Andrea Raspa1, Edoardo Bolla2, Claudia Cuscona2, Fabrizio Gelain1,2. 1. Opera di San Pio da Pietrelcina, IRCCS Casa Sollievo della Sofferenza, San Giovanni Rotondo (FG), Italy. 2. Center for Nanomedicine and Tissue Engineering (CNTE), A.O. Ospedale Niguarda Cà Granda, Piazza dell'Ospedale Maggiore, Milan, Italy.
Abstract
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.
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.
Authors: Katalin Bartus; Nicholas D James; Athanasios Didangelos; Karen D Bosch; Joost Verhaagen; Rafael J Yáñez-Muñoz; John H Rogers; Bernard L Schneider; Elizabeth M Muir; Elizabeth J Bradbury Journal: J Neurosci Date: 2014-04-02 Impact factor: 6.167
Authors: Shashank Kosuri; Carlos H Borca; Heloise Mugnier; Matthew Tamasi; Roshan A Patel; Isabel Perez; Suneel Kumar; Zachary Finkel; Rene Schloss; Li Cai; Martin L Yarmush; Michael A Webb; Adam J Gormley Journal: Adv Healthc Mater Date: 2022-02-21 Impact factor: 11.092