RESUMO
Currently, there is a lack of models representing the skin dermal heterogeneity for relevant research and skin engineering applications. This is the first study reporting production of dermal equivalents reproducing features of papillary and reticular dermal compartments. Inspired from our current knowledge on the architecture and composition differences between the papillary and reticular dermis, we evaluated different collagen-based porous materials to serve as scaffolds for the three-dimensional expansion of freshly isolated papillary and/or reticular fibroblasts. The scaffolds, composed of either collagen I or collagen I and III mixtures, were prepared by lyophilization. Pore size and hydrolytic stability were controlled by crosslinking with 1-ethyl-3-(3-dimethyl aminopropyl) carbodiimide (EDC) and N-hydroxysuccinimide (NHS) or EDC/NHS with covalently bound heparin. The evaluation of the resultant "papillary" and "reticular" dermal equivalents was based on the analysis of characteristic features of each dermal compartment, such as cell density and deposition of newly synthetized extracellular matrix components in histological sections. Crosslinking supported cell growth during dermal tissue formation independent on the fibroblast subpopulation. The presence of collagen III seemed to have some positive but non-specific effect only on the maintenance of the mechanical strength of the scaffolds during dermal formation. Histological analyses demonstrated a significant and specific effect of heparin on generating dermal equivalents reproducing the respective higher papillary than reticular cell densities and supporting distinct extracellular matrix components deposition (three to five times more carbohydrate material deposited by papillary fibroblasts in all scaffolds containing heparin, while higher collagen production was observed only in the presence of heparin).
Assuntos
Derme , Heparina , Colágeno/metabolismo , Colágeno Tipo I/metabolismo , Derme/patologia , Fibroblastos/metabolismo , Heparina/farmacologia , Humanos , Engenharia Tecidual/métodos , Alicerces TeciduaisRESUMO
OBJECTIVE: To study the aging of neat resins, prepared from bis-GMA, UDMA, D(3)MA or a mixture of bis-GMA/UDMA/D(3)MA (65/20/15 w/w/w), in water or 75% (v/v) ethanol/water 37 degrees C. Also the study of aging of Heliomolar RO, which is a radiopaque, microfilled, light cured composite, the resin matrix of which is the copolymer of bis-GMA/UDMA/D(3)MA (65/20/15 w/w/w). METHODS: Samples of neat resins and Heliomolar RO prepared by light curing were immersed in water or 75% (v/v) ethanol/water 37 degrees C, for 1, 7 or 30 days. Then the flexural and tensile strength were determined. The fractured surface of samples after the flexural tests was observed by scanning electron microscopy. RESULTS: Bis-GMA and copolymer resin did not showed any significant change in mechanical properties after immersion in water or 75% (v/v) ethanol/water 37 degrees C, for 30 days. On the contrary UDMA, D(3)MA and the composite Heliomolar RO showed a significant decrease. SIGNIFICANCE: The results obtained showed that the effect of aging in water or ethanol/water solution on mechanical properties of a light cured dimethacrylate resin depends on the chemical structure of resin. In the case of resin composite this effect depends on the filler-matrix bond strength.