Indian Journal of Animal Research

  • Chief EditorK.M.L. Pathak

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Indian Journal of Animal Research, volume 55 issue 5 (may 2021) : 530-535

In vitro Cytotoxicity Analysis of Hybrid Graphene Oxide (hGO) Nano Structures in Caprine Wharton’s Jelly Derived Mesenchymal Stem Cells (WJ-MSCs)

S.A. Dhenge, N.E. Gade, O.P. Mishra, Abinash Kumar, V.N. Khandait
1Department of Veterinary Physiology and Biochemistry, College of Veterinary Science and Animal Husbandry, Anjora, Durg-491 001, Chhattisgarh, India.
Cite article:- Dhenge S.A., Gade N.E., Mishra O.P., Kumar Abinash, Khandait V.N. (2020). In vitro Cytotoxicity Analysis of Hybrid Graphene Oxide (hGO) Nano Structures in Caprine Wharton’s Jelly Derived Mesenchymal Stem Cells (WJ-MSCs). Indian Journal of Animal Research. 55(5): 530-535. doi: 10.18805/ijar.B-3992.
Background: Nanotechnology is used in stem cell culture as well as in vivo delivery and tracking of stem cells. Graphene oxide (GO) is a carbon based nanomaterial and it has large surface area as well as good biocompatibility and heteroatoms doped GO exploit its properties. Hybrid GO (hGO) nano structures biocompatibility is depends on its size, dose and exposure time as well as in vitro cell models and hence, need thorough cytotoxicity studies in different species in vitro cell models. 
Methods: Caprine Wharton’s jelly derived mesenchymal stem cells (WJ-MSCs) were isolated, characterized and dose dependent (100, 50, 25, 10 and 0µg /ml) in vitro cytotoxicity of  three different hGO nano structures (phosphorus doped graphene oxide titanium oxide tubes, rods and sheets) were  analysed in caprine WJ-MSCs by studying cell cytotoxicity assays.  
Result: All three hGO nano structures were damaged cell morphology at 100 and 50 µg /ml doses, however, morphologically more good cells were observed in hGO tubes treated group than hGO rods and hGO sheets at 25 and 10 µg/ml doses as compared to control. Cell viability percentage was significantly (P<0.01) decreased at dose 100 µg/ml and it was significantly (P<0.01) increased at 25 µg/ml dose as compared to 50, 10 and 0 µg/ml doses. But, hGO tubes significantly (P<0.01) increased cell viability % as compared to hGO rods and hGO sheets. Cell population doubling time (PDT) was not altered significantly by all hGO nano structures, but 100 and 50 µg/ml doses significantly (P<0.01)increased cell PDT as compared to 25, 10 and 0 µg/ml doses. All hGO nano structures were non significantly altered growth curve, however, all hGO nano structures at 25 µg /ml dose altered (inclined) shape of growth curve, while 100 and 50 µg /ml doses significantly declined growth curve shape as compared to 10 and 0 µg /ml doses. Cell proliferation % was significantly (P<0.01) increased at 25 and 10 µg/ml doses, while, it was significantly (P<0.01) decreased at 100 µg /ml dose as compared to 50 and 0 µg /ml. However, there was no significance difference was observed in cell proliferation % in groups treated by different hGO nanostructures. In last, it was concluded as, hGO nano structures cytotoxicity was dose dependent and hGO nano tubes were least cytotoxic in caprine WJ-MSCs. 
  1. Babaei, H., Moshrefi, M., Golchin, M. and Mematollahi-Mahani, S. N. (2008). Assess the pluripotency of caprine umbilical cord Wharton’s jelly mesenchymal cells by RT-PCR analysis of early transcription factor nanog. Iranian Journal of Veterinary Surgery. 3(2): 57-65.
  2. Baghaban, E., M., Nazarian, H. and Taghiyar, L. (2008). Mesenchymal stem cell isolation from the removed medium of rat bone marrow primary culture and their differentiation skeletal cell lineages. Yakhteh Medical Journal. 10(1): 65-72.
  3. Bhat, I.A., Sivanarayanan, T.B., Somal, A., Pandey, S., Bharti, M.K., Bibhudatta S.K., Indu, B., Verma, M., Anand, J., Sonwane, A., Sai, G.K., Amarpal, Chandra, V. and Sharma, G.T. (2018). An allogenic therapeutic strategy for canine spinal cord injury using mesenchymal stem cells. Journal of Cell Physiology. 1-14.
  4. Bregoli, L., Chiarini, F., Gambarelli, A., Sighinolfi, G., Gatti, A.M., Santi, P., Martelli, A.M. and Cocco, L. (2009). Toxicity of antimony trioxide nanoparticles on human hematopoietic progenitor cells and comparison to cell lines. Toxicology. 262(2): 121-129. 
  5. Cardoso, T.C., Ferrari, H.F., Garcia, A.F., Novais, J.B., Frade, C.S., Ferrarezi, M.C., Andrade, A.L. and Gameiro, R. (2012). Isolation and characterization of Wharton’s jelly derived multipotent mesenchymal stromal cells obtained from bovine umbilical cord and maintained in a defined serum free three dimensional systems. BMC Biotechnology. 12: 18.
  6. Dar, R.M., Gade, N.E., Mishra, O.P., Khan, J.R., Vinod, K. and Patiyal, A. (2015). In vitro cytotoxicity assessment of graphene quantum dots in caprine Wharton’s jelly derived mesenchymal stem cells. Journal of Cell and Tissue Research. 15(1): 4703-4710.
  7. Deb, K.D., Griffith, M., Muinck, E.D. and Rafat, M. (2012) Nanotechnology in stem cells research: advances and applications. Frontiers in Bioscience. 1: 1747-1760.
  8. Elkhenany, H., Lisa, A., Andersen, L., Shawn, B., Mark C., Nancy N., Enkeleda, D., Oshin, D. Alexandru, S., Biris, D.A. and Dhar, M. (2015). Graphene support in vitro proliferation and osteogenic differentiation of goat adult mesenchymal stem cells: Potential for bone tissue engineering. Journal of Applied Toxicology. 35: 367-374.
  9. Eswari, S., Monisha. M., Vijayarani, K. and Kumanan, K. (2016). Expression of early transcription factors by mesenchymal stem cells derived from ovine umbilical cord Wharton’s jelly. Indian Journal of Animal Sciences. 86(10): 1132-1135.
  10. Figarol, A., Jeremie, P., Delphine, B., Valerie, F., Celine, A., Jean, M.T., Jean, P.L., Michele, C., Didier, B.A. and Philippe, G. (2015). In vitro toxicity of carbon nanotubes, nanographite and carbon black, similar impacts of acid functionalization. Journal of Toxicolology. In vitro. 30: 476-485.
  11. Gade, N.E., Dar, R.M., Mishra, O.P., Khan, J.R., Vinod, K. and Patyal, A. (2015). Evaluation of dose dependent cytotoxic effects of graphene oxide-iron oxide nanocomposite on caprine Wharton’s jelly derived mesenchymal stem cells. Journal of Animal Research. 5(3): 415-421.
  12. Guo, W., Qiu, J., Liu, J. and Liu, H. (2017). Graphene microfiber as a scaffold for regulation of neural stem cells differentiation. Scientific Reports. 7: 5678.
  13. Guo, Y.Y., Zhang, J., Zheng, Y.F., Yang, J. and Zhu, X.Q. (2011). Cytotoxic and genotoxic effects of multi wall carbon nanotubes on human umbilical vein endothelial cells in vitro. Mutation Research - Genetic Toxicology and Environmental Mutagenesis. 721(2): 184-191.
  14. Kim, J., Kim, H.D., Park, J., Lee, E.S., Kim, E., Lee, S.S., Yang, J.K., Lee, Y.K. and Hwang, N.S. (2018). Enhanced osteogenic commitment of murine mesenchymal stem cells on graphene oxide substrate. Biomaterials Research. 22: 1.
  15. La, W.G., Jin, M., Park, S., Yoon, H.H., Jeong, G.J. and Bhang, S.H. (2014). Delivery of bone morphogenetic protein-2 and substance P using graphene oxide for bone regeneration. International Journal of Nanomedicine. 9: 107-116.
  16. Liu, Q., Wei, L., Wang, J., Peng, F., Luo, D., Cui, R., Niu, Y., Qin, X., Liu, Y., Sun, H., Yang, J. and Li, Y. (2012). Cell imaging by graphene oxide based on surface enhanced Raman scattering. Nanoscale. 4(22): 7084-7089.
  17. Ming, G., Longwei, L., Feng, D., Tianxiao, N., Tong, C., Dandan, X., Wang, S., Zhao, X., Liu., X., Liu, Y., Xiong, C. and Zhou, Y. (2018). Effects of thermal treatment on the adhesion strength and osteoinductive activity of single layer graphene sheets on titanium substrates. Scientific Reports. 8: 8141.
  18. Mirza, E.H., Khan A.A., Khureif, A.A., Saadaldin, S.A., Mohamed, B.A., Fareedi, F., Khan, M.M., Alfayez, M., Fotawi, R.A., Pekka, K.V. and Mahmood, A. (2019). Characterization of osteogenic cells grown over modified graphene oxide biostable polymers. Biomedical Materials. 14: 065004
  19. Nair, M., Nancy, D., Krishnan, A.G., Anjusree, G.S., Vadukumpully, S. and Nair, S.V. (2015). Graphene oxide nanoflakes incorporated gelatin hydroxyapatite scaffolds enhance osteogenic differentiation of human mesenchymal stem cells. Nanotechnology. 26(16): 161001.
  20. Nishida, E., Miyaji, H., Takita, H., Kanayama, I., Tsuji, M., Akasaka, T., Sugaya, T., Sakagami, R. and Kawanami, M. (2014). Graphene oxide coating facilitates the bioactivity of scaffold material for tissue engineering. Japanese Journal of Applied Physics. 53(6S): 06JD04.
  21. Park, S.Y., Park J., Sim, S.H., Sung, M.G., Kim, K.S., Hong, B.H. and Hong, S. (2011). Enhanced differentiation of human neural stem cells into neurons on graphene. Advances in Materials. 23(36): 263-267.
  22. Qu, C., Salla, K., Kroger, H., Lappalainen, R. and Lammi, M.J. (2016). Behavior of human bone marrow derived mesenchymal stem cells on various titanium based coatings. Materials. 9(10): 827.
  23. Ravindran, N. A., Maiti, S.K., Palakkara, S., Kritaniya, D., Mahan, T. and Naveen, K. (2016). In vitro osteoinduction potential of a novel silica coated hydroxyapatite bioscaffold seeded with rabbit mesenchymal stem cell. Journal of Stem Cell Research and Therapeutics. 1(2): 00009.
  24. Rodriguez, L.N., Romero, P., Estivill, T.G., Guzma, V.R. and Aguirre, J.A. (2017). Cell survival and differentiation with nanocrystalline glass like carbon using substantia nigra dopaminergic cells derived from transgenic mouse embryos. PLoS ONE. 12(3): e0173978. 
  25. Sangeetha, P., Maiti, K., Singh, K., Gopinathan, A., Singh, K.P., Mohan, D., Ninu, A.R. and Kumar, N. (2017). Evaluation of bio-engineered corneal scaffold for the repair of corneal defect in rabbit model. Indian Journal of Animal Sciences. 87 (11): 1332-1339.
  26. Sohaebuddin, S.K., Thevenot, P.T., Baker, D., Eaton, J.W. and Tang, L. (2010). Nanomaterial cytotoxicity is composition, size and cell type dependent. Particle and Fiber Toxicology. 7:22.
  27. Somal, A., Bhat, I. A., Indu, B., Pandey. S., Panda, B.S.K., Thakur, N., Sarkar, M., Chandra, V., Saikumar, G. and Sharma, G.T. (2016). A comparative study of growth kinetics, in vitro differentiation potential and molecular characterization of fetal adnexa derived caprine Mesenchymal Stem Cells. PLoS ONE. 11(6): 1-17. e0156821.
  28. Szczypta, A.F., Menaszek, E., Syeda, T.M., Mishra, A., Alavijeh, M.,Adu, J. and Blazewicz, S. (2012). Effect of MWCNT surface and chemical modification on in vitro cellular response. Journal of Nanoparticle Research. 14: 1181. 
  29. Tilton, S.C., Karin, N.J., Tolic, A., Xie, Y., Lai, X., Hamilton, R.F., Waters, K.M., Holian, A., Witzmann, A. and Orr, G. (2014). Three human cell types respond to multi walled carbon nanotubes and titanium dioxide nanobelts with cell specific transcriptomic and proteomic expression patterns. Nanotoxicology. 8(5): 533-548.
  30. Wadhwa, S., Rea. C., Hare, P.O., Mathur, A., Roy, S.S., Dunlop, P.S., Byrne, J.A., Burke, G., Meenan, B. and McLaughlin, J.A. (2011). Comparative in vitro cytotoxicity study of carbon nanotubes and titanium nanostructures on human lung epithelial cells. Journal of Hazardous Materials. 191(1-3): 56-61.
  31. Wang, J., Sun, P., Bao, Y., Liu, J. and An, L. (2011). Cytotoxicity of single walled carbon nanotubes on PC12 cells. Toxicology in vitro. 25(1): 242-250.
  32. Zhuang, S., Singh, H., Nunna, B.B., Mandal, D., Anibal B. and Lee, E.S. (2018). Nitrogen-doped graphene-based catalyst with metal-reduced organic framework: Chemical analysis and structure control. Carbon. 139: 933-944.

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