Limits...
Diverse effects of lead nitrate on the proliferation, differentiation, and gene expression of stem cells isolated from a dental origin.

Abdullah M, Rahman FA, Gnanasegaran N, Govindasamy V, Abu Kasim NH, Musa S - ScientificWorldJournal (2014)

Bottom Line: We found that Pb(2+) treatment altered the morphology and adhesion of the cells in a dose-dependent manner.There were no significant changes in terms of cell surface phenotypes.Gene expression studies revealed a constant expression of key markers associated with stemness (Oct 4, Rex 1) and DNA repair enzyme markers, but downregulation occurred with some ectoderm and endoderm markers, demonstrating an irregular and untimely differentiation trail.

View Article: PubMed Central - PubMed

Affiliation: Department of Conservative Dentistry, Faculty of Dentistry, University of Malaya, 50603 Kuala Lumpur, Malaysia.

ABSTRACT
Lead (Pb(2+)) exposure continues to be a significant public health problem. Therefore, it is vital to have a continuous epidemiological dataset for a better understanding of Pb(2+) toxicity. In the present study, we have exposed stem cells isolated from deciduous and permanent teeth, periodontal ligament, and bone marrow to five different types of Pb(2+) concentrations (160, 80, 40, 20, and 10 µM) for 24 hours to identify the adverse effects of Pb(2+) on the proliferation, differentiation, and gene expression on these cell lines. We found that Pb(2+) treatment altered the morphology and adhesion of the cells in a dose-dependent manner. There were no significant changes in terms of cell surface phenotypes. Cells exposed to Pb(2+) continued to differentiate into chondrogenesis and adipogenesis, and a severe downregulation was observed in osteogenesis. Gene expression studies revealed a constant expression of key markers associated with stemness (Oct 4, Rex 1) and DNA repair enzyme markers, but downregulation occurred with some ectoderm and endoderm markers, demonstrating an irregular and untimely differentiation trail. Our study revealed for the first time that Pb(2+) exposure not only affects the phenotypic characteristics but also induces significant alteration in the differentiation and gene expression in the cells.

Show MeSH

Related in: MedlinePlus

(a) Conventional qualitative SA-β-gal assay by X-gal staining of bone marrow mesenchymal stem cells (BM-MSCs), deciduous (SCDs), permanent (DPSCs), and periodontal ligament (PDLs) upon exposure to various concentrations of Pb2+. A locally region of senescence cell is shown; (b) quantification of percentage (%) of SA-β-gal positive cells exposed to various concentrations of Pb2+. In all experiments, the results represent average of five culture replicates with standard deviation and a representative photomicrograph was given for each experiment.
© Copyright Policy - open-access
Related In: Results  -  Collection


getmorefigures.php?uid=PMC3927845&req=5

fig2: (a) Conventional qualitative SA-β-gal assay by X-gal staining of bone marrow mesenchymal stem cells (BM-MSCs), deciduous (SCDs), permanent (DPSCs), and periodontal ligament (PDLs) upon exposure to various concentrations of Pb2+. A locally region of senescence cell is shown; (b) quantification of percentage (%) of SA-β-gal positive cells exposed to various concentrations of Pb2+. In all experiments, the results represent average of five culture replicates with standard deviation and a representative photomicrograph was given for each experiment.

Mentions: The 10 μM Pb2+ treatment did not significantly (P > 0.05) inhibit the proliferation rate of any of the cell lines. However, a drastic inhibition of cell growth was observed in BM-MSCs exposed to 20 μM Pb2+ (cell count: 1.3 × 106 cells; MTT: 0.98 absorbance; LDH: 136%; P < 0.05) and up to 160 μM Pb2+ treatment (cell count: 0.6 × 106 cells; MTT: 0.24 absorbance; LDH: 164%; P < 0.05). While SCDs and DPSCs have constant rates of inhibition of cell growth, PDLs seem to be resistant to Pb2+ exposure. A small variation was observed in the control versus cells exposed to 160 μM (cell count: 7.10 × 105 cells; MTT: 0.78 absorbance; LDH: 146%; P > 0.05) (Table 1). This result was reflected in the aging of cells. The percentage of SA-βgal activity was increased in a concentration-dependent manner in all cell lines with the highest in BM-MSCs (ratio) followed by SCDs, DPSCs, and PDLs (Figure 2).


Diverse effects of lead nitrate on the proliferation, differentiation, and gene expression of stem cells isolated from a dental origin.

Abdullah M, Rahman FA, Gnanasegaran N, Govindasamy V, Abu Kasim NH, Musa S - ScientificWorldJournal (2014)

(a) Conventional qualitative SA-β-gal assay by X-gal staining of bone marrow mesenchymal stem cells (BM-MSCs), deciduous (SCDs), permanent (DPSCs), and periodontal ligament (PDLs) upon exposure to various concentrations of Pb2+. A locally region of senescence cell is shown; (b) quantification of percentage (%) of SA-β-gal positive cells exposed to various concentrations of Pb2+. In all experiments, the results represent average of five culture replicates with standard deviation and a representative photomicrograph was given for each experiment.
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3927845&req=5

fig2: (a) Conventional qualitative SA-β-gal assay by X-gal staining of bone marrow mesenchymal stem cells (BM-MSCs), deciduous (SCDs), permanent (DPSCs), and periodontal ligament (PDLs) upon exposure to various concentrations of Pb2+. A locally region of senescence cell is shown; (b) quantification of percentage (%) of SA-β-gal positive cells exposed to various concentrations of Pb2+. In all experiments, the results represent average of five culture replicates with standard deviation and a representative photomicrograph was given for each experiment.
Mentions: The 10 μM Pb2+ treatment did not significantly (P > 0.05) inhibit the proliferation rate of any of the cell lines. However, a drastic inhibition of cell growth was observed in BM-MSCs exposed to 20 μM Pb2+ (cell count: 1.3 × 106 cells; MTT: 0.98 absorbance; LDH: 136%; P < 0.05) and up to 160 μM Pb2+ treatment (cell count: 0.6 × 106 cells; MTT: 0.24 absorbance; LDH: 164%; P < 0.05). While SCDs and DPSCs have constant rates of inhibition of cell growth, PDLs seem to be resistant to Pb2+ exposure. A small variation was observed in the control versus cells exposed to 160 μM (cell count: 7.10 × 105 cells; MTT: 0.78 absorbance; LDH: 146%; P > 0.05) (Table 1). This result was reflected in the aging of cells. The percentage of SA-βgal activity was increased in a concentration-dependent manner in all cell lines with the highest in BM-MSCs (ratio) followed by SCDs, DPSCs, and PDLs (Figure 2).

Bottom Line: We found that Pb(2+) treatment altered the morphology and adhesion of the cells in a dose-dependent manner.There were no significant changes in terms of cell surface phenotypes.Gene expression studies revealed a constant expression of key markers associated with stemness (Oct 4, Rex 1) and DNA repair enzyme markers, but downregulation occurred with some ectoderm and endoderm markers, demonstrating an irregular and untimely differentiation trail.

View Article: PubMed Central - PubMed

Affiliation: Department of Conservative Dentistry, Faculty of Dentistry, University of Malaya, 50603 Kuala Lumpur, Malaysia.

ABSTRACT
Lead (Pb(2+)) exposure continues to be a significant public health problem. Therefore, it is vital to have a continuous epidemiological dataset for a better understanding of Pb(2+) toxicity. In the present study, we have exposed stem cells isolated from deciduous and permanent teeth, periodontal ligament, and bone marrow to five different types of Pb(2+) concentrations (160, 80, 40, 20, and 10 µM) for 24 hours to identify the adverse effects of Pb(2+) on the proliferation, differentiation, and gene expression on these cell lines. We found that Pb(2+) treatment altered the morphology and adhesion of the cells in a dose-dependent manner. There were no significant changes in terms of cell surface phenotypes. Cells exposed to Pb(2+) continued to differentiate into chondrogenesis and adipogenesis, and a severe downregulation was observed in osteogenesis. Gene expression studies revealed a constant expression of key markers associated with stemness (Oct 4, Rex 1) and DNA repair enzyme markers, but downregulation occurred with some ectoderm and endoderm markers, demonstrating an irregular and untimely differentiation trail. Our study revealed for the first time that Pb(2+) exposure not only affects the phenotypic characteristics but also induces significant alteration in the differentiation and gene expression in the cells.

Show MeSH
Related in: MedlinePlus