Modified gellan gum-based hydrogel with enhanced mechanical properties for application as a cell carrier for cornea endothelial cells.

Recently, the number of people suffering from visual loss due to eye diseases is increasing rapidly around the world. However, due to the severe donor shortage and the immune response, corneal replacement is needed. Gellan gum (GG) is biocompatible and widely used for cell delivery or drug delivery, but its strength is not suitable for the corneal substitute. In this study, a GM hydrogel was prepared by blending methacrylated gellan gum with GG (GM) to give suitable mechanical properties to the corneal tissue. In addition, lithium phenyl-2,4,6-trimethylbenzoylphosphinate (LAP), a crosslinking initiator, was added to the GM hydrogel. After the photo-crosslinking treatment, it was named GM/LAP hydrogel. GM and GM/LAP hydrogels were analyzed for physicochemical properties, mechanical characterization, and transparency tests to confirm their applicability as carriers for corneal endothelial cells (CEnCs). Also, in vitro studies were performed with cell viability tests, cell proliferation tests, cell morphology, cell-matrix remodeling analysis, and gene expression evaluation. The compressive strength of the GM/LAP hydrogel was improved compared to the GM hydrogel. The GM/LAP hydrogel showed excellent cell viability, proliferation, and cornea-specific gene expression than the GM hydrogel. Crosslinking-improved GM/LAP hydrogel can be applied as a promising cell carrier in corneal tissue engineering.

Preparation and characterization of a soluble eggshell membrane/agarose composite scaffold with possible applications in cartilage regeneration.

Articular hyaline cartilage is an extremely hydrated, not vascularized tissue with a low-cell density. The damage of this tissue can occur after injuries or gradual stress and tears (osteoarthritis), minor damages can be self-healed in several weeks, but major injuries may eventually require surgery. In fact, in this case, because of nature of the cartilage (the absence of cells and vascularization) it is difficult to expect its natural regeneration in a reasonable amount of time. In recent years, cell therapy, in which cells are directly transplanted, has attracted attention. In this study, a scaffold for implanting chondrocytes was prepared. The scaffold was made as a sponge using the eggshell membrane and agarose. The eggshell membrane is structurally similar to the extracellular matrix and nontoxic due to its many collagen components and has good biocompatibility and biodegradability. However, scaffolds made of collagen only has poor mechanical properties. For this reason, the disulfide bond of collagen extracted from the insoluble eggshell membrane was cut, converted into water-soluble, and then mixed with agarose to prepare a scaffold. Agarose is capable of controlling mechanical properties, has excellent biocompatibility, and is suitable for forming a hydrogel having a three-dimensional porosity. The scaffold was examined for Fourier-transform infrared, mechanical properties, biodegradability, and biocompatibility. In in vitro experiment, cytotoxicity, cell proliferation, and messenger RNA expression were investigated. The study demonstrated that the agarose/eggshell membrane scaffold can be used for chondrocyte transplantation.

Dopamine-Functionalized Gellan Gum Hydrogel as a Candidate Biomaterial for a Retinal Pigment Epithelium Cell Delivery System.

In this study, dopamine-functionalized gellan gum (DFG) hydrogel was prepared as a carrier for retinal pigment epithelium (RPE) cell delivery via a carbodiimide reaction. The carboxylic acid of gellan gum (GG) was replaced with catechol in a 21.3% yield, which was confirmed by NMR. Sol fraction and weight loss measurements revealed that dopamine improved degradability in the GG hydrogel. Measurements of the viscosity, injection force, and compressibility also showed that dopamine-functionalized GG hydrogels had more desirable rheological/mechanical properties for improving injectability. These characteristics were confirmed to arise from the GG's helix structure loosened by the dopamine's bulky nature. Moreover, dopamine's hydrophilic characteristics were confirmed to create a more favorable microenvironment for cell growth by promoting swelling capability and cell attachment. This improved biocompatibility became more pronounced when the hydrophilicity of dopamine was combined with a larger specific surface area stemming from the less porous structure of the dopamine-grafted hydrogels. This effect was apparent from the live/dead staining images of the as-prepared hydrogels. Meanwhile, the nonionic cross-linked DFG (DG) hydrogel showed the lowest protein expression in the immunofluorescence staining images obtained after 28 days of culture, supporting that it had the highest degradability and associated cell-releasing ability. That tendency was also observed in the gene expression data acquired by real-time polymerase chain reaction (RT-PCR) analysis. RT-PCR analysis also revealed that the DG hydrogel carrier could upregulate the visual function-related gene of RPE. Overall, the DG hydrogel system demonstrated its feasibility as a carrier of RPE cells and its potential as a means of improving visual function.