研究目的
The aim of this study is to evaluate the feasible effect of LLLT combined with phloroglucinol (PHL) on the inhibition of fibrosis in vitro.
研究成果
PHL‐assisted LLLT may be an effective treatment to inhibit fibrosis due to its additive effects. The combined treatment has a potential to be an alternative treatment for fibrosis.
研究不足
A limitation of the current study exists from the lack of identifying the cell signaling factors that underlie the therapeutic mechanism in the combined treatment. The heat effects of the LLLT from the photon‐cell interaction was not verified. Although 635 nm might be quite effective for LLLT among various wavelengths, high absorption of blood with wavelengths <700 nm would impede the efficacy of the 635 nm LLLT under in vivo conditions. Another limitation is a decrease in the percentage of wound closure. The combined therapy should be developed to promote the wound healing while minimizing overexpression of fibrotic markers. Another limitation is that TGF‐β was used as a sole biological factor for the current evaluation.
1:Experimental Design and Method Selection:
NIH/3T3 murine embryonic fibroblasts cells were cultured and transforming growth factor‐β1 (TGF‐β1) was treated for transition of fibroblasts. After TGF‐β1 treatment, LLLT and PHL were used, respectively, and in combination to suppress fibrosis.
2:Sample Selection and Data Sources:
NIH/3T3 murine embryonic fibroblasts cells obtained from the Korean Cell Line Bank.
3:List of Experimental Equipment and Materials:
Visible laser system (SD‐635‐HS‐1W; CNI laser), frontal light distributor (Medlight), power detector (PD‐300‐3W; Ophir), power meter (Nova Ⅱ; Ophir), microplate spectrophotometer (Multiskan Go; Thermo Fisher Scientific), confocal laser scanning microscope (LSM 700; ZEISS).
4:Experimental Procedures and Operational Workflow:
Cell viability and proliferation were evaluated using MTT and BrdU assays. Confocal immunofluorescence and western blot were used to evaluate the expression of fibrotic markers.
5:Data Analysis Methods:
Statistical analysis was performed using SPSS software 22, which calculated mean ± standard deviation for at least three independent experiments. The Mann–Whitney U test was used for nonparametric statistical analysis.
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