Red fluorescence of oral bacteria interacting with Porphyromonas gingivalis / 대한구강보건학회지

OBJECTIVES: Dental plaque is composed of 700 bacterial species. It is known that some oral microorganisms produce porphyrin, and thus, they emit red fluorescence when illuminated with blue light at a specific wavelength of <410 nm. Porphyromonas gingivalis belongs to the genus Porphyromonas, which is characterized by the production of porphyrin. The aim of this study was to evaluate red fluorescence emission of some oral microorganisms interacting with P. gingivalis. METHODS: Five bacterial strains (P. gingivalis, Streptococcus mutans, Lactobacillus casei, Actinomyces naeslundii, and Fusobacterium nucleatum) were used for this study. Tryptic soy agar medium supplemented with hemin, vitamin K3, and sheep blood was used as a growth medium. The fluorescence emission of bacterial colonies was evaluated under 405 nm-wavelength blue light using a Quantitative Light-induced Fluorescence Digital (QLF-D) camera system. Each bacterium was cultured alone and co-cultured in close proximity with P. gingivalis. The red/green (R/G) ratio of fluorescence image was calculated and the differences of R/G ratio according to each growth condition were compared using the Mann-Whitney test (P<0.05). RESULTS: Single cultured S. mutans, L. casei and A. naeslundii colonies emitted red fluorescence (R/G ratio=2.15±0.06, 4.31±0.17, 5.52±1.29, respectively). Fusobacterium nucleatum colonies emitted green fluorescence (R/G ratio=1.36±0.06). The R/G ratios of A. naeslundii and F. nucleatum were increased when P. gingivalis was co-cultured with each bacterium (P<0.05). In contrast, the R/G ratios of S. mutans and L. casei were decreased when P. gingivalis was co-cultured with each bacterium (P=0.002, 0.003). CONCLUSIONS: This study confirmed that P. gingivalis could affect the red fluorescence of other oral bacteria under 405 nm-wavelength blue light. Our findings concluded that P. gingivalis has an important role for red fluorescence emission of dental biofilm.

Red fluorescence of oral bacteria is affected by blood in the growth medium / 대한구강보건학회지

OBJECTIVES: Dental plaque emits red fluorescence under a visible blue light near the ultra-violet end of the light spectrum. The fluorescence characteristics of each microorganism have been reported in several studies. The aim of this study was to evaluate changes in red fluorescence of oral microorganisms that is affected by blood in the culture media. METHODS: The gram-positive Actinomyces naeslundii (AN, KCTC 5525) and Lactobacillus casei (LC, KCTC 3109) and gram negative Prevotella intermedia (PI, KCTC 3692) that are known to emit red fluorescence were used in this study. Each bacterium was activated in broth and cultivated in different agar media at 37℃ for 7 days. Tryptic soy agar with hemin and vitamin K3 (TSA), TSA with sheep blood (TSAB), basal medium mucin (BMM) medium, and BMM with sheep blood (BMMB) were used in this study. Fluorescence due to bacterial growth was observed under 405-nm wavelength blue light using the quantitative light-induced fluorescence-digital (QLF-D) device. The red, green, and blue fluorescence values of colonies were obtained using image-analysis software and the red to green ratio (R/G value) and red to total RGB ratio (R/RGB value) were calculated for quantitative comparison. RESULTS: The QLF-D images of the AN, LC, and PI colonies showed red fluorescence in all media, but the fluorescence of all bacteria was reduced in TSA and BMM media, compared with in TSAB and BMMB media. Both the R/G and the R/RGB values of all bacteria were significantly reduced in growth media without blood (P<0.001). CONCLUSIONS: Based on this in vitro study, it can be concluded that red fluorescence of oral bacteria can be affected by growth components, especially blood. Blood-containing medium could be a significant factor influencing red fluorescence of oral bacteria. It can be further hypothesized that bleeding in the oral cavity can increase the red fluorescence of dental plaque.

UV-induced red fluorescence on facial part of patients with acne vulgaris and seborrheic dermatitis: a fluorescence spectrum study / 中华医学美学美容杂志

Objective To measure the fluorescence spectral information of UVRF in the facial skin of patients with acne vulgaris and seborrheic dermatitis and to comprehend the optical properties of UVRF.Methods The lesions were squeezed to extract the sebaceous secretions containing UVRF in the facial skin.Ocean Optics USB2000 +F02243 Spectrometer and the 308nm excimer laser were used as an excitation light source to measure the signal of the LIF spectrum of UVRF in 6 acne vulgaris and 4 seborrheic dermatitis patients' facial part,compared with protoporphyrin Ⅸ.Results 1-4 peaks were detected in 10 patients' UVRF specimen respectively.The fluorescence spectra's characteristic peak of UVRF in 10 patients was at 680 nm.Protoporphyrin Ⅸ was at 688 nm and more gentle and generous than UVRF's characteristic peak.Conclusions LIF system can be used to detect the fluorescence spectroscopy of UVRF.UVRF has similar spectral characteristics to protoporphyrin Ⅸ,but it is not the same species.

Red fluorescent transplantation tumor model of mouse bladder carcinoma and fluoroscopic image study / 中华泌尿外科杂志

Objective To explore molecular fluorescence imaging features of the growth and metastasis of DsRed-marked mouse bladder carcinoma. Methods The study used lipofectamine 2000 transfection method,transferred on the BTT739 cells with plasmid chickenβ-actin-DsRed-Neo vector.The stably expressing BTT739-DsRed monoelonal cells were got with G418 selection.It randomly divided the 615 mouse of 24 into three groups,injected cell suspension on the hindlimb,the first and second group with BTT739-DsRed cell and the third group with BTT739 cell to found xenograft roodel.MAESTRO imager recorded fluorescence images of the growth and metastasis of the tumors in vivo and the fluorescence intensity was measured.The excitation wavelength was 560-580 nm,emission wavelength was 590-610 nm and exposure time was 5000 ms.After continuous observation of 4 weeks,every week killed the mouse of the second group and cut into image,made records of the red fluorescent mouse bladder cancer xenograft model,measured the tumor size and fluorescence sighal values; analyzed the relations between the tumor size and fluorescence signal values as well as between the whole image and cut image. Results DsRed tumor could be observed at the first week. Central local fluorescence loss could be detected at the second week, pathologically confirmed necrotic tumor tissue and a little connective tissue. At the fourth week, a local lymph node metastasis could be observed with no distant metastasis. The measured values of fluorescent signal were as follows: (89±18), (122±55), (133±69), (715±343)counts; the tumor size were as follows: (13±4), (45±22), (83±29), (253±67)mm2. The whole body image of tumor size were as follows: (12± 3),(50±23), (90±29), (290±74)mm2. The cut image of tumor size were as follows: (12±5), (72±30), (141±43), (524±237)mm2. The tumor size and fluorescent signal values reflect positive linear correlation with 0. 74 coefficient (t= 3. 97, P<0.05), whole body imaging and cut image reflect positive linear correlation with 0. 97 coefficient (t=10, P<0.05). The whole body image of tumor size was (70. 85±17.13) % of cut image. Conclusions Red fluorescent mouse bladder cancer xenograft model could observe the growth and metastasis of the tumor intuitively, continuously, and sensitively.As the tumor increased, the fluorescence range also increased, the fluorescence disappeared after tumor necrosis, the expression of the red fluorescent transferred after the metastasis of the tumor.

Effects of different dosages of BMSC on lung fibrosis in mice / 上海交通大学学报（医学版）

Objective To investigate the effects of different dosages of bone marrow mesenchymal stromal cells (BMSC) on lung fibrosis. Methods BMSCs with red fluorescence protein (RFP) from male FVB mice were cultured in vitro. Twenty-four female wild type FVB mice were randomly divided into four groups: normal group, model group, BMSC 1 group and BMSC 2 group (n = 6). Mouse pulmonary fibrosis models were induced by bleomycin via single intratracheal perfusion. Twenty-four h after model establishment, mice in BMSC 1 group and BMSC 2 group were injected with 1 × 10~6 BMSCs and 2 × 10~6 BMSCs, respectively through vena caudalis for each mouse. All the animals were sacrificed 21 d after model estalishment, and mouse lung tissue samples were obtained. The pathological changes were observed by light microscopy, the hydroxyproline ( Hyp) contents were measured by alkaline hydrolysis assay, the distribution of RFP( + ) BMSCs and quantitation of RFP were analysed by laser scanning confocal microscopy and immunohistochemistry, the expression of surfactant protein A (SP-A) was detected by immunohistochemistry, and the expression of transforming growth factor-β (TGF-β) and platelet-derived growth factor (PDGF) mRNA was detected by Real-time PCR. Results Compared with model group, the pulmonary fibrosis in BMSC 1 group was significantly alleviated, and that of BMSC 2 group became much more severe.A large number of RFP( +) BMSCs were found in fibrosis area of BMSC 2 group,which exhibited morphology similar to fibroblasts. As far as the expression of SP-A was concerned, normal group was higher than BMSC 1 group, BMSC 1 group was higher than BMSC 2 group and model group (P < 0. 05), while there was no significant difference between BMSC 2 group and model group (P >0. 05). Normal group, BMSC I group, model group and BMSC 2 group fell in the increase order by Hyp contents (P <0.01, P <0.05), and BMSC 2 group, BMSC 1 group, model group and normal group fell in the decrease order by expression of TCF-|$ and PDGF mRNA (P < 0.05). Conclusion Proper dose of BMSC has a favourable effect on bleomycin-induced lung fibrosis, while excessive dose of BMSC can aggravate the fibrosis.