Effect of in-vitro differentiated bone marrow mesenchymal stem cells in the treatment of peripheral nerve injury in rats

Abstract :

Peripheral nerves are more prone to damage during trauma. Though nerve grafts are used as an alternative method in treating it, the results are purely ambiguous. One such modern approach to treat peripheral nerve injury is bone marrow differentiated neuronal cells. Our present aim is to study the effect of in-vitro differentiated bone marrow mesenchymal stem cells (BMSCs) in the treatment of peripheral nerve injury in rats. Six weeks old rat weighing 80 gm was used for isolation and culture of BMSCs. The second passage cells were taken for neuronal differentiation. Flow cytometry and immunocytochemistry were performed with Anti goat IgG antibody indirectly conjugated with FITC to express nestin. In vitro differentiated BMSC along with PLGA Scaffold is injected into the site of peripheral nerve injury and the results were studied by ENMG, microdissection, and histopathology. The cells were expressed with Nestin goat polyclonal antibody. At the end of the second week, the rat reveals increased amplitude (8.3 mv) with decreased latency (0.8 ms) of the peripheral nerve. Micro dissection confirms the neuronal continuity of the injured peripheral nerve. Histopathology distinctly exhibit increased myelination and decreased endoneuronal space. In conclusion, neuronal differentiated BMSC, regenerate peripheral nerve injury faster than conventional methods and can be applied as an alternate therapy in peripheral nerve repair.

Introduction
Tissue engineering is multifaceted, involves the isolation, characterization and differentiation of bone marrow cells. Transplantation of neuronal differentiated bone marrow mesenchymal stem cells (BMSCs) onto the site of injury results in peripheral nerve repair. This method of clinical transplantation of differentiated neuronal cells along with bioscaffolds challenges regenerative medicine for faster and complete nerve repair. Autologous nerve grafts had been the gold standard for the treatment of the peripheral nerve injury that exceeded the critical gap length. Transplantation of differentiated BMSCs into Schwann cells along with a bioengineered scaffold holds promise for nerve regeneration because of the limited availability of donor nerves and donor morbidity [1]. BMSCs are supposed to be the ideal transplantable cell due to its easy accessibility, rapid expansion capacity when cultured, immune-competent and immune-privileged nature [2]. Recent transplantation studies suggest in-vitro differentiation of BMSCs into cells expressing Schwann cell antigen, followed by in-vivo transp-lantation, shown to have a faster regenerative effect on damaged sciatic nerve [3]. This finding suggests that in-vivo transplantation of BMSCs into a central and peripheral nervous system following ex-vivo differentiation makes it potential to fill nerve gap repair. The main purpose of this study is ex-vivo differentiation of BMSCs into neuronal cells and in-vivo transplantation of differentiated neuronal cells with poly lactic co glycolic acid (PLGA) scaffold aids faster regeneration of nerve gaps in trauma, gunshot and neurodegenerative diseases.

References

1. Monje PV (2020) Schwann cell cultures: biology, technology and therapeutics. Cells. 9: 1848. doi.org/10.3390 /cells9081848.
2. Azizi SA, Stokes D, Augelli BJ, DiGirolamo C, Prockop DJ (1998) Engraftment and migration of human bone marrow stromal cells implanted in the brains of albino rats - similarities to astrocyte grafts. Proceedings of the Notational Academy of Sciences. 95 (7): 3908-3913. doi.org/10.1073/pnas.95.7.3908.
3. Keilhoff G, Stang F, Goihl A, Wolf G, Fansa H (2006) Transdifferentiated mesenchymal stem cells as alternative therapy in supporting nerve regeneration and myelination. Cellular and Molecular Neurobiology. 26 (7-8): 1235-1252. doi: 10.1007/s10571-006-9029-9.
4. Kretlow JD, Jin Y-Q, Liu W, Zhang WJ, Hong T-H, Zhou G, Baggett LS, Mikos AG, Cao Y (2008) Donor age and cell passage affects differentiation potential of murine bone marrow derived stem cells. BMC Molecular and Cell Biology. 9: 60. doi.org/10.1186/1471-2121-9-60.
5. Morrison SJ, White PM, Zock C, Anderson DJ (1999) Prospective identification, isolation by flow cytometry, and in vivo self-renewal of multipotent mammalian neural crest stem cells. Cell. 96 (5): 737-749. doi: 10.1016/ s0092-8674(00)80583-8.
6. Höving AL, Windmöller BA, Knabbe C, Kaltschmidt B, Kaltschmidt C, Greiner JFW (2021) Between fate choice and self-renewal-heterogeneity of adult neural crest-derived stem cells. Frontiers in Cell and Developmental Biology. 9: 662754. doi: 10.3389/fcell.2021.662754.
7. Chen X, Wang XD, Chen G, Lin WW, Yao J, Gu XS (2006) Study of in vivo differentiation of rat bone marrow stromal cells into Schwann cell-like cells. Microsurgery. 26 (2): 111-115. doi: 10.1002/micr.20184.
8. Saraswathi P, Saravanakumar S (2010) A simple method of tooth regeneration by bone marrow mesenchymal stem cells in albino rats. European Journal of Anatomy. 14 (3): 121-126.
9. Torrente Y, Polli E (2008) Mesenchymal stem cell transplantation for neurodegenerative diseases. Cell Transplantation. 17 (10-11): 1103-1113. doi: 10.3727/09636890878236576.
10. Wakao S, Hayashi T, Kitada M, Kohama M, Matsue D, Teramoto N, Ose T, Itokazu Y, Koshino K, Watabe H, Iida H, Takamoto T, Tabata Y, Dezawa M (2010) Long-term observation of auto-cell transplantation in non-human primate reveals safety and efficiency of bone marrow stromal cell-derived Schwann cells in peripheral nerve regeneration. Experimental Neurology. 223 (2): 537-47. doi: 10.1016/ j.expneurol.2010.01.022.
11. Gocmen S, Sirin S, Kaan Oysul K, Ulas UH, Oztas E (2012) The effect of low dose radiation in the treatment of sciatic nerve injury in rats. Turkish Neurosurgery. 22 (2): 167-173. doi: 105137/1019-5149.JTN5039-11.1.
12. Gorulu A, Uzal C, Doganay L, Imer M, Eliuz K, Cobanoglu S (2013) The effect of low dose radiation external beam radiation on extra neural scarring after peripheral nerve surgery in rats. Neurosurgery. 53 (5):1389-1396. doi: 10.1227/01.neu.0000093827.05319.e5.

Citation :

Atmaram et al. (2023) Effect of in-vitro differentiated bone marrow mesenchymal stem cells in the treatment of peripheral nerve injury in rats. Mediterr J Pharm Pharm Sci. 3 (3): 9-15. https://doi.org/10.5281/zenodo.8190815.