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High glucose inhibits osteogenic differentiation through the BMP signaling pathway in bone mesenchymal stem cells in mice.

Wang J, Wang B, Li Y, Wang D, Lingling E, Bai Y, Liu H - EXCLI J (2013)

Bottom Line: The intracellular BMP-2 level in BMSCs cultured in a high-glucose microenvironment was significantly decreased and suppressed activation of the BMP signaling pathway.Consequently, expression of the osteogenic markers Runx2, alkaline phosphatase, and osteocalcin were decreased.Thus, it is possible that agents modifying this pathway could be used by BMSCs to promote bone regeneration in high-glucose microenvironments.

View Article: PubMed Central - PubMed

Affiliation: Department of Stomatology, Chinese PLA General Hospital and Postgraduate Military Medical School, Beijing 100853, China.

ABSTRACT
Patients with diabetes tend to have an increased risk of osteoporosis that may be related to hyperglycemia. In vitro evidence has shown that high glucose can affect the proliferation and osteogenic differentiation of mesenchymal stem cells (MSCs). Tissue regeneration depends mainly on MSCs. However, the exact mechanisms involved in high glucose-induced bone loss remain unknown. In this study, we investigated the effects of high glucose on the proliferation and osteogenic differentiation of mice bone MSCs (BMSCs) and determined the specific mechanism of bone morphogenetic protein 2 (BMP-2) in the osteogenic differentiation of mice BMSCs in a high-glucose microenvironment. High glucose (< 25 mM) promoted cell growth but suppressed mineralization. The intracellular BMP-2 level in BMSCs cultured in a high-glucose microenvironment was significantly decreased and suppressed activation of the BMP signaling pathway. Consequently, expression of the osteogenic markers Runx2, alkaline phosphatase, and osteocalcin were decreased. Meanwhile, supplementation with ectogenic BMP-2 reversed the cell osteogenic differentiation and osteogenic marker down-regulation under high glucose. Our data indicate that BMP-2 plays an important role in regulating the osteogenic differentiation of BMSCs in a high-glucose microenvironment. Thus, it is possible that agents modifying this pathway could be used by BMSCs to promote bone regeneration in high-glucose microenvironments.

No MeSH data available.


Related in: MedlinePlus

Figure 1: Different concentrations of high glucose had different effects on BMSC proliferation.A: High-glucose microenvironments (16.5 and 25 mM) promoted BMSC proliferation. Another high-glucose environment (35 mM) inhibited BMSC proliferation. Values are the mean ± SD for triplicate samples from a representative experiment. Student's t-test was performed to determine statistical significance. *P < 0.05B, D: Flow cytometry analysis of the cell cycle showing that 25 mM glucose had the best effects on BMSC proliferation than the other three glucose concentrations. One-way ANOVA was performed to determine statistical significance. *P < 0.05C, E: Annexin V-FITC conjugated to PI by flow cytometry showed that 35 mM glucose caused more significant cell apoptosis than the other three glucose concentrations. One-way ANOVA was performed to determine statistical significance. *P < 0.05
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Figure 1: Figure 1: Different concentrations of high glucose had different effects on BMSC proliferation.A: High-glucose microenvironments (16.5 and 25 mM) promoted BMSC proliferation. Another high-glucose environment (35 mM) inhibited BMSC proliferation. Values are the mean ± SD for triplicate samples from a representative experiment. Student's t-test was performed to determine statistical significance. *P < 0.05B, D: Flow cytometry analysis of the cell cycle showing that 25 mM glucose had the best effects on BMSC proliferation than the other three glucose concentrations. One-way ANOVA was performed to determine statistical significance. *P < 0.05C, E: Annexin V-FITC conjugated to PI by flow cytometry showed that 35 mM glucose caused more significant cell apoptosis than the other three glucose concentrations. One-way ANOVA was performed to determine statistical significance. *P < 0.05

Mentions: It was demonstrated that, based on the glucose content, high glucose had a contrary effect on BMSC proliferation. From day five, the growth of BMSCs cultured in DMEM containing 25 mM glucose was better than that of BMSCs cultured in DMEM containing 5.5 mM glucose (P < 0.05). From day seven, DMEM containing 16.5 and 25 mM glucose both promoted BMSC proliferation. Furthermore, the medium containing 25 mM glucose exerted the best effects on BMSC proliferation in the four groups (P < 0.05). The medium containing 35 mM glucose had a notable effect on BMSC proliferation. Initially, it promoted BMSC proliferation; at day three, it was evident that BMSCs cultured in the DMEM containing 35 mM glucose had the best growth compared to the other three groups (P < 0.05). However, the BMSC proliferation deteriorated and had worse growth than the other three groups after three days (P < 0.05; Figure 1A(Fig. 1)). The flow cytometric analysis showed that the BMSC proliferation indices were as follows: 25 mM group > 16.5 mM group > 5.5 mM group > 35 mM group (P < 0.05; Table 2(Tab. 2); Figure 1B, 1D(Fig. 1)). The annexin V-FITC/PI quantification assay demonstrated that the number of apoptotic cells cultured in DMEM containing 35 mM glucose was much greater (approximately 31.28 %) than that of the other three groups (< 16 %, P < 0.05; Figure 1C, 1E(Fig. 1)).


High glucose inhibits osteogenic differentiation through the BMP signaling pathway in bone mesenchymal stem cells in mice.

Wang J, Wang B, Li Y, Wang D, Lingling E, Bai Y, Liu H - EXCLI J (2013)

Figure 1: Different concentrations of high glucose had different effects on BMSC proliferation.A: High-glucose microenvironments (16.5 and 25 mM) promoted BMSC proliferation. Another high-glucose environment (35 mM) inhibited BMSC proliferation. Values are the mean ± SD for triplicate samples from a representative experiment. Student's t-test was performed to determine statistical significance. *P < 0.05B, D: Flow cytometry analysis of the cell cycle showing that 25 mM glucose had the best effects on BMSC proliferation than the other three glucose concentrations. One-way ANOVA was performed to determine statistical significance. *P < 0.05C, E: Annexin V-FITC conjugated to PI by flow cytometry showed that 35 mM glucose caused more significant cell apoptosis than the other three glucose concentrations. One-way ANOVA was performed to determine statistical significance. *P < 0.05
© Copyright Policy - open-access
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4834668&req=5

Figure 1: Figure 1: Different concentrations of high glucose had different effects on BMSC proliferation.A: High-glucose microenvironments (16.5 and 25 mM) promoted BMSC proliferation. Another high-glucose environment (35 mM) inhibited BMSC proliferation. Values are the mean ± SD for triplicate samples from a representative experiment. Student's t-test was performed to determine statistical significance. *P < 0.05B, D: Flow cytometry analysis of the cell cycle showing that 25 mM glucose had the best effects on BMSC proliferation than the other three glucose concentrations. One-way ANOVA was performed to determine statistical significance. *P < 0.05C, E: Annexin V-FITC conjugated to PI by flow cytometry showed that 35 mM glucose caused more significant cell apoptosis than the other three glucose concentrations. One-way ANOVA was performed to determine statistical significance. *P < 0.05
Mentions: It was demonstrated that, based on the glucose content, high glucose had a contrary effect on BMSC proliferation. From day five, the growth of BMSCs cultured in DMEM containing 25 mM glucose was better than that of BMSCs cultured in DMEM containing 5.5 mM glucose (P < 0.05). From day seven, DMEM containing 16.5 and 25 mM glucose both promoted BMSC proliferation. Furthermore, the medium containing 25 mM glucose exerted the best effects on BMSC proliferation in the four groups (P < 0.05). The medium containing 35 mM glucose had a notable effect on BMSC proliferation. Initially, it promoted BMSC proliferation; at day three, it was evident that BMSCs cultured in the DMEM containing 35 mM glucose had the best growth compared to the other three groups (P < 0.05). However, the BMSC proliferation deteriorated and had worse growth than the other three groups after three days (P < 0.05; Figure 1A(Fig. 1)). The flow cytometric analysis showed that the BMSC proliferation indices were as follows: 25 mM group > 16.5 mM group > 5.5 mM group > 35 mM group (P < 0.05; Table 2(Tab. 2); Figure 1B, 1D(Fig. 1)). The annexin V-FITC/PI quantification assay demonstrated that the number of apoptotic cells cultured in DMEM containing 35 mM glucose was much greater (approximately 31.28 %) than that of the other three groups (< 16 %, P < 0.05; Figure 1C, 1E(Fig. 1)).

Bottom Line: The intracellular BMP-2 level in BMSCs cultured in a high-glucose microenvironment was significantly decreased and suppressed activation of the BMP signaling pathway.Consequently, expression of the osteogenic markers Runx2, alkaline phosphatase, and osteocalcin were decreased.Thus, it is possible that agents modifying this pathway could be used by BMSCs to promote bone regeneration in high-glucose microenvironments.

View Article: PubMed Central - PubMed

Affiliation: Department of Stomatology, Chinese PLA General Hospital and Postgraduate Military Medical School, Beijing 100853, China.

ABSTRACT
Patients with diabetes tend to have an increased risk of osteoporosis that may be related to hyperglycemia. In vitro evidence has shown that high glucose can affect the proliferation and osteogenic differentiation of mesenchymal stem cells (MSCs). Tissue regeneration depends mainly on MSCs. However, the exact mechanisms involved in high glucose-induced bone loss remain unknown. In this study, we investigated the effects of high glucose on the proliferation and osteogenic differentiation of mice bone MSCs (BMSCs) and determined the specific mechanism of bone morphogenetic protein 2 (BMP-2) in the osteogenic differentiation of mice BMSCs in a high-glucose microenvironment. High glucose (< 25 mM) promoted cell growth but suppressed mineralization. The intracellular BMP-2 level in BMSCs cultured in a high-glucose microenvironment was significantly decreased and suppressed activation of the BMP signaling pathway. Consequently, expression of the osteogenic markers Runx2, alkaline phosphatase, and osteocalcin were decreased. Meanwhile, supplementation with ectogenic BMP-2 reversed the cell osteogenic differentiation and osteogenic marker down-regulation under high glucose. Our data indicate that BMP-2 plays an important role in regulating the osteogenic differentiation of BMSCs in a high-glucose microenvironment. Thus, it is possible that agents modifying this pathway could be used by BMSCs to promote bone regeneration in high-glucose microenvironments.

No MeSH data available.


Related in: MedlinePlus