Ⅰ型胶原/聚己内酯/凹凸棒石复合支架材料体外诱导成骨的研究

doi:10.13523/j.cb.20160504

中国生物工程杂志

2016, Vol. 36

Issue (5): 27-33 DOI: 10.13523/j.cb.20160504

研究报告

Ⅰ型胶原/聚己内酯/凹凸棒石复合支架材料体外诱导成骨的研究

张晓敏^1,2, 王世勇², 李根², 赵红斌^1,2

1. 甘肃农业大学生命科学技术学院兰州 730070;
2. 兰州军区兰州总医院骨科研究所兰州 730050

The Study of Osteogenic Induction of Type Ⅰ Collagen /Poly(caprolactone)/Attapulgite Composite Scaffold Materials in Vitro

ZHANG Xiao-min^1,2, WANG Shi-yong², LI Gen², ZHAO Hong-bin^1,2

1. School of Life Science and Technology, Gansu Agriculture University, Lanzhou 730070, China;
2. Institute of Orthopedics, General Hospital of Lanzhou Military Command of the PLA, Lanzhou 730050, China

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摘要：

目的:研究Ⅰ型胶原(ColⅠ)/聚己内酯(PCL)/凹凸棒石(ATP)复合支架材料的生物相容性及体外骨诱导性。方法:采用溶液浇铸-粒子滤沥法制备三种不同ATP含量(0% wt、10% wt、30% wt)的ColⅠ/PCL/ATP复合支架材料;将D1细胞与三种支架材料共培养,扫描电镜、鬼笔环肽和H&E染色、CCK-8法评价支架材料的生物相容性;D1细胞复合三种支架材料培养7天、14天、21天后RT-qPCR检测其成骨相关基因(Runx-2、Osterix、ALP、Col I、OPN、OC)的相对表达量,分别评价比较三种支架材料的成骨诱导效应。结果:SEM、鬼笔环肽和H&E染色显示D1细胞在三种支架材料表面均呈现良好的黏附;CCK-8结果显示,细胞在ATP含量30% wt的支架材料上增殖率显著高于其他两组,RT-qPCR检测结果显示,与0% wt、10% wt ATP相比,30% wt ATP组的Runx-2相对表达量在7天时显著升高, 14天、21天降低;ALP相对表达量在14天时显著升高,21天时显著降低;Osterix、Col I、OPN、OC的相对表达量随时间和ATP剂量的增加显著上调(P<0.05)。结论:ColⅠ/PCL/ATP复合支架材料具有良好的生物相容性及骨诱导性,有望成为一种理想的骨组织工程支架材料。

关键词： 凹凸棒石; 生物相容性; 复合支架材料; 骨诱导性

Abstract:

Objective:To study the biocompatibility and osteoinductivity of typeⅠcollagen/poly (caprolactone)/attapulgite composite scaffolds in vitro. Methods:The ColⅠ/PCL/ATP composite scaffolds materials of three different content (0%, 10%, and 30%wt) ATP were fabricated by the solution perfusion-salt-leaching process. D1 cells were co-cultured with the materials for 7d or 1~6d, the biocompatibility of scaffold was evaluated by scanning electron microscopy, Phalloidin and H&E staining and CCK-8, respectively. D1 cells were co-cultured with the materials for 7d, 14d, and 21d; the relative expression of bone-related genes including Runx-2, Osterix, ALP, Col I, OPN, and OC were measured by RT-qPCR, and evaluated osteogenic effect comparison of three scaffolds. Results:Three scaffolds had suitable cell adhesion and appreciation properties, especially 30%wt ATP materials. The results of RT-qPCR show that Runx-2 expression of 30%wt ATP was significant increased in 7d, but decreased in 14d, 21d; and ALP expression was significant increased in 14d, but significant decreased in 21d; and there were significant increased in the relative mRNA levels of Osterix, Col I, OPN, OC, compared with the 0% and 10%wt ATP materials (P<0.05), which had time and dose dependent manner. Conclusion:ColⅠ/PCL/ATP composite scaffolds may be used as an ideal scaffold for bone tissue engineering.

Key words: Attapulgite Osteoinductivity Biocompatibility Composite scaffold biomaterials

收稿日期: 2016-01-11 出版日期: 2016-05-25

ZTFLH:

R318.08

基金资助:

甘肃省重大科技专项资助项目(1203FKDA036)

通讯作者: 赵红斌 E-mail: zhao761032@163.com

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引用本文:

张晓敏, 王世勇, 李根, 赵红斌. Ⅰ型胶原/聚己内酯/凹凸棒石复合支架材料体外诱导成骨的研究[J]. 中国生物工程杂志, 2016, 36(5): 27-33.

ZHANG Xiao-min, WANG Shi-yong, LI Gen, ZHAO Hong-bin. The Study of Osteogenic Induction of Type Ⅰ Collagen /Poly(caprolactone)/Attapulgite Composite Scaffold Materials in Vitro. China Biotechnology, 2016, 36(5): 27-33.

链接本文:

https://manu60.magtech.com.cn/biotech/CN/10.13523/j.cb.20160504 或 https://manu60.magtech.com.cn/biotech/CN/Y2016/V36/I5/27

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