Gelzan cm gelrite

Methacrylated gellan gum (GG-MA) was obtained as previously described by Silva-Correia et al. [30 (link)]. Briefly, a solution of low-acyl gellan gum (Gelzan™ CM Gelrite^®, Sigma-Aldrich) reacted with glycidyl methacrylate (GMA, 97%, Sigma-Aldrich) overnight at room temperature, with constant control of pH at 8.5 and under vigorous stirring. The reaction products were precipitated by the addition of cold acetone and further purified by dialysis (cellulose membrane, molecular weight cut-off 12 kDa, Sigma-Aldrich) against distilled water, for one week. Then, the obtained GG-MA was frozen at −80 °C, freeze-dried, and the final dry material was stored protected from light, in a dry place, until further use. GG-MA solutions of desired concentrations were prepared by dissolving the dry material in Milli-Q water using gentle agitation. Manganese (II) chloride powder (MnCl₂ powder, Sigma-Aldrich) was used to prepare the MnCl₂ aqueous solutions used as a supplement for GG-MA hydrogels. Artificial cerebrospinal fluid (aCSF) was prepared following the composition listed in Table 1, and final pH adjustment to 7.3, with NaOH. For in vitro and in vivo assays, dry GG-MA was sterilized by UV light for 30 min in a laminar flow hood. All other materials and solutions were sterilized by filtration (0.22 µm filter).

Gelatin A from porcine skin, gellan gum (Gelzan CM Gelrite), adipic dihydrazide (ADH), dimethyl sulfoxide (DMSO), ethylene glycol, spermidine trihydrochloride (SPD), 1-ethyl-3-[3 dimethylamino)-propyl]–carbodiimide(EDC), hydroxylamine hydrochloride, N-hydroxybentzotriazole (HOBt), hydrochloric acid, sodium hydroxide, sodium chloride (NaCl), and sodium periodate were obtained from Sigma Aldrich. Geltrex was obtained from Thermo Fisher.