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Characterization of three washing/decellularization procedures for the production of bioactive human micronized neural tissue (hMINT)

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Abstract

Background

We developed a novel, injectable and decellularized human peripheral nerve-based scaffold, named Micronized Human Neural Tissue (hMINT), designed to be used as a supportive matrix for stem cell transplantation in the context of spinal cord injury (SCI).

Materials and methods

Human donated sciatic nerves were micronized at liquid nitrogen temperature prior to decellularization using 3 different procedures of various harshness. hMINT were characterized in terms of particle size, DNA, sulfated glycosaminoglycans (sGAG) and growth factors content. To test the biocompatibility and bioactivity of the various preparations, we used a type of mesenchymal stromal cells (MSCs), termed MIAMI cells, which were placed in contact with hMINT to monitor cell attachment by confocal microscopy and gene expression by RT-qPCR in vitro.

Results

The content of DNA, sGAG and growth factors left in the product after processing was highly dependent on the decellularization procedure used. We demonstrated that hMINT are biocompatible and promoted the attachment and long-term survival of MIAMI cells in vitro. Finally, combination with hMINT increased MIAMI cells mRNA expression of pro-survival and anti-inflammatory factors. Importantly, the strongest bioactivity on MIAMI cells was observed with the hMINT decellularized using the mildest decellularization procedure, therefore emphasizing the importance of achieving an adequate decellularization without losing the hMINT’s bioactivity.

Perspectives and clinical significance

The capacity of hMINT/stem cells to facilitate protection of injured neural tissue, promote axon re-growth and improve functional recovery will be tested in an animal model of SCI and other neurodegenerative disorders in the future.

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Data availability

The datasets generated during and/or analyzed during the current study are available from the corresponding author on reasonable request.

References

  • Bamber NI, Li H, Lu X, Oudega M, Aebischer P, Xu XM (2001) Neurotrophins BDNF and NT-3 promote axonal re-entry into the distal host spinal cord through Schwann cell-seeded mini-channels. Eur J Neurosci 13(2):257–268

    CAS  PubMed  Google Scholar 

  • Barzilay R, Kan I, Ben-Zur T, Bulvik S, Melamed E, Offen D (2008) Induction of human mesenchymal stem cells into dopamine-producing cells with different differentiation protocols. Stem Cells Dev 17(3):547–554

    Article  CAS  PubMed  Google Scholar 

  • Boido M, Garbossa D, Fontanella M, Ducati A, Vercelli A (2012) Mesenchymal stem cell transplantation reduces glial cyst and improves functional outcome following spinal cord compression. World Neurosurg 81(1):183–190

    Article  PubMed  Google Scholar 

  • Bregman BS, Coumans JV, Dai HN, Kuhn PL, Lynskey J, McAtee M, Sandhu F (2002) Transplants and neurotrophic factors increase regeneration and recovery of function after spinal cord injury. Prog Brain Res 137:257–273

    Article  CAS  PubMed  Google Scholar 

  • Brohlin M, Mahay D, Novikov LN, Terenghi G, Wiberg M, Shawcross SG, Novikova LN (2009) Characterisation of human mesenchymal stem cells following differentiation into Schwann cell-like cells. Neurosci Res 64(1):41–49

    Article  PubMed  Google Scholar 

  • Cho SR, Kim YR, Kang HS, Yim SH, Park CI, Min YH, Lee BH, Shin JC, Lim JB (2009) Functional recovery after the transplantation of neurally differentiated mesenchymal stem cells derived from bone barrow in a rat model of spinal cord injury. Cell Transplant 18(12):1359–1368

    Article  PubMed  Google Scholar 

  • Cizkova D, Rosocha J, Vanicky I, Jergova S, Cizek M (2006) Transplants of human mesenchymal stem cells improve functional recovery after spinal cord injury in the rat. Cell Mol Neurobiol 26(7–8):1167–1180

    PubMed  Google Scholar 

  • Curtis K, Gomez L, Rios C, Garbayo E, Raval A, Perez-Pinzon M, Schiller P (2010) EF1alpha and RPL13a represent normalization genes suitable for RT-qPCR analysis of bone marrow derived mesenchymal stem cells. BMC Mol Biol 11(1):1–15

    Article  Google Scholar 

  • Delcroix GJ, Curtis KM, Schiller PC, Montero-Menei CN (2010) EGF and bFGF pre-treatment enhances neural specification and the response to neuronal commitment of MIAMI cells. Differentiation 80(4–5):213–227

    Article  CAS  PubMed  Google Scholar 

  • Delcroix GJ, Garbayo E, Sindji L, Thomas O, Vanpouille-Box C, Schiller PC, Montero-Menei CN (2011) The therapeutic potential of human multipotent mesenchymal stromal cells combined with pharmacologically active microcarriers transplanted in hemi-parkinsonian rats. Biomaterials 32(6):1560–1573

    Article  CAS  PubMed  Google Scholar 

  • Delcroix GJ, Kaimrajh DN, Baria D, Cooper S, Reiner T, Latta L, D’Ippolito G, Schiller PC, Temple HT (2013) Histologic, biomechanical, and biological evaluation of fan-folded iliotibial band allografts for anterior cruciate ligament reconstruction. Arthroscopy 29:756–765

    Article  PubMed  Google Scholar 

  • D’Ippolito G, Diabira S, Howard GA, Menei P, Roos BA, Schiller PC (2004) Marrow-isolated adult multilineage inducible (MIAMI) cells, a unique population of postnatal young and old human cells with extensive expansion and differentiation potential. J Cell Sci 117(Pt 14):2971–2981

    Article  CAS  PubMed  Google Scholar 

  • Garbayo E, Raval AP, Curtis KM, Della-Morte D, Gomez LA, D’Ippolito G, Reiner T, Perez-Stable C, Howard GA, Perez-Pinzon MA, Montero-Menei CN, Schiller PC (2011) Neuroprotective properties of marrow-isolated adult multilineage-inducible cells in rat hippocampus following global cerebral ischemia are enhanced when complexed to biomimetic microcarriers. J Neurochem 119(5):972–988

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Guo JS, Zeng YS, Li HB, Huang WL, Liu RY, Li XB, Ding Y, Wu LZ, Cai DZ (2007) Cotransplant of neural stem cells and NT-3 gene modified Schwann cells promote the recovery of transected spinal cord injury. Spinal Cord 45(1):15–24

    Article  PubMed  Google Scholar 

  • Haggerty AE, Oudega M (2013) Biomaterials for spinal cord repair. Neurosci Bull 29:445–459

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Hudson TW, Liu SY, Schmidt CE (2004a) Engineering an improved acellular nerve graft via optimized chemical processing. Tissue Eng 10(9–10):1346–1358

    Article  CAS  PubMed  Google Scholar 

  • Hudson TW, Zawko S, Deister C, Lundy S, Hu CY, Lee K, Schmidt CE (2004b) Optimized acellular nerve graft is immunologically tolerated and supports regeneration. Tissue Eng 10(11–12):1641–1651

    Article  CAS  PubMed  Google Scholar 

  • Keilhoff G, Goihl A, Langnase K, Fansa H, Wolf G (2006a) Transdifferentiation of mesenchymal stem cells into Schwann cell-like myelinating cells. Eur J Cell Biol 85(1):11–24

    Article  CAS  PubMed  Google Scholar 

  • Keilhoff G, Stang F, Goihl A, Wolf G, Fansa H (2006b) Transdifferentiated mesenchymal stem cells as alternative therapy in supporting nerve regeneration and myelination. Cell Mol Neurobiol 26(7–8):1235–1252

    PubMed  Google Scholar 

  • Kubinova S, Sykova E (2012) Biomaterials combined with cell therapy for treatment of spinal cord injury. Regen Med 7(2):207–224

    Article  CAS  PubMed  Google Scholar 

  • Li C, Zhang X, Cao R, Yu B, Liang H, Zhou M, Li D, Wang Y, Liu E (2012) Allografts of the acellular sciatic nerve and brain-derived neurotrophic factor repair spinal cord injury in adult rats. PLoS ONE 7(8):e42813

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Novikova LN, Brohlin M, Kingham PJ, Novikov LN, Wiberg M (2011) Neuroprotective and growth-promoting effects of bone marrow stromal cells after cervical spinal cord injury in adult rats. Cytotherapy 13(7):873–887

    Article  CAS  PubMed  Google Scholar 

  • Onose G, Anghelescu A, Muresanu DF, Padure L, Haras MA, Chendreanu CO, Onose LV, Mirea A, Ciurea AV, El Masri WS, von Wild KR (2009) A review of published reports on neuroprotection in spinal cord injury. Spinal Cord 47(10):716–726

    Article  CAS  PubMed  Google Scholar 

  • Osaka M, Honmou O, Murakami T, Nonaka T, Houkin K, Hamada H, Kocsis JD (2010) Intravenous administration of mesenchymal stem cells derived from bone marrow after contusive spinal cord injury improves functional outcome. Brain Res 1343:226–235

    Article  CAS  PubMed  Google Scholar 

  • Park HW, Lim MJ, Jung H, Lee SP, Paik KS, Chang MS (2010) Human mesenchymal stem cell-derived Schwann cell-like cells exhibit neurotrophic effects, via distinct growth factor production, in a model of spinal cord injury. Glia 58(9):1118–1132

    Article  PubMed  Google Scholar 

  • Park JH, Kim DY, Sung IY, Choi GH, Jeon MH, Kim KK, Jeon SR (2012) Long-term results of spinal cord injury therapy using mesenchymal stem cells derived from bone marrow in humans. Neurosurgery 70(5):1238–1247

    Article  PubMed  Google Scholar 

  • Perale G, Giordano C, Bianco F, Rossi F, Tunesi M, Daniele F, Crivelli F, Matteoli M, Masi M (2011) Hydrogel for cell housing in the brain and in the spinal cord. Int J Artif Organs 34(3):295–303

    Article  CAS  PubMed  Google Scholar 

  • Sondell M, Lundborg G, Kanje M (1998) Regeneration of the rat sciatic nerve into allografts made acellular through chemical extraction. Brain Res 795(1–2):44–54

    Article  CAS  PubMed  Google Scholar 

  • Tatard VM, D’Ippolito G, Diabira S, Valeyev A, Hackman J, McCarthy M, Bouckenooghe T, Menei P, Montero-Menei CN, Schiller PC (2007) Neurotrophin-directed differentiation of human adult marrow stromal cells to dopaminergic-like neurons. Bone 40(2):360–373

    Article  CAS  PubMed  Google Scholar 

  • Vaquero J, Zurita M (2009) Bone marrow stromal cells for spinal cord repair: a challenge for contemporary neurobiology. Histol Histopathol 24(1):107–116

    CAS  PubMed  Google Scholar 

  • Volpato FZ, Fuhrmann T, Migliaresi C, Hutmacher DW, Dalton PD (2013) Using extracellular matrix for regenerative medicine in the spinal cord. Biomaterials 34:4945–4955

    Article  CAS  PubMed  Google Scholar 

  • Wislet-Gendebien S, Hans G, Leprince P, Rigo JM, Moonen G, Rogister B (2005a) Plasticity of cultured mesenchymal stem cells: switch from nestin-positive to excitable neuron-like phenotype. Stem Cells 23(3):392–402

    Article  CAS  PubMed  Google Scholar 

  • Wislet-Gendebien S, Wautier F, Leprince P, Rogister B (2005b) Astrocytic and neuronal fate of mesenchymal stem cells expressing nestin. Brain Res Bull 68(1–2):95–102

    Article  CAS  PubMed  Google Scholar 

  • Yang J, Lou Q, Huang R, Shen L, Chen Z (2008) Dorsal root ganglion neurons induce transdifferentiation of mesenchymal stem cells along a Schwann cell lineage. Neurosci Lett 445(3):246–251

    Article  CAS  PubMed  Google Scholar 

  • Zhou Z, Chen Y, Zhang H, Min S, Yu B, He B, Jin A (2013) Comparison of mesenchymal stem cells from human bone marrow and adipose tissue for the treatment of spinal cord injury. Cytotherapy 15:434–448

    Article  CAS  PubMed  Google Scholar 

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Acknowledgements

We are grateful to Laurence Sindji (MINT Inserm U1066, University of Angers, France) for particle sizing experiments as well as to Justin Kallman, Jeffrey Davila, Cassandra Anton and Laura Varela for the technical assistance provided during hMINT processing.

Funding

This work was funded by the University of Miami Tissue Bank. This study was also supported by the Miami VA Healthcare System Geriatric Research Education and Clinical Center and used resources and facilities of VINCI, VA HSR RES 13-457. The views expressed are those of the authors and not those of the Geriatric Research Education and Clinical Center or VINCI.

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Authors and Affiliations

  1. College of Allopathic Medicine, Nova Southeastern University, Fort Lauderdale, FL, USA

    Gaëtan J.-R. Delcroix

  2. Miami Project to Cure Paralysis, University of Miami Miller School of Medicine, Miami, FL, USA

    Amber Hackett

  3. Geriatric Research Education and Clinical Center, Miami VA Healthcare System, Miami, FL, USA

    Paul C. Schiller

  4. Vivex Biologics, Westview, FL, USA

    H. Thomas Temple

Authors
  1. Gaëtan J.-R. Delcroix
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  2. Amber Hackett
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  3. Paul C. Schiller
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  4. H. Thomas Temple
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Corresponding author

Correspondence to Gaëtan J.-R. Delcroix.

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Conflicts of interest

The authors have no conflict of interest to disclose. HTT is the medical director at Vivex Biologics, Inc. The other authors have no relevant financial or non-financial interests to disclose.

Human and animal Participants

All human tissues were recovered within the University of Miami Tissue Bank following rules and regulations set by the American Association for Tissue Banking (AATB).

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Delcroix, G.JR., Hackett, A., Schiller, P.C. et al. Characterization of three washing/decellularization procedures for the production of bioactive human micronized neural tissue (hMINT). Cell Tissue Bank 24, 693–703 (2023). https://doi.org/10.1007/s10561-023-10075-3

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  • DOI: https://doi.org/10.1007/s10561-023-10075-3

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