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Role of ox-PAPCs in the differentiation of mesenchymal stem cells (MSCs) and Runx2 and PPARγ2 expression in MSCs-like of osteoporotic patients.

Valenti MT, Garbin U, Pasini A, Zanatta M, Stranieri C, Manfro S, Zucal C, Dalle Carbonare L - PLoS ONE (2011)

Bottom Line: Finally, we examined the effects of sera obtained from OPs in hMSCs comparing the results with age-matched normal donors (NDs).Quantitative RT-PCR demonstrated that ox-PAPCs enhanced PPARγ2 and adipogenic gene expression and reduced Runx2 and osteoblast differentiation marker gene expression in differentiating hMSCs.In OPs, ox-PAPC levels and PPARγ2 expression were higher than in NDs, whereas Runx2 was lower than in ND circulant MSCs-like.

View Article: PubMed Central - PubMed

Affiliation: Department of Medicine, Clinica di Medicina Interna, Sezione D, University of Verona, Verona, Italy.

ABSTRACT

Background: Mesenchymal stem cells (MSCs) can differentiate into osteoblasts and adipocytes and conditions causing bone loss may induce a switch from the osteoblast to adipocyte lineage. In addition, the expression of Runx2 and the PPARγ2 transcription factor genes is essential for cellular commitment to an osteogenic and adipogenic differentiation, respectively. Modified lipoproteins derived from the oxidation of arachidonate-containing phospholipids (ox-PAPCs: POVPC, PGPC and PEIPC) are considered important factors in atherogenesis.

Methodology: We investigated the effect of ox-PAPCs on osteogenesis and adipogenesis in human mesenchymal stem cells (hMSCs). In particular, we analyzed the transcription factor Runx2 and the PPARγ2 gene expression during osteogenic and adipogenic differentiation in absence and in presence of ox-PAPCs. We also analyzed gene expression level in a panel of osteoblastic and adipogenic differentiation markers. In addition, as circulating blood cells can be used as a "sentinel" that responds to changes in the macro- or micro-environment, we analyzed the Runx2 and the PPARγ2 gene expression in MSCs-like and ox-PAPC levels in serum of osteoporotic patients (OPs). Finally, we examined the effects of sera obtained from OPs in hMSCs comparing the results with age-matched normal donors (NDs).

Principal findings: Quantitative RT-PCR demonstrated that ox-PAPCs enhanced PPARγ2 and adipogenic gene expression and reduced Runx2 and osteoblast differentiation marker gene expression in differentiating hMSCs. In OPs, ox-PAPC levels and PPARγ2 expression were higher than in NDs, whereas Runx2 was lower than in ND circulant MSCs-like.

Conclusions: Ox-PAPCs affect the osteogenic differentiation by promoting adipogenic differentiation and this effect may appear involved in bone loss in OPs.

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Related in: MedlinePlus

OIL Red O staining.When the hMSCs were cultured for 72 h in presence of ox-PAPCs during adipogenic differentiation, they showed a dose dependent significant increase of lipid-folled droplets (p<0.05). (original magnification 20×, insert 40×). The percentage of area stained with OIL Red O (red droplets) was quantified by using the IMAGE J Software.
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pone-0020363-g002: OIL Red O staining.When the hMSCs were cultured for 72 h in presence of ox-PAPCs during adipogenic differentiation, they showed a dose dependent significant increase of lipid-folled droplets (p<0.05). (original magnification 20×, insert 40×). The percentage of area stained with OIL Red O (red droplets) was quantified by using the IMAGE J Software.

Mentions: In addition, when cells were cultured for 72 h in presence of ox-PAPCs during adipogenic differentiation, OIL Red O staining showed an increase of intracellular lipid-filled droplets in a dose dependent manner (Figure 2). In particular, the area of oil red O staining was 0.1%±0.05 in the control and 1.2%±0.6, 1.8%±0.5 and 2.1%±0.5 at 5 µg/ml, 10 µg/ml and 20 µg/ml, respectively.


Role of ox-PAPCs in the differentiation of mesenchymal stem cells (MSCs) and Runx2 and PPARγ2 expression in MSCs-like of osteoporotic patients.

Valenti MT, Garbin U, Pasini A, Zanatta M, Stranieri C, Manfro S, Zucal C, Dalle Carbonare L - PLoS ONE (2011)

OIL Red O staining.When the hMSCs were cultured for 72 h in presence of ox-PAPCs during adipogenic differentiation, they showed a dose dependent significant increase of lipid-folled droplets (p<0.05). (original magnification 20×, insert 40×). The percentage of area stained with OIL Red O (red droplets) was quantified by using the IMAGE J Software.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3108593&req=5

pone-0020363-g002: OIL Red O staining.When the hMSCs were cultured for 72 h in presence of ox-PAPCs during adipogenic differentiation, they showed a dose dependent significant increase of lipid-folled droplets (p<0.05). (original magnification 20×, insert 40×). The percentage of area stained with OIL Red O (red droplets) was quantified by using the IMAGE J Software.
Mentions: In addition, when cells were cultured for 72 h in presence of ox-PAPCs during adipogenic differentiation, OIL Red O staining showed an increase of intracellular lipid-filled droplets in a dose dependent manner (Figure 2). In particular, the area of oil red O staining was 0.1%±0.05 in the control and 1.2%±0.6, 1.8%±0.5 and 2.1%±0.5 at 5 µg/ml, 10 µg/ml and 20 µg/ml, respectively.

Bottom Line: Finally, we examined the effects of sera obtained from OPs in hMSCs comparing the results with age-matched normal donors (NDs).Quantitative RT-PCR demonstrated that ox-PAPCs enhanced PPARγ2 and adipogenic gene expression and reduced Runx2 and osteoblast differentiation marker gene expression in differentiating hMSCs.In OPs, ox-PAPC levels and PPARγ2 expression were higher than in NDs, whereas Runx2 was lower than in ND circulant MSCs-like.

View Article: PubMed Central - PubMed

Affiliation: Department of Medicine, Clinica di Medicina Interna, Sezione D, University of Verona, Verona, Italy.

ABSTRACT

Background: Mesenchymal stem cells (MSCs) can differentiate into osteoblasts and adipocytes and conditions causing bone loss may induce a switch from the osteoblast to adipocyte lineage. In addition, the expression of Runx2 and the PPARγ2 transcription factor genes is essential for cellular commitment to an osteogenic and adipogenic differentiation, respectively. Modified lipoproteins derived from the oxidation of arachidonate-containing phospholipids (ox-PAPCs: POVPC, PGPC and PEIPC) are considered important factors in atherogenesis.

Methodology: We investigated the effect of ox-PAPCs on osteogenesis and adipogenesis in human mesenchymal stem cells (hMSCs). In particular, we analyzed the transcription factor Runx2 and the PPARγ2 gene expression during osteogenic and adipogenic differentiation in absence and in presence of ox-PAPCs. We also analyzed gene expression level in a panel of osteoblastic and adipogenic differentiation markers. In addition, as circulating blood cells can be used as a "sentinel" that responds to changes in the macro- or micro-environment, we analyzed the Runx2 and the PPARγ2 gene expression in MSCs-like and ox-PAPC levels in serum of osteoporotic patients (OPs). Finally, we examined the effects of sera obtained from OPs in hMSCs comparing the results with age-matched normal donors (NDs).

Principal findings: Quantitative RT-PCR demonstrated that ox-PAPCs enhanced PPARγ2 and adipogenic gene expression and reduced Runx2 and osteoblast differentiation marker gene expression in differentiating hMSCs. In OPs, ox-PAPC levels and PPARγ2 expression were higher than in NDs, whereas Runx2 was lower than in ND circulant MSCs-like.

Conclusions: Ox-PAPCs affect the osteogenic differentiation by promoting adipogenic differentiation and this effect may appear involved in bone loss in OPs.

Show MeSH
Related in: MedlinePlus