IADR Abstract Archives
Viability and proliferation of mesenchymal stem cell seeded onto two xenograft scaffolds - a pilot study
Objectives: Since cell viability and proliferation onto a scaffold are critical for successful bone formation, the aim of the present study was to compare viability and proliferation of mesenchymal stem cells (MSCs) onto two commercial xenografts: Bio-Oss (BO) and Bioactive Bone Bovine BB)- a new natural mineral matrix reinforced with biodegradable synthetic polymers).
Methods: Monolayers of Bioactive Bone Bovine or Bio-Oss granules (0.25-1mm granules) were placed to cover the bottom of 96 well plates. 3X103 bone-marrow derived human MSCs suspended in DMEM were dripped (50µl) over the granules layer and incubated for 48 hours. XTT – Cell Proliferation Kit was used to quantify cell growth, viability and proliferation. This in vitro assay was used to investigate cell population’s response to the examined scaffolds. The assay was performed in triplicates after 0,2,20 and 48 hours of incubation. Absorbance of the wells containing the cells (indicating on cell metabolic activity) and the blank background control wells were investigated at 450-500 nm wavelength using a microtiter plate reader.
Results: Absorbance levels were similar among the two scaffolds at zero time point (0.268± 0.036 vs. 0.226 ± 0.017, p=0.15, BO and BB respectively). Two hours after incubation, metabolic activity was not changed compared to zero time point in the BO wells (0.277± 0.06, p=0.84). However, metabolic activity was elevated compared to zero time point in the BB (0.359± 0.06, p=0.046). A similar trend was observed after 22 hours of incubation: the metabolic activity of MSCs seeded onto BO was decreasing (0.144± 0.01, p=0.005) while cell activity was not changed in the BB group (0.289±0.1, p=0.356). Intergroup comparison revealed higher metabolic activity of MSCs seeded on BB compared to BO after 20 hours (p=0.08).
Conclusions: the results of this pilot study demonstrating different metabolic activity of MSCs seeded onto two xenograft scaffolds. Metabolic activity of the cells is an acceptable assay to evaluate cell vitality and proliferation. Based on the present study, vitality and proliferation of MSCs seeded onto BB was higher compared with BO. Furthermore, reduction in the metabolic activity of MSCs that were seeded onto BO suggest apoptosis of the cells.
Methods: Monolayers of Bioactive Bone Bovine or Bio-Oss granules (0.25-1mm granules) were placed to cover the bottom of 96 well plates. 3X103 bone-marrow derived human MSCs suspended in DMEM were dripped (50µl) over the granules layer and incubated for 48 hours. XTT – Cell Proliferation Kit was used to quantify cell growth, viability and proliferation. This in vitro assay was used to investigate cell population’s response to the examined scaffolds. The assay was performed in triplicates after 0,2,20 and 48 hours of incubation. Absorbance of the wells containing the cells (indicating on cell metabolic activity) and the blank background control wells were investigated at 450-500 nm wavelength using a microtiter plate reader.
Results: Absorbance levels were similar among the two scaffolds at zero time point (0.268± 0.036 vs. 0.226 ± 0.017, p=0.15, BO and BB respectively). Two hours after incubation, metabolic activity was not changed compared to zero time point in the BO wells (0.277± 0.06, p=0.84). However, metabolic activity was elevated compared to zero time point in the BB (0.359± 0.06, p=0.046). A similar trend was observed after 22 hours of incubation: the metabolic activity of MSCs seeded onto BO was decreasing (0.144± 0.01, p=0.005) while cell activity was not changed in the BB group (0.289±0.1, p=0.356). Intergroup comparison revealed higher metabolic activity of MSCs seeded on BB compared to BO after 20 hours (p=0.08).
Conclusions: the results of this pilot study demonstrating different metabolic activity of MSCs seeded onto two xenograft scaffolds. Metabolic activity of the cells is an acceptable assay to evaluate cell vitality and proliferation. Based on the present study, vitality and proliferation of MSCs seeded onto BB was higher compared with BO. Furthermore, reduction in the metabolic activity of MSCs that were seeded onto BO suggest apoptosis of the cells.