Soft hydrazone crosslinked hyaluronan- and alginate-based hydrogels as 3D supportive matrices for human pluripotent stem cell-derived neuronal cells

Abstract

Regenerative medicine, especially cell therapy combined with a supportive biomaterial scaffold, is considered to be a potential treatment for various deficits in humans. Here, we have produced and investigated the detailed properties of injectable hydrazone crosslinked hyaluronanpolyvinyl alcohol (HA-PVA) and alginate-polyvinyl alcohol (AL-PVA) hydrogels to be used as a supportive biomaterial for 3D neural cell cultures. To the best of our knowledge, this is the first time the polymerization and properties of hydrazone crosslinked AL-PVA hydrogel have been reported. The effect of the degree of substitution and molecular weight of the polymer components as well as the polymer concentration of the hydrogel on the swelling, degradation and mechanical properties of the hydrogels is reported. Furthermore, we studied the effect of the above parameters on the growth of human pluripotent stem cell-derived neuronal cells. The most neural cell supportive HA-PVA hydrogel was composed of high molecular weight HA component with brain-mimicking mechanical properties and decreased polymer concentration. AL-PVA hydrogel, with stiffness quite similar to brain tissue, was also shown to be similarly supportive. Neuronal spreading and 3D network formation was enhanced inside the softest hydrogels.

Screening of Hydrogels for Human Pluripotent Stem Cell–Derived Neural Cells: Hyaluronan‐Polyvinyl Alcohol‐Collagen‐Based Interpenetrating Polymer Network Provides an Improved Hydrogel Scaffold

“…As the stiffness of the 3D scaffold affects neuronal differentiation and maturation, using scaffolds with stiffness values near those of natural brain tissue is reasonable. Mechanical tests performed with the compression method showed that all the studied hydrogels had compression moduli under 10 kPa, which is in the range of brain tissue . The highest measured modulus of the hydrogels used in PCA analysis, shown in Figure 4 , was the modulus of GG (1.25% SPD) at 9.4 ± 2.7 kPa.…”

“…Aldehyde groups were introduced to HA either by using periodate oxidation (HA1) or selective oxidation of diol‐modified HA (HA2). The syntheses and determinations of the degree of substitution (DS%) of modified HA1 and HA2 components were carried out according to previously reported procedures . Hydrazide groups were introduced to PVA using glycine ethyl ester and hydrazine as a source of the hydrazide unit.…”

“…Hydrazide groups were introduced to PVA using glycine ethyl ester and hydrazine as a source of the hydrazide unit. The synthesis and determination of the DS% of the modified PVA component were carried out according to the previously reported procedure . The modified components are presented in Table 1 .…”

“…Two types of HA‐PVA hydrogels (HA1‐PVA and HA2‐PVA) were prepared as previously described . Freeze‐dried HA1 or HA2 were dissolved in 10% sucrose to a concentration of 20 mg mL −1 , and PVA was dissolved to a final concentration of 10 mg mL −1 .…”

Glycosaminoglycan-Inspired Biomaterials for the Development of Bioactive Hydrogel Networks

“…Different types of GAG-based biomaterials have been described and characterized as potential candidates for biomedical applications, particularly for cartilage [37,38] and neural [39,40] tissues. While non-modified GAG can be used [41,42], most strategies resort to chemical modifications to add specific moieties to GAG.…”