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Abstract
Background:
Near-infrared light (NIR, 0.8-1.5 μm light) has been used in therapeutic devices for various injuries such as infected, ischemic and hypoxic wound. NIR-emitting technology has been developed recently in Korea. We hypothesized that NIR may have an anti-inflammatory effect and investigated the effect of NIR-irradiated media on cell culture.
Methods:
Three kinds of cell lines, CAPE (vascular endothelial cell), NIH3T3 (fibroblast), and RD (smooth muscle cell) cells were cultured for 4 days in 10% FBS-containing media (1×104 cells/well), which were irradiated or not irradiated (control) by Eco-NFIR Drive (Model #0210, Ecowavetech, Korea). The cells were stimulated by 10 mcg/mL of bacterial lipopolysaccharide (LPS) or sodium nitroprusside (SNP). Cellular proliferation was measured by methylthiazol tetrazolium assay. Expression of interleukin (IL)-1 beta and nitric oxide was measured by ELISA. Expression of cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS) was measured by immunofluorescence staining.
Results:
NIR-irradiated medium was favorable for CAPE cell proliferation (N=8, P=0.000). IL-1 beta secretion from LPS-stimulated NIH3T3 cells incubated in the NIR medium was below that of control medium (N=4, P=0.026). Nitrate production seemed to be low in NIR-irradiated medium although statistically insignificant (N=4, P=0.076). Expression of iNOS of the LPS-stimulated cells was decreased in NIR medium, however, Cox-2 expression was not different between the two media.
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Fig. 1.
Methylthiazol tetrazolium test result of the cells. NIR-irradiated medium provided favorable condition for CAPE cells in terms of cellular mitochondrial enzyme (succinate dehydrogenase) actvity (mean±SE, N=8, P=0.000). However, there was no significant difference of cellular proliferation of NIH3T3 cell (mean±SE, N=16, P=0.170) and RD cell (mean±SE, N=8, P=0.295) between NIR-irradiated medium (N, –Δ–) and control medium (C, –○–).
Abbreviations: C, control medium; N, NIR-irradiated medium; MTT, methylthiazol tetrazolium; NIR, near-infrared irradiated.
Fig. 2.
Methylthiazol tetrazolium test result of SNP-stimulated NIH3T3 cells. The SNP-stimulated NIH3T3 cells incubated in the NIR-irradiated medium (–Δ–) proliferated more than the cells incubated in the control medium (–○–) at day 2 (mean±SE, N=16, P=0.000). Abbreviations: See Fig. 1.
Fig. 3.
IL-1 beta concentration of the supernatant of NIH3T3 cells which were stimulated with LPS (10 mcg/mL). The IL-1 beta secretion of NIH3T3 cells incubated in NIR-irradiated medium (–Δ–) was below that in the control medium (–○–) (mean±SE, N=4, P=0.026).
Abbreviations: See Fig. 1.
Fig. 4.
Nitric oxide production of NIH3T3 cells which were stimulated with LPS (10 mcg/mL). The NO level of NIH3T3 cells incubated in NIR-irradiated medium (NIR) was lower than that in the control medium (control), although statistically insignificant (mean±SE, N=4, P=0.076).
Abbreviations: See Fig. 1.
Fig. 5.
LPS (10 mcg/mL) stimulated NIH3T3 cells show green fluorescence after Immunofluorescence staining of iNOS-FITC antibody (epi-fluoresence microscopy, NiKon, micro-FXA ×400). Abbreviations: See Table 1.
Table 1.
Result of immunofluorescence staining of inducible nitric oxide synthase expression of the NIH3T3 cells
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