In Vivo and In Vitro Evaluation of a Novel Hyaluronic Acid–Laminin Hydrogel as Luminal Filler and Carrier System for Genetically Engineered Schwann Cells in Critical Gap Length Tubular Peripheral Nerve Graft in Rats
In the current study we investigated the suitability of a novel hyaluronic acid–laminin hydrogel (HAL) as luminal filler and carrier system for co-transplanted cells within a composite chitosan-based nerve graft (CNG) in a rat critical nerve defect model. The HAL was meant to improve the performance...
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doaj-9c13f93bd8a046479b406624743d75542020-11-25T03:01:39ZengSAGE PublishingCell Transplantation1555-38922020-03-012910.1177/0963689720910095In Vivo and In Vitro Evaluation of a Novel Hyaluronic Acid–Laminin Hydrogel as Luminal Filler and Carrier System for Genetically Engineered Schwann Cells in Critical Gap Length Tubular Peripheral Nerve Graft in RatsNina Dietzmeyer0Zhong Huang1Tobias Schüning2Shimon Rochkind3Mara Almog4Zvi Nevo5Thorsten Lieke6Svenja Kankowski7Kirsten Haastert-Talini8 Center for Systems Neuroscience, Hannover, Germany Center for Systems Neuroscience, Hannover, Germany Center for Systems Neuroscience, Hannover, Germany Research Center for Nerve Reconstruction, Department of Neurosurgery, Tel-Aviv Sourasky Medical Center, Tel Aviv University, Tel Aviv, Israel Research Center for Nerve Reconstruction, Department of Neurosurgery, Tel-Aviv Sourasky Medical Center, Tel Aviv University, Tel Aviv, Israel Department of Human Molecular Genetics and Biochemistry, Sackler School of Medicine, Tel Aviv University, Tel Aviv, Israel Transplant Laboratory, Department of General-, Visceral-, and Transplantation Surgery, Hannover Medical School, Hannover, Germany Institute of Neuroanatomy and Cell Biology, Hannover Medical School, Hannover, Germany Center for Systems Neuroscience, Hannover, GermanyIn the current study we investigated the suitability of a novel hyaluronic acid–laminin hydrogel (HAL) as luminal filler and carrier system for co-transplanted cells within a composite chitosan-based nerve graft (CNG) in a rat critical nerve defect model. The HAL was meant to improve the performance of our artificial nerve guides by giving additional structural and molecular support to regrowing axons. We filled hollow CNGs or two-chambered nerve guides with an inserted longitudinal chitosan film (CNG[F]s), with cell-free HAL or cell-free HA or additionally suspended either naïve Schwann cells (SCs) or fibroblast growth factor 2-overexpressing Schwann cells (FGF2-SCs) within the gels. We subjected female Lewis rats to immediate 15 mm sciatic nerve gap reconstruction and comprehensively compared axonal and functional regeneration parameters with the gold standard autologous nerve graft (ANG) repair. Motor recovery was surveyed by means of electrodiagnostic measurements at 60, 90, and 120 days post-reconstruction. Upon explantation after 120 days, lower limb target muscles were harvested for calculation of muscle-weight ratios. Semi-thin cross-sections of nerve segments distal to the grafts were evaluated histomorphometrically. After 120 days of recovery, only ANG treatment led to full motor recovery. Surprisingly, regeneration outcomes revealed no regeneration-supportive effect of HAL alone and even an impairment of peripheral nerve regeneration when combined with SCs and FGF2-SCs. Furthermore, complementary in vitro studies, conducted to elucidate the reason for this unexpected negative result, revealed that SCs and FGF2-SCs suspended within the hydrogel relatively downregulated gene expression of regeneration-supporting neurotrophic factors. In conclusion, cell-free HAL in its current formulation did not qualify for optimizing regeneration outcome through CNG[F]s. In addition, we demonstrate that our HAL, when used as a carrier system for co-transplanted SCs, changed their gene expression profile and deteriorated the pro-regenerative milieu within the nerve guides.https://doi.org/10.1177/0963689720910095 |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Nina Dietzmeyer Zhong Huang Tobias Schüning Shimon Rochkind Mara Almog Zvi Nevo Thorsten Lieke Svenja Kankowski Kirsten Haastert-Talini |
spellingShingle |
Nina Dietzmeyer Zhong Huang Tobias Schüning Shimon Rochkind Mara Almog Zvi Nevo Thorsten Lieke Svenja Kankowski Kirsten Haastert-Talini In Vivo and In Vitro Evaluation of a Novel Hyaluronic Acid–Laminin Hydrogel as Luminal Filler and Carrier System for Genetically Engineered Schwann Cells in Critical Gap Length Tubular Peripheral Nerve Graft in Rats Cell Transplantation |
author_facet |
Nina Dietzmeyer Zhong Huang Tobias Schüning Shimon Rochkind Mara Almog Zvi Nevo Thorsten Lieke Svenja Kankowski Kirsten Haastert-Talini |
author_sort |
Nina Dietzmeyer |
title |
In Vivo and In Vitro Evaluation of a Novel Hyaluronic Acid–Laminin Hydrogel as Luminal Filler and Carrier System for Genetically Engineered Schwann Cells in Critical Gap Length Tubular Peripheral Nerve Graft in Rats |
title_short |
In Vivo and In Vitro Evaluation of a Novel Hyaluronic Acid–Laminin Hydrogel as Luminal Filler and Carrier System for Genetically Engineered Schwann Cells in Critical Gap Length Tubular Peripheral Nerve Graft in Rats |
title_full |
In Vivo and In Vitro Evaluation of a Novel Hyaluronic Acid–Laminin Hydrogel as Luminal Filler and Carrier System for Genetically Engineered Schwann Cells in Critical Gap Length Tubular Peripheral Nerve Graft in Rats |
title_fullStr |
In Vivo and In Vitro Evaluation of a Novel Hyaluronic Acid–Laminin Hydrogel as Luminal Filler and Carrier System for Genetically Engineered Schwann Cells in Critical Gap Length Tubular Peripheral Nerve Graft in Rats |
title_full_unstemmed |
In Vivo and In Vitro Evaluation of a Novel Hyaluronic Acid–Laminin Hydrogel as Luminal Filler and Carrier System for Genetically Engineered Schwann Cells in Critical Gap Length Tubular Peripheral Nerve Graft in Rats |
title_sort |
in vivo and in vitro evaluation of a novel hyaluronic acid–laminin hydrogel as luminal filler and carrier system for genetically engineered schwann cells in critical gap length tubular peripheral nerve graft in rats |
publisher |
SAGE Publishing |
series |
Cell Transplantation |
issn |
1555-3892 |
publishDate |
2020-03-01 |
description |
In the current study we investigated the suitability of a novel hyaluronic acid–laminin hydrogel (HAL) as luminal filler and carrier system for co-transplanted cells within a composite chitosan-based nerve graft (CNG) in a rat critical nerve defect model. The HAL was meant to improve the performance of our artificial nerve guides by giving additional structural and molecular support to regrowing axons. We filled hollow CNGs or two-chambered nerve guides with an inserted longitudinal chitosan film (CNG[F]s), with cell-free HAL or cell-free HA or additionally suspended either naïve Schwann cells (SCs) or fibroblast growth factor 2-overexpressing Schwann cells (FGF2-SCs) within the gels. We subjected female Lewis rats to immediate 15 mm sciatic nerve gap reconstruction and comprehensively compared axonal and functional regeneration parameters with the gold standard autologous nerve graft (ANG) repair. Motor recovery was surveyed by means of electrodiagnostic measurements at 60, 90, and 120 days post-reconstruction. Upon explantation after 120 days, lower limb target muscles were harvested for calculation of muscle-weight ratios. Semi-thin cross-sections of nerve segments distal to the grafts were evaluated histomorphometrically. After 120 days of recovery, only ANG treatment led to full motor recovery. Surprisingly, regeneration outcomes revealed no regeneration-supportive effect of HAL alone and even an impairment of peripheral nerve regeneration when combined with SCs and FGF2-SCs. Furthermore, complementary in vitro studies, conducted to elucidate the reason for this unexpected negative result, revealed that SCs and FGF2-SCs suspended within the hydrogel relatively downregulated gene expression of regeneration-supporting neurotrophic factors. In conclusion, cell-free HAL in its current formulation did not qualify for optimizing regeneration outcome through CNG[F]s. In addition, we demonstrate that our HAL, when used as a carrier system for co-transplanted SCs, changed their gene expression profile and deteriorated the pro-regenerative milieu within the nerve guides. |
url |
https://doi.org/10.1177/0963689720910095 |
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