中国组织工程研究 ›› 2019, Vol. 23 ›› Issue (1): 47-54.doi: 10.3969/j.issn.2095-4344.1525

• 骨髓干细胞 bone marrow stem cells • 上一篇    下一篇

聚多巴胺仿生法制备羟基磷灰石涂层及对骨髓间质干细胞生物学特性的影响

徐炎安1,阳淇名1,李 鸿2,蒋电明3,谯 波1   

  1. 1重庆医科大学附属第一医院骨科,重庆市 400016;2四川大学物理科学技术学院,四川省成都市 610015;3重庆医科大学附属第三医院骨科,重庆市 401120
  • 修回日期:2018-07-17 出版日期:2019-01-08 发布日期:2018-11-28
  • 通讯作者: 谯波,主治医师,重庆医科大学附属第一医院骨科,重庆市 400016
  • 作者简介:徐炎安,男,1983年生,湖北省监利县人,汉族,重庆医科大学在读博士,主治医师,主要从事脊柱研究。
  • 基金资助:

    国家青年自然科学基金(81501876),项目负责人:谯波;重庆市教委科学与技术项目(KJ1702031),项目负责人:谯波;重庆市教委成果转换项目(KJZH17110),项目负责人:蒋电明

Dopamine-induced biomimetic hydroxyapatite coating: preparation and biological effect on bone marrow mesenchym stem cells

Xu Yanan1, Yang Qiming1, Li Hong2, Jiang Dianming3, Qiao Bo1   

  1. 1Department of Orthopedics, the First Affiliated Hospital of Chongqing Medical University, Chongqing 400016, China; 2College of Physical Science and Technology, Sichuan University, Chengdu 610015, Sichuan Province, China; 3Department of Orthopedics, the Third Affiliated Hospital of Chongqing Medical University, Chongqing 401120, China
  • Revised:2018-07-17 Online:2019-01-08 Published:2018-11-28
  • Contact: Qiao Bo, Attending physician, Department of Orthopedics, the First Affiliated Hospital of Chongqing Medical University, Chongqing 400016, China
  • About author:Xu Yanan, Doctorate candidate, Attending physician, Department of Orthopedics, the First Affiliated Hospital of Chongqing Medical University, Chongqing 400016, China
  • Supported by:

    the National Natural Science Foundation of China (for the Youth), No. 81501876 (to QB); the Science and Technology Project of Chongqing Education Committee, No. KJ1702031 (to QB); the Achievement Transformation Project of Chongqing Education Committee, No. KJZH17110 (to JDM)

摘要:

文章快速阅读:

文题释义:
聚多巴胺:
是受海洋贻贝在潮湿环境中具有超强黏附能力启发,由德国学者Messersmith等发现多巴胺在弱碱性环境下通过相互氧化聚合而成,其几乎可形成于任何种类及任何形状的材料表面,但目前其形成机制尚不清楚。同时还发现聚多巴胺具有黏附能力,能够吸附如药物、离子及生物大分子等,实现不同的生物功效。
羟基磷灰石:是脊椎动物骨骼和牙齿的主要无机组成成分,其具有优良的生物相容性和生物活性,常被应用于骨组织再生工程。研究发现利用羟基磷灰石对材料进行表面修饰,可提高生物材料的生物相容性和生物活性,提高生物材料的骨整合能力。目前制备羟基磷灰石涂层方法有等离子喷涂、溶胶凝胶法、电化学沉积和仿生法等。

 

摘要
背景:
聚多巴胺仿生法被证实可在多种不同金属表面制备羟基磷灰石涂层,但目前缺乏利用其在人工合成材料表面制备羟基磷灰石涂层的研究。
目的:利用聚多巴胺仿生法在纳米羟基磷灰石/聚酰胺66表面制备羟基磷灰石涂层,评价其对骨髓间质干细胞生物学的影响。
方法:采用聚多巴胺仿生法在纳米羟基磷灰石/聚酰胺66表面制备羟基磷灰石涂层,获得羟基磷灰石-聚多巴胺-纳米羟基磷灰石/聚酰胺66(hydroxyapatite-polydopamine-nano-hydroxyapatite/polyamide66,HA-PDA-HA/ P66)材料,通过X射线光电子能谱分析和扫描电镜检测羟基磷灰石涂层,并检测材料的表面粗糙度及亲水性。将骨髓间质干细胞C3H10T1/2分别与纳米羟基磷灰石/聚酰胺66、聚多巴胺-纳米羟基磷灰石/聚酰胺66、HA-PDA-HA/P66材料共培养,培养6,12 h后,采用CCK-8法检测细胞黏附情况;培养1 d,通过扫描电镜观察细胞形态;培养1,3 d,DAPI染色检测细胞增殖;成骨诱导培养7,10 d,检测细胞碱性磷酸酶活性;成骨诱导培养14 d,茜素红染色检测细胞矿化能力。
结果与结论:①X射线衍射和扫描电镜证实成功制备了羟基磷灰石涂层,且羟基磷灰石涂层明显提高了材料亲水性和表面粗糙度;②培养6,12 h,HA-PDA-HA/P66材料表面黏附的细胞数明显多于其余两种材料(P < 0.05);培养1 d,在HA-PDA-HA/P66及聚多巴胺-纳米羟基磷灰石/聚酰胺66材料上的细胞呈多边形,且有较多伪足;培养1,3 d,HA-PDA-HA/P66材料表面的细胞增殖能力高于其余两种材料(P < 0.05);④成骨诱导7,10 d,HA-PDA-HA/P66材料表面细胞内碱性磷酸酶活性高于其余两种材料(P < 0.05);成骨诱导14 d,HA-PDA-HA/P66材料周围钙结节形成多于其余两种材料;⑤结果表明,利用聚多巴胺仿生法可在纳米羟基磷灰石/聚酰胺66表面制备羟基磷灰石涂层,且涂层具有良好的生物活性与生物相容性。

中国组织工程研究杂志出版内容重点:干细胞;骨髓干细胞;造血干细胞;脂肪干细胞;肿瘤干细胞;胚胎干细胞;脐带脐血干细胞;干细胞诱导;干细胞分化;组织工程
ORCID:
0000-0001-8884-6665(徐炎安)

关键词: 纳米羟基磷灰石/聚酰胺66, 骨髓间质干细胞, 生物相容性, 生物活性, 成骨分化, 干细胞

Abstract:

BACKGROUND: Hydroxyapatite (HA) coating can be formed on different kinds of metals assisted by polydopamine (PDA). However, the preparation of HA coating with biomimetic method assisted by PDA on artificial synthetic materials is rarely reported.
OBJECTIVE: To prepare the HA coating on HA/P66 surface assisted by PDA and to evaluate its effect on bone marrow mesenchymal stem cells.
METHODS: First HA coating was prepared on the HA/P66 surface using the biomimetic method assisted by PDA to obtain HA-PDA-HA/P66 substrate and then verified by X-ray photoelectron spectroscopy and scanning electron microscopy. Its surface properties such as hydrophilicity and surface roughness were also characterized. Then C3H10T1/2 cells were co-cultured with HA/P66, PDA-HA/P66 and HA-PDA-HA/P66. After 6 and 12 hours of cultivation, the initial cell adhesion was observed by cell counting kit-8. After 1 day of cultivation, cell morphology was observed by scanning electron microscope. After 1 and 3 days of cultivation, cell proliferation was assessed by 4,6-diamino-2-phenyl indole staining. After osteogenic induction for 7 and 10 days, intracellular alkaline phosphatase activity was detected. After osteogenic induction for 14 days, mineralization of the extracellular matrix was detected by alizarin red staining.
RESULTS AND CONCLUSION: (1) HA coating was successfully prepared on the HA/P66 substrate which was verified by X-ray photoelectron spectroscopy and scanning electron microscope. HA coating greatly increased the hydrophilicity and surface roughness of HA/P66. (2) After 6 and 12 hours of cultivation, the number of adherent cells on the surface of HA-PDA-HA/P66 was significantly higher than that in the other groups (P < 0.05). After 1 day of cultivation, the cells on the surface of HA-PDA-HA/P66 and PDA-HA/P66 showed polygonal shape with a large number of pseudopodia. (3) After 1 and 3 days of cultivation, the cell proliferation on the surface of HA-PDA-HA/P66 was significantly higher than that in the other two groups (P < 0.05). (4) After 7 and 10 days of osteogenic induction, the alkaline phosphatase activity of cells on the surface of HA-PDA-HA/P66 was significantly higher than that of the other groups. After 14 days of osteogenic induction, there were more calcium nodules on the surface of HA-PDA-HA/P66 than in the other groups. These findings indicate that HA coating can be successfully prepared on the surface of HA/P66 through the biomimetic process assisted by PDA and has good biocompatibility and bioactivity. 

中国组织工程研究杂志出版内容重点:干细胞;骨髓干细胞;造血干细胞;脂肪干细胞;肿瘤干细胞;胚胎干细胞;脐带脐血干细胞;干细胞诱导;干细胞分化;组织工程

Key words: Hydroxyapatites, Nylons, Cell Biology, Tissue Engineering

中图分类号: