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Effects of a discoloration-resistant calcium aluminosilicate cement on the viability and proliferation of undifferentiated human dental pulp stem cells.

Niu LN, Watson D, Thames K, Primus CM, Bergeron BE, Jiao K, Bortoluzzi EA, Cutler CW, Chen JH, Pashley DH, Tay FR - Sci Rep (2015)

Bottom Line: Discoloration-resistant calcium aluminosilicate cement has been formulated to overcome the timely problem of tooth discoloration reported in the clinical application of bismuth oxide-containing hydraulic cements.The present study examined the effects of this experimental cement (Quick-Set2) on the viability and proliferation of human dental pulp stem cells (hDPSCs) by comparing the cellular responses with commercially available calcium silicate cement (white mineral trioxide aggregate; WMTA) after different aging periods.Based on these results, it is envisaged that any potential beneficial effect of the discoloration-resistant calcium aluminosilicate cement on osteogenesis by differentiated hDPSCs is more likely to be revealed after outward diffusion and removal of its cytotoxic components.

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory of Military Stomatology, School of Stomatology, Department of Prosthodontics, Fourth Military Medical University, Xi'an, Shaanxi, China.

ABSTRACT
Discoloration-resistant calcium aluminosilicate cement has been formulated to overcome the timely problem of tooth discoloration reported in the clinical application of bismuth oxide-containing hydraulic cements. The present study examined the effects of this experimental cement (Quick-Set2) on the viability and proliferation of human dental pulp stem cells (hDPSCs) by comparing the cellular responses with commercially available calcium silicate cement (white mineral trioxide aggregate; WMTA) after different aging periods. Cell viability and proliferation were examined using assays that examined plasma membrane integrity, leakage of cytosolic enzyme, caspase-3 activity for early apoptosis, oxidative stress, mitochondrial metabolic activity and intracellular DNA content. Results of the six assays indicated that both Quick-Set2 and WMTA were initially cytotoxic to hDPSCs after setting for 24 h, with Quick-Set2 being comparatively less cytotoxic than WMTA at this stage. After two aging cycles, the cytotoxicity profiles of the two hydraulic cements were not significantly different and were much less cytotoxic than the positive control (zinc oxide-eugenol cement). Based on these results, it is envisaged that any potential beneficial effect of the discoloration-resistant calcium aluminosilicate cement on osteogenesis by differentiated hDPSCs is more likely to be revealed after outward diffusion and removal of its cytotoxic components.

No MeSH data available.


Related in: MedlinePlus

Results of cell viability assays after the hDPSCs were exposed to materials derived from the 3 aging cycles.(A) Membrane integrity of hDPSCs after the cells were stained with FITC-Annexin V and ethidium homodimer III. Chart represents the percentages of healthy hDPSCs that were not stained by Annexin V and ethidium homodimer III. (B) Leakage of lactate dehydrogenase from hDPSCs that had compromised plasma membrane permeability. (C) Caspase-3 activity of hDPSCs as an indicator of cell apoptosis. (D) Expression of reactive oxygen species from hDPSCs as an indicator of intracellular oxidative stress. Statistical analyses were only conducted for hDPSCs exposed to the two hydraulic cements and unexposed hDPSCs (negative control). For the factor “material” in each chart, groups labeled with the same designators (numerals for 1st cycle, upper case letters for 2nd cycle and lower case letters for 3rd cycle) are not significantly different (P > 0.05). For the factor “aging cycle” in each chart, cycles from the same hydraulic cement that are connected with a horizontal bar are not significantly different (P > 0.05). For unexposed hDPSCs, there are no differences in activities among the 3 cycles (P > 0.05; horizontal bar not shown).
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f3: Results of cell viability assays after the hDPSCs were exposed to materials derived from the 3 aging cycles.(A) Membrane integrity of hDPSCs after the cells were stained with FITC-Annexin V and ethidium homodimer III. Chart represents the percentages of healthy hDPSCs that were not stained by Annexin V and ethidium homodimer III. (B) Leakage of lactate dehydrogenase from hDPSCs that had compromised plasma membrane permeability. (C) Caspase-3 activity of hDPSCs as an indicator of cell apoptosis. (D) Expression of reactive oxygen species from hDPSCs as an indicator of intracellular oxidative stress. Statistical analyses were only conducted for hDPSCs exposed to the two hydraulic cements and unexposed hDPSCs (negative control). For the factor “material” in each chart, groups labeled with the same designators (numerals for 1st cycle, upper case letters for 2nd cycle and lower case letters for 3rd cycle) are not significantly different (P > 0.05). For the factor “aging cycle” in each chart, cycles from the same hydraulic cement that are connected with a horizontal bar are not significantly different (P > 0.05). For unexposed hDPSCs, there are no differences in activities among the 3 cycles (P > 0.05; horizontal bar not shown).

Mentions: Cells exposed to toxic materials may result in a variety of cell fates. Depending on the toxicity level, the cells may undergo necrosis, in which they lose membrane integrity and die from cell lysis. Alternatively, the cells may activate a genetic program of controlled cell death (apoptosis). They can also stop active growth and cell division (decrease in cell viability and proliferation). In addition, cytotoxic materials may reduce intracellular production of antioxidants or enhance mitochondrial production of ROS, which, in turn, augment the level of intracellular oxidative stress that adversely affect cell proliferation. Results of the four cell viability assays are shown in Fig. 3. Figure 3A represents the percentage of healthy cells with intact plasma membranes that were present within a consortium of hDPSCs after their exposure to different materials that had been aged for 3 cycles. Two-factor repeated measures ANOVA comparing hDPSCs exposed to the two hydraulic cements with unexposed hDPSCs indicated that the type of material (P < 0.001), aging cycle (P < 0.001) and the interaction of these two factors (P < 0.001) all had significant influences on the percentage of healthy hDPSCs that were non-permeable to the two fluorescence stains for cytoplasmic membrane phospholipids and nucleic acids. Post-hoc pairwise comparisons (only comparisons with significant differences are described) showed that for the factor “aging cycle” within materials, the numbers of healthy cells in the 1st cycle of Quick-Set2 was lower than those in the subsequent two cycles. For WMTA, the number of healthy hDPSCs in each cycle was higher than the subsequent cycle, in the order: 1st cycle < 2nd cycle < 3rd cycle. For the factor “material” within the 1st cycle, the number of healthy unexposed hDPSCs was higher than those exposed to Quick-Set2 or WMTA, while the number of healthy cells in Quick-Set2 was higher than WMTA. For the factor “material” within the 2nd cycle, the number of healthy unexposed hDPSCs was higher than Quick-Set2 or WMTA. For the factor “material” within the 3rd cycle, significant difference in the number of healthy cells was only observed between unexposed hDPSCs and hDPSCs exposed to Quick-Set2.


Effects of a discoloration-resistant calcium aluminosilicate cement on the viability and proliferation of undifferentiated human dental pulp stem cells.

Niu LN, Watson D, Thames K, Primus CM, Bergeron BE, Jiao K, Bortoluzzi EA, Cutler CW, Chen JH, Pashley DH, Tay FR - Sci Rep (2015)

Results of cell viability assays after the hDPSCs were exposed to materials derived from the 3 aging cycles.(A) Membrane integrity of hDPSCs after the cells were stained with FITC-Annexin V and ethidium homodimer III. Chart represents the percentages of healthy hDPSCs that were not stained by Annexin V and ethidium homodimer III. (B) Leakage of lactate dehydrogenase from hDPSCs that had compromised plasma membrane permeability. (C) Caspase-3 activity of hDPSCs as an indicator of cell apoptosis. (D) Expression of reactive oxygen species from hDPSCs as an indicator of intracellular oxidative stress. Statistical analyses were only conducted for hDPSCs exposed to the two hydraulic cements and unexposed hDPSCs (negative control). For the factor “material” in each chart, groups labeled with the same designators (numerals for 1st cycle, upper case letters for 2nd cycle and lower case letters for 3rd cycle) are not significantly different (P > 0.05). For the factor “aging cycle” in each chart, cycles from the same hydraulic cement that are connected with a horizontal bar are not significantly different (P > 0.05). For unexposed hDPSCs, there are no differences in activities among the 3 cycles (P > 0.05; horizontal bar not shown).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4663481&req=5

f3: Results of cell viability assays after the hDPSCs were exposed to materials derived from the 3 aging cycles.(A) Membrane integrity of hDPSCs after the cells were stained with FITC-Annexin V and ethidium homodimer III. Chart represents the percentages of healthy hDPSCs that were not stained by Annexin V and ethidium homodimer III. (B) Leakage of lactate dehydrogenase from hDPSCs that had compromised plasma membrane permeability. (C) Caspase-3 activity of hDPSCs as an indicator of cell apoptosis. (D) Expression of reactive oxygen species from hDPSCs as an indicator of intracellular oxidative stress. Statistical analyses were only conducted for hDPSCs exposed to the two hydraulic cements and unexposed hDPSCs (negative control). For the factor “material” in each chart, groups labeled with the same designators (numerals for 1st cycle, upper case letters for 2nd cycle and lower case letters for 3rd cycle) are not significantly different (P > 0.05). For the factor “aging cycle” in each chart, cycles from the same hydraulic cement that are connected with a horizontal bar are not significantly different (P > 0.05). For unexposed hDPSCs, there are no differences in activities among the 3 cycles (P > 0.05; horizontal bar not shown).
Mentions: Cells exposed to toxic materials may result in a variety of cell fates. Depending on the toxicity level, the cells may undergo necrosis, in which they lose membrane integrity and die from cell lysis. Alternatively, the cells may activate a genetic program of controlled cell death (apoptosis). They can also stop active growth and cell division (decrease in cell viability and proliferation). In addition, cytotoxic materials may reduce intracellular production of antioxidants or enhance mitochondrial production of ROS, which, in turn, augment the level of intracellular oxidative stress that adversely affect cell proliferation. Results of the four cell viability assays are shown in Fig. 3. Figure 3A represents the percentage of healthy cells with intact plasma membranes that were present within a consortium of hDPSCs after their exposure to different materials that had been aged for 3 cycles. Two-factor repeated measures ANOVA comparing hDPSCs exposed to the two hydraulic cements with unexposed hDPSCs indicated that the type of material (P < 0.001), aging cycle (P < 0.001) and the interaction of these two factors (P < 0.001) all had significant influences on the percentage of healthy hDPSCs that were non-permeable to the two fluorescence stains for cytoplasmic membrane phospholipids and nucleic acids. Post-hoc pairwise comparisons (only comparisons with significant differences are described) showed that for the factor “aging cycle” within materials, the numbers of healthy cells in the 1st cycle of Quick-Set2 was lower than those in the subsequent two cycles. For WMTA, the number of healthy hDPSCs in each cycle was higher than the subsequent cycle, in the order: 1st cycle < 2nd cycle < 3rd cycle. For the factor “material” within the 1st cycle, the number of healthy unexposed hDPSCs was higher than those exposed to Quick-Set2 or WMTA, while the number of healthy cells in Quick-Set2 was higher than WMTA. For the factor “material” within the 2nd cycle, the number of healthy unexposed hDPSCs was higher than Quick-Set2 or WMTA. For the factor “material” within the 3rd cycle, significant difference in the number of healthy cells was only observed between unexposed hDPSCs and hDPSCs exposed to Quick-Set2.

Bottom Line: Discoloration-resistant calcium aluminosilicate cement has been formulated to overcome the timely problem of tooth discoloration reported in the clinical application of bismuth oxide-containing hydraulic cements.The present study examined the effects of this experimental cement (Quick-Set2) on the viability and proliferation of human dental pulp stem cells (hDPSCs) by comparing the cellular responses with commercially available calcium silicate cement (white mineral trioxide aggregate; WMTA) after different aging periods.Based on these results, it is envisaged that any potential beneficial effect of the discoloration-resistant calcium aluminosilicate cement on osteogenesis by differentiated hDPSCs is more likely to be revealed after outward diffusion and removal of its cytotoxic components.

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory of Military Stomatology, School of Stomatology, Department of Prosthodontics, Fourth Military Medical University, Xi'an, Shaanxi, China.

ABSTRACT
Discoloration-resistant calcium aluminosilicate cement has been formulated to overcome the timely problem of tooth discoloration reported in the clinical application of bismuth oxide-containing hydraulic cements. The present study examined the effects of this experimental cement (Quick-Set2) on the viability and proliferation of human dental pulp stem cells (hDPSCs) by comparing the cellular responses with commercially available calcium silicate cement (white mineral trioxide aggregate; WMTA) after different aging periods. Cell viability and proliferation were examined using assays that examined plasma membrane integrity, leakage of cytosolic enzyme, caspase-3 activity for early apoptosis, oxidative stress, mitochondrial metabolic activity and intracellular DNA content. Results of the six assays indicated that both Quick-Set2 and WMTA were initially cytotoxic to hDPSCs after setting for 24 h, with Quick-Set2 being comparatively less cytotoxic than WMTA at this stage. After two aging cycles, the cytotoxicity profiles of the two hydraulic cements were not significantly different and were much less cytotoxic than the positive control (zinc oxide-eugenol cement). Based on these results, it is envisaged that any potential beneficial effect of the discoloration-resistant calcium aluminosilicate cement on osteogenesis by differentiated hDPSCs is more likely to be revealed after outward diffusion and removal of its cytotoxic components.

No MeSH data available.


Related in: MedlinePlus