RESUMEN
Xenogeneic extracellular matrices (ECMs) have been developed as off-the-shelf biologic scaffolds that have been effectively used in preclinical and clinical applications for tissue reconstruction. Such materials must be suitable for terminal sterilization and capable of storage for extended periods of time without significant changes in material properties and bioactivity. Material properties of interest for ECM scaffolds include hydrostatic permeability index (PI), uniaxial maximum load and elongation, maximum tangential stiffness (MTS), suture retention strength (SRS), and ball-burst strength (BBS). The present study evaluated these material properties for lyophilized forms of an ECM scaffold derived from the porcine urinary bladder, termed urinary bladder matrix (UBM), that was terminally sterilized by e-beam irradiation at 22 kGy and stored at room temperature (RT; 20-24 degrees C) or refrigerated temperature (REFT; 4-8 degrees C) for up to 12 months. UBM devices showed no change in SRS, BBS, and hydrostatic PI after the evaluation period. Lyophilized devices stored at RT showed an increase in maximum load and MTS while devices stored at REFT showed an increase in maximum elongation after 1 year of storage (p < 0.05). These results indicate that structural changes in the UBM device may slowly occur as a function of prolonged storage and storage temperature.
Asunto(s)
Bioprótesis , Matriz Extracelular , Ensayo de Materiales , Vejiga Urinaria , Animales , Liofilización , Porcinos , Factores de TiempoRESUMEN
Extracellular matrices (ECM) derived from porcine tissue are associated with rapid and extensive repopulation with host cells when used as scaffolds for in vivo tissue repair. Cell adhesion to substrates used for tissue engineering has been studied extensively but the factors that mediate this phenomenon in ECM scaffolds following treatment with oxidants and sterilants have not been examined. Cell adhesion assays were used to examine human microvascular endothelial cell (HMEC) attachment to ECM graft materials harvested from small intestinal submucosa (SIS) and urinary bladder matrix (UBM) following decellularization and sterilization procedures designed to render the ECM safe for clinical use. HMECs were able to attach directly to these ECM scaffolds via several attachment proteins present within the ECM, including type I collagen, type IV collagen, and fibronectin. The ability of the SIS ECM and UBM ECM to support the growth and proliferation of HMEC was also examined. HMEC were able to grow to single-layer confluence on both surfaces of SIS and UBM sheets. The endothelial cells were also able to penetrate the SIS and UBM at later time points if they were seeded on the abluminal side of the ECM sheets. The ability of the processed ECM to support HMEC attachment and proliferation is similar to that reported for unprocessed ECM and may therefore play a role in the rapid remodeling response observed when these matrices are implanted in vivo as scaffolds for wound repair.
Asunto(s)
Endotelio Vascular/citología , Endotelio Vascular/fisiología , Matriz Extracelular/fisiología , Ingeniería de Tejidos/métodos , Animales , Adhesión Celular , Colágeno Tipo I/fisiología , Colágeno Tipo IV/fisiología , Desinfección , Ácido Edético , Fibronectinas/fisiología , Humanos , Mucosa Intestinal/citología , Mucosa Intestinal/fisiología , Esterilización , Porcinos , Vejiga Urinaria/citología , Cicatrización de HeridasRESUMEN
Biologic scaffolds composed of naturally occurring extracellular matrix (ECM) are currently in clinical use for the repair and reconstruction of damaged or missing tissues. The material and structural properties of the ECM scaffold are important determinants of the potential clinical applications and these properties may be affected by manufacturing steps, processing steps, and storage conditions. The present study compared the structural properties of hydrated and lyophilized forms of a biologic scaffold derived from the porcine urinary bladder (urinary bladder matrix or UBM). The structural properties evaluated include: maximum load and elongation, maximum tangential stiffness, energy absorbed, suture retention strength, ball-burst strength, and the hydrostatic permeability index. Other properties that were investigated include changes in the water content, structural morphology, and thickness and the ability to support in vitro growth of NIH 3T3 cells. Lyophilization caused no changes in the structural properties evaluated with the exception of a decrease in maximum elongation. NIH 3T3 cells showed invasion of the scaffold when seeded on the abluminal side of both hydrated and lyophilized UBM, and there were more cells present on lyophilized UBM when compared to hydrated UBM devices after the 7-days culture period. Irreversible changes were observed in the microstructure and ultrastructure of lyophilized UBM devices. We conclude that lyophilization affects the overall in vitro cell growth of NIH 3T3 cells and the ultrastructural morphology of UBM devices, but does not result in significant changes in structural properties.