Seeding decellularized nerve allografts with adipose-derived mesenchymal stromal cells

An in vitro analysis of the gene expression and growth factors produced

Nadia Rbia, Liselotte F. Bulstra, Eric A. Lewallen, Steven E.R. Hovius, Andre J van Wijnen, Alexander Yong-Shik Shin

Research output: Contribution to journalArticle

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.

Original languageEnglish (US)
JournalJournal of Plastic, Reconstructive and Aesthetic Surgery
DOIs
StatePublished - Jan 1 2019

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Mesenchymal Stromal Cells
Allografts
Intercellular Signaling Peptides and Proteins
Gene Expression
Nerve Regeneration
Brain-Derived Neurotrophic Factor
Vascular Endothelial Growth Factor A
Nerve Growth Factor
Enzyme-Linked Immunosorbent Assay
In Vitro Techniques
Polymerase Chain Reaction
Angiogenesis Inducing Agents
Nerve Growth Factors
Peripheral Nerves
Proteins
Up-Regulation
RNA
Transplants
Messenger RNA
Wounds and Injuries

Keywords

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

ASJC Scopus subject areas

  • Surgery

Cite this

Seeding decellularized nerve allografts with adipose-derived mesenchymal stromal cells : An in vitro analysis of the gene expression and growth factors produced. / Rbia, Nadia; Bulstra, Liselotte F.; Lewallen, Eric A.; Hovius, Steven E.R.; van Wijnen, Andre J; Shin, Alexander Yong-Shik.

In: Journal of Plastic, Reconstructive and Aesthetic Surgery, 01.01.2019.

Research output: Contribution to journalArticle

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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.",
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AU - Lewallen, Eric A.

AU - Hovius, Steven E.R.

AU - van Wijnen, Andre J

AU - Shin, Alexander Yong-Shik

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