钛表面氧化石墨烯涂层对施旺氏细胞粘附与增殖的影响

doi:10.3969/j.issn.1005-4979.2019.02.001

摘要/Abstract

摘要： 目的:研究纯钛表面氧化石墨烯修饰对施旺氏细胞粘附与增殖的影响。方法:在纯钛片表面制备氧化石墨烯涂层，检测复合材料的表面形貌、接触角等特征。以未处理的光滑钛片和喷砂酸蚀后的钛片为对照组，0.25mg/mL和0.50mg/mL氧化石墨烯溶液制备的钛片为实验组，将大鼠施旺氏细胞接种于材料表面。通过CCK8实验检测不同处理组钛片表面施旺氏细胞的增殖水平，活死细胞染色检测材料的细胞毒性，鬼笔环肽和DAPI染色、扫描电镜观察施旺氏细胞接种24小时后的表面粘附状态，Real-time PCR检测施旺氏细胞中神经生长因子相关基因的表达水平。结果:氧化石墨烯涂层修饰后材料表面呈现典型的皱褶样结构。细胞培养第3、5天，实验组施旺氏细胞的增殖能力高于对照组，以0.25mg/mL氧化石墨烯溶液涂层组最为显著。实验组材料表面，施旺氏细胞的胞体伸展较对照组更加充分，有多个细胞突起。结论:氧化石墨烯修饰的钛表面可以促进施旺氏细胞的粘附与增殖。

关键词: 氧化石墨烯, 纯钛, 施旺氏细胞

Abstract: Objective: To explore the effects of graphene oxide modified titanium on the adhesion and proliferation of Schwann cells. Methods：Graphene oxide coating was prepared on the surface of pure titanium sheets by spin-coating method. The surface morphology and hydrophilic of the materials were examined. The untreated smooth titanium sheets and the sandblasted titanium sheets were used as control groups and the coating prepared through 0.25mg/mL and 0.50mg/mL graphene oxide solution were used as the experimental groups. Schwann cells were inoculated on the surface of the materials. The proliferation of Schwann cells on the surface of titanium slices was observed by CCK8 assay. The live/dead cell staining was used to observe the cytotoxicity of these materials.Phalloidin and DAPI staining was used to observe the adhesion morphology of Schwann cells after 24 hours of inoculation. Real-time PCR was used to detect the expression of neurofactors related genes in Schwann cells. Results: The graphene oxide modified surfaces showed a typical wrinkle-like structure. Compared with the control groups, the experimental groups could apparently promote the proliferation of Schwann cells, and the coating group with 0.25mg/mL graphene oxide solution showed the most significant. The Schwann cells in the experimental groups were more fully stretched and had more processes than those in the control groups. Conclusion: Graphene oxide modified titanium surface can promote the adhesion and proliferation of Schwann cells.

Key words: Graphene oxide, pure titanium, Schwann cell

中图分类号:

冯曦, 王佐林. 钛表面氧化石墨烯涂层对施旺氏细胞粘附与增殖的影响[J]. 《口腔颌面外科杂志》, doi: 10.3969/j.issn.1005-4979.2019.02.001.

FENG Xi, WANG Zuo-lin. Graphene Oxide Modified Titanium Promote Adhesion and Proliferation of Schwann Cells[J]. 《Journal of Oral and Maxillofacial Surgery》, doi: 10.3969/j.issn.1005-4979.2019.02.001.

https://journal06.magtech.org.cn/Jweb_joms/CN/Y2019/V29/I2/61

参考文献

[1] Enkling N, Nicolay C, Utz KH, et al. Tactile sensibility of single-tooth implants and natural teeth[J]. Clin Oral Implants Res, 2007, 18(2):231-236.

[2] 朱一博, 林野, 邱立新, 等. 牙种植体骨感知外周神经机制的动物实验[J]. 中华口腔医学杂志, 2009, 44(8):460-463.

[3] Lee C, Wei X, Li Q, et al. Elastic and frictional properties of graphene [J]. Phys. Status Solidi B, 2009, 246:2562-2567.

[4] Singh DP, Herrera CE, Singh B, et al. Graphene oxide: an efficient material and recent approach for biotechnological and biomedical applications[J]. Mater Sci Eng C Mater Biol Appl, 2018, 86:173-197.

[5] Zancanela DC, Simão AM, Francisco CG, et al. Graphene oxide and titanium: synergistic effects on the biomineralization ability of osteoblast cultures[J]. J Mater Sci Mater Med， 2016, 27(4):71.

[6] Wang Q, Chen J, Niu Q, et al. The application of graphene oxidized combining with decellularized scaffold to repair of sciatic nerve injury in rats[J]. Saudi Pharm J， 2017, 25(4):469-476.

[7] Bunge MB, Williams AK, Wood PM. Neuron-Schwann cell interaction in basal lamina formation[J]. Dev Biol, 1982, 92(2):449-60.

[8] Jalani G, Jeyachandran D, Bertram Church R, et al. Graphene oxide-stabilized perfluorocarbon emulsions for controlled oxygen delivery[J]. Nanoscale. 2017， 9(29):10161-10166.

[9] Durán N, Martinez DS, Silveira CP, et al. Graphene oxide: a carrier for Pharmaceuticals and a scaffold for cell interactions[J]. Curr Top Med Chem, 2015, 15(4):309-327.

[10] Lee WC, Lim CH, 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.

[11] Pan CJ, Pang LQ, Gao F, et al. Anticoagulation and endothelial cell behaviors of heparin-loaded graphene oxide coating on titanium surface[J]. Mater Sci Eng C Mater Biol Appl, 2016, 63:333-340.

[12] Zhao Y, Wang Y, Niu C, et al. Construction of polyacrylamide/graphene oxide/gelatin/sodium alginate composite hydrogel with bioactivity for promoting schwann cells growth[J]. J Biomed Mater Res A. 2018, 106(7):1951-1964.

[13] Li G, Zhao Y, Zhang L, et al. Preparation of graphene oxide/polyacrylamide composite hydrogel and its effect on schwann cells attachment and proliferation[J]. Colloids Surf B Biointerfaces. 2016, 143:547-556.

[14] Mosahebi A, Fuller P, Wiberg M, et al. Effect of allogeneic schwann cell transplantation on peripheral nerve regeneration[J]. Exp Neurol. 2002, 173(2):213-223.

[15] Lee DY, Khatun Z, Lee JH, et al. Blood compatible graphene/heparin conjugate through noncovalent chemistry[J]. Biomacromolecules. 2011, 12(2):336-341.

[16] Fu C, Yang X, Tan S, et al. Enhancing cell proliferation and osteogenic differentiation of MC3T3-E1 pre-osteoblasts by BMP-2 delivery in graphene oxide-incorporated PLGA/HA biodegradable microcarriers[J]. Sci Rep. 2017, 7(1):12549.

[17] Upadhyay R, Naskar S, Bhaskar N, et al. Modulation of protein adsorption and cell proliferation on polyethylene immobilized graphene oxide reinforced HDPE bionanocomposites[J]. ACS Appl Mater Interfaces. 2016, 8(19):11954-11968.

[18] Nishida E, Miyaji H, Kato A, et al. Graphene oxide scaffold accelerates cellular proliferative response and alveolar bone healing of tooth extraction socket[J]. Int J Nanomedicine. 2016, 11:2265-2277.

[19] Luo Y, Shen H, Fang Y, et al. Enhanced proliferation and osteogenic differentiation of mesenchymal stem cells on graphene oxide-incorporated electrospun poly (lactic-co-glycolic acid) nanofibrous mats[J]. ACS Appl Mater Interfaces. 2015, 7(11):6331-6339.

[20] Rajaraman R, Rounds DE, Yen SP, et al. A scanning electron microscope study of cell adhesion and spreading in vitro[J]. Exp Cell Res. 1974, 88(2):327-339.

[21] Guo S, Zhu X, Li M, et al. Parallel control over surface charge and wettability using polyelectrolyte architecture: effect on protein adsorption and cell adhesion.[J]. ACS Appl Mater Interfaces， 2016, 8(44):30552-30563.

[22] Nalluri SM, Krishnan GR, Cheah C, et al. Hydrophilic polyurethane matrix promotes chondrogenesis of mesenchymal stem cells[J]. Mater Sci Eng C Mater Biol Appl， 2015, 54:182-195.

[23] Yang D, Li T, Xu M, et al. Graphene oxide promotes the differentiation of mouse embryonic stem cells to dopamine neurons[J]. Nanomedicine (Lond). 2014, 9(16):2445-2255.

[24] Lee WC, Lim CH, 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.

选择文件类型/文献管理软件名称

选择包含的内容