Application of adipose-derived mesenchymal stem cells in an in vivo model of peripheral nerve damage

Elsa González-Cubero; María Luisa González-Fernández; María Rodríguez-Díaz; Marta Palomo-Irigoyen; Ashwin Woodhoo; Vega Villar-Suárez

doi:10.3389/fncel.2022.992221

Application of adipose-derived mesenchymal stem cells in an in vivo model of peripheral nerve damage

Front Cell Neurosci. 2022 Sep 8:16:992221. doi: 10.3389/fncel.2022.992221. eCollection 2022.

Authors

Elsa González-Cubero¹, María Luisa González-Fernández¹, María Rodríguez-Díaz¹, Marta Palomo-Irigoyen^{2

3}, Ashwin Woodhoo^{2

4

5}, Vega Villar-Suárez^{1

6}

Affiliations

¹ Department of Anatomy, Faculty of Veterinary Sciences, University of León-Universidad de León, León, Spain.
² Center for Cooperative Research in Biosciences (CIC bioGUNE), Basque Research and Technology Alliance (BRTA), Bizkaia Technology Park, Derio, Spain.
³ Genes and Disease Group, Department of Dermatology, Medical University of Vienna, Anna Spiegel Center of Translational Research, Vienna, Austria.
⁴ IKERBASQUE, Basque Foundation for Science, Bilbao, Spain.
⁵ Gene Regulatory Control in Disease Group, Center for Research in Molecular Medicine and Chronic Diseases (CIMUS), Health Research Institute of Santiago de Compostela (IDIS), University of Santiago de Compostela, Santiago de Compostela, Spain.
⁶ Institute of Biomedicine (IBIOMED), University of León-Universidad de León, León, Spain.

Abstract

Background: Neuropathic pain is one of the most difficult to treat chronic pain syndromes. It has significant effects on patients' quality of life and substantially adds to the burden of direct and indirect medical costs. There is a critical need to improve therapies for peripheral nerve regeneration. The aim of this study is to address this issue by performing a detailed analysis of the therapeutic benefits of two treatment options: adipose tissue derived-mesenchymal stem cells (ASCs) and ASC-conditioned medium (CM).

Methods: To this end, we used an in vivo rat sciatic nerve damage model to investigate the molecular mechanisms involved in the myelinating capacity of ASCs and CM. Furthermore, effect of TNF and CM on Schwann cells (SCs) was evaluated. For our in vivo model, biomaterial surgical implants containing TNF were used to induce peripheral neuropathy in rats. Damaged nerves were also treated with either ASCs or CM and molecular methods were used to collect evidence of nerve regeneration. Post-operatively, rats were subjected to walking track analysis and their sciatic functional index was evaluated. Morphological data was gathered through transmission electron microscopy (TEM) of sciatic nerves harvested from the experimental rats. We also evaluated the effect of TNF on Schwann cells (SCs) in vitro. Genes and their correspondent proteins associated with nerve regeneration were analyzed by qPCR, western blot, and confocal microscopy.

Results: Our data suggests that both ASCs and CM are potentially beneficial treatments for promoting myelination and axonal regeneration. After TNF-induced nerve damage we observed an upregulation of c-Jun along with a downregulation of Krox-20 myelin-associated transcription factor. However, when CM was added to TNF-treated nerves the opposite effect occurred and also resulted in increased expression of myelin-related genes and their corresponding proteins.

Conclusion: Findings from our in vivo model showed that both ASCs and CM aided the regeneration of axonal myelin sheaths and the remodeling of peripheral nerve morphology.

Keywords: Schwann cells; adipose tissue derived-mesenchymal stem cells; conditioned medium; nerve regeneration using mesenchymal stem cells; peripheral nerve regeneration; peripheral neuropathy.