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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

Immunophenotypic and multipotency characteristics of hDPSCs.(A) Immunophenotyping of hDPSCs using FITC dye-conjugated antibodies to identify different Cluster of Differentiation (CD) cell-surface molecules. FITC dye-conjugated mouse IgM antibody was used as isotype control. The weak staining of the isotype IgM control antibody is indicative of the specificity of binding of the CD antibodies. (B) Cartilage extracellular matrix formed by the differentiated cells after incubation in chondrogenic medium (stained with Alcian blue). (C) Intracellular lipid vacuoles that were found in differentiated cells after incubation in adipogenic medium (stained with Oil Red O). (D) Mineralized nodules formed by differentiated cells after incubation in osteogenic medium (stained with Alizarin red S). For (B–D)bar = 50 μm.
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f2: Immunophenotypic and multipotency characteristics of hDPSCs.(A) Immunophenotyping of hDPSCs using FITC dye-conjugated antibodies to identify different Cluster of Differentiation (CD) cell-surface molecules. FITC dye-conjugated mouse IgM antibody was used as isotype control. The weak staining of the isotype IgM control antibody is indicative of the specificity of binding of the CD antibodies. (B) Cartilage extracellular matrix formed by the differentiated cells after incubation in chondrogenic medium (stained with Alcian blue). (C) Intracellular lipid vacuoles that were found in differentiated cells after incubation in adipogenic medium (stained with Oil Red O). (D) Mineralized nodules formed by differentiated cells after incubation in osteogenic medium (stained with Alizarin red S). For (B–D)bar = 50 μm.

Mentions: Consistent with other mesenchymal stem cell populations, the majority of hDPSCs exhibited intense expressions of mesenchymal surface molecular markers (CD29–98.9%, CD44–98.5%, CD90–99.5% and CD105–96.4%). The hDPSCs also exhibited weak expressions of surface markers for hematopoietic system-derived cells (CD34–0.9% and CD45–0.9%) (Fig. 2A). Weak staining of the mouse IgM isotype control antibody confirmed the specificity of primary antibody binding (Fig. 2A). The multipotent nature of the hDPSCs was confirmed by the observation that these cells have the potential to develop into chondrocytes after chondrogenic induction, with the formation of a cartilaginous-like extracellular matrix (Fig. 2B). Following adipogenic induction, intracellular lipid vacuoles could be identified within the differentiated adipocytes (Fig. 2C). Mineralized nodules that were stained with Alizarin red S appeared in the extracellular milieu after the hDPSCs were cultured in osteogenic differentiation medium (Fig. 2D).


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)

Immunophenotypic and multipotency characteristics of hDPSCs.(A) Immunophenotyping of hDPSCs using FITC dye-conjugated antibodies to identify different Cluster of Differentiation (CD) cell-surface molecules. FITC dye-conjugated mouse IgM antibody was used as isotype control. The weak staining of the isotype IgM control antibody is indicative of the specificity of binding of the CD antibodies. (B) Cartilage extracellular matrix formed by the differentiated cells after incubation in chondrogenic medium (stained with Alcian blue). (C) Intracellular lipid vacuoles that were found in differentiated cells after incubation in adipogenic medium (stained with Oil Red O). (D) Mineralized nodules formed by differentiated cells after incubation in osteogenic medium (stained with Alizarin red S). For (B–D)bar = 50 μm.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

f2: Immunophenotypic and multipotency characteristics of hDPSCs.(A) Immunophenotyping of hDPSCs using FITC dye-conjugated antibodies to identify different Cluster of Differentiation (CD) cell-surface molecules. FITC dye-conjugated mouse IgM antibody was used as isotype control. The weak staining of the isotype IgM control antibody is indicative of the specificity of binding of the CD antibodies. (B) Cartilage extracellular matrix formed by the differentiated cells after incubation in chondrogenic medium (stained with Alcian blue). (C) Intracellular lipid vacuoles that were found in differentiated cells after incubation in adipogenic medium (stained with Oil Red O). (D) Mineralized nodules formed by differentiated cells after incubation in osteogenic medium (stained with Alizarin red S). For (B–D)bar = 50 μm.
Mentions: Consistent with other mesenchymal stem cell populations, the majority of hDPSCs exhibited intense expressions of mesenchymal surface molecular markers (CD29–98.9%, CD44–98.5%, CD90–99.5% and CD105–96.4%). The hDPSCs also exhibited weak expressions of surface markers for hematopoietic system-derived cells (CD34–0.9% and CD45–0.9%) (Fig. 2A). Weak staining of the mouse IgM isotype control antibody confirmed the specificity of primary antibody binding (Fig. 2A). The multipotent nature of the hDPSCs was confirmed by the observation that these cells have the potential to develop into chondrocytes after chondrogenic induction, with the formation of a cartilaginous-like extracellular matrix (Fig. 2B). Following adipogenic induction, intracellular lipid vacuoles could be identified within the differentiated adipocytes (Fig. 2C). Mineralized nodules that were stained with Alizarin red S appeared in the extracellular milieu after the hDPSCs were cultured in osteogenic differentiation medium (Fig. 2D).

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