Application of novel anodized titanium for enhanced recruitment of H9C2 cardiac myoblast

Document Type : Original Article

Authors

1 Cardiovascular Research Center, Isfahan Cardiovascular Research Institute, Isfahan University of Medical Sciences, Isfahan, Iran

2 Heart failure Research Center, Isfahan Cardiovascular Research Institute, Isfahan, Isfahan University of Medical Sciences, Isfahan, Iran

3 Nanotechnology Consultancy and Development Center (NCDC), Padova, Italy

4 Department of Genetics, Isfahan University of Medical Sciences, Isfahan, Iran

Abstract

Objective(s):Anodized treated titanium surfaces, have been proposed as potential surfaces with better cell attachment capacities. We have investigated the adhesion and proliferation properties of H9C2 cardiac myoblasts on anodized treated titanium surface. 
Materials and Methods: Surface topography and anodized tubules were examined by high-resolution scanning electron microscopy (SEM). Control and test substrates were inserted to the bottom of 24-well tissue culture plates. Culture media including H9C2 cells were loaded on the surface of substrate and control wells at the second passage. Evaluation of cell growth, proliferation, viability and surface cytotoxicity was performed using MTT test. After 48 hr, some samples were inspected by SEM. DAPI-staining was used to count attached cells.
Results: MTT results for cells cultured on anodized titanium  and unanodized titanium surfaces  was equal to 1.56 and 0.55 fold change compared to tissue culture polystyrene (TCPS). The surface had no cytotoxic effects on cells. The average cell attachment to TCPS, unanodized and anodized titanium surface was 2497±40.16, 1250±20.11 and 4859.5±54.173, respectively. Cell adhesion to anodized titanium was showed 1.95 and 3.89 fold increase compared to TCPS and unanodized titanium, respectively (P<0.05).
Conclusion: Anodized titanium surfaces can be potentially applied for enhanced recruitment of H9C2 cells. This unique property makes these inexpensive anodized surfaces as a candidate surface for attachment of cardiac cells and consequently for cardiac regeneration purposes.

Keywords


1. Shachar M, Cohen S. Cardiac tissue engineering, ex-vivo: design principles in biomaterials and bioreactors. Heart Fail Rev 2003; 8:271-276.
2. Mercola M, Ruiz-Lozano P, Schneider MD. Cardiac muscle regeneration: lessons from development. Genes Dev 2011; 25:299-309.
3. Lin Z, Pu WT. Strategies for Cardiac Regeneration and Repair. Sci Transl Med 2014; 6:239rv1.
4. Cardiac Regenerative Capacity and Mechanisms. Annual Review of Cell and Developmental Biology. 2012. Vol. 28.p.719-741.
5. Mardani M, Hashemibeni B, Ansar MM, Zarkesh Esfahani SH, 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; 16:763-773.
6. Mahdavishahri N, Moghatam Matin M, Fereidoni M, Yarjanli Z, Banihashem Rad SA, Khajeh Ahmadi S.            In vitro assay of human gingival scaffold in differentiation of rat's bone marrow mesenchymal stem cells to keratinocystes. Iran J Basic Med Sci 2012; 15:1185-1190.
7. Bursac N. Cardiac tissue engineering using stem cells. IEEE Eng Med Biol Mag 2009; 28:80, 82, 84-86, 88-89.
8. Wu YC, Lee TM, Lin JC, Shaw SY, Yang CY. Argon-plasma-treated chitosan: surface characterization and initial attachment of osteoblasts. J Biomater Sci Polym Ed 2010; 21:563-579.
9. Bohl A, Rohm HW, Ceschi P, Paasche G, Hahn A, Barcikowski S, et al. Development of a specially tailored local drug delivery system for the prevention of fibrosis after insertion of cochlear implants into the inner ear. J Mater Sci Mater Med 2012; 23:2151-2162.
10. Djordjevic I, Choudhury NR, Dutta NK, Kumar S, Szili EJ, Steele DA. Polyoctanediol citrate/sebacate bioelastomer films: surface morphology, chemistry and functionality. J Biomater Sci Polym Ed 2010; 21:237-251.
11. Petrulyte S. Advanced textile materials and biopolymers in wound management. Dan Med Bull 2008; 55:72-77.
12. Machado LG, Savi MA. Medical applications of shape memory alloys. Braz J Med Biol Res 2003; 36:683-691.
13. Barthélémy B, Devillers S, Minet I, Delhalle J, Mekhalif Z. Induction heating for surface triggering styrene polymerization on titanium modified with ATRP initiator. J Colloid Interface Sci 2011; 354:873-879.
14. Wang XX, Xie L, Wang R. Biological fabrication of nacreous coating on titanium dental implant. Biomaterials 2005; 26:6229-6232.
15. Moradi I, Behjati M, Kazemi M. Application of anodized titanium for enhanced recruitment of endothelial progenitor cells. Nanoscale Res Lett 2012; 7:298.
16. Pun DK, Berzins DW. Corrosion behavior of shape memory, superelastic, and nonsuperelastic nickel-titanium-based orthodontic wires at various temperatures. Dent Mater 2008; 24:221-227.
17. Park IS, Yang EJ, Bae TS, “Effect of Cyclic Precalcification of Nanotubular TiO2 Layer on the Bioactivity of Titanium Implant,” BioMed Research International, vol. 2013, Article ID 293627, 7 pages, 2013. doi:10.1155/2013/293627.
 18. Hamlekhan A, Butt A, Patel S, Royhman D, Takoudis C, Sukotjo C, et al. Fabrication of anti-aging TiO2 nanotubes on biomedical Ti alloys. PLoS One 2014 ; 9:e96213.
19. Alpaslan E, Ercan B, Webster TJ. Anodized 20 nm diameter nanotubular titanium for improved bladder stent applications. Int J Nanomed 2011; 6:219–225.
20. Rajyalakshmi A, Ercan B, Balasubramanian K, Webster TJ. Reduced adhesion of macrophages on anodized titanium with select nanotube surface features. Int J Nanomed 2011; 2011:6:1765 – 1771.
21. Burns K, Yao C, Webster TJ. Increased chondrocyte adhesion on nanotubular anodized titanium. J Biomed Mater Res A 2009; 88:561-568.
22. Kimes BW, Brandt BL. Properties of a clonal muscle cell line from rat heart. Exp Cell Res 1976; 98:367-381.
23. Feridooni T, Mac Donald C, Shao D, Yeung P, Agu RU. Cytoprotective potential of anti-ischemic drugs against chemotherapy-induced cardiotoxicity in H9c2 myoblast cell line. Acta Pharm 2013; 63:493-503.
24. Chen C, Zhang X, Dai Y, Zhang X. [Effect of pulsed electrical stimulation on the proliferation and differentiation of H9c2 cells]. Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi 2013; 29:337-340.
25. Prathapan A, Vineetha VP, Abhilash PA, Raghu KG. Boerhaavia diffusa L. attenuates angiotensin II-induced hypertrophy in H9c2 cardiac myoblast cells via modulating oxidative stress and down-regulating NF-κβ and transforming growth factor β1. Br J Nutr 2013; 110:1201-1210.
26. Chang W, Zhang M, Li J, Meng Z, Wei S, Du H, et al. Berberine improves insulin resistance in cardio-myocytes via activation of 5'-adenosine monophos-phate-activated protein kinase. Metabolism 2013; 62:1159-1167.
28. Chang YM, Tsai CT, Wang CC, Chen YS, Lin YM, Kuo CH, et al. Alpinate oxyphyllae fructus (Alpinia Oxyphylla Miq) extracts inhibit angiotensin-II induced cardiac apoptosis in H9c2 cardiomyoblast cells. Biosci Biotechnol Biochem 2013; 77:229-234.