Few studies have evaluated the apoptosis-inducing efficacy of NaF on cancer cells in vitro but there has been no previous investigation of the apoptotic effects of NaF on human oral squamous cell carcinoma cells. In this study, we have investigated the mechanisms underlying the apoptotic response to NaF treatment in the YD9 human squamous cell carcinoma cell line. The viability of YD9 cells and their growth inhibition were assessed by MTT and clonogenic assays, respectively. Hoechst staining, DNA electrophoresis and TUNEL staining were conducted to detect apoptosis. YD9 cells were treated with NaF, and western blotting, immunocytochemistry, confocal microscopy, FACScan flow cytometry, and MMP and proteasome activity assays were performed sequentially. The NaF treatment resulted in a time- and dose-dependent decrease in YD9 cell viability, a dose-dependent inhibition of cell growth, and the induction of apoptotic cell death. The apoptotic response of these cells was manifested by nuclear condensation, DNA fragmentation, the reduction of MMP and proteasome activity, a decreased DNA content, the release of cytochrome c into the cytosol, the translocation of AIF and DFF40 (CAD) into the nucleus, a significant shift of the Bax/Bcl-2 ratio, and the activation of caspase-9, caspase-3, PARP, Lamin A/C and DFF45 (ICAD). Furthermore, NaF treatment resulted in the downregulation of G1 cell cyclerelated proteins, and upregulation of p53 and the Cdk inhibitor $p27^{KIP1}$. Taken collectively, our present findings demonstrate that NaF strongly inhibits YD9 cell proliferation by modulating the expression of G1 cell cycle-related proteins and inducing apoptosis via mitochondrial and caspase pathways.
Kim, Young-Youn;Kim, Myung-Jin;Choi, Keum-Kang;Hong, Seong-Doo;Myoung, Hoon
Journal of the Korean Association of Oral and Maxillofacial Surgeons
/
v.34
no.1
/
pp.71-82
/
2008
Survivin is a member of the inhibitors of apoptosis (IAP) family that have been known to inhibit activated caspases in apoptosis. In contrast to most IAP family members, survivin mRNA is expressed during fetal development, is not found in normal adult tissues and is overexpressed again in the cancer. Though survivin expression has been documented in most human cancers, little is known about its expression in OSCC and its potential value as a predictor of cancer survival. The purpose of this study was to investigate survivin expression in OSCC and to evaluate its value as a prognostic marker. We evaluated survivin expressions in cancer lines and OSCC samples and investigated the relationships between survivin expressions and clini-co-pathological parameters including stage, differentiation, proliferation, lymph node metastasis, blood vessel density, and gelatinolytic activity. With immunohistochemistry, we analyzed survivin expression in 38 OSCCs. Patients' clinico-pathological parameters and their survival rate were analyzed to reveal their correlations with Survivin expressions. We cultured oral cancer cell lines and evaluated the correlation between gelatinolytic activities and survivin expressions of them. Survivin protein was observed both in nuclei and cytoplasm of tumor specimens while little or not observed in normal gingival mucosal tissues. Additionally, survivin expressions were correlated with lymph node metastasis, tumor proliferation and survival rate. Survivin expression was observed in 100% of 38 samples of OSCC and its expression levels are statistically associated with the proliferative activity of the tumors, lymph node metastasis and the survival of the patients. Based on these results, survivin is commonly expressed in OSCC and may thus provide valuable prognostic information related with lymph node metastasis, proliferation and survival rate as well as a potential therapeutic target in OSCC.
Oleanolic acid is a natural triterpenoid that exists widely in foods and some medicinal herbs. The purpose of this study was to determine the antimicrobial activity of oleanolic acid against Streptococcus mutans strains isolated from a Korean population. Antimicrobial activity against these bacteria was evaluated by minimal inhibitory concentration (MIC) and time kill curves. The tolerance of human gingival fibroblasts and human periodontal ligaments to oleanolic acid was tested using a methyl thiazolyl tetrazolium (MTT) assay. The $MIC_{90}$ value of oleanolic acid for both S. mutans and S. sobrinus isolated from Koreans was 8 ${\mu}g/ml$. Oleanolic acid showed bactericidal effects against S. mutans ATCC $25175^T$ and S. sobrinus ATCC $33478^T$ at $1\;{\times}\;MIC$ ($8{\mu}g/ml$) and had no cytotoxic effects against KB cells at this dose. The results suggest that oleanolic acid could be useful in the future development of oral hygiene products for the prevention of dental caries.
Cis-3-O-p-hydroxycinnamoyl ursolic acid (HCUA), a triterpenoid compound, was purified from Elaeagnus oldhamii Maxim. This traditional medicinal plant has been used for treating rheumatoid arthritis and lung disorders as well as for its anti-inflammation and anticancer activities. This study aimed to investigate the anti-proliferative and apoptotic-inducing activities of HCUA in oral cancer cells. HCUA exhibited anti-proliferative activity in oral cancer cell lines (Ca9-22 and SAS cells), but not in normal oral fibroblasts. The inhibitory concentration of HCUA that resulted in 50% viability was $24.0{\mu}M$ and $17.8{\mu}M$ for Ca9-22 and SAS cells, respectively. Moreover, HCUA increased the number of cells in the sub-G1 arrest phase and apoptosis in a concentration-dependent manner in both oral cancer cell lines, but not in normal oral fibroblasts. Importantly, HCUA induced p53-mediated transcriptional regulation of pro-apoptotic proteins (Bax, Bak, Bim, Noxa, and PUMA), which are associated with mitochondrial apoptosis in oral cancer cells via the loss of mitochondrial membrane potential. HCUA triggered the production of intracellular reactive oxygen species (ROS) that was ascertained to be involved in HCUA-induced apoptosis by the ROS inhibitors YCG063 and N-acetyl-L-cysteine. As a result, HCUA had potential antitumor activity to oral cancer cells through eliciting ROS-dependent and p53-mediated mitochondrial apoptosis. Overall, HCUA could be applicable for the development of anticancer agents against human oral cancer.
Journal of the Korean Association of Oral and Maxillofacial Surgeons
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v.34
no.3
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pp.293-299
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2008
Bite force is created by the force of adjacent teeth accompanied with tension of masticatory muscle. The bite force value is greater in male than in female and ha maximum value at first molar. Masseter muscle is associated with bite force and during muscle contraction the electric signal is expressed in EMG form. The aim of the study is to assess recovery time for masseter muscle activity and according to each part of bite force after open reduction with internal fixation when mandibular angle fracture and subcondyle fracture occurred. And to determine the appropriate period for mandibular fracture patients to have normal masticatory activity. 30 patients with normal bite condition was selected for control group and from April, 2007 to September, 2007, 20 patients who visited our department of oral and maxillofacial surgery of Dankook University, were selected for the study and were diagnosed as mandibular angle fracture and subcondyle fracture. For control group, the bite force for incisors, canine, premolars and molars and activity of the masseter muscle was measured and compared for 1, 2, 3, 4, 6 and 8 weeks. That was divided as fracture side and normal side. Mann-Whitney U test was performed for significant difference and the following result was obtained. 1. The maximum voluntary bite force for incisors, canine, premolars and molars portion were 0.113 kN, 0.182kN, 0.295kN and 0.486kN and the masseter muscle activity was 0.192 volts in the control group. 2. The maximum bite force at fracture side was recovered by 4th weeks for incisors, 6th weeks for canine and premolars and 8th weeks for molars and the masseter muscle activity was recovered by 6th weeks in the experimental group. 2. The maximum bite force at normal side was recovered by 4th weeks for incisors, 6th weeks for canine, premolars and molars and the masseter muscle activity was recovered by 3rd weeks in the experimental group. 3. The method for internal fixation by 2.0mm miniplates at both superior and inferior border had no complications according for twenty patients and had a satisfactory recovery. According to the result, patient with mandibular angle fracture and subcondyle fracture, 8 weeks was required for bite force recovery. Therefore, patients with open reduction and internal fixation under general anesthesis, it can be assumed that 8 weeks was needed after operation in order to have normal bite force and masseter muscle recovery.
The purpose of this study was to investigate the quality of life of clients who treated and equipped prosthesis in Eulji University Hospital in Dae-Jun City. The results of this study were as follows. The oral status of Female had more carious tooth than male had. Carious tooth(p<0.05) and losed tooth(p<0.001) was more popular in older female. The frequency of treated tooth was higher under 59 years old female than over 60 years old(p<0.01). In education category, carious tooth(p<0.05) and losed tooth(p<0.000) was more popular in low education than high education In the score of relation of OHIP-14 category and general characteristic, female was significantly higher than male in dysfunction( p<0.05), physical disorder(p<0.001) and activity disorder(p<0.05). The score was significantly higher in older than younger in dysfunction(p<0.05), physical disorder(p<0.001) and activity disorder(p<0.001). The score of education was significantly higher in low education group than high education group in physical pain (p<0.001), mental discomfort (p<0.001). The score of marriage status was significantly higher in marriaged group than other group in dysfunction (p<0.05) and mental discomfort (p<0.05). In the score of relation of OHIP-14 category and oral status, the score of carious tooth group was significantly higher in dysfunction(p<0.05), physical disorder(p<0.05) and activity disorder(p<0.05). The score of losed tooth group was significantly higher in dysfunction(p<0.05), physical pain (p<0.05), mental discomfort(p<0.05), physical disorder(p<0.05) and activity disorder(p<0.01). The score of non treated tooth group was significantly higher in dysfunction(p<0.05), mental discomfort(p<0.05), social disorder(p<0.05) and activity disorder(p<0.05). In the score of relation of OHIP-14 category and the sort of prosthesis, the score of being bridge group was significantly higher than being crown group in activity disorder(p<0.05). The OHIP-14 category and the number of prosthesis was not significant relationship. These results suggest that the information and services of the process of dental prosthesis based on subjective evaluation should be provided to clients rather than based on clinical evaluation. The continued system of oral management should be developed and provided.
Background: The purpose of this study was to investigate the anti-oral microbial activity and anti-inflammatory effects of rosmarinic acid (RA) in lipopolysaccharide (LPS)-stimulated MC3T3-E1 osteoblastic cells on a titanium (Ti) surface during osseointegration, and to confirm the possibility of using RA as a safe natural substance for the control of peri-implantitis (PI) in Ti-based dental implants. Methods: A disk diffusion test was conducted to confirm the antimicrobial activity of RA against oral microorganisms. In order to confirm the anti-inflammatory effects of RA, inflammatory conditions were induced with 100 ng/ml of LPS in MC3T3-E1 osteoblastic cells on the Ti surface treated with or without 14 ㎍/ml of RA. The production of nitric oxide (NO) and prostaglandin E2 (PGE2) in LPS-stimulated MC3T3-E1 osteoblastic cells on the Ti surface was confirmed using an NO assay kit and PGE2 enzyme-linked immunosorbent assay kit. Reverse transcription polymerase chain reaction and western blot analysis were performed to confirm the expression of interleukin (IL)-1β and tumor necrosis factor (TNF)-α in total RNA and protein. Results: RA showed weak antimicrobial effects against Streptococcus mutans and Escherichia coli, but no antimicrobial activity against the bacteria Aggregatibacter actinomycetemcomitans and the fungus Candida albicans. RA reduced the production of pro-inflammatory mediators, NO and PGE2, and proinflammatory cytokines, TNF-α and IL-1β, in LPS-stimulated MC3T3-E1 osteoblastic cells on the Ti surface at the protein and mRNA levels. Conclusion: RA not only has anti-oral microbial activity, but also anti-inflammatory effects in LPS-stimulated MC3T3-E1 osteoblasts on the Ti surface, therefore, it can be used as a safe functional substance derived from plants for the prevention and control of PI for successful Ti-based implants.
In this inverstigation, the various flavonoid aglycones were evaluated for their inhibitory activities against croton-oil or arachidonic acid induced mouse ear edema by oral or topical administration. The compounds tested were thirteen derivatives of flavan-3-ol(catechin and epicatechin), flavanone (flavanone and naringenin), flavone (flavone, chrysin and apigenin), flavonol(favonol, galangin, quercetin and morin) and isoflavone (biochanin A and 2-carbethoxy-5,7-dihydroxy-4'-methoxyisoflavone), along with hydrocortisone, indomethacin, 4-bormophenacyl bromide, nordihydroguaiaretic acid and phenidone as positive controls. A(isoflavone) were found to show broad inhibitoty activities (14-52%) against croton-oil or arachidonic acid induced ear edema by oral or topical application at the dose of 2 mg/mouse, although they showed less activity than hydrocortisone (26-88%) or indomethacin (36-80%). Flavonoid agtlycones tested showed higher activity when aplied topically than by the oral administration. It was also found that they inhibited arachidonic acid induced edema more profoundly than croton-oil induced edema by topical application. In arachidonic acid induced edema when applied topically, flavone derivatives such as flavone, chrysin and apigenin were revealed to be the good inhibitory agents in addition to flavonols and isoflavones. When quercetin and biochanin. A were selected for evaluating in carrageenan induced rat pleurisy and biochanin both flavonoids showed antiinflammatory activity at the dose of 70 mg/kg by the oral adminis-tration. All of these results revealed that flavonoid aglycones, especially 5,7-dihydroxy-flavonols having hydroxyl group(s) in B-ring and biochanin A (isoflavone) possessed in vivo antiinflammatory activity.
Purpose To observe the histopathological changes in the periodontal tissues of mandibular molars in streptozotocin-induced diabetic rats after irradiation. Materials and Methods : The male Sprague-Dawley rats weighing approximately 250 gm were divided into four groups: control, diabetes, irradiation, and diabetes- irradiation groups. Diabetes mellitus was induced in the rats by injecting streptozotocin. Rats in the control and irradiation groups were injected with citrate buffer only After 5days, the head and neck region of the rats in irradiation and diabetes-irradiation groups were irradiated with a single absorbed dose of 10Gy. All the rats were sacrificed at 3, 7, 14, 21, and 28 days after irradiation. The specimen including the mandibular molars were sectioned and observed using a histopathological method. Results In the diabetes group, osteoclastic activity was observed in the alveolar bone and the root throughout the period of experiment. Also, osteoblastic and fibroblastic activities were markedly decreased. In the irradiation group, the osteoclasts were observed in the alveolar bone and the dilated capillaries were increased in the early experimental phases. However, vigorous osteoblastic activity was noted in the late experimental phases. In the diabetes-irradiation group, osteoblastic activity in the alveolar bone and the root was observed in the early experimental phases. However, there were no resorption and osteoblastic activity in the alveolar bone and the root in the late experimental phases, and obvious atrophic change of fibrous tissues was noted. Conclusion : This experiment suggests that osteoblastic activity was caused by irradiation in the late experimental phases, but atrophic change of the periodontal ligament tissues was induced after irradiation in diabetic state.
Endothelin-1 (ET-1) is a recently discovered potent vasoconstrictive peptide. It was first identified in vascular endothelial cells. ET-1 is a 21-amino acid peptide and elicits systemic effects such as stimulation of the production of atrial natriuretic peptide and release of aldosterone and corticosterone. In this study, to examine the role of ET-1 in the bone metabolism, effect of ET-1 on the proliferation and activity of osteoblastic cells was studied using HOS cells as osteoblast model. ET-1 dose-dependently increased the cell proliferation as determined by cell counting and MTT reduction assay after 48hr treatment. Alkaline phosphatase activity was inhibited by ET-1 and showed significant inhibition by 50 and 100 nM ET-1. ET-1 increased NBT reduction by HOS cells dose-dependently showing that ET-1 may increase the superoxide production by osteoblasts. Nitrite concentration in the media of HOS cell culture without cytokine stimulation was negligible and unaffected by ET-1 after 48hr treatment. Finally, after collection and concentration of conditioned media, gelatinase activity produced by HOS cells was determined by zymography. HOS cells can produce and secrete the gelatinase (gelatinase A type as determined by molecular weight of about 65,000) into culture media, however, ET-1 had no effect on the gelatinase activity. These findings suggest that ET-1 may have diverse effects on the proliferation and differentiation of osteoblasts, therefore, it may play an important role in bone metabolism.
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