自撑式石墨烯水凝胶诱导人脂肪干细胞成骨分化的体外研究

›› 2014, Vol. 34 ›› Issue (7): 486-491.

自撑式石墨烯水凝胶诱导人脂肪干细胞成骨分化的体外研究

吕成奇¹,陆家瑜²,³,于佳⁴,邹德荣²

1. 上海交通大学附属第六人民医院
2. 上海交通大学附属第六人民医院口腔科
3.
4. 上海交通大学附属上海第六人民医院

收稿日期:2014-01-24 修回日期:2014-03-17 出版日期:2014-07-28 发布日期:2014-08-04
通讯作者: 邹德荣 E-mail:derongzou@gmail.com
基金资助:
上海市科委基础研究重点项目;上海市自然科学基金青年项目

In vitro effects of self-sustaining graphene hydrogel film on the osteogenic differentiation of human adipose-derived stem cells

Received:2014-01-24 Revised:2014-03-17 Online:2014-07-28 Published:2014-08-04

摘要/Abstract

摘要： 　目的　通过体外实验评价本课题组制备的自撑式石墨烯水凝胶（self-sustaining graphene hydrogel,SGH）的骨诱导性能。方法　（1）将人脂肪干细胞（hADSC）接种于SGH和玻片表面，采用荧光显微镜图像计数检测ADSC在材料表面粘附能力；通过扫描电镜观察材料本身的表征及细胞在SGH表面生长的形态；（2）SGH组加入普通培养液作为实验组，玻片组加入成骨培养基作为诱导组，玻片组加入普通培养液作为对照；通过实时定量荧光PCR、ALP活性检测和茜素红半定量测定钙沉积评价各组hADSC的分化及成骨能力。结果　hADSC能以等同于在玻片表面的粘附效率和正常的形态在SGH表面生长。实验组细胞在不同时间点成骨相关基因的表达水平、ALP活性以及钙盐沉积量均显著高于对照组而略低于诱导组（P<0.05）。结论　自撑式石墨烯水凝胶作为一种生物相容性材料，具有较强的诱导人脂肪干细胞成骨分化的能力。

关键词: 自撑式石墨烯水凝胶, 人脂肪干细胞, 成骨分化

Abstract: Objective　In vitro assessment of the excellent osteoinductive ability of a self-sustaining graphene hydrogel on human adipose-derived stem cells (hADSCs).Methods　（1）The hADSCs were cultured on both SGH films and control glass, and cell adhesion and morphology were examined by scanning electron microscopy (SEM) and immunofluorescence analysis;（2）The in vitro experiments were divided into three groups, SGH films group, glass group (osteogenic medium) and control glass group (normal growth medium). In addition, in vitro experiments for osteogenic potential by hADSCs were performed for comparison among the three sample groups. Results　The hADSCs, as on control glass, also attached tightly on the surface of the SGH film and appeared normal cell morphology. The in vitro study showed better osteogenic differentiation of hADSCs on self-sustaining graphene hydrogel group scaffolds than in the control group（P<0.05）, due to the higher ALP activity, osteocalcin mRNA expression and mineralization, while its osteogenic ability was lower than in an osteogenic medium. Conclusion　This self-sustaining graphene hydrogel film appears to be highly biocompatible and osteoinductive for hADSCs.

Key words: self-sustaining graphene hydrogel, human adipose-derived stem cells, osteoinductive differentiation

中图分类号:

R329.29

吕成奇陆家瑜于佳邹德荣. 自撑式石墨烯水凝胶诱导人脂肪干细胞成骨分化的体外研究[J]. 口腔医学, 2014, 34(7): 486-491.

参考文献

[1]Liao H T,Chen J P,Lee M Y.Bone T11 Engineering with Adipose-Derived Stem Cells in Bioactive Composites of Laser-Sintered Porous Polycaprolactone Scaffolds and Platelet-Rich Plasma[J].Materials,2013,6(11):4911-4929
[2]Wang S,Zhao P,Lin C,et al.In Vitro and In Vivo Osteoconductivity of Bone Marrow Stromal Cells in Biomimetic Polycaprolactone/Calcium Phosphate Cement Composites[J].Journal of Biomaterials and T5 Engineering,2013,3(5):512-522
[3]吴广新,汪建国,杜晓岩.纳米晶胶原基骨复合骨髓单个核细胞修复下颌骨缺损[J].中国组织工程研究,2013,17(38):6791-6796
[4]Dupont K M,Boerckel J D,Stevens H Y,et al.Synthetic scaffold coating with adeno-associated virus encoding BMP2 to promote endogenous bone repair[J].Cell and T3 Research,2012,347(3):575-588
[5]Novoselov K S,Geim A K,Morozov S V,et al.Electric field effect in atomically thin carbon films[J].Science,2004,306(5696):666-669
[6]Nayak T R,Andersen H,Makam V S,et al.Graphene for controlled and accelerated osteogenic differentiation of human mesenchymal stem cells[J].ACS nano,2011,5(6):4670-4678
[7]Lee W C,Lim C H Y X,Shi H,et al.Origin of enhanced stem cell growth and differentiation on graphene and graphene oxide[J].ACS nano,2011,5(9):7334-7341
[8]Yang X,Qiu L,Cheng C,et al.Ordered Gelation of Chemically Converted Graphene for Next‐Generation Electroconductive Hydrogel Films[J].Angewandte Chemie International Edition,2011,50(32):7325-7328
[9]Lu J,He Y S,Cheng C,et al.Self‐Supporting Graphene Hydrogel Film as an Experimental Platform to Evaluate the Potential of Graphene for Bone Regeneration[J].Advanced Functional Materials,2013,23(28):3494-
[10]HUMMERS W S,OFFEMAN R E.Preparation of graphitic oxide[J].J Am Chem Soc,1958,80(6):1339-
[11]Chen H,Müller M B,Gilmore K J,et al.Mechanically strong,electrically conductive,and biocompatible graphene paper[J].Advanced Materials,2008,20(18):3557-3561
[12]Q Liu,LCen,S. Yin,et al.. A comparative study of proliferation and osteogenic differentiation of adipose-derived stem cells on akermanite and beta-TCP ceramics[J]..Biomaterials,2008,29(36):4792-4799
[13]Fan H,Wang L,Zhao K,et al.Fabrication,mechanical properties,and biocompatibility of graphene-reinforced chitosan composites[J].Biomacromolecules,2010,11(9):2345-2351
[14]Kalbacova M,Broz A,Kong J,et al.Graphene substrates promote adherence of human osteoblasts and mesenchymal stromal cells[J].Carbon,2010,48(15):4323-4329
[15]Deepachitra R,Chamundeeswari M,Krithiga G,et al.Osteo mineralization of fibrin-decorated graphene oxide[J].Carbon,2013,56:64-76
[16]Niemeyer P,Kornacker M,Mehlhorn A,et al.Comparison of immunological properties of bone marrow stromal cells and adipose t1-derived stem cells before and after osteogenic differentiation in vitro[J].T1 engineering,2007,13(1):111-121
[17]McBeath R,Pirone D M,Nelson C M,et al.Cell shape,cytoskeletal tension,and RhoA regulate stem cell lineage commitment[J].Developmental cell,2004,6(4):483-495
[18]Brammer K S,Choi C,Frandsen C J,et al.Hydrophobic nanopillars initiate mesenchymal stem cell aggregation and osteo-differentiation[J].Acta biomaterialia,2011,7(2):683-690