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Gene-activated fat grafts for the repair of spinal cord injury: a pilot study

  • Experimental research - Spine
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Abstract

Background

Spinal cord injury (SCI) is a complex disease requiring a concerted multi-target approach. The most appropriate combination of therapeutic gene, cellular vehicle, and space filling scaffold still has to be determined. We present an approach that employs syngeneic adipose tissue serving as a three-dimensional biological implant, source of progenitor cells, and delivery system for therapeutic genes. In this pilot experiment, we evaluated the feasibility and short-term effects using gene-activated autologous fat grafts after SCI.

Methods

An experimental SCI model was established in syngeneic Fischer 344 rats by a T9-T10 hemimyelonectomy. Fat tissue was harvested from two donor rats. Animals were divided into four groups and treated with either (i) fat grafts activated by an adenoviral vector carrying the human NT-3 cDNA, (ii) or BDNF, (iii) or with untreated fat grafts or (iv) remained untreated. Animals were euthanized either 7 or 21 days after surgery, and spinal cord tissue was investigated by histological and immunohistochemical methods.

Results

NT-3 and BDNF were produced by gene-activated fat grafts for at least 21 days in vitro and in vivo. Fat tissue grafts remained stable at the site of implantation at 7 days and at 21 days. Neither BDNF-activated nor NT-3-activated fat graft had a detectable limiting effect on the neuronal degeneration. BDNF recruited microglia to perilesional site and attenuated their inflammatory response.

Conclusions

Gene-activated syngeneic fat tissue serves as a three-dimensional biological material delivering therapeutic molecules to the site of SCI over an extended period of time. The BDNF-fat graft attenuated the inflammatory response. Whether these findings translate into functional recovery will require extended observation times.

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Authors’ contributions

VMB, KHSF, OBB, and MK designed the study. VBM, KHSF, EL, AI, CT, and MS performed experiments. VMB, KHSF, MK, OU, OBB, GS, and HZ analyzed data and wrote the manuscript. MK coordinated the study. All authors commented on and revised the manuscript.

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    Authors and Affiliations

    1. Department of Trauma and Reconstructive Surgery, University Hospital Carl Gustav Carus, Technische Universität Dresden, Dresden, Germany

      Volker M. Betz & Hans Zwipp

    2. Department of Neurosurgery, University Hospital Carl Gustav Carus, Technische Universität Dresden, Fetscherstrasse 74, 01307, Dresden, Germany

      K. Hakan Sitoci-Ficici, Ortrud Uckermann, Elke Leipnitz, Anne Iltzsche, Gabriele Schackert & Matthias Kirsch

    3. SIRION Biotech GmbH, Martinsried, Germany

      Christian Thirion & Michael Salomon

    4. Department of Orthopedic Surgery, University Hospital Grosshadern, University of Munich, Munich, Germany

      Oliver B. Betz

    5. DFG-Center for Regenerative Therapies Dresden (CRTD), Technische Universität Dresden, Dresden, Germany

      Matthias Kirsch

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    1. Volker M. Betz
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    Correspondence to Oliver B. Betz or Matthias Kirsch.

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    All authors certify that they have no affiliations with or involvement in any organization or entity with any financial interest (such as honoraria; educational grants; participation in speakers’ bureaus; membership, employment, consultancies, stock ownership, or other equity interest; and expert testimony or patent-licensing arrangements), or non-financial interest (such as personal or professional relationships, affiliations, knowledge or beliefs) in the subject matter or materials discussed in this manuscript.

    Animal experiments

    All applicable international, national, and/or institutional guidelines for the care and use of animals were followed. The study was approved by the Dresden University of Technology and the federal government of Saxony (24D-9168.11-1-20078), and all procedures involving animals were in accordance with the ethical standards of the Dresden University of Technology and the federal government of Saxony.

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    Volker M. Betz and K. Hakan Sitoci-Ficici contributed equally to this work.

    Oliver B. Betz and Matthias Kirsch are co-corresponding authors and contributed equally to this work.

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    Betz, V.M., Sitoci-Ficici, K.H., Uckermann, O. et al. Gene-activated fat grafts for the repair of spinal cord injury: a pilot study. Acta Neurochir 158, 367–378 (2016). https://doi.org/10.1007/s00701-015-2626-y

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