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Far infra-red therapy promotes ischemia-induced angiogenesis in diabetic mice and restores high glucose-suppressed endothelial progenitor cell functions.

Huang PH, Chen JW, Lin CP, Chen YH, Wang CH, Leu HB, Lin SJ - Cardiovasc Diabetol (2012)

Bottom Line: Far infra-red (IFR) therapy was shown to exert beneficial effects in cardiovascular system, but effects of IFR on endothelial progenitor cell (EPC) and EPC-related vasculogenesis remain unclear.Doppler perfusion imaging demonstrated that the ischemic limb/normal side blood perfusion ratio in the thermal therapy group was significantly increased beyond that in controls, and significantly greater capillary density was seen in the IFR therapy group.In in-vitro studies, cultured EPCs treated with IFR radiation markedly augmented high glucose-impaired EPC functions, inhibited high glucose-induced EPC senescence and reduced H(2)O(2) production.

View Article: PubMed Central - HTML - PubMed

Affiliation: Division of Cardiology, Taipei Veterans General Hospital, Taipei, Taiwan.

ABSTRACT

Background: Far infra-red (IFR) therapy was shown to exert beneficial effects in cardiovascular system, but effects of IFR on endothelial progenitor cell (EPC) and EPC-related vasculogenesis remain unclear. We hypothesized that IFR radiation can restore blood flow recovery in ischemic hindlimb in diabetic mice by enhancement of EPCs functions and homing process.

Materials and methods: Starting at 4 weeks after the onset of diabetes, unilateral hindlimb ischemia was induced in streptozotocin (STZ)-induced diabetic mice, which were divided into control and IFR therapy groups (n = 6 per group). The latter mice were placed in an IFR dry sauna at 34°C for 30 min once per day for 5 weeks.

Results: Doppler perfusion imaging demonstrated that the ischemic limb/normal side blood perfusion ratio in the thermal therapy group was significantly increased beyond that in controls, and significantly greater capillary density was seen in the IFR therapy group. Flow cytometry analysis showed impaired EPCs (Sca-1(+)/Flk-1(+)) mobilization after ischemia surgery in diabetic mice with or without IFR therapy (n = 6 per group). However, as compared to those in the control group, bone marrow-derived EPCs differentiated into endothelial cells defined as GFP(+)/CD31(+) double-positive cells were significantly increased in ischemic tissue around the vessels in diabetic mice that received IFR radiation. In in-vitro studies, cultured EPCs treated with IFR radiation markedly augmented high glucose-impaired EPC functions, inhibited high glucose-induced EPC senescence and reduced H(2)O(2) production. Nude mice received human EPCs treated with IFR in high glucose medium showed a significant improvement in blood flow recovery in ischemic limb compared to those without IFR therapy. IFR therapy promoted blood flow recovery and new vessel formation in STZ-induced diabetic mice.

Conclusions: Administration of IFR therapy promoted collateral flow recovery and new vessel formation in STZ-induced diabetic mice, and these beneficial effects may derive from enhancement of EPC functions and homing process.

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Effects of IFR therapy on EPC migration, tube formation, and senescence in vitro. (A,B) Scratch test and modified Boyden chamber assay were used to assess the migratory function of ECFCs in high glucose conditions. Boyden chamber assay using VEGF as a chemoattracting factor was used to evaluate the effects of IFR radiation on EPC migration. (C) An in vitro angiogenesis assay for late ECFCs used ECMatrix gel. Representative photos for in vitro angiogenesis are shown. Cells were stained with crystal violet, and the averages of the total area of complete tubes formed by cells were compared by using computer software. (D) To determine the onset of cellular aging, acidic ß-galactosidase was used as a biochemical marker for acidification, typical for ECFCs senescence. Data are means ± SEM; n = 4 in each experiment.
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Figure 6: Effects of IFR therapy on EPC migration, tube formation, and senescence in vitro. (A,B) Scratch test and modified Boyden chamber assay were used to assess the migratory function of ECFCs in high glucose conditions. Boyden chamber assay using VEGF as a chemoattracting factor was used to evaluate the effects of IFR radiation on EPC migration. (C) An in vitro angiogenesis assay for late ECFCs used ECMatrix gel. Representative photos for in vitro angiogenesis are shown. Cells were stained with crystal violet, and the averages of the total area of complete tubes formed by cells were compared by using computer software. (D) To determine the onset of cellular aging, acidic ß-galactosidase was used as a biochemical marker for acidification, typical for ECFCs senescence. Data are means ± SEM; n = 4 in each experiment.

Mentions: To investigate the effects of IFR radiation on EPCs, we used both scratch test and modified Boyden chamber assay to assess the migratory function of EPCs in high glucose conditions. Compared with the control group, incubation of EPCs with high glucose (4 days) significantly decreased EPC migration (100 ± 9 versus 63 ± 11/HPF, p = 0.031). However, treatment with IFR radiation for 30 minutes significantly recovered high glucose-suppressed late EPC migratory function (high glucose versus high glucose + IFR, 63 ± 11 versus 174 ± 10 cells/HPF, p = 0.005; Figure 6A). Similar findings were observed by modified Boyden chamber assay (Figure 6B).


Far infra-red therapy promotes ischemia-induced angiogenesis in diabetic mice and restores high glucose-suppressed endothelial progenitor cell functions.

Huang PH, Chen JW, Lin CP, Chen YH, Wang CH, Leu HB, Lin SJ - Cardiovasc Diabetol (2012)

Effects of IFR therapy on EPC migration, tube formation, and senescence in vitro. (A,B) Scratch test and modified Boyden chamber assay were used to assess the migratory function of ECFCs in high glucose conditions. Boyden chamber assay using VEGF as a chemoattracting factor was used to evaluate the effects of IFR radiation on EPC migration. (C) An in vitro angiogenesis assay for late ECFCs used ECMatrix gel. Representative photos for in vitro angiogenesis are shown. Cells were stained with crystal violet, and the averages of the total area of complete tubes formed by cells were compared by using computer software. (D) To determine the onset of cellular aging, acidic ß-galactosidase was used as a biochemical marker for acidification, typical for ECFCs senescence. Data are means ± SEM; n = 4 in each experiment.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3472269&req=5

Figure 6: Effects of IFR therapy on EPC migration, tube formation, and senescence in vitro. (A,B) Scratch test and modified Boyden chamber assay were used to assess the migratory function of ECFCs in high glucose conditions. Boyden chamber assay using VEGF as a chemoattracting factor was used to evaluate the effects of IFR radiation on EPC migration. (C) An in vitro angiogenesis assay for late ECFCs used ECMatrix gel. Representative photos for in vitro angiogenesis are shown. Cells were stained with crystal violet, and the averages of the total area of complete tubes formed by cells were compared by using computer software. (D) To determine the onset of cellular aging, acidic ß-galactosidase was used as a biochemical marker for acidification, typical for ECFCs senescence. Data are means ± SEM; n = 4 in each experiment.
Mentions: To investigate the effects of IFR radiation on EPCs, we used both scratch test and modified Boyden chamber assay to assess the migratory function of EPCs in high glucose conditions. Compared with the control group, incubation of EPCs with high glucose (4 days) significantly decreased EPC migration (100 ± 9 versus 63 ± 11/HPF, p = 0.031). However, treatment with IFR radiation for 30 minutes significantly recovered high glucose-suppressed late EPC migratory function (high glucose versus high glucose + IFR, 63 ± 11 versus 174 ± 10 cells/HPF, p = 0.005; Figure 6A). Similar findings were observed by modified Boyden chamber assay (Figure 6B).

Bottom Line: Far infra-red (IFR) therapy was shown to exert beneficial effects in cardiovascular system, but effects of IFR on endothelial progenitor cell (EPC) and EPC-related vasculogenesis remain unclear.Doppler perfusion imaging demonstrated that the ischemic limb/normal side blood perfusion ratio in the thermal therapy group was significantly increased beyond that in controls, and significantly greater capillary density was seen in the IFR therapy group.In in-vitro studies, cultured EPCs treated with IFR radiation markedly augmented high glucose-impaired EPC functions, inhibited high glucose-induced EPC senescence and reduced H(2)O(2) production.

View Article: PubMed Central - HTML - PubMed

Affiliation: Division of Cardiology, Taipei Veterans General Hospital, Taipei, Taiwan.

ABSTRACT

Background: Far infra-red (IFR) therapy was shown to exert beneficial effects in cardiovascular system, but effects of IFR on endothelial progenitor cell (EPC) and EPC-related vasculogenesis remain unclear. We hypothesized that IFR radiation can restore blood flow recovery in ischemic hindlimb in diabetic mice by enhancement of EPCs functions and homing process.

Materials and methods: Starting at 4 weeks after the onset of diabetes, unilateral hindlimb ischemia was induced in streptozotocin (STZ)-induced diabetic mice, which were divided into control and IFR therapy groups (n = 6 per group). The latter mice were placed in an IFR dry sauna at 34°C for 30 min once per day for 5 weeks.

Results: Doppler perfusion imaging demonstrated that the ischemic limb/normal side blood perfusion ratio in the thermal therapy group was significantly increased beyond that in controls, and significantly greater capillary density was seen in the IFR therapy group. Flow cytometry analysis showed impaired EPCs (Sca-1(+)/Flk-1(+)) mobilization after ischemia surgery in diabetic mice with or without IFR therapy (n = 6 per group). However, as compared to those in the control group, bone marrow-derived EPCs differentiated into endothelial cells defined as GFP(+)/CD31(+) double-positive cells were significantly increased in ischemic tissue around the vessels in diabetic mice that received IFR radiation. In in-vitro studies, cultured EPCs treated with IFR radiation markedly augmented high glucose-impaired EPC functions, inhibited high glucose-induced EPC senescence and reduced H(2)O(2) production. Nude mice received human EPCs treated with IFR in high glucose medium showed a significant improvement in blood flow recovery in ischemic limb compared to those without IFR therapy. IFR therapy promoted blood flow recovery and new vessel formation in STZ-induced diabetic mice.

Conclusions: Administration of IFR therapy promoted collateral flow recovery and new vessel formation in STZ-induced diabetic mice, and these beneficial effects may derive from enhancement of EPC functions and homing process.

Show MeSH
Related in: MedlinePlus