IADR Abstract Archives
Therapeutic Ultrasound on Proliferation, Desinflamatory Effect and Differentiation in Culture-hDPSC
Objectives: To evaluate the in vitro effects of therapeutic ultrasound (TU) on proliferation, desinflamatory effect in culture of human dental pulp stem cells (hDPSC) and differentiation to muscle in a co-culture model of hDSPC-skeletal muscle (C1C12).
Methods: hDPSC culture were isolated and seeded under international guidelines. The hDPSC were subcultured and TU were applied in different modes [1Mhz, (Mode,W/cm2, minutes); G1: Continuous/0.5/5; G2: Continuous/0.5/10; G3: Continuous/1.0/5; G4: Continuous/1.0/10; G5: Pulsed/0.5/5; G6: Pulsed/0.5/10; G7: Pulsed/1.0/5; G8: Pulsed/1.0/10; GC: control group)] and incubated during 48 hours, cell proliferation were determined by MTT method. hDPSC were induced to a proinflammatory state with IL-1 beta huma, G8 parameters were applied and cuantificated the expresión of PGE2 by ELISA assay. A co-culture of hDPSC and C2C12 were cultured in specific differentiation media and exposed to G8 parameters, skeletal muscle differentiation potential was evaluated by morphological characteristics with fluorescence microscopy and with inmunihistoshamestry. The assay was conducted in three independent experiments (n=9) and data were analyzed by Shapiro-Wilk normality test and ANOVA post-hoc de Tukey test.
Results: hDPSC showed fibroblast morphology, attachment to the plate, differentiation to chondrogenic, adipogenic and osteogenic lineage, positive markers to CD105+, CD90+, CD73+ and negatives to CD14-, CD34-, CD45-. TU treatment was significant increase (p<0.05) in cell proliferation (%) in order: GC=100±14, G8=141±21< G5=139±33< G7=217±20< G4=121±25< G6=118±32< G2=97±28< G1=88±25< G3=23±8. TU reduced the expression of PGE2 with G8 parameters and enhanced muscle differentiation in the co-culture showing morphological characteristic and positive immunochemistry by integration in muscular fibres.
Conclusions: TU have an in vitro proliferative and desinflamatory effects on hDPSC and increased and promote the skeletal muscle differentiation moreover hDPSC represent a potential source of stem cells alone and together with TU an exceptional option for wound healing process.
Methods: hDPSC culture were isolated and seeded under international guidelines. The hDPSC were subcultured and TU were applied in different modes [1Mhz, (Mode,W/cm2, minutes); G1: Continuous/0.5/5; G2: Continuous/0.5/10; G3: Continuous/1.0/5; G4: Continuous/1.0/10; G5: Pulsed/0.5/5; G6: Pulsed/0.5/10; G7: Pulsed/1.0/5; G8: Pulsed/1.0/10; GC: control group)] and incubated during 48 hours, cell proliferation were determined by MTT method. hDPSC were induced to a proinflammatory state with IL-1 beta huma, G8 parameters were applied and cuantificated the expresión of PGE2 by ELISA assay. A co-culture of hDPSC and C2C12 were cultured in specific differentiation media and exposed to G8 parameters, skeletal muscle differentiation potential was evaluated by morphological characteristics with fluorescence microscopy and with inmunihistoshamestry. The assay was conducted in three independent experiments (n=9) and data were analyzed by Shapiro-Wilk normality test and ANOVA post-hoc de Tukey test.
Results: hDPSC showed fibroblast morphology, attachment to the plate, differentiation to chondrogenic, adipogenic and osteogenic lineage, positive markers to CD105+, CD90+, CD73+ and negatives to CD14-, CD34-, CD45-. TU treatment was significant increase (p<0.05) in cell proliferation (%) in order: GC=100±14, G8=141±21< G5=139±33< G7=217±20< G4=121±25< G6=118±32< G2=97±28< G1=88±25< G3=23±8. TU reduced the expression of PGE2 with G8 parameters and enhanced muscle differentiation in the co-culture showing morphological characteristic and positive immunochemistry by integration in muscular fibres.
Conclusions: TU have an in vitro proliferative and desinflamatory effects on hDPSC and increased and promote the skeletal muscle differentiation moreover hDPSC represent a potential source of stem cells alone and together with TU an exceptional option for wound healing process.