中国神经再生研究(英文版) ›› 2021, Vol. 16 ›› Issue (8): 1613-1621.doi: 10.4103/1673-5374.303031

• 原著:周围神经损伤修复保护与再生 • 上一篇    下一篇

接种于神经同种异体移植物或胶原神经导管的人脂肪间充质干细胞的基因表达谱

  

  • 出版日期:2021-08-15 发布日期:2021-01-13
  • 基金资助:

    NIH资助项目

Gene expression profiles of human adipose-derived mesenchymal stem cells dynamically seeded on clinically available processed nerve allografts and collagen nerve guides

Femke Mathot1, 2, Nadia Rbia1, 3, Roman Thaler1, 4, Allan B. Dietz5, Andre J. van Wijnen1, 4, Allen T. Bishop1, Alexander Y. Shin1, *#br#   

  1. 1Department of Orthopedic Surgery, Mayo Clinic, Rochester, MN, USA; 2Department of Plastic Surgery, Radboudumc, Nijmegen, The Netherlands; 3Department of Dermatology, Erasmus Medical Center, Rotterdam, The Netherlands; 4Department of Biochemistry and Molecular Biology, Mayo Clinic, Rochester, MN, USA; 5Department of Laboratory Medicine and Pathology, Mayo Clinic, Rochester, MN, USA
  • Online:2021-08-15 Published:2021-01-13
  • Contact: Alexander Y. Shin, MD, shin.alexander@mayo.edu.
  • Supported by:
    The study was supported by the National Institute of Neurological Disorders and Stroke of the National Institutes of Health (No. R01NS102360). The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health.

摘要:

接种于神经移植物表面的间充质干细胞可提供必要的营养因子。(1)为了解Avance®神经移植物或NeuraGen®神经导管对接种的人脂肪间充质干细胞基因表达的影响,实验将人脂肪间充质干细胞接种到Avance®神经移植物和NeuraGen®神经导管中,然后 应用qPCR检测接种后1-21d人脂肪间充质干细胞中神经营养因子 和细胞表面标志物的基因表达。以未接种于Avance®神经移植和NeuraGen®神经导管的干细胞基因表达为基线;(2)人脂肪间充质干细胞与Avance®神经移植物的相互作用导致神经营养因子因子神经生长因子,神经胶质细胞系神经营养因子和脑源性神经营养因子,髓鞘形成蛋白周围神经髓鞘蛋白22和髓磷脂蛋白零,和血管生成基因CD31和血管内皮细胞生长因子α短期上调以及脑源性神经营养因子,血管内皮细胞生长因子α和COL1A1长期上调;(3) 人脂肪间充质干细胞与NeuraGen®神经导管之间的相互作用导致神经营养因子神经生长因子,神经胶质细胞系神经营养因子和脑源性神经营养因子,髓鞘形成蛋白周围神经髓鞘蛋白22和髓磷脂蛋白零,血管生成基因CD31和血管内皮细胞生长因子α,细胞外基质(胶原1型1 )和细胞表面标志物CD96的短期上调基因和神经营养(神经胶质细胞系神经营养因子和脑源性神经营养因子),血管生成(CD31和血管内皮细胞生长因子α),细胞外基质基因(胶原1型1 ,胶原蛋白αIII ,纤维蛋白1)和细胞表面(CD96)基因的长期上调。(3)分析表明,与接种到Avance®神经移植物的人脂肪间充质干细胞相比,接种到NeuraGen®神经导管的人脂肪间充质干细胞的神经营养(多营养蛋白),血管生成(血管内皮细胞生长因子α)和细胞外基质(胶原蛋白αIII和纤维蛋白1)长期基因表达水平更高;(4)总之,人脂肪间充质干细胞与两种神经移植替代物之间的相互作用导致大量重要的神经再生基因表达明显上调。人脂肪间充质干细胞与NeuraGen®神经导管的体外相互作用更加明显,尤其是长期作用更为明显。

https://orcid.org/0000-0001-9658-8192 (Alexander Y. Shin) 

Abstract: It was hypothesized that mesenchymal stem cells (MSCs) could provide necessary trophic factors when seeded onto the surfaces of commonly used nerve graft substitutes. We aimed to determine the gene expression of MSCs when influenced by Avance® Nerve Grafts or NeuraGen® Nerve Guides. Human adipose-derived MSCs were cultured and dynamically seeded onto 30 Avance® Nerve Grafts and 30 NeuraGen® Nerve Guides for 12 hours. At six time points after seeding, quantitative polymerase chain reaction analyses were performed for five samples per group. Neurotrophic [nerve growth factor (NGF), glial cell line-derived neurotrophic factor (GDNF), pleiotrophin (PTN), growth associated protein 43 (GAP43) and brain-derived neurotrophic factor (BDNF)], myelination [peripheral myelin protein 22 (PMP22) and myelin protein zero (MPZ)], angiogenic [platelet endothelial cell adhesion molecule 1 (PECAM1/CD31) and vascular endothelial cell growth factor alpha (VEGFA)], extracellular matrix (ECM) [collagen type alpha I (COL1A1), collagen type alpha III (COL3A1), Fibulin 1 (FBLN1) and laminin subunit beta 2 (LAMB2)] and cell surface marker cluster of differentiation 96 (CD96) gene expression was quantified. Unseeded Avance® Nerve Grafts and NeuraGen® Nerve Guides were used to evaluate the baseline gene expression, and unseeded MSCs provided the baseline gene expression of MSCs. The interaction of MSCs with the Avance® Nerve Grafts led to a short-term upregulation of neurotrophic (NGF, GDNF and BDNF), myelination (PMP22 and MPZ) and angiogenic genes (CD31 and VEGFA) and a long-term upregulation of BDNF, VEGFA and COL1A1. The interaction between MSCs and the NeuraGen® Nerve Guide led to short term upregulation of neurotrophic (NGF, GDNF and BDNF) myelination (PMP22 and MPZ), angiogenic (CD31 and VEGFA), ECM (COL1A1) and cell surface (CD96) genes and long-term upregulation of neurotrophic (GDNF and BDNF), angiogenic (CD31 and VEGFA), ECM genes (COL1A1, COL3A1, and FBLN1) and cell surface (CD96) genes. Analysis demonstrated MSCs seeded onto NeuraGen® Nerve Guides expressed significantly higher levels of neurotrophic (PTN), angiogenic (VEGFA) and ECM (COL3A1, FBLN1) genes in the long term period compared to MSCs seeded onto Avance® Nerve Grafts. Overall, the interaction between human MSCs and both nerve graft substitutes resulted in a significant upregulation of the expression of numerous genes important for nerve regeneration over time. The in vitro interaction of MSCs with the NeuraGen® Nerve Guide was more pronounced, particularly in the long term period (> 14 days after seeding). These results suggest that MSC-seeding has potential to be applied in a clinical setting, which needs to be confirmed in future in vitro and in vivo research.

Key words: Avance? Nerve Grafts, dynamic seeding, mesenchymal stem cell, NeuraGen? Nerve Guides, peripheral nerve repair, qPCR