Photodynamic therapy(PDT) is recently developed as an effective treatment for malignant disease. Carboplatin, a less nephrotoxic analog of cisplatin, has been widely used for the treatment of multiple malignancies. In this study, we investigated the cytotoxic and apoptotic effect of combined modality of photofrin mediated PDT with cisplatin and carboplatin on KB cell human oral cancer cell line in vitro. The a ttached KB cells were incu bated with c isplatin(0.04mg/ml) and carboplatin(0.02mg/ml) for 24h at 37℃ and followed by photosensitization with photofrin for 6h and laser irradiation with 630nm LED at an intensity of 2.0 J/cm2 for activating photofrin for 15min. Then MTT assay and SYTO 16 green & Propidium iodide (PI) double staining were used respectively to measure the cytotoxicity and nuclear morphology at 24h after PDT. This study demonstrates that the combined modality with carbopaltin resulted in enhanced apoptotic cell death as well as cytotoxic e ffect on KB c ells in vitro, which s uggests the feasibility of combined modality and the possibility o f reducing the effective dosage of photofrin and carboplatin and lowering the side effects on normal cells

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.

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.

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.

Laser is used to prevent the early dental caries in dental f ield and to apply for treatment of stomatitis and hyper sens it ivity , and laser mass Recently it is reported that laser i1'r adiation affect on soft tissue treatment and bone 1'emodelling after dental implantation. The purpose of this study was to examine laser irradia ti on effect on activity of normal human osteoblast on titanium plate in vitro by various laser wave length, and to observe morphologic change of NHost on LiLa nium plate and to a nalysis concentration of Ca"++ , IP and ALP NHost were cultured in DMEM containing 10% FBS, and observed by in verted microscope 1"or attatchment to the surface of titanium plate. Ca ++, I.P. , and a lkaline phosphat ase(ALP) concent ration in medium was calculated during 4 weeks, which was treated with Wilcoxon rank, Anova test and linear regression. The obtained results were as follows Morphologic changes showed rapid growth rate of NHost ++ at 3 days of laser lrradiatlOn ln spite of laser wave type, Ca" and P concentration was decreased at 2 weeks and was the hig hest at 3 weeks, but decreased at 4 weeks In spite of laser wave type, ALP concentration was decreased at 2 weeks but was increased at 3-4 weeks, From the aboving results, in spite of laser wave type, there were rapid growth rat e of NHost a nd no significant of Ca"++ , IP and P concen tr ation but these concentration showed predominant change than that of control

PDT is an establi shed cancer treatment modali ty , This can be attributed to the attractive basic concept of PDT; the combina ti on 0[' two ther a peut ic agents, a photosensitizing drug and light, which are r elatively harmless by themselves but combined ultimately ca use more 0 1' less selective tumor destruction, The bacteriochlorophyll - derivatived photosensitizers are known to be s ta ble and hi ghly efficient‘ In this s tudy, we conducted a series of experiments to develope the ligh t induced anticancer drugs against oral cancer cell ‘ We tested the cytotoxicity of the hydroxybact eriochlorine by MTT a ssay and observed the cell death pattern (apoptosis or necrosis) after PDT by hoechst 33342 and propidium iodide s taining methods , IC50 value of the hydroxybacteriochlorine was 31,3ngjm.Q, At higher doses of hydroxybacteriochlorine () 60 ng/ 뼈) ， cancer cells died exc lus ively by nec rosis after PDT By contrast, at IC50 value, h ydroxybacteri ochlorine in duced ca ncel' cell to undergo a poptotic cell death The induction begins approximately 6 hours after PDT We investigates int racellu la r localizati on of hydroxybact eri ochl orine by ora l cancer cell via confocal laser scanning microscopy, Oral can cer cells dual-stained with hydroxybactel' iochlorine and organelle-specific fluoresc ence probes (Mi totracker, Lysotracker , ER- Trac ker) revealed an int l'acellula l' flu orescence distribution restrict ed to cytoplasmic compartments with no detectable fl uoresce nce in the nucleus Confocal images of cells containing hydroxybacteriochlorine were never overlap to mi tochondria, lysosome, endoplasmic l'eticulum when digitally overlapped with the organelle-specific flu orescence probe images of the same cells , These resul ts demonstrat ed that the hydroxybacteriochlorine may have a function as a photosens it izer and cytotoxicity hydroxybactel' ioc hlorine for oral ca ncer cell is more sensitive than head & neck cancer cell or cervical cance l' cell Ther efore PDT using hydroxybact eriochlorine is suitable treatment for oral cavity car cinoma patients.

This experiment was performed to study the biocompatibility of xenograft materials (ABBM. coralline HA). Both autogenous bone grafts and allogenic banked bone were frequently and successfully used to promote regeneration of parts of skeleton. The use of these types of grafts were limited by the cost of donor site operation for autogenous boneor by fear of the risk of infection of allogenic materials. Another type of graft is xenograft which include ABBM and coralline HA. For investigating the biocompatibility, generally many investigators used cancer cell lines or animal cell lines. But cancer cell lines and animal cell lines had functioned different metabolism from normal human cell. So the experiment used normal human osteoblast for compare the biocompatibility of ABBM with coralline HA which were fixed in 24 well base contained culture medium. After 1st, 3rd, 7th, 14th, 28th days, the culture medium were taken out and checked the concentrations ofcalcium( Ca), inorganic phosphate(IP) and alkaline phosphatase(ALP). In another method, histologic samples were investigated after 8weeks of xenograft materials implantated on rabbit's tibia, the bone was cut and made undecalcified ground samples and checked with fluorecent microscope, polarizing microscope, reflection electron microscope and electron probe microanalysis. The statistical results of concentrations (Ca, IP, ALP) of materials in the culture medium have decreasedby day's, which meant that xenograft materials were effective for bone remodelling. The concentrations in the culture medium of ABBM were lower than that of coralline HA, that meant that biocompatibility of ABBM were superior than that of coralline HA. Histologic samples showed that ABBM had better bone remodelling effect than coralline HA. ABBM showed good alizarin red marking lines, more deposition of Ca, IP, and dense color of bone around newly formed osteon and bone trabeculae. it was concluded that ABBM was more biocompatible than corallineHA in vivo and in vitro test

Since ancient Eygypt, various dental materials were used for lost teeth including gold. The key point of this materials were nontoxic to human body. Since early of 1990's, dental implant was done for recovery of maxillofacial defects. From middle of 1970's, osseointergation concept of implant was introduced and performed in dental field. Biocompatibility of titanium showed good effect for osseointergration but had some problems (Galvance current and toxic corrosion) with suprastructures such as gold crowns. This study was performed to make safe dental implants which have reduced Galvanic currents and corrosion. 3 kind of dental casting gold alloys (different Gold contents, 1㎝×1㎝×0.1㎝ plates.) were used as experimental group, while Titanium were used as control group. Normal human osteoblasts(NHOsts)were cultured during 1-4weeks for histologic study. For analysing the calcium(Ca), Phosphorus(P) and alkaline phosphatase(ALP), NHosts were cultred during 2-23days. After experiments, histologic finding were observed by LSM and SEM. Ca, P, ALP concentration by automatic biochemical analyzer were analyzed by ANOVA test and linear regression method. The results were as follows. Biocompatibility of dental casting gold alloys were similar to titianium alloys histolgically. Biochemical analysis of dental casting gold alloys had no significant difference to titianium alloy except AIGIS-Fine. We could conclude that biocompatibility of dental casting gold alloys with high contents of gold were superior to that of low contents and alloys with high contents of gold had no significant difference from titanium on NHost culture. Gold dental implant might be better than titanium implant due to similar biocompatibility and no galvanic currency.

In order to investigate stresses of dental patients in dental clinic by dental-analogous stimulations, we selected 23 women and 36 men who are students of dental college in D university. This experiment was performed to compare and analyze the changes of Galvanic skin resistance and heart rate by dental-analogous stimulations. In paired t-test of male group's GSR average, there were significant differences between sound and touch, and among pain and other stimulations. And in paired t-test of female group's GSR average, there were significant differences among pain and other stimulations. In paired t-test of male group's heart rate average, there were significant differences between vision and pain, and between touch and pain. And in paired t-test of female group's heart rate average, there were significant differences between smell and pain. In unpaired t-test of male and female group's GSR average, there were no significant differences in smell, sound, vision, touch, and pain. In unpaired t-test of male and female group's heart rate average, there were no significant differences in smell, sound, vision, touch, and pain. It seemed that stresses during dental treatment could be changed by surrounding circumstances and sex distinction.

1'hi s s t u dy was pe rformed to investigate the osseointegration on the interface between bone and dental casting gold alloy's impla nts . Some inves tigators prese nted that only fi brous integratioJ1 in implant interface of precious met al im plants was occu red. 1'he mate ri a ls of dental implant must have biocompatibility and have no phY5ical 01' chemical s ide effect lmpla nts whi ch we re made of pure t itanium or titanium alloys had 50me chemical problem such as Galvanic current wi th s uprastru ctu re cur re ntly. So. we t hought that there was no problem if dental casting gold al1 0y’s impla nts had 5ucceed osseoin tegration. Thi s s tudy used 99%. 86%. 70% gold composition ìmplant as experimental group and tita niuITI implant as cont ral g rOllp. After 4th a nd 8th weeks of implant impLantation. experimental animals were saC l 나 i ce d and hi stologic sections we re made. All sections were examined by 8EM. PM. L8M. and EPMA. lmplants made of dental casting gold a ll oys occllrred good osseointegration as titanium implants. Osseointegr ation were not depend on the gold composit ion in implant groups ’ denta l casting gold al1oys. Both groll ps‘ osseointegration were 1TI0re completed in 8th weeks than 4th weeks . From t he a boving results. dental casting gold alloy's implants had induced similar osseointegrat ion to t h at 01" titan iu rn irnplants . So if we make implant and suprastructllre of dental casting gold alloys. Galvanic current and chemical co rrosio ns wOllld be sllppressed

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.

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.

In this study, we tried to offer the possibility of White Hibiscus syriacus L. (WHS) flower extracts as a preven-tive and improving agent of osteoporosis that bone mass reduction is induced by an decrease of osteoblast involved in boneformation and increase of bone resorption by osteoclast activity. As a results, it was found to have antioxidant activity andcontain a flavonoid contents (47.74㎎/g) of the WHS flower. There was cytotoxicity at more than 250㎍/㎖ concentration ofWHS flower extract of RANKL-induced osteoclast in RAW264.7. There were no significant inhibited TRAP activity in theWHS leaf and stem. However, it was confirmed that it is significantly inhibited the differentiation activity of osteoclasts in 50and 100㎍/㎖ concentration of cells of stability levels of only WHS flower extracts (p<0.01). The WHS flower prominentlyinhibited RANKL-induced osteoclast differentiation activity by decreased calcitonin receptor and TRAP mRNA (p<0.01).These results indicate that of osteoclasts differentiation activity is inhibited by protection of oxidative stress due to the anti-oxidant activity of the WHS flower. Therefore, suggesting the WHS flower may be a presents the possibility as a preventiveand therapeutic agents for osteoporosis.

Osteoporosis induces a bone mineral density loss due to imbalance of bone homeostasis that is achieved by osteoclasts (which are involved in bone resorption) and osteoblasts (which are involved in bone formation). Thus, this study was performed to evaluate the effects of hot water extract of the Achyranthes bidentata Blume (ABB) and Panax ginseng (Gin) on osteoclast and osteoblast differentiation. In this study, there was no cytotoxicity by ABB, 50 and 100 μg/ml of Gin significantly decreased cell viability of RANKL-induced osteoclast in RAW264.7 cell (p < 0.01). But, it was 50 μg/ml of ABB and Gin mixtures increased due to protective action of ABB. Furthermore, Gin contained groups (Gin, ABB and Gin mixtures) were inhibitory effects on osteoclast differentiation and bone resorption, and increased in osteoblast differentiation activity. Gin clearly inhibited RANKL-induced osteoclast differentiation by decreased calcitonin and TRAP (p < 0.01). Also, these extracts significantly increased calcium accumulation formation of osteoblastic differentiation reagents-induced osteoblast in MC3T3-E1 cell (p < 0.05). These results suggest that ABB and Gin mixtures may be a potential as drug for the treatment of osteoporosis.