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Evaluation of a novel detergent-based method for decellularization of peripheral nerve allografts

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Abstract
Introduction: Within the field of peripheral nerve regeneration, there is a need for efficient alternatives to nerve autografts in cases of severe nerve damage. A promising option is the use of decellularized peripheral nerve allografts (DPNAs). These DPNAs provide a natural nerve architecture without the risk of immune response and thus create a favorable micro-environment for peripheral nerve regeneration. The aim of this study was to develop an improved DPNA and compare this to other, previously described DPNAs. Methods: Sciatic nerves of Wistar rats were harvested and randomly divided into four groups. Group A contained native nerves; group B consisted of nerves decellularized by Triton X-100 and sodium deoxycholate as described by Sondell et al.; nerves in group C were decellularized with sulfobetaine (SB)-10 and SB-16, based on the protocol of Hudson et al. and finally, nerves in group D were decellularized with Triton X- 100 and enzymes as recently described by Roosens et al. Histological, biochemical and tensile analyzes were carried out to evaluate the impact of the different detergent treatments on tissue structure and extracellular matrix (ECM) composition. Results: While nerves of group B seemed to have undergone severe structural damage with loss of glycosaminoglycans, the nerves of group C had better preservation of the ECM, but were not completely cell-free. On the other hand, nerves of group D showed a good balance between conservation of the nerve structure and removal of cellular components. Interestingly, HE and DAPI staining confirmed the absence of nuclear remnants in the nerves decellularized in group D. In addition, important ECM components such as laminin and collagen I were well preserved, maintaining a homogeneous tissue distribution. Discussion: Finally, Roosens’ combined decellularization method showed superior results in the decellularization of rat sciatic nerves as compared to Sondell’s and Hudson’s methods. However, in order to determine the potential clinical usefulness of these decellularized nerves, it is still necessary to perform future in vivo studies.
Keywords
neural tissue engineering, decellularization techniques, allograft, histological techniques, biochemical methods, biomechanical evaluation

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Chicago
Philips, Charlot, Fernando Campos, Annelies Roosens, Maria Cornelissen, and Víctor Carriel. 2017. “Evaluation of a Novel Detergent-based Method for Decellularization of Peripheral Nerve Allografts.” In eCM Conferences Open Access Online Periodical.
APA
Philips, Charlot, Campos, F., Roosens, A., Cornelissen, M., & Carriel, V. (2017). Evaluation of a novel detergent-based method for decellularization of peripheral nerve allografts. eCM Conferences Open Access Online Periodical. Presented at the European Chapter Meeting of the Tissue Engineering and Regenerative Medicine International Society 2017.
Vancouver
1.
Philips C, Campos F, Roosens A, Cornelissen M, Carriel V. Evaluation of a novel detergent-based method for decellularization of peripheral nerve allografts. eCM Conferences Open Access Online Periodical. 2017.
MLA
Philips, Charlot, Fernando Campos, Annelies Roosens, et al. “Evaluation of a Novel Detergent-based Method for Decellularization of Peripheral Nerve Allografts.” eCM Conferences Open Access Online Periodical. 2017. Print.
@inproceedings{8551207,
  abstract     = {Introduction: Within the field of peripheral nerve regeneration, there is a need for efficient alternatives to nerve autografts in cases of severe nerve damage. A promising option is the use of decellularized peripheral nerve allografts (DPNAs). These DPNAs provide a natural nerve architecture without the risk of immune response and thus create a favorable micro-environment for peripheral nerve regeneration. The aim of this study was to develop an improved DPNA and compare this to other, previously described DPNAs.
Methods: Sciatic nerves of Wistar rats were harvested and randomly divided into four groups. Group A contained native nerves; group B consisted of nerves decellularized by Triton X-100 and sodium deoxycholate as described by Sondell et al.; nerves in group C were decellularized with sulfobetaine (SB)-10 and SB-16, based on the protocol of Hudson et al. and finally, nerves in group D were decellularized with Triton X- 100 and enzymes as recently described by Roosens et al. Histological, biochemical and tensile analyzes were carried out to evaluate the impact of the different detergent treatments on tissue structure and extracellular matrix (ECM) composition.
Results: While nerves of group B seemed to have undergone severe structural damage with loss of glycosaminoglycans, the nerves of group C had better preservation of the ECM, but were not completely cell-free. On the other hand, nerves of group D showed a good balance between conservation of the nerve structure and removal of cellular components. Interestingly, HE and DAPI staining confirmed the absence of nuclear remnants in the nerves decellularized in group D. In addition, important ECM components such as laminin and collagen I were well preserved, maintaining a homogeneous tissue distribution.
Discussion: Finally, Roosens{\textquoteright} combined decellularization method showed superior results in the decellularization of rat sciatic nerves as compared to Sondell{\textquoteright}s and Hudson{\textquoteright}s methods. However, in order to determine the potential clinical usefulness of these decellularized nerves, it is still necessary to perform future in vivo studies.},
  articleno    = {abstract P811},
  author       = {Philips, Charlot and Campos, Fernando and Roosens, Annelies and Cornelissen, Maria and Carriel, V{\'i}ctor},
  booktitle    = {eCM Conferences Open Access Online Periodical},
  issn         = {2522-235X},
  keyword      = {neural tissue engineering,decellularization techniques,allograft,histological techniques,biochemical methods,biomechanical evaluation},
  language     = {eng},
  location     = {Davos, Switzerland},
  title        = {Evaluation of a novel detergent-based method for decellularization of peripheral nerve allografts},
  year         = {2017},
}