The effect of high frequency electric field on enhancement of chondrogenesis in human adipose-derived stem cells

Document Type: Original Article

Authors

1 Department of Anatomical Sciences, Medical School, Isfahan University of Medical Sciences, Isfahan, Iran

2 Department of Electrical Engineering, Engineering School, Isfahan University, Isfahan, Iran

3 Department of Molecular Biology, Medical School, Isfahan University of Medical Sciences, Isfahan, Iran

4 Department of Anatomical Sciences, Medical School, Kermanshah University of Medical Sciences, Kermanshah, Iran

Abstract

Objective(s):Osteoarthritis (OA) is globally one of the most common diseases from the middle age onwards. Cartilage is an avascular tissue therefore it cannot be repaired in the body. Conservative treatments have failed as a good remedy and cell therapy as a decisive cure is needed. One of the best and easily accessible cell sources for this purpose is adipose-derived stem cells which can be differentiated into chondrocytes by tissue engineering techniques. Chemical and physical inducers have a key role in stem cell – chondrocyte differentiation. We have tried to determine the role of electric fields (EF) in promoting this kind of chondrogenesis process.
Materials and Methods: Human adipose derived stem cells (ADSCs) were extracted from subcutaneous abdominal adipose tissue during cesarean section. A high frequency (60 KHz) EF (20 mv/cm), as a physical inducer for chondrogenesis in a 3D micromass culture system of ADSCs was utilized. Also, MTT, ELISA, flow cytometry, and real-time PCR techniques were used for this study.
Results: We found that using physical electric fields leads to chondrogenesis. Furthermore, results show that using both physical (EF) and chemical (TGFβ3) inducers simultaneously, has best outcomes in chondrogenesis, and expression of SOX9 andtype II collagen genes. It also causes significant decreased expression of type I and X collagen genes in pure EF group compared with control group.
Conclusion:The EF was found as a proper effective inducer in chondrogenic differentiation of human ADSCs micromass culture.

Keywords


1. Marlovits P, Zeller P, Singer C, Resinger V. Cartilage repair: generations of autologous chondrocyte transplantation. Eur J Radiol 2006; 57:24–31.

2. Kuo K, LiWan J, Mauck L, Tuan S. Cartilage tissue engineering: it’s potential and uses. Curr Opin Rheumatol 2006; 18:64-73.

3. Kurtz S, Ong K, Lau E, Mowat F, Halpern M. Projections of primary and revision hip and knee arthroplasty in the United States from 2005 to 2030. J Bone Joint Surg Am 2007; 89:780-785.

4. Ahmed N, Stanford WL, Kandel RA. Mesenchymal stem and progenitor cells for cartilage repair. Skeletal Radiol 2007;36:909-12.

5. Brittberg M, Lindahl A, Nilsson A, Ohlsson C, Isaksson O, Peterson L. Treatment of deep cartilage defects in the knee with autologous chondrocyte transplantation. N Engl J Med 1994; 331:889-895.

6. Esfandiari E, Nazem Kh, Safdarian A,  Fesharaki M,  Moulavi F, Shakibaei M, et al . Treatment of full thickness cartilage defects in human knees with Autologous Chondrocyte Transplantation. Res Med Sci 2011; 16:855–861.

7. Esfandiary E, Shakibaei M, Amirpour N, Razavi Sh, Nasresfahani M, Moulavi F, et al. Study of human chondrocyte redifferntiation capacity in three-dimensional hydrogel culture. Iran J Basic Med Sci 2008; 11:152-158.

8. Raghunath J, Salacinski J, Sales M, Butler E, Seifalian M. Advancing cartilage tissue engineering: the application of stem cell technology. Curr Opin Biotechnol 2005; 16:503-509.

9. Song H, Chang W, Song BW, Hwang KC. Specific differentiation of mesenchymal stem cells by small molecules. Am J Stem Cells 2012; 1:22-30.

10. Zuk PA, Zhu M, Ashjian P, De Ugarte DA, Huang JI, Mizuno H, et al . Human adipose tissue is a source of multipotent stem cells. Mol Biol Cell 2002; 13:4279-4295.

11. Zuk PA, Zhu M, Ashjian P, De Ugarte DA, Huang JI, Mizuno H, et al . Multilineage cells from human adipose tissue: implications for cell-based therapies. Tissue Eng 2001; 7:211-28.

12. Chung C, Burdick JA.   Engineering cartilage tissue. Adv Drug Deliv Rev. 2008; 60: 243-62.

13. Hashemibeni B, Razavi S, Esfandiary E, Karbasi S, Mardani M, Nasresfahani M. Induction of chondrogenic differentiation of human adipose-derived stem cells with TGF-β3 in pellet culture system. Iran J Basic Med Sci 2008; 11:10-17.

14. Ansar M, Esfandiariy E, Mardani M, Hashemibeni B, Zarkesh H, Hatef M, et al. A comparative study of aggrecan synthesis between natural articular chondrocytes and differentiated chondrocytes from adipose derived stem cells in 3D culture. Adv Biomed Res 2012; 1:30-37.

15. Yue X, Balooch G, Chio M, Bekerman E, Ritchie R, Longaker M. Analysis of the material properties of early chondrogenic differentiated adipose-derived stromal cells using an in in vitro three dimentional micromass culture system. Biochem Biophys Res Commun 2007; 359:311-316.

16. Xu J, Wang W, Clark C, Brighton T. Signal transduction in electrically stimulated articular chondrocytes involves translocation of extracellular calcium through voltage-gated channels. Osteoarthritis Cartilage 2009; 17:397-405.

17. Esfandiary E, Valiani A, Hashemibeni B, Moradi I, Narimani M. The evaluation of toxicity of carbon nanotubes on the human adipose-derived-stem cells in vitro. Adv Biomed Res 2013; 4:17-25.

18. Yan J, Dong L, Zhang B, Qi N. Effects of extremely low-frequency magnetic field on growth and differentiation of human mesenchymal stem cells. Electromagn Biol Med 2010; 29:165-176.

19.Mardani M, Hashemibeni B, Ansar M, Zarkesh Esfahani H, Kazemi M, Goharian V, et al . Comparison between chondrogenic markers of differentiated chondrocytes from adipose derived stem cells and articular chondrocytes In vitro. Iran J Basic Med Sci 2013; 16:763-771. 

20. Creecy C, Neill C, Arulanandam B, Sylvia V, Navara C, Bizios R. Mesenchymal stem cell osteodifferentiation in response to alternating electric current. Tissue Eng 2012; 19:10-17.

21. Seth M, McQuilling J, Grossfeld R, Lubischer J, Clarke L, Loboa E. Application of low-frequency alternating current electric fields Via interdigitated electrodes: effects on cellular viability, cytoplasmic calcium, and osteogenic differentiation of human adipose-derived stem cells. Tissue eng 2010; 16:1377-1366.

22. Hronik-Tupaj M, Rice W, Cronin-Golomb M, Kaplan D, Georgakoudi I. Osteoblastic differentiation and stress response of human mesenchymal stem cells exposed to alternating current electric fields. Biomed Eng 2011; 10:9-15.

23. Hammerick K, James A, Huang Z, Prinz F, Longaker F. Pulsed direct current electric fields enhance osteogenesis in adipose-derived stromal cells. Tissue eng 2010; 16:917-931.

24. Mayer-Wagner S, Passberger A, Sievers B, Aigner J, Summer B, Schiergens T, et al . Effects of low frequency electromagnetic fields on the chondrogenic differentiationof human mesenchymal stem cells. Bioelectromagnetics 2011; 32:283-290.

25. Wang W, Wang Z, Zhang G, Clark C, Brighton C. Up-regulation of chondrocytes matrix genes and products by electric fields. Clin Orthop Relat Res 2004; 427:5163-5173.

26. Gan J, Fredericks D, Glazer P. Direct current and capacitive coupling electrical stimulation upregulates osteopromotive factors for spinal fusions. Spine 2005; 15:15-22.

27. Goodwin C, Brighton C, Guyer R. A double blind study of capacitively coupled electrical stimulation as an adjunct to lumbar spinal fusions. Spine 1999; 24:1349-1357