丝素改性聚乳酸-羟基乙酸共聚物多孔微球作为牙龈间充质干细胞递送载体的研究

doi:10.7503/cjcu20190193

高等学校化学学报 ›› 2019, Vol. 40 ›› Issue (11): 2419.doi: 10.7503/cjcu20190193

• 高分子化学 • 上一篇

丝素改性聚乳酸-羟基乙酸共聚物多孔微球作为牙龈间充质干细胞递送载体的研究

刘垚杉¹,吴海林¹,贾智¹,杜博²,刘大勇^1,^*(),周志敏^2,^*()

^1. 天津医科大学口腔医院, 天津 300070
^2. 中国医学科学院北京协和医学院生物医学工程研究所屏障医学研究中心, 天津市生物医学材料重点实验室, 天津 300192

收稿日期:2019-04-02 出版日期:2019-11-10 发布日期:2019-10-15
通讯作者: 刘大勇,周志敏 E-mail:dyliuperio@tmu.edu.cn;zhouzm@bme.cams.cn
基金资助:
国家自然科学基金(81371109);国家自然科学基金(81670953);中国医学科学院医学与健康科技创新工程项目(2017-I2M-1-007);人力资源和社会保障部留学人员科技活动项目择优资助项目资助.(201519205)

Silk Fibroin-modified Ploylactic Acid-glycolic Acid Copolymer Porous Microspheres as Gingival Mesenchymal Stem Cells Delivery Carrier ^†

LIU Yaoshan¹,WU Hailin¹,JIA Zhi¹,DU Bo²,LIU Dayong^1,^*(),ZHOU Zhimin^2,^*()

^1. Stomatological Hospital of Tianjin Medical University, Tianjin 300070, China
^2. Tianjin Key Laboratory of Biomedical Materials, Biomedical Barriers Research Center, Institute of Biomedical Engineering, Chinese Academy of Medical Sciences & Peking Union Medical College, Tianjin 300192, China

Received:2019-04-02 Online:2019-11-10 Published:2019-10-15
Contact: LIU Dayong,ZHOU Zhimin E-mail:dyliuperio@tmu.edu.cn;zhouzm@bme.cams.cn
Supported by:
? Supported by the National Natural Science Foundation of China(81371109);? Supported by the National Natural Science Foundation of China(81670953);the CAMS Innovation Fund for Medical Sciences, China(2017-I2M-1-007);the Technology Foundation for Selected Overseas Chinese Scholar, Ministry of Human Resources and Social Security, China(201519205)

摘要/Abstract

摘要：

利用复乳-溶剂挥发法合成适合细胞三维培养的聚乳酸-羟基乙酸共聚物(PLGA)多孔微球, 并对其表面进行丝素改性, 利用扫描电子显微镜、能谱、红外光谱和X射线衍射等对改性前后PLGA多孔微球的理化特性进行表征. 原代培养人牙龈间充质干细胞并进行成骨(茜素红染色)成脂(油红O染色)分化鉴定. 通过负压混悬法将牙龈干细胞负载于丝素改性的PLGA多孔微球上进行5-乙炔基-2'-脱氧尿嘧啶核苷(EdU)细胞增殖及成骨分化研究. 结果表明, 原代培养的牙龈干细胞具有多向分化潜能, 负载在丝素改性的PLGA多孔微球上的细胞有利于细胞增殖. 丝素改性的PLGA多孔微球是良好的细胞递送载体, 为进一步修复牙槽骨缺损提供了科学依据.

关键词: 牙龈间充质干细胞, 聚乳酸-羟基乙酸共聚物多孔微球, 丝素蛋白改性, 组织修复

Abstract:

The polylactic acid-glycolic acid copolymer(PLGA) porous microspheres suitable for three-dimensional cell culture were prepared by double emulsion-solvent evaporation method, which were then subjected to silk fibroin modification, and characterized by scanning electron microscopy, energy dispersive spectroscopy, infrared spectroscopy and X-ray diffraction. Primary cultured human gingival mesenchymal stem cells were used for osteogenic(alizarin red staining) and adipogenic(oil red O staining) differentiation identification experiments. The gingival stem cells were seeded on silk fibroin-modified PLGA porous microspheres by negative pressure suspension method, and the 5-ethynyl-2'-deoxyuridine(EdU) cell proliferation experiment and osteogenic differentiation experiment were performed. The result confirmed that primary cultured gingival stem cells had multi-directional differentiation potential, and those loaded on silk fibroin-modified PLGA porous microspheres were beneficial to cell proliferation. Silk fibroin modified PLGA porous microspheres can be good cell delivery carriers, providing a scientific basis for further repair of alveolar bone defects.

Key words: Gingival mesenchymal stem cell, Polylactic acid-glycolic acid copolymer porous microspheres, Silk fibroin-modification, Tissue repairing

中图分类号:

O631

TrendMD:

刘垚杉,吴海林,贾智,杜博,刘大勇,周志敏. 丝素改性聚乳酸-羟基乙酸共聚物多孔微球作为牙龈间充质干细胞递送载体的研究. 高等学校化学学报, 2019, 40(11): 2419.

LIU Yaoshan,WU Hailin,JIA Zhi,DU Bo,LIU Dayong,ZHOU Zhimin. Silk Fibroin-modified Ploylactic Acid-glycolic Acid Copolymer Porous Microspheres as Gingival Mesenchymal Stem Cells Delivery Carrier ^†. Chem. J. Chinese Universities, 2019, 40(11): 2419.

图/表 9

Fig.1 SEM images of PLGA porous microspheres without(A―C) and with(D) NaOH treatment PLGA concentration(mg/mL): (A) 16; (B, C, D) 25.

Fig.2 SEM image of PLGA(A) and SEM image(B), EDS(C) and FTIR spectra(D) of SF-PLGA porous microspheres

Fig.3 Optical microscopy images of primary(A) and the third-generation(B―D) gingival mesenchymal stem cells (C) Dyed by alizarin red;(D) dyed by oil red O.

Fig.4 Confocal laser scanning microscopy images of PLGA(A) and SF-PLGA(B) porous microspheres

Fig.5 DNA concentration quantitative analysis

Fig.6 Alkaline phosphatase activity assay

Fig.7 Optical microscopy images of PLGA(A) and SF-PLGA(B) after Alizarin red staining

Fig.8 SEM images(A,B) and EDS(C,D) of calcium nodules on PLAG(A, C) and SF-PLGA(B, D) porous microspheres

Fig.9 XRD patterns of PLGA(a, b) and SF-PLGA(c) porous microspheres a. Without cells; b,c. after osteogenic induction for 16 d.

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丝素改性聚乳酸-羟基乙酸共聚物多孔微球作为牙龈间充质干细胞递送载体的研究

Silk Fibroin-modified Ploylactic Acid-glycolic Acid Copolymer Porous Microspheres as Gingival Mesenchymal Stem Cells Delivery Carrier ^†

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丝素改性聚乳酸-羟基乙酸共聚物多孔微球作为牙龈间充质干细胞递送载体的研究

Silk Fibroin-modified Ploylactic Acid-glycolic Acid Copolymer Porous Microspheres as Gingival Mesenchymal Stem Cells Delivery Carrier †

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Silk Fibroin-modified Ploylactic Acid-glycolic Acid Copolymer Porous Microspheres as Gingival Mesenchymal Stem Cells Delivery Carrier ^†