dong Zhiheng, Master, Department of Children’s Stomatology, Affiliated Hospital of Binzhou Medical College, Binzhou 256600, Shandong Province, China
  • About author:Liu Xiaolin, Master candidate, Department of Prosthodontics, Affiliated Hospital of Binzhou Medical College, Binzhou 256600, Shandong Province, China
  • Supported by:
    Shandong Provincial Medical and Health Science and Technology Development Project, No. ZR2019PH075 (to WWL); Science and Technology Planning Project of Binzhou Medical College, No. BY2019KJ15 (to HXQ)

摘要:

文题释义:
辛伐他汀:属于他汀类药物,临床中主要应用于降低患者胆固醇水平。近年的研究发现辛伐他汀能够促进骨的再生,抑制牙龈链球菌的生物活性,从而达到抑制牙周炎、促进牙周骨组织形成的目的。
水凝胶:具有良好的三维结构,能够为细胞生长提供空间;具有控释作用,能够缓释药物;具有流动性能,够更加贴合缺损部位的形态;具有可注射性,能够方便操作和储存;具有良好的力学性能、稳定性和抗扩张性。

背景:牙周炎患者逐年增加,采用传统的牙周治疗方法并不能恢复牙周软硬组织,因此需要制备出一种药物缓释材料辅助治疗牙周炎,恢复牙周破坏的软硬组织。
目的:制备装载辛伐他汀的牛血清白蛋白微球复合水凝胶材料,检测其对辛伐他汀的双重缓释作用,并进一步研究此释药复合材料对成骨细胞黏附、增殖的影响。
方法:①采用去溶剂法制备载辛伐他汀的牛血清白蛋白微球,采用透射电镜、扫描电镜观察微球的形态,测量其粒径,采用酶标仪检测载药微球的包封率、载药率及体外药物释放;②将载药微球负载到明胶水凝胶中,采用酶标仪检测水凝胶的体外药物释放,扫描电镜观察复合材料的形态;利用该水凝胶浸提液培养MC3T3-E1细胞,采用CCK-8法检测细胞增殖,碱性磷酸酶试剂盒检测细胞碱性磷酸酶的表达。
结果与结论:①透射电镜显示,载药微球呈光滑的圆球形,较为分散,未见明显聚集,微球粒径也较为均匀,80%的微球粒径在0.2-
0.8 µm之间;扫描电镜显示,载药微球光滑,呈圆球形,分散性较好,粒径分布较为均匀,粒径在0.2-0.8 µm之间;②载药微球的包封率为68.9%-87.5%,载药率为0.95%-1.21%;③载药微球中辛伐他汀的释放曲线是温和的持续释放过程,载药水凝胶中辛伐他汀的释放曲线为前期释放速度快后期缓慢的持续释放过程,其中载药水凝胶比载药微球能更快释放并达到药物的作用浓度,在后期缓慢释放维持药物的作用浓度;④扫描电镜显示,载药水凝胶呈多孔条状结构,适合成骨细胞的黏附和生长,在水凝胶表面可以看到圆球形的载药微球;⑤载药水凝胶可促进MC3T3-E1细胞的增殖与碱性磷酸酶表达;⑥结果表明,缓释辛伐他汀微球水凝胶复合材料具有良好的生物相容性,能够促进成骨细胞的增殖与成骨分化。

https://orcid.org/0000-0002-2885-891x (柳晓琳)

中国组织工程研究杂志出版内容重点:生物材料;骨生物材料口腔生物材料纳米材料缓释材料材料相容性组织工程

关键词: 辛伐他汀, 药物缓释, 微球, 水凝胶, 成骨细胞, 增殖与分化

Abstract: BACKGROUND: The number of patients with periodontitis increases year by year, and the use of traditional periodontal treatment cannot restore the periodontal soft and hard tissue. Therefore, it is necessary to prepare a drug sustained-release material to assist the treatment of periodontitis and restore the damaged soft and hard tissues of periodontitis.
OBJECTIVE: To prepare bovine serum albumin microspheres composite hydrogel material, detect the dual sustained release effect on simvastatin, and further study the effect of the composite material on the adhesion and proliferation of osteoblasts. 
METHODS: (1) Bovine serum albumin microspheres loaded with simvastatin were prepared by solvent removal method. The morphology of microspheres was observed by transmission electron microscopy and scanning electron microscopy and the particle size of microspheres was measured. The encapsulation rate, drug loading rate and in vitro release of microspheres were detected by a microplate analyzer. (2) Simvastatin albumin microspheres were loaded into the prepared hydrogel, and in vitro drug release from hydrogels was detected using a microplate reader. The morphology of the composite material was observed by scanning electron microscopy. The hydrogel extract was used to culture MC3T3-E1 cells. The proliferation of osteoblasts was detected by CCK-8 assay. The alkaline phosphatase expression in osteoblasts was detected by alkaline phosphatase kit. 
RESULTS AND CONCLUSION: (1) Transmission electron microscopy showed that bovine serum albumin microspheres and blank microspheres loaded with simvastatin were smooth, spherical and dispersed, with no obvious aggregation. The particle size of bovine serum albumin microspheres was uniform, and 80% of the microspheres had a particle size between 0.2 μm and 0.8 μm. Scanning electron microscopy showed that the microspheres were smooth and spherical with good dispersion and uniform particle size distribution, ranging from 0.2 μm to 0.8 μm. (2) The encapsulation rate and drug loading rate of simvastatin microspheres were 68.9%-87.5% and 0.95%-1.21%, respectively. (3) Drug-loaded microspheres had the performance of gentle and sustained release of simvastatin. The release curve of simvastatin in the drug-loaded hydrogel is a fast release in the early stage and a slow sustained release process in the later stage, in which the drug-loaded hydrogel can release faster than the drug-loaded microspheres and reach the effective concentration of the drug. The slow release in the later stage maintains the concentration of the drug. (4) Scanning electron microscopy showed that the drug-loaded hydrogel had a porous strip-like structure, which was suitable for the adhesion and growth of osteoblasts, and spherical drug-loaded microspheres could be seen on the surface of the hydrogel. (5) The drug-loaded hydrogel can promote the proliferation and alkaline phosphatase expression of MC3T3-E1 cells. (6) Simvastatin microspheres hydrogel composite had good biocompatibility and could promote the adhesion and proliferation of osteoblasts. 

Key words: simvastatin, sustained drug release, microsphere, hydrogel, osteoblasts, proliferation and differentiation

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