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Collagen Coating (collagen + coating)
Selected AbstractsCryopreservation of Fibroblasts Immobilized Within a Porous Scaffold: Effects of Preculture and Collagen Coating of Scaffold on Performance of Three-Dimensional CryopreservationARTIFICIAL ORGANS, Issue 7 2010Hirotoshi Miyoshi Abstract As a preliminary investigation to establish a cryopreservation method suited for bioartificial livers (BALs), three-dimensional (3-D) cryopreservation experiments with fibroblasts were performed, in which the cells were firstly seeded into a porous scaffold, and the scaffold containing the cells was then cryopreserved. After thawing, 65% of the initially applied cells were still attached to the scaffold, and this efficiency was significantly higher than that in the control experiments (39%), in which fibroblasts cryopreserved in a suspension were seeded into the scaffold. This higher efficiency was mainly caused by higher immobilization efficiency at the time of cell seeding (83%) than in the controls (54%). Collagen coating of the scaffold in the 3-D cryopreservation enhanced immobilization efficiency at the time of cell seeding, and 1-day precultures before the 3-D cryopreservation considerably improved cell growth after thawing. From these favorable results, this 3-D cryopreservation method may become useful for developing BALs. [source] Hepatic differentiation of human bone marrow-derived UE7T-13 cells: Effects of cytokines and CCN family gene expressionHEPATOLOGY RESEARCH, Issue 12 2007Takashi Shimomura Aim:, Bone marrow-derived mesenchymal stem cells (MSC) are expected to be an excellent source of cells for transplantation. We aimed to study the culture conditions and involved genes to differentiate MSC into hepatocytes. Methods:, The culture conditions to induce the efficient differentiation of human bone marrow-derived UE7T-13 cells were examined using cytokines, hormones, 5-azacytidine and type IV collagen. Results:, We found that combination of acidic fibroblast growth factor (aFGF), basic fibroblast growth factor (bFGF) and hepatocyte growth factor (HGF) with type IV collagen coating induced hepatic differentiation of UE7T-13 cells at over 30% frequency, where expression of albumin mRNA was increased over 20-fold. The differentiated cells had functions of albumin production, glycogen synthesis and urea secretion as well as expressing hepatocyte-specific genes. In addition, these cellshave binuclear and cuboidal morphology, which is a characteristic feature of hepatocytes. During hepatic differentiation, UE7T-13 cells showed depressed expression of WISP1 and WISP2 genes, members of the CCN family. Conversely, knockdown of WISP1 or WISP2 gene by siRNA stimulated hepatic differentiation. The effect of aFGF/bFGF/HGF/type IV collagen coating and WISP1-siRNA on hepatic differentiation was additive. Conclusion:, The present study suggests that aFGF/bFGF/HGF/type IV collagen coating is the efficient condition for hepatic differentiation of UE7T-13 cells, and that WISP1 and WISP2 play an important role in hepatic transdifferentiation of these cells. [source] Hepatocyte dynamics in a three-dimensional rotating bioreactorJOURNAL OF GASTROENTEROLOGY AND HEPATOLOGY, Issue 11 2007Mitsuo Miyazawa Abstract Background and Aims:, The use of an artificial liver system with extracorporeal circulation or a three-dimensional bioreactor perfused with liquid culture medium inevitably exposes hepatocytes to fluid mechanical stress (MS). The expression of liver-specific hepatocyte functions seems to be modulated by the magnitude of MS. Nonetheless, few studies have focused on the direct effects of MS on hepatocytes. We subjected hepatocytes to MS using an MS loading device and investigated the effects on the cytoskeleton and hepatocyte dynamics inside three-dimensional scaffolds by monitoring the changes in actin fiber, one of the components of the cytoskeleton. We also assessed the influence of MS on specific hepatocyte functions. Methods:, We subjected hepatocytes to MS by a rotating radial flow bioreactor (RRFB) and examined the effects by comparing the MS-loaded culture cells with cells cultured under stationary conditions without MS loading. The hepatocytes (1 × 106/cm3) were seeded on gauze without collagen coating and examined to determine morphological changes after 60 h incubation. Actin filaments in samples from the MS-loaded hepatocyte culture were stained by fluorescein isothiocyanate-labeled phalloidin. Results:, Hepatocyte aggregation was observed in the MS-loaded culture, but not in the unloaded stationary culture. Better albumin products were observed in the MS-loaded group than in the stationary culture group at all measurement points. Actin filaments extended toward the scaffold after the start of MS loading incubation and polymerized around the hepatocytes. The hepatocyte aggregation eventually advanced to the formation of spheroids. Conclusion:, These results suggest that MS-induced polymerization of actin filaments stimulate hepatocyte aggregation and thereby improve hepatocyte-specific function. [source] Effect of RGD peptide coating of titanium implants on periimplant bone formation in the alveolar crestCLINICAL ORAL IMPLANTS RESEARCH, Issue 3 2002An experimental pilot study in dogs Abstract: The aim of the present study was to analyse the effect of organic coating of titanium implants on periimplant bone formation and bone/implant contact. Three types of implants were used: (i) Ti6Al4V implants with polished surface (control 1) (ii) Ti6Al4V implants with collagen coating (control 2) (iii) Ti6Al4V implants with collagen coating and covalently bound RGD peptides. All implants had square cross-sections with an oblique diameter of 4.6 mm and were inserted press fit into trephine burr holes of 4.6 mm in the mandibles of 10 beagle dogs. The implants of five animals each were evaluated after a healing period of 1 month and 3 months, during which sequential fluorochrome labelling of bone formation was performed. Bone formation was evaluated by morphometric measurement of the newly formed bone around the implant and the percentage of implant bone contact. After 1 month there was only little bone/implant contact, varying between 2.6 and 6.7% in the cortical bone and 4.4 and 5.7% in the cancellous bone, with no significant differences between the three types of implants. After 3 months, implants with polished surfaces exhibited 26.5 and 31.2% contact in the cortical and cancellous bone, respectively, while collagen-coated implants had 19.5 and 28.4% bone contact in these areas. Implants with RGD coating showed the highest values with 42.1% and 49.7%, respectively. Differences between the surface types as such were not significant, but the increase in bone/implant contact from 1 to 3 months postoperatively was significant only in the group of RGD-coated implants (P = 0.008 and P = 0.000). The results of this pilot study thus provide only weak evidence that coating of titanium implants with RGD peptides in the present form and dosage may increase periimplant bone formation in the alveolar process. The results therefore require further verification in a modified experimental setting. Résumé Le but de l'étude présente a été d'évaluer l'effet d'un recouvrement organique des implants en titane sur la formation osseuse paro?mplantaire et le contact os/implant. Trois types d'implants ont été utilisés: 1) implants Ti6AI4V avec surface polie (contrôle 1), 2) implants Ti6AI4V avec recouvrement de collagène (contrôle 2), 3) implants Ti6aI4V avec recouvrement de collagène et des peptides RGD accrochés de manière covalente. Tous les implants avaient une section carrée avec un diamètre oblique de 4.6 mm et étaient insérés dans des trous percés à l'aide de trépans de 4.6 mm dans la mandibule de dix chiens beagle. Les implants de cinq animaux ont étéévalués après une période de guérison d'un mois et de trois mois durant laquelle un marquage fluochrome séquentiel de la néoformation osseuse a été effectué. La formation osseuse a étéévaluée par mesure morphométrique de l'os néoformé autour de l'implant et par pourcentage de contact os/implant. Après un mois, il n'y avait que peu de contact os/implant, variant entre 2.6 et 6.7% dans l'os cortical et 4.4 et 5.7% dans l'os spongieux, sans aucune différence significative entre les trois types d'implants. Après trois mois, les implants avec les surfaces polies montraient respectivement 26.5 et 31.2% d'os cortical et spongieux, tandis que les implants recouverts de collagène avaient 19.5 et 28.4% de contact osseux dans ces zones. Les implants avec le recouvrement RGD avaient les valeurs les plus importantes avec respectivement 42.1% et 49.7%. Les différences entre les types de surface n'étaient pas significatives mais l'augmentation du contact os/implant de 1 à 3 mois après l'opération n'était significative que dans le groupe RGD (P=0.008 et P=0.0000). Les résultats de cette ewtude pilote n'ont donc mis en évidence qu'une maigre preuve que les recouvrements des implants en titane par les petpides RGD sous la forme présente et le dosage présent, pouvaient augmenter la formation osseuse paro?mplantaire dans le processus alvéolaire. Ces résultats requièrent donc davantage de vérifications dans un système expérimental modifié. Zusammenfassung Es war das Ziel dieser Untersuchung, den Einfluss einer organischen Beschichtung von Titanimplantaten auf die periimplantäre Knochenformation und auf den KnochenImplantatkontakt zu analysieren. Drei Typen von Implantaten wurden verwendet: i) Ti6AI4V Implantate mit polierter Oberfläche (Kontrolle 1), ii) Ti6AI4V Implantate mit Kollagenbeschichtung (Kontrolle 2), iii) Ti6AI4V Implantate mit Kollagenbeschichtung und kovalent gebundenen RGD-Peptiden. Alle Implantate hatten einen quadratischen Querschnitt mit einem Querdurchmesser von 4.6 mm. Sie wurden in die Unterkiefer von zehn Beaglehunden in zylindrische Bohrungen mit einem Durchmesser von 4.6 mm mit Pressitz eingesetzt. Die Implantate von je funf Tieren wurden nach einer Helungszeit von einem bzw. drei Monaten ausgewertet. Während der gesamten Zeit wurden sequentielle Fluorochrommarkierungen der Knochenbildung durchgeführt. Die Knochenbildung wurde durch morphometrische Messungen des neugebildeten Knochens um die Implantate und durch Messungen der prozentualen Anteile des Knochen-/Implantatkontakts ausgewertet. Nach 1 Monat war nur wenig Knochen-/Implantatkontakt vorhanden. Er variierte zwischen 2.6 und 6.7% im kortikalen Knochen und zwischen 4.4 und 5.7% im spongiösen Knochen. Zwischen den drei Implantattypen bestanden keine signifikanten Unterschiede. Nach drei Monaten zeigten die Implantate mit polierten Oberflächen 26.5% Knochenkontakt im kortikalen bzw. 31.2% im spongiösen Knochen, während die kollagenbeschichteten Implantate 19.5% und 28.4% Knochenkontakt in diesen Regionen zeigten. Implantate mit RGD-Beschichtung zeigten mit 42.1% bzw. 49.7% die höchsten Werte. Zwischen den Oberflächentypen bestanden keine signifikanten Unterschiede. Nur in der Gruppe mit RGD-beschichteten Implantaten nahm der Knochen-/Implantatkontakt zwischen 1 bis 3 Monaten nach Chirurgie signifikant zu (P=0.008 und P=0.000). Die Resultate dieser zeigen nur mit schwacher Evidenz, dass die Beschichtung von Titanimplantaten mit RGD-Peptiden in der vorliegenden Form und Dosierung die periimplantäre Knochenformation im Alveolarkamm erhöht. Die Resultate müssen daher in einem modifizerten experimentellen Aufbau verifiziert werden. Resumen La intención del presente estudio fue analizar el efecto de la cobertura orgánica de implantes de titanio en la formación de hueso y contacto hueso/implante. Se usaron tres tipos de implantes: i) implantes de Ti6AI4V con superficie pulida (control 1), ii) implantes Ti6AI4V con cubierta de colágeno (control 2), iii) implantes de Ti6AI4V con cubierta de colágeno y péptidos RGD con unión convalente. Todos los implantes tenían secciones cuadradas con un diámetro oblicuo de 4.6 mm y se insertaron a presión en orificios trepanados de 4.6 mm en las mandíbulas de 10 perros beagle. Se evaluaron los implantes de 5 animales tras un periodo de cicatrización de un mes y tres meses los otros cinco, durante este tiempo se llevó a cabo marcado secuencial con fluorocromo de la formación de hueso. La formación de hueso se evaluó mediante mediciones morfométricas del hueso neoformado alrededor del implante y el porcentaje de contacto hueso implante. Tras 1 mes existió muy poco contacto hueso/implante variando entre 2.6 y 6.7% en el hueso cortical y 4.4 y 5.7% en el hueso esponjoso sin diferencias significativas entre los tres tipos de implantes. Tras tres meses, los implantes con superficies pulidas exhibieron un 26.5 y 31.2% en el hueso cortical y esponjoso, respectivamente, mientras que los implantes con cubiertas de colágeno tuvieron un 19.5 y un 28.4% de contacto óseo en estas áreas. Los implantes con cubiertas de RGD mostraron los valores más altos con un 42.1 y un 49.7% respectivamente. Las diferencias ente tipos de superficie como tales no fueron significativas pero el incremento de contacto hueso/implante de 1 a 3 meses tras la operación fue significativo solo en el grupo de implantes con cubierta RGD (P=0.008 y P=0.000). Los resultados de este estudio piloto aunque han mostrado solo una débil evidencia, que la cobertura de implantes de titanio con péptidos RGD en la forma y dosis presente pueden incrementar la formación de hueso periimplantario en el proceso alveolar. Los resultados, por lo tanto, requieren una posterior verificación en una situación experimental modificada. [source] | ||||||||||