Enzymatically crosslinked tyramine-gellan gum hydrogels as drug delivery system for rheumatoid arthritis treatment.

Rheumatoid arthritis (RA) is a chronic inflammatory disease characterized by joint synovial inflammation, as well as cartilage and bone tissue destruction. Current strategies for the treatment of RA can reduce joint inflammation, but the treatment options still represent stability concerns since they are not sufficient and present a fast clearing. Thus, several drug delivery systems (DDS) have been advanced to tackle this limitation. Injectable gellan gum (GG) hydrogels, reduced by physical crosslinking methods, also being proposed as DDS, but this kind of crosslinking can produce hydrogels that become weaker in physiological conditions. Nevertheless, enzymatic crosslinking emerged as an alternative to increase mechanical strength, which can be adjusted by the degree of enzymatic crosslinking. In this study, tyramine-modified gellan gum (Ty-GG) hydrogels were developed via horseradish peroxidase (HRP) crosslinking; and betamethasone was encapsulated within, to increase the specificity and safety in the treatment of patients with RA. Physicochemical results showed that it was possible to modify GG with tyramine, with a degree of substitution of approximately 30%. They showed high mechanical strength and resistance, presenting a controlled betamethasone release profile over time. Ty-GG hydrogels also exhibited no cytotoxic effects and do not negatively affected the metabolic activity and proliferation of chondrogenic primary cells. Furthermore, the main goal was achieved since betamethasone-loaded Ty-GG hydrogels demonstrated to have a more effective therapeutic effect when compared with the administration of betamethasone alone. Therefore, the developed Ty-GG hydrogels represent a promising DDS and a reliable alternative to traditional treatments in patients with RA.

Eggshell Membrane/Gellan Gum Composite Hydrogels with Increased Degradability, Biocompatibility, and Anti-Swelling Properties for Effective Regeneration of Retinal Pigment Epithelium.

A gellan gum (GG) hydrogel must demonstrate a number of critical qualities-low viscosity, degradability, desirable mechanical properties, anti-swelling properties, and biocompatibility-in order to be regarded as suitable for retinal pigment epithelium (RPE) regeneration. In this study, we investigated whether the application of an eggshell membrane (ESM) to a GG hydrogel improved these critical attributes. The crosslinking of the ESM/GG hydrogels was most effectively reduced, when a 4 w/v% ESM was used, leading to a 40% less viscosity and a 30% higher degradation efficiency than a pure GG hydrogel. The compressive moduli of the ESM/GG hydrogels were maintained, as the smaller pores formed by the addition of the ESM compensated for the slightly weakened mechanical properties of the ESM/GG hydrogels. Meanwhile, due to the relatively low hydrophilicity of ESM, a 4 w/v% ESM enabled an ESM/GG hydrogel to swell 30% less than a pure GG hydrogel. Finally, the similarity in components between the ESM and RPE cells facilitated the proliferation of the latter without any significant cytotoxicity.

Anti-Inflammatory Properties of Injectable Betamethasone-Loaded Tyramine-Modified Gellan Gum/Silk Fibroin Hydrogels.

Rheumatoid arthritis is a rheumatic disease for which a healing treatment does not presently exist. Silk fibroin has been extensively studied for use in drug delivery systems due to its uniqueness, versatility and strong clinical track record in medicine. However, in general, natural polymeric materials are not mechanically stable enough, and have high rates of biodegradation. Thus, synthetic materials such as gellan gum can be used to produce composite structures with biological signals to promote tissue-specific interactions while providing the desired mechanical properties. In this work, we aimed to produce hydrogels of tyramine-modified gellan gum with silk fibroin (Ty-GG/SF) via horseradish peroxidase (HRP), with encapsulated betamethasone, to improve the biocompatibility and mechanical properties, and further increase therapeutic efficacy to treat rheumatoid arthritis (RA). The Ty-GG/SF hydrogels presented a ß-sheet secondary structure, with gelation time around 2-5 min, good resistance to enzymatic degradation, a suitable injectability profile, viscoelastic capacity with a significant solid component and a betamethasone-controlled release profile over time. In vitro studies showed that Ty-GG/SF hydrogels did not produce a deleterious effect on cellular metabolic activity, morphology or proliferation. Furthermore, Ty-GG/SF hydrogels with encapsulated betamethasone revealed greater therapeutic efficacy than the drug applied alone. Therefore, this strategy can provide an improvement in therapeutic efficacy when compared to the traditional use of drugs for the treatment of rheumatoid arthritis.

Accelerating bone defects healing in calvarial defect model using 3D cultured bone marrow-derived mesenchymal stem cells on demineralized bone particle scaffold.

Bone defects are usually difficult to be regenerated due to pathological states or the size of the injury. Researchers are focusing on tissue engineering approaches in order to drive the regenerative events, using stem cells to regenerate bone. The purpose of this study is to evaluate the osteogenic differentiation of bone marrow-derived mesenchymal stem cells (BMSCs) on biologically derived Gallus gallus domesticus-derived demineralized bone particle (GDD) sponge. The sponges were prepared by freeze-drying method using 1, 2, and 3 wt% GDD and cross-linked with glutaraldehyde. The GDD sponge was characterized using scanning electron microscopy, compressive strength, porosity, and Fourier transform infrared. The potential bioactivity of the sponge was evaluated by osteogenic differentiation of BMSCs using 3(4, dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide assay and quantifying alkaline phosphatase (ALP) activity. in vivo experiments were evaluated through a micro-computerized tomography (µ-CT) and histological assays. The analysis confirmed that an increase in the concentration of the GDD in the sponge leads to a higher bone formation and deposition in rat calvarial defects. Histological assay results were in line with µ-CT. The results reported in this study demonstrated the potential application of GDD sponges as osteoinductor in bone tissue engineering in pathological or nonunion bone defects.

Evaluation of cartilage regeneration of chondrocyte encapsulated gellan gum-based hyaluronic acid blended hydrogel.

Hydrogels have shown to be advantageous in supporting damaged cartilage because of its analogous to the extracellular matrix (ECM) of cartilage tissue. However, problems such as infection and inflammation are still a challenge to be solved. In terms of tissue engineering, natural materials are more advantageous than synthetic materials in biocompatibility and biodegradability status. Herein, physically blended nature-derived gellan gum (GG) hydrogel and hyaluronic acid (HA) hydrogel is suggested as a one of solution for cartilage tissue engineering material. The purpose of this study is to determine the effect of GG/HA hydrogel in vitro and in vivo. The chemical and mechanical properties were measured to confirm the compatibility of hydrogels for cartilage tissue engineering. The viability, proliferation, morphology, and gene expression of chondrocytes encapsulated in hydrogels were examined in vitro. Furthermore, the beneficial effect of the blended hydrogel was confirmed by performing the in vivo experiment. The chemical properties of hydrogels confirmed the well physically blended hydrogels. The mechanical studies of hydrogels displayed that as the content of HA increases, the swelling ratio was higher, compressive strength decreased and degradation was faster. Therefore, to use the hydrogel of GG and HA network, the proper amount must be blended. The in vitro study of chondrocytes encapsulated GG/HA hydrogel showed that the proper amount of HA enhanced the cell growth, attachment, and gene expression. The in vivo examination verified the advantageous effect of GG/HA hydrogel. Overall results demonstrate that GG/HA hydrogel is suitable for culturing chondrocyte and can be further applied for the treatment of cartilage defects.

Injectable taurine-loaded alginate hydrogels for retinal pigment epithelium (RPE) regeneration.

The purpose of this study is to produce injectable taurine (Tr)-loaded alginate (Agn) hydrogel for age-related macular degeneration (AMD) treatment by inducing the regeneration of RPE (retinal pigment epithelium) cells. Porosity and swelling ratio were measured to evaluate the mechanical properties of the hydrogels, and Fourier transform infrared spectroscopy (FTIR) was used to evaluate the physical and chemical properties. RPE cells extracted from the pigmented epithelium of rabbits were encapsulated in the Tr/Agn hydrogels. Cells proliferation and migration were improved in Tr/Agn hydrogels with an enhanced expression of RPE-specific genes including RPE65, CRALBP, NPR-A, MITF and collagen type I and II. In vivo tests demonstrated the excellent biocompatibility and biodegradability without inflammatory response by the host when implanted with the hydrogel. Moreover, when the Tr/Agn hydrogels were injected into the sub-retinal space, high adhesion of RPE cells and retinal regeneration were confirmed. These results demonstrated a potential role of injectable Tr/Agn hydrogels as potential therapeutic tools for the treatment of retinal diseases, including AMD.

Enhanced retinal pigment epithelium (RPE) regeneration using curcumin/alginate hydrogels: In vitro evaluation.

The retinal pigment epithelium (RPE) plays a significant role in retaining structural integrity of eye. Factors such as reduction in cell regeneration due to aging and physical injury pose a major hurdle in RPE regeneration. In this study, we exploited the use of alginate (AGT) incorporated with Curcumin (CCI) forming a hydrogel based system CCI/AGT. The fabricated hydrogel could anchor RPE cell in it. In vitro cell analysis revealed that the CCI/AGT hydrogel shows good biocompatibility, enhanced cell growth ability and higher ECM formation compared to the pure AGT hydrogel. In particular, the presence of CCI in the hydrogels enhances the cells proliferation of the 23% respect to the pure alginate. Also the expression of crucial genes for retina functions and matrix production were positively affected by CCI presence, with an increment of 45% for RPE65, 32% for CRALBP and 26% for Collagen type 1. In vitro tests demonstrated the potential application of CCI/AGT hydrogels for transplantation under the sub-retinal space acting as a cell delivery vehicle and also their capability to provide an appropriate environment for RPE regeneration. These results suggest that CCI/AGT hydrogel could be translated into a potential surgical graft for biological implantation of retinal tissue engineering.