Development of 3D Printable Gelatin Methacryloyl/Chondroitin Sulfate/Hyaluronic Acid Hydrogels as Implantable Scaffolds.

The development of biomaterials tailored for various tissue engineering applications has been increasingly researched in recent years; however, stimulating cells to synthesise the extracellular matrix (ECM) is still a significant challenge. In this study, we investigate the use of ECM-like hydrogel materials composed of Gelatin methacryloyl (GelMA) and glycosaminoglycans (GAG), such as hyaluronic acid (HA) and chondroitin sulphate (CS), to provide a biomimetic environment for tissue repair. These hydrogels are fully characterised in terms of physico-chemical properties, including compression, swelling behaviour, rheological behaviour and via 3D printing trials. Furthermore, porous scaffolds were developed through freeze drying, producing a scaffold morphology that better promotes cell proliferation, as shown by in vitro analysis with fibroblast cells. We show that after cell seeding, freeze-dried hydrogels resulted in significantly greater amounts of DNA by day 7 compared to the GelMA hydrogel. Furthermore, freeze-dried constructs containing HA or HA/CS were found to have a significantly higher metabolic activity than GelMA alone.

Synthesis and evaluation of alginate, gelatin, and hyaluronic acid hybrid hydrogels for tissue engineering applications.

Tissue engineering (TE) has been proposed extensively as a potential solution to the worldwide shortages of donor organs needed for transplantation. Over the years, numerous hydrogel formulations have been studied for various TE endeavours, including bone, cardiac or neural TE treatment strategies. Amongst the materials used, organic and biocompatible materials which aim to mimic the natural extracellular matrix of the native tissue have been investigated to create biomimicry regenerative environments. As such, the comparison between studies using the same materials is often difficult to accomplish due to varying material concentrations, preparation strategies, and laboratory settings, and as such these variables have a huge impact on the physio-chemical properties of the hydrogel systems. The purpose of the current study is to investigate popular biomaterials such as alginate, hyaluronic acid and gelatin in a variety of concentrations and hydrogel formulations. This aims to provide a clear and comprehensive understanding of their behaviours and provide a rational approach as to the appropriate selection of natural polysaccharides in specific targeted TE strategies.

Electroconductive PEDOT nanoparticle integrated scaffolds for spinal cord tissue repair.

BACKGROUND: Hostile environment around the lesion site following spinal cord injury (SCI) prevents the re-establishment of neuronal tracks, thus significantly limiting the regenerative capability. Electroconductive scaffolds are emerging as a promising option for SCI repair, though currently available conductive polymers such as polymer poly(3,4-ethylenedioxythiophene) polystyrene sulfonate (PEDOT:PSS) present poor biofunctionality and biocompatibility, thus limiting their effective use in SCI tissue engineering (TE) treatment strategies. METHODS: PEDOT NPs were synthesized via chemical oxidation polymerization in miniemulsion. The conductive PEDOT NPs were incorporated with gelatin and hyaluronic acid (HA) to create gel:HA:PEDOT-NPs scaffolds. Morphological analysis of both PEDOT NPs and scaffolds was conducted via SEM. Further characterisation included dielectric constant and permittivity variances mapped against morphological changes after crosslinking, Young's modulus, FTIR, DLS, swelling studies, rheology, in-vitro, and in-vivo biocompatibility studies were also conducted. RESULTS: Incorporation of PEDOT NPs increased the conductivity of scaffolds to 8.3 × 10-4 ± 8.1 × 10-5 S/cm. The compressive modulus of the scaffold was tailored to match the native spinal cord at 1.2 ± 0.2 MPa, along with controlled porosity. Rheological studies of the hydrogel showed excellent 3D shear-thinning printing capabilities and shape fidelity post-printing. In-vitro studies showed the scaffolds are cytocompatible and an in-vivo assessment in a rat SCI lesion model shows glial fibrillary acidic protein (GFAP) upregulation not directly in contact with the lesion/implantation site, with diminished astrocyte reactivity. Decreased levels of macrophage and microglia reactivity at the implant site is also observed. This positively influences the re-establishment of signals and initiation of healing mechanisms. Observation of axon migration towards the scaffold can be attributed to immunomodulatory properties of HA in the scaffold caused by a controlled inflammatory response. HA limits astrocyte activation through its CD44 receptors and therefore limits scar formation. This allows for a superior axonal migration and growth towards the targeted implantation site through the provision of a stimulating microenvironment for regeneration. CONCLUSIONS: Based on these results, the incorporation of PEDOT NPs into Gel:HA biomaterial scaffolds enhances not only the conductive capabilities of the material, but also the provision of a healing environment around lesions in SCI. Hence, gel:HA:PEDOT-NPs scaffolds are a promising TE option for stimulating regeneration for SCI.

Printable alginate/gelatin hydrogel reinforced with carbon nanofibers as electrically conductive scaffolds for tissue engineering.

Shortages of organs and damaged tissues for transplantation have prompted improvements in biomaterials within the field of tissue engineering (TE). The rise of hybrid hydrogels as electro-conductive biomaterials offers promise in numerous challenging biomedical applications. In this work, hybrid printable biomaterials comprised of alginate and gelatin hydrogel systems filled with carbon nanofibers (CNFs) were developed to create electroconductive and printable 3-D scaffolds. Importantly, the preparation method allows the formation of hydrogels with homogenously dispersed CNFs. These hybrid composite hydrogels were evaluated in terms of mechanical, chemical and cellular response. They display excellent mechanical performance, which is augmented by the CNFs, with Young's moduli and conductivity reaching 534.7 ± 2.7 kPa and 4.1 × 10-4 ± 2 × 10-5 S/cm respectively. CNF incorporation enhances shear-thinning behaviour, allowing ease of 3-D printing. In-vitro studies indicate improved cellular proliferation compared to controls. These conductive hydrogels have the potential to be used in a myriad of TE strategies, particularly for those focused on the incorporation of electroconductive components for applications such as cardiac or neuronal TE strategies.