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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

Fluorescent microscopic evaluation of the plasma membrane integrity and oxidative stress after the hDPSCs were exposed to materials derived from the 1st and 3rd aging cycle.(A) Fluorescent microscopy images of hDPSCs that were triple-stained with Hoechst 33342 (blue fluorescent nuclear counterstain), ethidium homodimer III (red fluorescent non-vital DNA dye) and FITC-Annexin V (green fluorescent phosphatidylserine-binding cytoplasmic dye). Healthy cell nuclei were stained blue. Apoptotic cells showed green cytoplasm and blue nuclei. Necrotic cells showed red or pink nuclei. Dead cells progressing from the apoptotic cell population were stained green, red and blue. Bars = 25 μm. (B) Fluorescent microscopy images of hDPSCs double-stained with CellROX® Orange and Hoechst 33342. Cytoplasms of cells exhibiting oxidative stress were stained orange. Bars = 50 μm.
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f4: Fluorescent microscopic evaluation of the plasma membrane integrity and oxidative stress after the hDPSCs were exposed to materials derived from the 1st and 3rd aging cycle.(A) Fluorescent microscopy images of hDPSCs that were triple-stained with Hoechst 33342 (blue fluorescent nuclear counterstain), ethidium homodimer III (red fluorescent non-vital DNA dye) and FITC-Annexin V (green fluorescent phosphatidylserine-binding cytoplasmic dye). Healthy cell nuclei were stained blue. Apoptotic cells showed green cytoplasm and blue nuclei. Necrotic cells showed red or pink nuclei. Dead cells progressing from the apoptotic cell population were stained green, red and blue. Bars = 25 μm. (B) Fluorescent microscopy images of hDPSCs double-stained with CellROX® Orange and Hoechst 33342. Cytoplasms of cells exhibiting oxidative stress were stained orange. Bars = 50 μm.

Mentions: Images acquired by fluorescent microscopy were complementary of the flow cytometry results (Fig. 4a). Unexposed hDPSCs from the 1st or 3rd cycle were predominantly healthy and exhibited blue-fluorescent nuclei with minimal signs of apoptosis or necrosis. Cells exposed to the IRM positive control were mostly apoptotic or necrotic, with prevalence of green fluorescent cytoplasm that could be attributed to apoptosis. Occasionally, the cells exhibited partially red-fluorescent cytoplasm (caused by leaching of nucleic acid components) or pink nuclei (merging of blue and red fluorescence) that are characteristic of necrosis or cell death progressing from apoptosis. Stem cells that were exposed to Quick-Set2 or WMTA in the 1st cycle were predominantly healthy; nevertheless, cells with green-fluorescent cytoplasm could be observed. The number of apoptotic cells that exhibited green-fluorescent cytoplasm was substantially reduced after hDPSCs were exposed to Quick-Set2 or WMTA from the 3rd cycle.


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)

Fluorescent microscopic evaluation of the plasma membrane integrity and oxidative stress after the hDPSCs were exposed to materials derived from the 1st and 3rd aging cycle.(A) Fluorescent microscopy images of hDPSCs that were triple-stained with Hoechst 33342 (blue fluorescent nuclear counterstain), ethidium homodimer III (red fluorescent non-vital DNA dye) and FITC-Annexin V (green fluorescent phosphatidylserine-binding cytoplasmic dye). Healthy cell nuclei were stained blue. Apoptotic cells showed green cytoplasm and blue nuclei. Necrotic cells showed red or pink nuclei. Dead cells progressing from the apoptotic cell population were stained green, red and blue. Bars = 25 μm. (B) Fluorescent microscopy images of hDPSCs double-stained with CellROX® Orange and Hoechst 33342. Cytoplasms of cells exhibiting oxidative stress were stained orange. Bars = 50 μm.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

f4: Fluorescent microscopic evaluation of the plasma membrane integrity and oxidative stress after the hDPSCs were exposed to materials derived from the 1st and 3rd aging cycle.(A) Fluorescent microscopy images of hDPSCs that were triple-stained with Hoechst 33342 (blue fluorescent nuclear counterstain), ethidium homodimer III (red fluorescent non-vital DNA dye) and FITC-Annexin V (green fluorescent phosphatidylserine-binding cytoplasmic dye). Healthy cell nuclei were stained blue. Apoptotic cells showed green cytoplasm and blue nuclei. Necrotic cells showed red or pink nuclei. Dead cells progressing from the apoptotic cell population were stained green, red and blue. Bars = 25 μm. (B) Fluorescent microscopy images of hDPSCs double-stained with CellROX® Orange and Hoechst 33342. Cytoplasms of cells exhibiting oxidative stress were stained orange. Bars = 50 μm.
Mentions: Images acquired by fluorescent microscopy were complementary of the flow cytometry results (Fig. 4a). Unexposed hDPSCs from the 1st or 3rd cycle were predominantly healthy and exhibited blue-fluorescent nuclei with minimal signs of apoptosis or necrosis. Cells exposed to the IRM positive control were mostly apoptotic or necrotic, with prevalence of green fluorescent cytoplasm that could be attributed to apoptosis. Occasionally, the cells exhibited partially red-fluorescent cytoplasm (caused by leaching of nucleic acid components) or pink nuclei (merging of blue and red fluorescence) that are characteristic of necrosis or cell death progressing from apoptosis. Stem cells that were exposed to Quick-Set2 or WMTA in the 1st cycle were predominantly healthy; nevertheless, cells with green-fluorescent cytoplasm could be observed. The number of apoptotic cells that exhibited green-fluorescent cytoplasm was substantially reduced after hDPSCs were exposed to Quick-Set2 or WMTA from the 3rd cycle.

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