Preparation of hybrid meniscal constructs using hydrogels and acellular matrices

Abstract

To search for a suitable meniscus repair material, acellular hybrid scaffolds consisting of in situ cross-linkable 3-D interpenetrating network structures were obtained by decellularization of the meniscus tissues followed by integration of the gel system. Decellularization efficiency was confirmed using a DNA quantification assay (82% decrease in DNA content) and histological stainings. In the second part of the study, the gelatin molecule was functionalized by adding methacrylic anhydride and the degree of functionalization was found to be 75% by (Proton-Nuclear Magnetic Resonance) 1H-NMR. Using this, a series of hybrid constructs named GelMA-Hybrid (G-Hybrid), GELMA/PEGDMA-Hybrid (PG-Hybrid), and GelMA/PEGDMA/HAMA-Hybrid (PGH-Hybrid) were prepared by cross-linking with UVA. Changes in the chemical structure were determined with Fourier Transform Infrared Spectrophotometer (FTIR). Water uptake capacities of cross-linked hybrid structures were measured in swelling studies, and it was found that hybrid scaffolds showed similar swelling properties compared to native counterparts. By compressive mechanical tests, enhanced mechanical properties were revealed in cross-linked scaffolds with PGH-Hybrid having the highest cross-link density. Protein denaturation and decomposition transition temperatures were improved by adding hydrogels to acellular scaffolds according to thermal gravimetric analyses (TGA). Cross-linked acellular scaffolds have exhibited a behavior close to native tissues with below 25% mass loss in phosphate buffer saline (PBS) and enzymatic solution. Cell viability was examined through Alamar Blue on the first day and cell viability in hybrid constructs was found to be above 80% while it was closer to the control group on the 7th day. It was concluded that the developed biomaterials could be used in meniscus tissue engineering with their tunable physicochemical and mechanical properties.