Effect of transforming growth factor beta1 on proliferation and secretion of osteoblasts on porous tantalum/MG63 osteoblast-like cell composites

doi:10.3969/j.issn.2095-4344.2016.25.005

Abstract

Abstract:

BACKGROUND: Previous studies have demonstrated that the Chinese porous tantalum made in China has non-toxicity and good biocompatibility, which can promote osteogenesis.

OBJECTIVE: To investigate the effects of transforming growth factor β1 on proliferation, cell cycle and secretion of osteoblasts on porous tantalum/MG63 osteoblast-like cell composites.

METHODS: Passage 3 MG63 osteoblast-like cell suspension (1×10⁹/L) was seeded onto the porous tantalum, then the cell composites were inoculated in the medium with 0, 0.5, 5 and 10 µg/L transforming growth factor β1, respectively. The proliferation of osteoblasts was detected by cell counting kit-8 assay at 1-13 days after inoculation; the cell morphology and ultrastructure observed by scanning electron microscope and transmission electron microscopy; and level of collagen type I detected by enzyme-linked immunosorbent assay.

RESULTS AND CONCLUSON: 0.5, 5, 10 µg/L transforming growth factor β1 could promote the osteoblast proliferation, and cell proliferation in the 5 µg/L transforming growth factor β1 group was higher than that in the other groups; in the 5 µg/L transforming growth factor β1 group, laminated osteoblasts adhered on the surface and grew into inner of porous tantalum, which extended more pseudopodia toward the scaffold; osteoblasts-secreted matrix could cover the scaffold and numerous rough endoplasmic reticulum, free ribosomes, dense mitochondria, Golgi apparatus as well as matrix vesicles could be found in the cytoplasm. In addition, the level of collagen type I in the 5 µg/L transforming growth factor β1 group was significantly higher than that in the other groups (P < 0.05). These results indicate that transforming growth factor β1 can promote proliferation, and collagen type I secretion of osteoblasts on porous tantalum/MG63 osteoblast-like cell composites, and the optimum mass concentration of transforming growth factor β1 is 5 µg/L.

中国组织工程研究杂志出版内容重点：生物材料；骨生物材料; 口腔生物材料; 纳米材料; 缓释材料; 材料相容性；组织工程

Key words: Osteoblasts, Tantalum, Transforming Growth Factor beta1, Tissue Engineering

CLC Number:

R318

Pang Hai-tao, Gan Hong-quan, Wang Qian, Cui Yi-shuang, Lai Zhen-quan, Zhou Guo-long, Pan Xiang-yu, Wang Zhi-qiang, Li Qi-jia. Effect of transforming growth factor beta1 on proliferation and secretion of osteoblasts on porous tantalum/MG63 osteoblast-like cell composites[J]. Chinese Journal of Tissue Engineering Research, 2016, 20(25): 3680-3686.

Figures/Tables 7

References

[1] Mrosek EH,Schagemann JC,Chung HW,et al.Porous tantalum and poly-epsilon -caprolactone biocomposites for osteochondral defect repair: preliminary studies in rabbits.J Orthop Res. 2010;28(2):141-148.

[2] 郑凯,于秀淳,郭征,等.生物金属材料在骨科的应用及发展[J].生物骨科材料与临床研究,2013,10(2):31-33.

[3] Dabrowski B,Swieszkowski W,Godlinski D,et al.Highly porous titanium scaffolds for orthopaedic applications.J Biomed Mater Res B A ppl Biomater. 2010;95(1):53-61.

[4] Cardaropoli F,Alfieri V,Caiazzo F,et al.Manufacturing of porous biomaterials for dental implant applications through selective laser melting.Adv Mater Res.2012; 6(535-537):1222-1229.

[5] 李琪佳,王茜,甘洪全,等.多孔钽材料细胞毒性、生物相容性及体内成骨性研究[J].中华骨科杂志, 2014,34(9):954-961.

[6] 陈良建,汪瑞芳,郭小平,等.载 TGF-β1 明胶微球涂层多孔钛对MG63 细胞功能的影响[J].中南大学学报(自然科学版),2011,42(8):2266-2273.

[7] Crane JL,Cao X.Bone marrow mesenchymal stem cells and TGF-β signaling in bone remodeling.J Clin Invest.2014;124(2):466-472.

[8] 张岭,李琪佳,赵季华,等.医用多孔钽材料复合大鼠软骨细胞的生长特性及功能变化[J].解放军医学杂志, 2014, 39(6): 464-469.

[9] Bobyn JD,Stackpool GJ,Hacking SA,et al. Characteristics of bone ingrowth and interface mechanics of a new porous tantalum biomaterial.J Bone Joint Surg Br.1999;81(5):907-914.

[10] de Wild M,Schumacher R,Mayer K,et al.Bone regeneration by the osteoconductivity of porous titanium implants manufactured by selective laser melting：a histological and microCT study in the rabbit. Tissue Eng Part A 2013,19(23-24):2645-2654．

[11] Hao L,Wei Y,Li J,et a1.Initial osteoblast functions on Ti-5Zr-3Sn-5Mo-15Nb titanium alloy surfaces modified by microarc oxidation.J Biomed Mater Res A. 2010; 92(2):432-440．

[12] Tang Z,Xie Y,Yang F,et al.Porous tantalum coatings prepared by vacuum plasma spraying enhance bmscs osteogenic differentiation and bone regeneration in vitro and in vivo.PLos One.2013;8(6):e66263.

[13] Sagomonyants KB,Hakim-Zargar M,Jhaveri A,et al.Porous tantalum stimulates the proliferation and osteogenesis of osteoblasts from elderly female patients.J Orthop Res.2011;29(4):609-616.

[14] Christie MJ.Clinical applications of trabecular metal. Am J Orthop.2002;31(4):219-220.

[15] 耿丽鑫,甘洪全,王茜,等.国产多孔钽对成骨细胞生物相容性及其相关成骨基因表达的影响[J].第三军医大学学报, 2014,36(11):1163-1167.

[16] 甘洪全,李琪佳,王茜,等.国产多孔钽材料兔髌腱内植入形态学特点及生物相容性评价[J].中国修复重建外科杂志, 2014,28(4):452-456.

[17] 甘洪全,刘鑫,赵济华,等.国产多孔钽材料兔竖脊肌植入组织学及生物学效应[J].南京医科大学学报, 2014,34(5): 611-616.

[18] Fox P,Pogson S,Sutcliffe CJ,et al.Interface interactions between porous titanium/tantalum coatings, produced by Selective Laser Melting(SLM), on a cobalt-chromium alloy.Surf Coat Techonl. 2008;202(20):5001-5007.

[19] Welldon KJ,Atkins GJ,Howie DW,et al.Primary human osteoblasts grow into porous tantalum and maintain an osteoblastic phenotype.J Biomed Mater Res A.2008; 84(3):691-701.

[20] Chen G,Deng C,Li YP.TGF-β and BMP signaling in osteoblast differentiation and bone formation.Int J Biol Sci.2012;8(2):272-288.

[21] 郑纪伟,郭宏亮,孟尧,等.转化生长因子β１对钛表面成骨细胞中骨钙素基因表达的影响[J].口腔医学研究, 2012, 28(9):868-871.

[22] 卢卫忠,唐康来,杨柳,等.TGF-β对成骨细胞的作用[J].第三军医大学学报, 2000,22(1):99-102.

[23] 郑纪伟,韩斐斐,王瑾,等.TGF-β1对钛表面成骨相关蛋白基因表达的影响[J].中华老年口腔医学杂志, 2011,9(2): 65-69.

[24] 朱奇,樊明文,边专,等.转化生长因子β1mRNA在鼠磨牙形态发生过程中表达的研究[J] .中华口腔医学杂志, 2001, 36(2):130-132.

[25] 张小恒,余占海,张国英,等.转化生长因子β１对大鼠牙周组织中白细胞介素，骨钙素水平的影响[J].口腔医学研究,2010,26(2):171-174.