Effect of low-frequency pulsed electromagnetic fields on proliferation and osteogenic ability of human adipose-derived stem cells in a three-dimensional scaffold

doi:10.3969/j.issn.2095-4344.2017.18.007

Abstract

Abstract:

BACKGROUND: Nowadays increasing experimental findings have proved that the low-frequency pulsed electromagnetic fields (LPEMF) can induce osteogenic differentiation of a variety of stem cells in the two-dimensional scaffold. However, little is reported on the LPEMF effect on the proliferation and osteogenic differentiation of stem cells in the three-dimensional scaffold.
OBJECTIVE: To investigate the effect of LPEMF on the proliferation and osteogenic differentiation of human adipose-derived stem cells (hASCs) in the 3D Insert-PCL scaffold.
METHODS: Passage 3 hASCs were directly cultured in the 3D Insert-PCL scaffolds followed by LPEMF (50 Hz, 1 mT) exposure, 2 hours per day, for continuous 14 days (experimental group) or no intervention (control group). After 7 days of culture, Live/Dead staining was used to observe cell survival. After 1, 3, 5, 7 days of culture, MTT assay was used to detect cell proliferation. After 7 and 14 days of culture, the osteogenic differentiation of hASCs was assessed through the alkaline phosphatase (ALP) staining and qRT-PCR.
RESULTS AND CONCLUSION: Live/dead cell staining proved that the hASCs had a good growth in the 3D Insert-PCL scaffolds as well as a high survival rate. The absorbance values of hASCs in the two groups were increased gradually with time, and the absorbance value in the experimental group was significantly higher than that in the control group at 1 and 3 days after culture (P < 0.05). The ALP activity in the experimental group was stronger than that in the control group at 7 and 14 days after culture. qRT-PCR findings showed that at 7 days after culture, the mRNA levels of ALP and type I collagen were significantly higher in the experimental group than the control group (P < 0.01), while at 14 days after culture, the mRNA levels of osteopontin, Runt-related transcription factor, ALP and type I collagen were significantly higher in the experimental group than the control group (P < 0.05). To conclude, the LPEMF exposure can promote the proliferation and osteogenic differentiation of hASCs cultured on the the 3D Insert-PCL scaffold.

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: Electromagnetic Fields, Stem Cells, Cell Proliferation, Tissue Engineering

CLC Number:

R318

Chen Yu-xiong, Chen Xian-zhe, Ni Meng-shan, Guo Wen-jie, Tian Jing.

Effect of low-frequency pulsed electromagnetic fields on proliferation and osteogenic ability of human adipose-derived stem cells in a three-dimensional scaffold [J]. Chinese Journal of Tissue Engineering Research, 2017, 21(18): 2828-2833.

Figures/Tables 4

References

[1]郭颖,李玉宝. 骨修复材料的研究进展[J].世界科技研究与发展, 2001,23(1):33-37.

[2]郝建原,邓先模.复合生物材料的研究进展[J].高分子通报,2002, 15(5):1-8.

[3]胡金龙,王静成,颜连启.组织工程学技术治疗骨缺损的最新研究进展[J].中国矫形外科杂志,2013,21(2):150-153.

[4]Hutmacher DW.Scaffolds in tissue engineering bone and cartilage. Biomaterials.2001;21(24):2529-2543.

[5]赵娜.脂肪干细胞诱导分化的现状及前景[J].中国组织工程研究, 2015,19(6):969-974.

[6]Minear S,Leucht P,Miller S,et al. rBMP Represses Wnt Signaling and Influences Skeletal Progenitor Cell Fate Specification During Bone Repair.J Bone Miner Res.2010; 25(6):1196-1207.

[7]张志达,江晓兵,沈耿杨,等.磷酸钙及硫酸钙支架在骨组织工程中的研究进展[J].中国组织工程研究,2016,20(8):1203-1209.

[8]刘越,赵艳梅,田学忠,等.明胶/纳米羟基磷灰石三维多孔骨支架的制备及其生物安全性评价[J].中国矫形外科杂志, 2013, 21(16):1634-1640.

[9]Flautre B,Descamps M,Delecourt C,et al.Porous HA ceramic for bone replacement: Role of the pores and interconnections -experimental study in the rabbit.J Mater Sci Mater Med.2001; 12(8):679-682.

[10]Murugan R,Ramakrishna S.Bioresorbable composite bone paste using polysaccharide based nano hydroxyapatite. Biomaterials.2004;25(17):3829-3835.

[11]赵文婧,刘玲珑,吴丽情,等.骨髓间充质干细胞的二维及三维培养实验研究[J].右江民族医学院学报,2014,36(3):355-356.

[12]滕伟,郭志坤.细胞三维培养的研究进展[J].医学综述,2008, 14(12):1767-1770.

[13]胡康洪,姚颖. 三维细胞培养技术的研究与应用[J].医学分子生物学杂志,2008,5(2):185-188.

[14]肖登.低频脉冲电磁场的生物学作用[J].中华物理医学与康复杂志,2010,32(11):871-873.

[15]宋先舟,陈继革,吴华,等.低频脉冲电磁场诱导成骨的研究进展[J].中华实验外科杂志,2008,25(10):1359-1360.

[16]许伟东,刘家富,钱红,等.脉冲电磁场治疗骨延迟愈合的临床分析[J].中国康复医学杂志,2006,21(5):440-441.

[17]许伟东.脉冲电磁场治疗骨不连的研究进展[J].中华物理医学与康复杂志,2009,31(10):707-708.

[18]Zuk PA,Zhu M,Ashjian P,et al. Human Adipose Tissue Is a Source of Multipotent Stem Cells.MolBiol Cell.2003; 13(12): 4279-4295.

[19]Frese L,Dijkman PE,Hoerstrup SP.Adipose Tissue-Derived Stem Cells in Regenerative Medicine. Transfus Med Hemother. 2016;43(4):268-274.

[20]Beane OS,Fonseca VC,Cooper LL,et al.Impact of aging on the regenerative properties of bone marrow-, muscle-, and adipose-derived mesenchymal stem/stromal cells.Plos One. 2014;9(12):e115963.

[21]Zuk PA,Zhu M,Mizuno H,et al.Multilineage cells from human adipose tissue: implications for cell-based therapies.Tissue Eng.2001;7(2):211-228.

[22]Zuk PA,Zhu M,Ashjian P,et al.Human adipose tissue is a source of multipotent stem cells.Mol Biol Cell.2002;13(12): 4279-4295.

[23]Lyu CQ,Lu JY,Cao CH,et al.Induction of Osteogenic Differentiation of Human Adipose-Derived Stem Cells by a Novel Self-Supporting Graphene Hydrogel Film and the Possible Underlying Mechanism. ACS Appl Mater Interfaces. 2015;7(36):20245-20254.

[24]陈犹白,陈聪慧,Qixu Zhang,等.脂肪干细胞成骨分化的研究进展[J].中华损伤与修复杂志电子版,2016,11(2):126-134.

[25]Cheng G,Chen K,Li Z,et al. Enhancement of osteoblastic differentiation of bone marrow mesenchymal stem cells in rats by sinusoidal electromagnetic fields.Sheng Wu Yi Xue Gong Cheng Xue Za Zhi.2011;28(4):683-688.

[26]Luo F,Hou T,Zhang Z,et al.Effects of pulsed electromagnetic field frequencies on the osteogenic differentiation of human mesenchymal stem cells.Orthopedics.2012;35(4):e526-e531.

[27]Liu C,Yu J,Yang Y,et al. Effect of 1 mT sinusoidal electromagnetic fields on proliferation and osteogenic differentiation of rat bone marrow mesenchymal stromal cells.Bioelectromagnetics.2013;34(6):453-464.

[28]Jansen JH,Op VDJ,Punt BJ,et al.Stimulation of osteogenic differentiation in human osteoprogenitor cells by pulsed electromagnetic fields: an in vitro study.Bmc Musculoskeletal Disorders.2010;11(1):1-11.

[29]Yang Y,Tao C,Zhao D,et al.EMF acts on rat bone marrow mesenchymal stem cells to promote differentiation to osteoblasts and to inhibit differentiation to adipocytes. Bioelectromagnetics.2010;31(4):277-285.

[30]Tsai MT,Li WJ,Tuan RS,et al. Modulation of osteogenesis in human mesenchymal stem cells by specific pulsed electromagnetic field stimulation.J Orthop Res.2009;27(9): 1169-1174.

[31]Lin CC,Lin RW,Chang CW,et al.Single-pulsed electromagnetic field therapy increases osteogenic differentiationthrough Wnt signaling pathway and sclerostin downregulation. Bioelectromagnetics.2015;36(7):494-505.

[32]Zhou YJ,Wang P,Chen HY,et al.Effect of Pulsed Electromagnetic Fields on Osteogenic Differentiation and Wnt/beta-catenin Signaling Pathway in Rat Bone Marrow Mesenchymal Stem Cells. Sichuan Da Xue Xue Bao Yi Xue Ban.2015;46(3):347-353.

[33]Garg P,Mazur MM,Buck AC,et al.Prospective Review of Mesenchymal Stem Cells Differentiation into Osteoblasts. Orthop Surg.2017;9(1):13-19.

[34]Petecchia L,Sbrana F,Utzeri R,et al. Electro-magnetic field promotes osteogenic differentiation of BM-hMSCs through a selective action on Ca(2+)-related mechanisms.Sci Rep.2015; 5:13856.

[35]Roskies M,Jordan JO,Fang D,et al.Improving PEEK bioactivity for craniofacial reconstruction using a 3D printed scaffold embedded with mesenchymal stem cells.J Biomater Appl.2016;31(1):132-139.

[36]Wang Z,Tian X,Bai S.Research progress of adipose-derived stem cells compound with three dimensional printing scaffold for engineered tissue.Zhongguo Xiu Fu Chong Jian Wai Ke Za Zhi.2016;30(3):320-322.