Chitosan-g-oligo(L,L-lactide) copolymer hydrogel for nervous tissue regeneration in glutamate excitotoxicity: in vitro feasibility evaluation

Abstract

Over the last decade, a number of hydrogels attracted great attention in the area of brain tissue engineering. The hydrogels are composed of hydrophilic polymers forming 3D network in water. Their function is promoting structural and functional restoration of damaged brain tissues by providing mechanical support and navigating cell fate. This paper reports on the neurocompatibility of chitosan-g-oligo(L,L-lactide) copolymer hydrogel with primary rat cortical neuron culture. The hydrogel was produced by a molding technique on the base of photocurable composition consisting of chitosan-g-oligo(L,L-lactide) copolymer, poly(ethylene glycol) diacrylate and photosensitizer Irgacure 2959. The influence of the hydrogel on cell viability, phenotype and calcium homeostasis, mitochondrial potential and oxygen consumption rate in glutamate excitotoxicity was analyzed using primary neuron cultures obtained from a neonatal rat cortex. This study revealed that the hydrogel is non-cytotoxic. Dissociated neonatal rat cortical cells were actively attaching to the hydrogel surface and exhibited the phenotype, calcium homeostasis and mitochondrial function in both standard conditions and glutamate excitotoxicity (100 μM) similar to the control cells cultured without the hydrogel. To conclude, in this study we assessed the feasibility of the application of chitosan-g-oligo(L,L-lactide) copolymer hydrogel for tissue engineering therapy of brain injury in an in vitro model. The results support that the hydrogel is able to sustain realization of the functional metabolic activity of neonatal rat cortical cells in response to glutamate excitotoxicity.

Viscoelasticity and Volume of Cortical Neurons under Glutamate Excitotoxicity and Osmotic Challenges

“…Glu was added in a concentration of 50 mM together with 10 mM of glycine in the N-buffer in which Mg 2þ was eliminated. This Glu concentration, as was established in the previous works on primary neuron cultures (23,25,26,(30)(31)(32), results in a fast and pronounced increase in the concentration of calcium and sodium in the soma of neurons and a high cytotoxicity level. After the Glu treatment, the neurons were washed with a nominally Ca 2þ -free N-buffer containing 2 mM MgCl 2 and 100 mM EGTA.…”

“…199n of 04/01/2016 and approved by the Ethics Committee of National Medical Research Center for Children's Health of the Russian Federation Ministry of Health. Primary cultures of rat brain cortical neurons were prepared from the cortex of 1-or 2-day-old Wistar rats according to the previously described protocol (25,26). Briefly, the rats were anesthetized and decapitated, and the cortex was removed and separated from the meninges.…”

Acute and Delayed Effects of Mechanical Injury on Calcium Homeostasis and Mitochondrial Potential of Primary Neuroglial Cell Culture: Potential Causal Contributions to Post-Traumatic Syndrome

“…This effect may be related, at least partially, to the primary cell culture maturation. Many studies used a primary neuronal cortical culture no older than 9–15 DIV [ 19 , 20 , 21 , 22 , 23 , 24 ], which were different from the long-term neural cultures differentiated from induced pluripotent stem cells [ 25 , 26 ].…”

Solid-State Synthesis of Water-Soluble Chitosan-g-Hydroxyethyl Cellulose Copolymers

“…Other forms of scaffolds for tissue engineering (such as hydrogels) are of great importance and open for polysaccharide-based materials. Among a variety of polymers used for the fabrication of hydrogels for neuroregeneration, chitosan and cellulose derivatives are widely recognized [38,39].…”