TY - JOUR
T1 - Feasible stabilization of chondroitinase abc enables reduced astrogliosis in a chronic model of spinal cord injury
AU - Raspa, Andrea
AU - Bolla, Edoardo
AU - Cuscona, Claudia
AU - Gelain, Fabrizio
N1 - Funding Information:
We thank Drs. M.Copetti and A. Fontana for their helpful assistance in statistical analysis. This work was supported in part by Fondazione Cariplo (No. 2011-0352), by the “Ricerca Corrente 2017” funding granted by the Italian Ministry of Health, by Revert Onlus, by the “5 × 1000” voluntary contributions and by Fondazione Banca Del Monte di Lombardia.
Publisher Copyright:
© 2018 John Wiley & Sons Ltd
Copyright:
Copyright 2019 Elsevier B.V., All rights reserved.
PY - 2019/1
Y1 - 2019/1
N2 - Aims: Usually, spinal cord injury (SCI) develops into a glial scar containing extracellular matrix molecules including chondroitin sulfate proteoglycans (CSPGs). Chondroitinase ABC (ChABC), from Proteus vulgaris degrading the glycosaminoglycan (GAG) side chains of CSPGs, offers the opportunity to improve the final outcome of SCI. However, ChABC usage is limited by its thermal instability, requiring protein structure modifications, consecutive injections at the lesion site, or implantation of infusion pumps. Methods: Aiming at more feasible strategy to preserve ChABC catalytic activity, we assessed various stabilizing agents in different solutions and demonstrated, via a spectrophotometric protocol, that the 2.5 mol/L Sucrose solution best stabilized ChABC as far as 14 days in vitro. Results: ChABC activity was improved in both stabilizing and diluted solutions at +37°C, that is, mimicking their usage in vivo. We also verified the safety of the proposed aqueous sucrose solution in terms of viability/cytotoxicity of mouse neural stem cells (NSCs) in both proliferating and differentiating conditions in vitro. Furthermore, we showed that a single intraspinal treatment with ChABC and sucrose reduced reactive gliosis at the injury site in chronic contusive SCI in rats and slightly enhanced their locomotor recovery. Conclusion: Usage of aqueous sucrose solutions may be a feasible strategy, in combination with rehabilitation, to ameliorate ChABC-based treatments to promote the regeneration of central nervous system injuries.
AB - Aims: Usually, spinal cord injury (SCI) develops into a glial scar containing extracellular matrix molecules including chondroitin sulfate proteoglycans (CSPGs). Chondroitinase ABC (ChABC), from Proteus vulgaris degrading the glycosaminoglycan (GAG) side chains of CSPGs, offers the opportunity to improve the final outcome of SCI. However, ChABC usage is limited by its thermal instability, requiring protein structure modifications, consecutive injections at the lesion site, or implantation of infusion pumps. Methods: Aiming at more feasible strategy to preserve ChABC catalytic activity, we assessed various stabilizing agents in different solutions and demonstrated, via a spectrophotometric protocol, that the 2.5 mol/L Sucrose solution best stabilized ChABC as far as 14 days in vitro. Results: ChABC activity was improved in both stabilizing and diluted solutions at +37°C, that is, mimicking their usage in vivo. We also verified the safety of the proposed aqueous sucrose solution in terms of viability/cytotoxicity of mouse neural stem cells (NSCs) in both proliferating and differentiating conditions in vitro. Furthermore, we showed that a single intraspinal treatment with ChABC and sucrose reduced reactive gliosis at the injury site in chronic contusive SCI in rats and slightly enhanced their locomotor recovery. Conclusion: Usage of aqueous sucrose solutions may be a feasible strategy, in combination with rehabilitation, to ameliorate ChABC-based treatments to promote the regeneration of central nervous system injuries.
KW - axonal regeneration
KW - chondroitinase ABC
KW - chronic spinal cord injury
KW - locomotor rehabilitation
KW - thermal stabilization
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U2 - 10.1111/cns.12984
DO - 10.1111/cns.12984
M3 - Article
C2 - 29855151
AN - SCOPUS:85047794110
VL - 25
SP - 86
EP - 100
JO - CNS Neuroscience and Therapeutics
JF - CNS Neuroscience and Therapeutics
SN - 1755-5930
IS - 1
ER -