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J Plast Reconstr Aesthet Surg. 2019 Aug;72(8):1316-1325. doi: 10.1016/j.bjps.2019.04.014. Epub 2019 May 9.

Seeding decellularized nerve allografts with adipose-derived mesenchymal stromal cells: An in vitro analysis of the gene expression and growth factors produced.

Author information

1
Department of Orthopedic Surgery, Division of Hand and Microvascular Surgery, Mayo Clinic, 200 First Street S.W., Rochester, MN 55905, USA; Department of Plastic, Reconstructive and Hand Surgery, Erasmus Medical Center, 's Gravendijkwal 230, 3015 CE Rotterdam, the Netherlands.
2
Department of Orthopedic Surgery, Division of Hand and Microvascular Surgery, Mayo Clinic, 200 First Street S.W., Rochester, MN 55905, USA; Department of Biological Sciences, Hampton University, 100 E Queen St, Hampton, VA 23668, USA.
3
Department of Plastic, Reconstructive and Hand Surgery, Erasmus Medical Center, 's Gravendijkwal 230, 3015 CE Rotterdam, the Netherlands; Xpert Clinic, Hand and Wrist Surgery, Jan Leentvaarlaan 14-24, 3065 DC Rotterdam, the Netherlands.
4
Department of Orthopedic Surgery, Division of Hand and Microvascular Surgery, Mayo Clinic, 200 First Street S.W., Rochester, MN 55905, USA; Department of Biochemistry and Molecular Biology, Mayo Clinic, 200 First Street S.W., Rochester, MN 55905, USA.
5
Department of Orthopedic Surgery, Division of Hand and Microvascular Surgery, Mayo Clinic, 200 First Street S.W., Rochester, MN 55905, USA. Electronic address: shin.alexander@mayo.edu.

Abstract

Mesenchymal stromal cells (MSCs) secrete many soluble growth factors and have previously been shown to stimulate nerve regeneration. MSC-seeded processed nerve allografts could potentially be a promising method for large segmental motor nerve injuries. Further progress in our understanding of how the functions of MSCs can be leveraged for peripheral nerve repair is required before making clinical translation. The present study, therefore, investigated whether interactions of adipose-derived MSCs with decellularized nerve allografts can improve gene and protein expression of growth factors that may support nerve regeneration. Human nerve allografts (n = 30) were decellularized and seeded with undifferentiated human adipose-derived MSCs. Subsequently, the MSCs and MSC-seeded grafts were isolated on days 3, 7, 14, and 21 in culture for RNA expression analysis by qRT-PCR. Evaluated genes included NGF, BDNF, PTN, GAP43, MBP, PMP22, VEGF, and CD31. Growth factor production was evaluated and quantified using enzyme-linked immunosorbent assay (ELISA). On day 21, semi-quantitative RT-PCR analysis showed that adherence of MSCs to nerve allografts significantly enhances mRNA expression of neurotrophic, angiogenic, endothelial, and myelination markers (e.g., BDNF, VEGF, CD31, and MBP). ELISA results revealed an upregulation of BDNF and reduction of both VEGF and NGF protein levels. This study demonstrates that seeding of undifferentiated adipose-derived MSCs onto processed nerve allografts permits the secretion of neurotrophic and angiogenic factors that can stimulate nerve regeneration. These favorable molecular changes suggest that MSC supplementation of nerve allografts may have potential in improving nerve regeneration.

KEYWORDS:

Adipose-derived stem cells; Gene expression; MSCs; Peripheral nerve; Processed nerve allograft; qPCR

PMID:
31175032
DOI:
10.1016/j.bjps.2019.04.014

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