Mouse 유발창상에서 저출력레이저치료의 창상치유 증진효과

오충훈,정필상,이상준,이진구,정신근,안진철,명나혜 대한이비인후과학회 2003 대한이비인후과학회지 두경부외과학 Vol.46 No.10

Background and Objectives:This study was conducted to investigate the healing effect of the low-level laser iradiation on wound healing in vivo using DPSS laser (532 nm) and Diode laser (660 nm). Materials and Method:Each mouse received dorsal, full-thicknes round incision (= 2 cm) and daily laser irradiation (4 J/cm2) was done before sacrifice. On sacrifice at 3, 7, 10 days, the wound was excised, then wound closure and histologic stages were measured, and standardized. Results:The percentages of wound closure in DPSS laser, Diode laser, control were 33.2± 2.4, 34.2± 3.5, 24.0± 2.7 at day 3, 64.8± 3.5, 72.2± 2.8, 42.8± 5.0 at day 7 and 82.2± 7.9, 87.2± 3.7, 71.4± 4.0 at day 10, respectively, with p<0.05. Histological evaluation showed that laser irradiation enhanced wound epithelialization, celular content deposition, granulation tissue formation, colagen deposition and neovascularization in the laser-treated wounds as compared to the control group. Conclusion:Low-level laser irradiation at 532 nm and 660 nm significantly enhanced cutaneous wound healing effect in the wounded mouse model. Further investigation of the mechanism of low-level laser therapy in primary wound healing is waranted. (Korean J Otolaryngol 203 ; 46 :851-5)

S . Viridochromogenes 의 배양 pH 와 지방산 조성의 변화

오충훈,정상운,엄태한,김재헌 한국미생물생명공학회 1993 한국미생물생명공학회 학술발표초록집 Vol.1993 No.1

레이저와 조직의 상호작용

오충훈,안진철 대한광역학학회 2007 대한광역학학회지 = Journal of Korean Photodynamic Associat Vol.4 No.2

포토딘을 이용한 구강암에 대한 in vitro 항암효과연구

오충훈,임현주 대한구강악안면병리학회 2010 대한구강악안면병리학회지 Vol.34 No.1

PDT is an established cancer treatment modality. This can be attributed to the attractive basic concept of PDT; Combination of two therapeutic agents, a photosensitizing drug and light, which are relatively harmless by themselves but when combined, cause more or less selective tumor destruction. Hematoporphyrin-derived photosensitizers are known to be stable and highly efficient. In this study, we conducted a series of experiments to develop light-induced anticancer drugs against oral cancer cells. We tested the cytotoxicity of photodin by MTT assay and observed cell death pattern (apoptosis or necrosis) by hoechst 33342 and propidium iodide staining methods after PDT. IC50 value of photodin was 0.65 ug/ml. At higher doses of photodin (>7.8 ug/ml), cancer cells died exclusively from necrosis after PDT. By contrast, at IC50 value, photodin induced cancer cell to undergo apoptotic cell death. The induction begins approximately 6 hours after PDT. We investigated intracellular localization of photodin by oral cancer cell via confocal laser scanning microscopy. Oral cancer cells dual-stained with photodin and organelle-specific fluorescence probes (Mitotracker, Lysotracker, ER-Tracker) revealed that an intracellular fluorescence distribution was restricted to cytoplasmic compartments with no detectable fluorescence in the nucleus. Confocal images of cells containing photodin were overlapped with the mitochondria-specific fluorescence probe images of the same cells. These results demonstrated that photodin may play the role of a photosensitizer for oral squamous cancer cells without swelling and inflammation. Therefore, photodin-based PDT is a suitable treatment for oral cavity carcinoma patients.

두경부암세포에서 hydroxibacteriochlorin 을 이용한 in vitro 광역학치료

오충훈,이정구,주수경,천재식 대한구강악안면병리학회 2005 대한구강악안면병리학회지 Vol.29 No.2

We conducted a series of in vitro experiments to evaluate the efficiency of photodynamic therapy on head and neck cancer cell using hydroxybacteriochlorine from photosynthetic bacteria. We tested the cytotoxicity of the hydroxybacteriochlorine by MTI assay and observed the cell death pattern(apoptosis or necrosis) after PDT by hoechst 33342 and propidium iodide staining methods IC50 value of the hydroxybacteriochlorine was 0.22μg/rrúi. At higher doses of hydroxybacteriochlorine () 0.6μg/rrúi) , cancer cells died exclusively by necrosis after PDT. By contrast, at IC50 value, hydroxybacteriochlorine induced cancer cell to undergo apoptotic cell death. The induction begins approximately 6 hours after PDT. We investigates intracellular localization of hydroxybacteriochlorine by head & neck cancer cell via confocal laser scanning microscopy. Head & neck cancer cells dual-stained with hydroxybacteriochlorine and a panel of organelle- specific fluorescence probes (Mitotracker, Lysotracker, ER-Tracker) revealed an intracellular fluorescence distribution restricted to cytoplasmic compartments with no detectable fluorescence in the nucleus Confocal images of cells containing hydroxybacteriochlorine were never overlap in subcellular organelle fluorescence when digitally over layed with the organelle-specific fluorescence probe images of the same cells. These results demonstrated that the hydroxybacteriochlorine may have a function as a photosensitizer.

인체편평세포암세포주에서 9-HpbD-a 과 632 nm 다이오드 레이저를 이용한 광역학치료의 항암효과

오충훈,정필상,안진철,천재식 대한구강악안면병리학회 2005 대한구강악안면병리학회지 Vol.29 No.2

This study was conducted to test the anticancer effect of photodynamic therapy using chlorophyll derivative (9-HpbD-a) and 632nm diode laser. Human SNU 1041 cells were seeded into 96 well plate of 104cells/well and cultured for 24 hours. Cells were washed with media containing various concentration of 9-HpbD-a ranging from Oug/ml to 3.75ug/ml. Then 932 nm diode laser was given at various lasering time setting, and at various starting time after ini tial 24 hours of culture. The treated cells were incubated 48 hours and tetrazolium-based colorimetric(M'IT) assay was done to measure the viability of cells For in vivo study, SNU- 1041 cells were xenografted into the back of nude mouse. When the xenografted tumors grew up to 400-600 mm3, the animals were randomly placed into 4 groups: Group 1 (n=20) , PDT group, interstitial injection of 9-HpbD- a (47 ug/kg) followed by irradiation with 3.2 J/c야 of light 6 hours after then i띠 ection; Group II (n=lO) , irradiation with 3.2 J/crrf of light using diode laser; Group III (n=lO), in terstitial injection of 9-HpbD- a only(47 ug/kg); Group IV (n=lO), normal control group. The viability of cells was de creased with increasing lasering time No significant difference of cell viability was noted by variously delayed starting time of lasering. PDT effects were observed in the xenografted nude mouse model Group IV (no 9-HpbD-a, no laser irradiation) was a control group which showed a continuous tumor growth. Group III (9-HpbD-a i띠 ection only) showed no response, Group II (laser irradiation only) sho￦ed 1 complete remission out of 10 (10%) , Group 1 (9-HpbD-a and laser irradiation) showed 13 cpmplete remission out of 20 (65%) , Group 1 showed significant remission rate, comparing to other groups (p<0.05). This study demonstrated anticancer effect of photodynamic therapy using 9-HpbD-a and 632nm diode laser on human squamous cell carcinoma cell line.

사람의 암 세포주에서 녹색 형광단백질 유전자의 발현 양상

오충훈,홍승희 대한구강악안면병리학회 2010 대한구강악안면병리학회지 Vol.34 No.5

Many methods have been developed for more efficient gene delivery and expression in human cells. A number of studies have been performed in achieving successful gene delivery and expression conditions. We investigated differential gene expression patterns after delivery adenoviral vector containing green fluorescent protein(GFP) gene into human cancer cell lines. We constructed recombinant adenoviral Ad-CMV-GFP containing CMV promoter and GFP gene. The efficiency of gene expression was assessed by observation GFP expressing cells using fluorescent microscopy after transfer of Ad-CMV-GFP in concentrations of 0.1μl. 1μl. 10μl. At first, we evaluated expression patterns of gene in several human cancer cell lines, gastric adenocarcinoma cell line AGS was showed high level of GFP expression compared with colorectal adenocarcinoma cell line HT-29. After transfer 0.1μl of Ad-CMV-GFP in AGS, we could found that GFP expression cells were observed in next day and highly increased 2 days. While, small number of GFP expressing cells were examined in HT-29 and SNU-C4. Therefore, these data showed that AGS was expressed the highest level of GFP and almost AGS cells seems to express GFP in concentration of 1μl of Ad-CMV-GFP. GFP expression pattern in HT-29 reveal that expression was low in next day after gene transfer but significantly increase expression level in 2 days. In case of SNU-C4, GFP expression increased with increasing concentration of Ad-CMV-GFP and transfer times. For examine effects of transfer times in small amount gene, we transfer in concentration of 0.1μl Ad-CMV-GFP and detected GFP expression patterns after 2 days or 4 days. As a result, expression level of GFP in AGS was increase about 2 fold after 4 days compared with 2 days, but any difference of GFP expression levels were not showed in HT-29 and SNU-C4. Our study suggested that adenovirus was very efficient gene transfer vector for gene expression in human cancer cell lines. In addition to, we also demonstrated that gene expression patterns was dependent on each human cell lines. Therefore, further studies will be needed to confirm the optimum conditions for efficient gene delivery and expression in each target cell lines with consideration to cellular properties.

구강암세포에서 hydroxybacteriochlorine을 이용한 in vitro 광역학치료

오충훈(Chung Hun Oh) 대한구강악안면병리학회 2007 대한구강악안면병리학회지 Vol.31 No.2

사람의 암 세포주에서 녹색 형광단백질 유전자의 발현 양상

오충훈,홍승희 대한구강악안면병리학회 2010 대한구강악안면병리학회지 Vol.34 No.5

Many methods have been developed for more efficient gene delivery and expression in human cells. A number of studies have been performed in achieving successful gene delivery and expression conditions. We investigated differential gene expression patterns after delivery adenoviral vector containing green fluorescent protein(GFP) gene into human cancer cell lines. We constructed recombinant adenoviral Ad-CMV-GFP containing CMV promoter and GFP gene. The efficiency of gene expression was assessed by observation GFP expressing cells using fluorescent microscopy after transfer of Ad-CMV-GFP in concentrations of 0.1μl. 1μl. 10μl. At first, we evaluated expression patterns of gene in several human cancer cell lines, gastric adenocarcinoma cell line AGS was showed high level of GFP expression compared with colorectal adenocarcinoma cell line HT-29. After transfer 0.1μl of Ad-CMV-GFP in AGS, we could found that GFP expression cells were observed in next day and highly increased 2 days. While, small number of GFP expressing cells were examined in HT-29 and SNU-C4. Therefore, these data showed that AGS was expressed the highest level of GFP and almost AGS cells seems to express GFP in concentration of 1μl of Ad-CMV-GFP. GFP expression pattern in HT-29 reveal that expression was low in next day after gene transfer but significantly increase expression level in 2 days. In case of SNU-C4, GFP expression increased with increasing concentration of Ad-CMV-GFP and t ransfer times. For examine effects of transfer times in small amount gene, we transfer in concentration of 0.1μl Ad-CMV-GFP and detected GFP expression patterns after 2 days or 4 days. As a result, expression level of GFP in AGS was increase about 2 fold after 4 days compared with 2 days, but any difference of GFP expression levels were not showed in HT-29 and SNU-C4. Our study suggested that adenovirus was very efficient gene transfer vector for gene expression in human cancer cell lines. In addition to, we also demonstrated that gene expression patterns was dependent on each human cell lines. Therefore, further studies will be needed to confirm the optimum conditions for efficient gene delivery and expression in each target cell lines with consideration to cellular properties.

티타늄 판상에 배양된 정상인 조골세포에 대한 LED 세기별 효과에 관한 연구

최민철,오충훈,천재식 대한구강악안면병리학회 2008 대한구강악안면병리학회지 Vol.32 No.1

Low energy photon irradiation by light in the far red to near infrared spectral range(630~1000nm) using low energy lasers or light emitting diode arrays has been found to modulate various biological processes in cell culture and animal models. The purpose of this study was to examine the light emitting diode irradiation effect on activity of normal human osteoblast on titanium plate in vitro by various energy density, and to observe morphologic change of NHost on titanium plate and to analysis concentration of Ca++, IP and ALP. NHost were cultured in DMEM containing 10% FBS, and observed by inverted microscope for attatchment to the surface of titanium plate. Ca++, I.P., and alkaline phosphatase( ALP) concentration in medium was calculated during 4 weeks, which was treated with Wilcoxon rank, Anova test and linear regression. Morphologic changes showed LED produced in vitro increases of cell growth of 144~256% in NHost. During a culture period, Ca++ concentration was decreased. LED treatment(>3J/cm2) stimulate calcium consumption in NHost. Statistically, a significant difference was not found between LED power density. LED treated group(>3J/cm2) had higher total inorganic phosphate concentrations than control group in NHost. Statistically, a significant difference was not found between LED power density. No significant changes were observed between ALP acitivity and LED treatment. In spite of LED power density, there were rapid growth rate of NHost and no significant of Ca++, IP and P concentration but these concentration showed predominant change than that of control.