ABSTRACT
Foetal mouse cortical cells were cultured on 2D films and within 3D thermally responsive chitosan/glycerophosphate salt (GP) hydrogels. The biocompatibility of chitosan/GP 2D films was assessed in terms of cell number and neurites per cell. Osmolarity of the hydrogel was a critical factor in promoting cell survival with isotonic GP concentrations providing optimal conditions. To improve cell adhesion and neurite outgrowth, poly-D-lysine (PDL) was immobilised onto chitosan via azidoaniline photocoupling. Increase in PDL concentrations did not alter cell survival in 2D cultures but neurite outgrowth was significantly inhibited. Neurons exhibited a star-like morphology typical of 2D culture systems. The effects of PDL attachment on cell number, cell morphology and neurite outgrowth were more distinct in 3D culture conditions. Neurones exhibited larger cell bodies and sent out single neurites within the macroporous gel. Immobilised PDL improved cell survival up to an optimum concentration of 0.1%, however, further increases resulted in drops in cell number and neurite outgrowth. This was attributed to a higher cell interaction with PDL within a 3D hydrogel compared to the corresponding 2D surface. The results show that thermally responsive chitosan/GP hydrogels provide a suitable 3D scaffolding environment for neural tissue engineering.
Subject(s)
Biocompatible Materials , Chitosan/chemistry , Hydrogel, Polyethylene Glycol Dimethacrylate/chemistry , Neurons/metabolism , Polylysine/chemistry , Tissue Engineering/methods , Animals , Cell Culture Techniques/methods , Cell Survival , Glycerophosphates/pharmacology , Hot Temperature , Hydrogels , Mice , Neurites/metabolismABSTRACT
Chitosan is a well-known biomaterial that, with the addition of glycerophosphate salt (GP), gels at physiological temperatures and therefore is useful for tissue engineering purposes. This study examines the procedure of injecting chitosan/ GP to the brain in order to form a gel track. The gel system and surgical technique were successful in this endeavour; however, on examining the inflammatory response to the material it was found that the chitosan/GP was wholly engulfed by macrophages within 7 days. This was determined by staining for both the gel and the macrophages, an important technique for localising injected material. The chitosan/GP-containing macrophages formed a neat tract at the lesion site, but after 45 days no chitosan/GP was found. It was concluded that, although chitosan/GP is present after implantation, it is not available for direct scaffolding in the brain.
Subject(s)
Biocompatible Materials/adverse effects , Chitosan/adverse effects , Glycerophosphates/adverse effects , Inflammation/pathology , Animals , Biocompatible Materials/chemistry , Chitosan/administration & dosage , Chitosan/chemistry , Gels/metabolism , Glycerophosphates/administration & dosage , Glycerophosphates/chemistry , Immunohistochemistry , Inflammation/chemically induced , Injections, Intraventricular , Macrophage Activation , Macrophages/metabolism , Male , Microscopy, Electron, Scanning , Molecular Structure , Molecular Weight , Rats , Rats, Wistar , Time Factors , Tissue Engineering/methodsABSTRACT
Galactose modified xyloglucan is a thermally reversible hydrogel that is increasingly used in the biomedical field due to the ease of altering the gelation time and temperature by modifying the galactose removal ratio. However there is little information concerning the morphology and rheological properties of the hydrogel under physiological conditions. Differential scanning microcalorimetry (DSmicroC) showed the thermal gelation process to occur over a broad temperature range (5-50 degrees C). The rheological properties of the hydrogels were investigated as a function of concentration, temperature and ionic strength. The final elastic moduli of the hydrogels increased with increases in concentration. Isothermal rheology suggests that the gelation occurred in two distinct stages, which was influenced by the solution media. Scanning electron microscopy (SEM) was used to characterize the morphology of the xyloglucan which were thermally gelled at 37 degrees C. The resultant morphology was strongly dependent on the concentration of the hydrogel. Strong hydrogels were only obtained at 3 wt.% at 37 degrees C, and the morphology characterized by an open 3-dimensional network, comprised of thin membranes. It is proposed that the first stage of the isothermal gelation is the formation and growth of the thin membranes, followed by the formation of a three dimensional network.
Subject(s)
Glucans/chemistry , Hydrogels/chemistry , Xylans/chemistry , Calorimetry, Differential Scanning , Glucans/ultrastructure , Molecular Structure , Rheology , Temperature , Xylans/ultrastructureABSTRACT
The morphology of physical hydrogels is often difficult to examine due to the delicate nature of the system and therefore has not been studied in detail. Chitosan/GP (glycerophosphate salt) is a significant hydrogel in the biomedical and cosmetic fields as it is thermosensitive and contains less than 5% polysaccharide. The morphology of this system was examined with laser scanning confocal microscopy (LSCM) to image the gel morphology. The images indicate that the gel is quite heterogeneous, and power spectra reveal a fractal-like morphology. A study of composition found that increasing chitosan concentration increased the amount of polymer-rich phase present in the gel, and that the smallest aggregates decreased in size.
