载VEGF/万古霉素的多层缓释微球对人胎盘源间充质干细胞增殖与成骨分化的影响

doi:10.13481/j.1671-587x.20170208

吉林大学学报(医学版) ›› 2017, Vol. 43 ›› Issue (02): 250-254.doi: 10.13481/j.1671-587x.20170208

载VEGF/万古霉素的多层缓释微球对人胎盘源间充质干细胞增殖与成骨分化的影响

柳康¹, 王敬龙^1,2, 杨军星¹, 宋立杰¹, 徐一驰¹, 赵楚翘¹, 王瑶¹, 刘志辉¹, 王博蔚³

1. 吉林大学口腔医院修复科, 吉林长春 130021;
2. 温州医科大学口腔医学院附属口腔医院牙周病科, 浙江温州 325000;
3. 吉林大学第二医院妇产科, 吉林长春 130041

收稿日期:2016-10-25 出版日期:2017-03-28 发布日期:2017-03-31
通讯作者: 王博蔚,副教授,硕士研究生导师(Tel:0431-88796018,E-mail:liuzhihui1975@sina.com) E-mail:liuzhihui1975@sina.com
作者简介:柳康(1988-),女,山东省德州市人,在读医学硕士,主要从事干细胞与组织再生修复方面的研究。
基金资助:
吉林省科技厅重点科技攻关项目资助课题（20140204066SF）；吉林省科技厅医药产业推进计划项目资助课题（20140311088YY）；吉林省长春市科技局重大科技攻关计划资助课题（14KG049）；吉林省长春市科技局"双十工程"项目资助课题（16ss12）

Influence of multilayersustained-release microspheres loading VEGF and vancomycin in proliferation and osteogenic differentiation of HPMSCs

LIU Kang¹, WANG Jinglong^1,2, YANG Junxing¹, SONG Lijie¹, XU Yichi¹, ZHAO Chuqiao¹, WANG Yao¹, LIU Zhihui¹, WANG Bowei³

1. Department of Prosthodontics, Stomatology Hospital, Jilin University, Changchun 130021, China;
2. Department of Periodontics, Affiliated Stomatology Hospital, School of Stomatology, Wenzhou Medical Univensity, Wenzhou 325000, China;
3. Department of Obstetrics and Gynecology, Second Hospital, Jilin University, Changchun 130041, China

Received:2016-10-25 Online:2017-03-28 Published:2017-03-31

摘要/Abstract

摘要： 目的：观察载血管内皮生长因子（VEGF）/万古霉素的多层海藻酸盐壳聚糖缓释微球对人胎盘源间充质干细胞（HPMSCs）增殖与成骨分化的影响，为其在组织工程修复骨缺损中的临床应用提供理论基础。方法：根据前期基础制备缓释微球，将从人胎盘组织中分离培养的一部分HPMSCs分为载药微球组（载药微球+ HPMSCs）、空载微球组（空载微球+HPMSCs）和无球组（仅HPMSCs）。采用CCK-8试剂盒检测HPMSCs增殖率。取另一部分HPMSCs分为载药微球诱导组（载药微球+HPMSCs+成骨诱导液）、空载微球诱导组（空载微球+HPMSCs+成骨诱导液）、无球诱导组（HPMSCs+成骨诱导液）和非诱导组（HPMSCs+PBS）。孵育21d后分别采用茜素红染色及碱性磷酸酶（ALP）试剂盒检测HPMSCs钙盐沉积与ALP活性。结果：与无球组比较，载药微球组和空载微球组共培养后HPMSCs增殖率均无明显改变（P>0.05）。成骨诱导后茜素红染色，载药微球诱导组钙盐沉积明显多于空载微球诱导组和无球诱导组；载药微球诱导组细胞内ALP活性明显高于空载微球诱导组和无球诱导组（P<0.05），空载微球诱导组细胞ALP活性高于无球诱导组（P<0.05）。结论：载VEGF/万古霉素的多层海藻酸盐壳聚糖缓释微球对HPMSCs增殖活性无明显影响，且能提高HPMSCs的成骨分化能力。

关键词: 微球, 细胞增殖, 干细胞, 万古霉素, 成骨, 血管内皮生长因子, 碱性磷酸酶

Abstract: Objective: To investigate the influence of multilayer alginate chitosan sustained-release microspheres loading vascular endothelial growth factor(VEGF) and vancomycin in the proliferation and osteogenic differentiation of human plaacenta-derived mesen chymal stem cells(HPMSCs), and to provide theoretical basis for its clinical application in the repair of bone defect. Methods: The microspheres were prepared based on the previous research and HPMSCs were co-cultured with drug (VEGF/vancomycin)-loaded microspheres (drug-loaded microspheres+HPMSCs group), non-drug loaded microspheres (microspheres+HPMSCs group) and without any microspheres (HPMSCs group). Then the proliferation rate of HPMSCs was identified by CCK-8 kit. The osteogenic differentiation potential of HPMSCs was detected by Alizarin red staining and alkaline phosphatase (ALP) kit when the HPMSCs had been co-cultured with drug loaded microspheres in osteogenic medium (HPMSCs+ drug-loaded microspheres+induction group), non-drug loaded microspheres in osteogenic medium (HPMSCs+microspheres+induction group), without any microspheres in osteogenic medium (HPMSCs+induction group) and without any micropheres in normal medium (HPMSCs+PBS group) for 21 d. Results: Compared with HPMSCs group,the proliferation rates of HPMSCs in drug-loaded microspheres+HPMSCs and microspheres+HPMSCs groups had no significant changes (P>0.05).The calcium deposition in HPMSCs+drug-loaded microspheres+induction group was more than those in microspheress+HPMSCs+indution group and HPMSCs+induction group after Aalizarin red staining; the ALP activity in drug-loaded microspheres+HPMSCs+indution group was higher than those in microspheres+HPMSCs+indution group and HPMSCs+induction group (P<0.05),and the ALP activity in microspheres+HPMSCs+induction group was higher than that in HPMSCs+induction group(P<0.05). Conclusion: The sustained-release microspheres loading VEGF and vancomycin have no significant effect on the proliferation activity of HPMSCs and the microspheres could stimulate the osteogenic differentiation of HPMSCs.

Key words: vascular endothelial growth factor, alkalin phosphatase, vancomycin, stem cells, microsphere, osteogenesis, cell proliferation

中图分类号:

R318

柳康, 王敬龙, 杨军星, 宋立杰, 徐一驰, 赵楚翘, 王瑶, 刘志辉, 王博蔚. 载VEGF/万古霉素的多层缓释微球对人胎盘源间充质干细胞增殖与成骨分化的影响[J]. 吉林大学学报(医学版), 2017, 43(02): 250-254.

LIU Kang, WANG Jinglong, YANG Junxing, SONG Lijie, XU Yichi, ZHAO Chuqiao, WANG Yao, LIU Zhihui, WANG Bowei. Influence of multilayersustained-release microspheres loading VEGF and vancomycin in proliferation and osteogenic differentiation of HPMSCs[J]. Journal of Jilin University Medicine Edition, 2017, 43(02): 250-254.

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载VEGF/万古霉素的多层缓释微球对人胎盘源间充质干细胞增殖与成骨分化的影响

Influence of multilayersustained-release microspheres loading VEGF and vancomycin in proliferation and osteogenic differentiation of HPMSCs

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