Composite Hydrogels Based on Cross-Linked Chitosan and Low Molecular Weight Hyaluronic Acid for Tissue Engineering

The objectives of the study were as follows: (1) to develop two methods for the preparation of macroporous composite chitosan/hyaluronic acid (Ch/HA) hydrogels based on covalently crosslinked Ch and low molecular weight (Mw) HA (5 and 30 kDa); (2) to investigate some properties (swelling and in vitro degradation) and structures of the hydrogels; (3) to evaluate the hydrogels in vitro as potential biodegradable matrices for tissue engineering. Chitosan was cross-linked with either genipin (Gen) or glutaraldehyde (GA). Method 1 allowed the distribution of HA macromolecules within the hydrogel (bulk modification). In Method 2, hyaluronic acid formed a polyelectrolyte complex with Ch over the hydrogel surface (surface modification). By varying compositions of the Ch/HA hydrogels, highly porous interconnected structures (with mean pore sizes of 50-450 mu m) were fabricated and studied using confocal laser scanning microscopy (CLSM). Mouse fibroblasts (L929) were cultured in the hydrogels for 7 days. Cell growth and proliferation within the hydrogel samples were studied via MTT-assay. The entrapment of low molecular weight HA was found to result in an enhancement of cell growth in the Ch/HA hydrogels compared to that in the Ch matrices. The Ch/HA hydrogels after bulk modification promoted better cell adhesion, growth and proliferation than the samples prepared by using Method 2 (surface modification).

Automated summary

The study aimed to develop two methods for preparing macroporous composite chitosan/hyaluronic acid (Ch/HA) hydrogels and evaluate their potential as biodegradable matrices for tissue engineering. The hydrogels were prepared by crosslinking chitosan with genipin or glutaraldehyde, and the distribution of hyaluronic acid within the hydrogel was controlled by different modification methods. The results showed that the Ch/HA hydrogels with low molecular weight HA exhibited enhanced cell growth compared to the Ch matrices, and the hydrogels prepared by bulk modification promoted better cell adhesion, growth, and proliferation.