Macroporous Hyaluronic Acid/Chitosan Polyelectrolyte Complex-Based Hydrogels Loaded with Hydroxyapatite Nanoparticles: Preparation, Characterization and In Vitro Evaluation

• Published in: Polysaccharides Vol. 3; no. 4; pp. 745 - 760
• Main Authors: Drozdova, Maria G., Demina, Tatiana S., Dregval, Ostap A., Gaidar, Anna I., Andreeva, Elena R., Zelenetskii, Alexander N., Akopova, Tatiana A., Markvicheva, Elena
• Format: Journal Article
• Language: English
• Published: Basel MDPI AG 01.12.2022
• Subjects: Biocompatibility; Bones; Cell morphology; Cell number; Cell viability; Chitosan; composite hydrogel; Composite materials; Confocal microscopy; Cytology; Fibroblasts; human mesenchymal stromal cells; Hyaluronic acid; Hydrogels; Hydroxyapatite; hydroxyapatite nanoparticles; Ionic strength; Lasers; Localization; Mechanical loading; Mesenchymal stem cells; Mesenchyme; Morphology; mouse fibroblasts L929; Nanoparticles; Polymers; Pore size; Scanning electron microscopy; Stromal cells; Tissue engineering; United States > US; Japan; Russia
• ISSN: 2673-4176; 2673-4176
• DOI: 10.3390/polysaccharides3040043

The aim of the study was to fabricate and characterize composite macroporous hydrogels based on a hyaluronic acid/chitosan (Hyal/Ch) polyelectrolyte complex (PEC) loaded with homogeneously distributed hydroxyapatite nanoparticles (nHAp), and to evaluate them in vitro using mouse fibroblasts (L929), osteoblast-like cells (HOS) and human mesenchymal stromal cells (hMSC). Hydrogel morphology as a function of the hydroxyapatite nanoparticle content was studied using scanning electron microscopy (SEM) and confocal laser scanning microscopy (CLSM). The mean pore size in the Hyal/Ch hydrogel was 204 ± 25 μm. The entrapment of nHAp (1 and 5 wt. %) into the Hyal/Ch hydrogel led to a mean pore size decrease (94 ± 2 and 77 ± 9 μm, relatively). Swelling ratio and weight loss of the hydrogels in various aqueous media were found to increase with an enhancement of a medium ionic strength. Cell morphology and localization within the hydrogels was studied by CLSM. Cell viability depended upon the nHAp content and was evaluated by MTT-assay after 7 days of cultivation in the hydrogels. An increase of the hydroxyapatite nanoparticles loading in a range of 1–10 wt. % resulted in an enhancement of cell growth and proliferation for all hydrogels. Maximum cell viability was obtained in case of the Hyal/Ch/nHAp-10 sample (10 wt. % nHAp), while a minimal cell number was found for the Hyal/Ch/nHAp-1 hydrogel (1 wt. % nHAp). Thus, the proposed simple original technique and the design of PEC hydrogels could be promising for tissue engineering, in particular for bone tissue repair.