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Short-term effects of calcium ions on the apoptosis and onset of mineralization of human dental pulp cells in vitro and in vivo.

An S, Gao Y, Huang Y, Jiang X, Ma K, Ling J - Int. J. Mol. Med. (2015)

Bottom Line: An animal experiment and scanning electron microscopic observation of ceramic graft implants were applied to measure the mineralization in vivo.The results showed that 5.4 and 9.0 mM Ca2+ accelerated the onset of mineralized matrix nodule formation, promoted osteopontin mRNA expression and induced marked cell apoptosis and necrosis, but had no obvious effect on cell proliferation.These findings indicated a positive association between cell apoptosis and/or death and the timing of formation as well as the quantity of extracellular mineralization induced by Ca2+ in short-term cultured hDPCs.

View Article: PubMed Central - PubMed

Affiliation: Department of Operative Dentistry and Endodontics, Guanghua School of Stomatology, Hospital of Stomatology, Sun Yat‑sen University; Guangdong Provincial Key Laboratory of Stomatology, Guangzhou, Guangdong 510055, P.R. China.

ABSTRACT
Calcium ions (Ca2+) are a major constituent of most pulp-capping materials and have an important role in the mineralization of human dental pulp cells (hDPCs). A previous study by our group has shown that increased levels of Ca2+ can promote hDPC-mediated mineralization in long-term cultures (21 days). However, the initiation of mineralization occurs in the early stage of osteogenic inductive culture, and the effects of Ca2+ on the mineralization of hDPCs in short-term cultures (five days) have not been studied in detail. Furthermore, the underlying mechanism by which Ca2+ stimulates the mineralization of hDPCs has remained controversial. A strong correlation between mineralization and cell apoptosis and/or death has been identified. Thus, the present study hypothesized that Ca2+ may promote the onset of hDPC-mediated mineralization through inducing their apoptosis and/or death. To verify this hypothesis, Ca2+ was added to the growth culture medium and osteogenic culture medium at various concentrations. Alizarin Red S staining and reverse transcription-polymerase chain reaction analysis were used to evaluate the onset of mineralization. Furthermore, the cell counting kit-8 and fluorescein isothiocyanate-Annexin V/propidium iodide double-staining method were adopted to detect the proliferation and apoptosis of hDPCs in the growth culture medium. An animal experiment and scanning electron microscopic observation of ceramic graft implants were applied to measure the mineralization in vivo. The results showed that 5.4 and 9.0 mM Ca2+ accelerated the onset of mineralized matrix nodule formation, promoted osteopontin mRNA expression and induced marked cell apoptosis and necrosis, but had no obvious effect on cell proliferation. These findings indicated a positive association between cell apoptosis and/or death and the timing of formation as well as the quantity of extracellular mineralization induced by Ca2+ in short-term cultured hDPCs.

No MeSH data available.


Related in: MedlinePlus

Scanning electron microscopy images of hDPC-HA/TCP complexes harvested from SCID mice (scale bar: Top row, 100 µm; bottom row, 50 µm). In the control Ca1.8 mM HA/TCP scaffolds, a large number of hDPCs proliferated significantly and aggregated to form a dense multilayer with clumping. By contrast, hDPCs cultured in medium containing 5.4 mM Ca2+ displayed undefined cell borders and intercellular fusion, and numerous extracellular matrixes on the scaffolds’ surface were formed. hDPC; human dental pulp cell; HA/TCP, hydroxyapatite/tri-calcium phosphate.
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f6-ijmm-36-01-0215: Scanning electron microscopy images of hDPC-HA/TCP complexes harvested from SCID mice (scale bar: Top row, 100 µm; bottom row, 50 µm). In the control Ca1.8 mM HA/TCP scaffolds, a large number of hDPCs proliferated significantly and aggregated to form a dense multilayer with clumping. By contrast, hDPCs cultured in medium containing 5.4 mM Ca2+ displayed undefined cell borders and intercellular fusion, and numerous extracellular matrixes on the scaffolds’ surface were formed. hDPC; human dental pulp cell; HA/TCP, hydroxyapatite/tri-calcium phosphate.

Mentions: At 7 days following implantation of HA/TCP scaffolds containing hDPCs incubated with 5.4 mM Ca2+ into SCID mice (Fig. 5), the transplants were analyzed using SEM. In the control group, a large number of hDPCs successfully adhered to the HA/TCP scaffolds and had proliferated, resulting in a dense multilayer and spread with clumping of the cells into the three-dimensional network of the scaffold. However, the scaffolds containing hDPCs cultured in medium supplemented with 5.4 mM Ca2+ displayed intercellular fusions and produced an extracellular matrix on the surface (Fig. 6).


Short-term effects of calcium ions on the apoptosis and onset of mineralization of human dental pulp cells in vitro and in vivo.

An S, Gao Y, Huang Y, Jiang X, Ma K, Ling J - Int. J. Mol. Med. (2015)

Scanning electron microscopy images of hDPC-HA/TCP complexes harvested from SCID mice (scale bar: Top row, 100 µm; bottom row, 50 µm). In the control Ca1.8 mM HA/TCP scaffolds, a large number of hDPCs proliferated significantly and aggregated to form a dense multilayer with clumping. By contrast, hDPCs cultured in medium containing 5.4 mM Ca2+ displayed undefined cell borders and intercellular fusion, and numerous extracellular matrixes on the scaffolds’ surface were formed. hDPC; human dental pulp cell; HA/TCP, hydroxyapatite/tri-calcium phosphate.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4494572&req=5

f6-ijmm-36-01-0215: Scanning electron microscopy images of hDPC-HA/TCP complexes harvested from SCID mice (scale bar: Top row, 100 µm; bottom row, 50 µm). In the control Ca1.8 mM HA/TCP scaffolds, a large number of hDPCs proliferated significantly and aggregated to form a dense multilayer with clumping. By contrast, hDPCs cultured in medium containing 5.4 mM Ca2+ displayed undefined cell borders and intercellular fusion, and numerous extracellular matrixes on the scaffolds’ surface were formed. hDPC; human dental pulp cell; HA/TCP, hydroxyapatite/tri-calcium phosphate.
Mentions: At 7 days following implantation of HA/TCP scaffolds containing hDPCs incubated with 5.4 mM Ca2+ into SCID mice (Fig. 5), the transplants were analyzed using SEM. In the control group, a large number of hDPCs successfully adhered to the HA/TCP scaffolds and had proliferated, resulting in a dense multilayer and spread with clumping of the cells into the three-dimensional network of the scaffold. However, the scaffolds containing hDPCs cultured in medium supplemented with 5.4 mM Ca2+ displayed intercellular fusions and produced an extracellular matrix on the surface (Fig. 6).

Bottom Line: An animal experiment and scanning electron microscopic observation of ceramic graft implants were applied to measure the mineralization in vivo.The results showed that 5.4 and 9.0 mM Ca2+ accelerated the onset of mineralized matrix nodule formation, promoted osteopontin mRNA expression and induced marked cell apoptosis and necrosis, but had no obvious effect on cell proliferation.These findings indicated a positive association between cell apoptosis and/or death and the timing of formation as well as the quantity of extracellular mineralization induced by Ca2+ in short-term cultured hDPCs.

View Article: PubMed Central - PubMed

Affiliation: Department of Operative Dentistry and Endodontics, Guanghua School of Stomatology, Hospital of Stomatology, Sun Yat‑sen University; Guangdong Provincial Key Laboratory of Stomatology, Guangzhou, Guangdong 510055, P.R. China.

ABSTRACT
Calcium ions (Ca2+) are a major constituent of most pulp-capping materials and have an important role in the mineralization of human dental pulp cells (hDPCs). A previous study by our group has shown that increased levels of Ca2+ can promote hDPC-mediated mineralization in long-term cultures (21 days). However, the initiation of mineralization occurs in the early stage of osteogenic inductive culture, and the effects of Ca2+ on the mineralization of hDPCs in short-term cultures (five days) have not been studied in detail. Furthermore, the underlying mechanism by which Ca2+ stimulates the mineralization of hDPCs has remained controversial. A strong correlation between mineralization and cell apoptosis and/or death has been identified. Thus, the present study hypothesized that Ca2+ may promote the onset of hDPC-mediated mineralization through inducing their apoptosis and/or death. To verify this hypothesis, Ca2+ was added to the growth culture medium and osteogenic culture medium at various concentrations. Alizarin Red S staining and reverse transcription-polymerase chain reaction analysis were used to evaluate the onset of mineralization. Furthermore, the cell counting kit-8 and fluorescein isothiocyanate-Annexin V/propidium iodide double-staining method were adopted to detect the proliferation and apoptosis of hDPCs in the growth culture medium. An animal experiment and scanning electron microscopic observation of ceramic graft implants were applied to measure the mineralization in vivo. The results showed that 5.4 and 9.0 mM Ca2+ accelerated the onset of mineralized matrix nodule formation, promoted osteopontin mRNA expression and induced marked cell apoptosis and necrosis, but had no obvious effect on cell proliferation. These findings indicated a positive association between cell apoptosis and/or death and the timing of formation as well as the quantity of extracellular mineralization induced by Ca2+ in short-term cultured hDPCs.

No MeSH data available.


Related in: MedlinePlus