Cytotoxicity and arecoline mechanisms in human gingival fibroblasts in vitro

Betel nut chewing, like cigarette smoking, is a popular oral habit which impinges on the daily lives of a population of approximately 200 million. People who chew betel nuts have a higher prevalence of periodontal diseases than those who do not. Many of the undesirable effects of betel nuts have been attributed to arecoline, a major component of the particular alkaloid in betel nuts. In this in vitro study, we have focused on the effects of arecoline and the role it could play in periodontal breakdown via its direct effects on human gingival fibroblasts. Human gingival fibroblasts were derived from three healthy individuals undergoing crown-lengthening procedures. We found that arecoline is cytotoxic to human gingival fibroblasts at a concentration higher than 50 μg/ml by depleting intracellular thiols and inhibiting mitochondrial activity (P<0.05). In addition, the cells displayed a marked arrest at G₂/M phase in a dose-dependent manner. Repeated and long-term exposure to arecoline could impair the gingival fibroblast functions. As they are cytotoxic, the use of betel nut products in conjunction with periodontal therapy may interfere with optimal healing and/or lead to further periodontal breakdown. Received: 16 February 2000 / Accepted: 25 July 2000     

Adverse effects of arecoline and nicotine on human periodontal ligament fibroblasts in vitro

BACKGROUND, aims: The habit of betel nut chewing impinges on the daily lives of approximately 200 million people. Betel quid chewers have a higher prevalence of periodontal diseases than non-chewers. This study examined the pathobiological effects of arecoline, a major component of the betel nut alkaloids, on human periodontal ligament fibroblasts (PDLF) in vitro. METHOD: Cell viability, proliferation, protein synthesis, and cellular thiol levels were used to investigate the effects of human PDLF exposed to arecoline levels of 0 to 200 microg/ml. In addition, nicotine was added to test how it modulated the effects of arecoline. RESULTS: Arecoline significantly inhibited cell proliferation in a dose-dependent manner. At concentrations of 10 and 30 microg/ml, arecoline suppressed the growth of PDLF by 20% and 50% (p < 0.05), respectively. Arecoline also decreased protein synthesis in a dose-dependent manner during a 24-h culture period. A 100 microg/ ml concentration level of arecoline significantly inhibited protein synthesis to only 50% of that in the untreated control (p < 0.05). Moreover, arecoline significantly depleted intracellular thiols in a dose-dependent manner. At concentrations of 25 microg/ml and 100 microg/ml, arecoline depleted about 18% and 56% of thiols (p < 0.05), respectively. This suggests that arecoline itself might augment the destruction of periodontium associated with betel nut use. Furthermore, the addition of nicotine acted with a synergistic effect on the arecoline-induced cytotoxicity. At a concentration of 60 microg/ml, arecoline suppressed the growth of PDLF by about 33% and 5 mM nicotine enhanced the arecoline-induced cytotoxic response to cause about 66% cell death. CONCLUSION: During thiol depletion, arecoline may render human PDLF more vulnerable to reactive agents within cigarettes. Taken together, people who combine habits of betel nut chewing with cigarette smoking could be more susceptible to periodontium damage than betel nut chewing alone.     

Cytotoxic and non-genotoxic effects of arecoline on human buccal fibroblasts in vitro

Chang YC, Tai KW, Cheng MH, Chou LSS, Chou MY: Cytotoxic and non-genotoxic effects of arecoline on human buccal fibroblasts in vitro. J Oral Pathol Med 1998; 27: 68–71. © Munksgaard, 1998.
Betel quid chewing has been linked to oral submucous fibrosis and oral cancer. Cytotoxicity and genotoxicity assays were used to investigate the pathobiologi-cal effects of arecoline on cultured human buccal fibroblasts. Arecoline increased double-stranded polynucleic acid at the concentration of 0.1 to 10 μg/ml in a concentration-dependent manner. At a concentration higher than 50 μg/ml, arecoline was cytotoxic to cultured fibroblasts and the cytotoxicity was dose-dependent. No genotoxicity for arecoline was found even at a concentration of 400 μg/ml. On the other hand, 600 μg/ml glutathione (GSH) and 200 μg/ml glycyrrhizin could prevent the arecoline-induced cytotoxicity. These results indicate that arecoline is a cytotoxic agent and no genotoxicity was found to human buccal fibroblasts. Furthermore, increasing consumption of GSH- and glycyrrhizin-rich foods may reduce the oral diseases associated with betel quid chewing.     

Inhibition of the migration, attachment, spreading, growth and collagen synthesis of human gingival fibroblasts by arecoline, a major areca alkaloid, in vitro

Because betel quid (BQ) chewing has been linked to a higher prevalence of periodontal diseases, the pathobiological effects of arecoline, a main alkaloid found in areca nut, were investigated in cultured human gingival fibroblasts. At concentrations higher than 0.4 mM, arecoline inhibits cell attachment, cell spreading and cell migration in a dose-dependent manner. These inhibitory effects were associated with intracellular depletion of glutathione (GSH). At concentrations of 0.4 mM and 1 mM. arecoline depleted about 26% and 45% of GSH after 2 h incubation. Exposure of cells to areeoline at concentrations lower than 0.4 mM for 2 h showed no significant decrease in either cell viability or intracellular GSH. However, incubation of cells for 24 h in 1 mM arecoline decreased the cell numbers to only 35% of those in the untreated control. Arecoline also decreased cell growth and collagen synthesis in a dose-dependent manner. Because of repeated and long-term exposure to arecoline. BQ chewers could be more susceptible to periodontal damage and less responsive to new attachment procedures.     

Salivary arecoline levels during areca nut chewing in human volunteers

J Oral Pathol Med (2010) 39 : 465–469 Background: Arecoline stimulates cultured cells above 0.1 μg/ml and is cytotoxic above 10 μg/ml. Although this alkaloid seems important for areca nut induced oral carcinogenesis, little is known of the levels achieved during chewing. Materials and methods: Saliva was collected in 3‐ to 5‐min intervals over 50 min in 32 habitual chewers: before, for 25 min during, and for 20 min after chewing areca nut (0.5 g) without any other additives. Salivary arecoline was quantitated by HPLC‐MS. Controls comprised six subjects who denied areca nut use, and who were given rubber‐base material to chew during experiments instead. Results: Arecoline was detected before chewing in 22 subjects, exceeding the 0.1 μg/ml threshold in 20 cases. Salivary arecoline exceeded either the 0.1 or 10 μg/ml thresholds in all participants during chewing (P < 0.001). Maximum concentrations ranged from 5.66 to 97.39 μg/ml. All subjects reached 0.1 μg/ml salivary arecoline in at least 85% of time points studied (P < 0.0001), whereas 10 μg/ml was reached in 11 participants in at least 30% of the time points (P < 0.003). Arecoline concentrations varied greatly over time between individuals, and levels were much lower when peak concentrations were reached before 3 min, than in cases where arecoline peaked later (P < 0.02). No salivary arecoline was found in control saliva. Conclusions: Areca nut users have persistent background salivary arecoline levels long after chewing, whereas concentrations achieved are highly variable and consistent with a role in oral pre‐malignancy and malignancy.     

Effects of arecoline, safrole, and nicotine on collagen phagocytosis by human buccal mucosal fibroblasts as a possible mechanism for oral submucous fibrosis in Taiwan

BACKGROUND: Oral submucous fibrosis (OSF) is associated with the betel quid chewing habit, and 86% of betel quid chewers in Taiwan are also smokers. Arecoline and safrole are major principles in the composition of betel quid, and nicotine is the main toxic ingredient of cigarettes. METHODS: To explore the pathogenesis of OSF, flow cytometry was used to compare collagen phagocytosis by fibroblasts from the normal and the OSF region of the same 15 OSF patients. RESULTS: The results indicated that heterogeneity of fibroblasts existed because collagen phagocytosis by fibroblasts from the normal region was higher than from the OSF region in the same patient. The percentage of phagocytic cells was significantly inhibited by 10, 25 and 50 microg/ml arecoline, safrole and nicotine in normal fibroblast cultures, respectively, and the percentage of phagocytic cells was significantly reduced by 25, 25 and 50 microg/ml arecoline, safrole and nicotine in OSF fibroblast cultures, respectively. Collagen phagocytosis by fibroblasts exhibited prominent dose-dependent inhibition as the concentration of arecoline, safrole, and nicotine increased. Besides, nicotine had a synergistic effect on arecoline- or safrole-inhibited collagen phagocytosis. CONCLUSIONS: The present study concludes that even in the same person, the collagen phagocytosis by fibroblasts is different between normal and OSF region. The deficiency in collagen phagocytosis by fibroblasts of the lesion might participate in the pathogenesis of OSF. Arecoline, safrole and nicotine, which are released in saliva during BQ chewing plus cigarette smoking, inhibit collagen phagocytosis by fibroblasts in a dose-dependent manner and may induce OSF formation in Taiwan's patients.     

Synergistic effects of nicotine on arecoline-induced cytotoxicity in human buccal mucosal fibroblasts

Areca quid chewing has been linked to oral submucous fibrosis and oral cancer. Arecoline, a major areca nut alkaloid, is considered to be the most important etiologic factor in the areca nut. In order to elucidate the pathobiological effects of arecoline, cytotoxicity assays, cellular glutathione S-transferase (GST) activity and lipid peroxidation assay were employed to investigate cultured human buccal mucosal fibroblasts. To date, there is a large proportion of areca quid chewers who are also smokers. Furthermore, nicotine, the major product of cigarette smoking, was added to test how it modulated the cytotoxicity of arecoline. At a concentration higher than 50 microg/ml, arecoline was shown to be cytotoxic to human buccal fibroblasts in a dose-dependent manner by the alamar blue dye colorimetric assay (P<0.05). In addition, arecoline significantly decreased GST activity in a dose-dependent manner (P<0.05). At concentrations of 100 microg/ml and 400 microg/ml, arecoline reduced GST activity about 21% and 46%, respectively, during a 24 h incubation period. However, arecoline at any test dose did not increase lipid peroxidation in the present human buccal fibroblast test system. The addition of extracellular nicotine acted synergistically on the arecoline-induced cytotoxicity. Arecoline at a concentration of 50 microg/ml caused about 30% of cell death over the 24 h incubation period. However, 2.5 mM nicotine enhanced the cytotoxic response and caused about 50% of cell death on 50 microg/ml arecoline-induced cytotoxicity. Taken together, arecoline may render human buccal mucosal fibroblasts more vulnerable to other reactive agents in cigarettes via GST reduction. The compounds of tobacco products may act synergistically in the pathogenesis of oral mucosal lesions in areca quid chewers. The data presented here may partly explain why patients who combined the habits of areca quid chewing and cigarette smoking are at greater risk of contracting oral cancer.     

Growth of oral and skin fibroblasts from patients with oral submucous fibrosis

The purpose of the investigation was to evaluate and compare the proliferation (growth) of mouth fibroblasts and skin fibroblasts from patients with oral submucous fibrosis (OSF). Material comprised fibroblasts from fibrous bands situated in the buccal mucosa and from the inner aspect of the forearm of 8 patients with classic features of OSF as well as fibroblasts from 6 buccal mucosa and 8 skin biopsy specimens from healthy non-areca nut chewing individuals. Cells were cultured for 8 days according to standard techniques. Their growth was monitored daily, under optimal conditions as well as exposure to concentrations of arecoline. The data were analyzed using regression analysis, analysis of variance and the Kruskal-Wallis test. We found no statistically significant differences between the proliferation patterns of oral and skin fibroblasts from patients or between those from patients and controls. The reaction of the cells exposed to concentrations of arecoline was similar; at low concentrations (0.1–10 μg/ml) normal growth was maintained, while 100 μg/ml inhibited growth. It is concluded that fibroblasts from mouths affected by OSF have proliferation patterns which fall within normal parameters, that the excessive collagen formation in established OSF is not due to increased fibroblast proliferation and that arecoline does not stimulate fibroblast proliferation.     

Effects of areca nut, inflorescence piper betle extracts and arecoline on cytotoxicity, total and unscheduled DNA synthesis in cultured gingival keratinocytes

Betel quid (BQ) chewing has a strong correlation with oral leukoplakia, submucous fibrosis and oral cancer. For elucidation of its pathogenesis, we investigated the effects of areca nut (AN) and inflorescence piper betle (IPB) extracts and arecoline on the growth, total DNA synthesis (TDS) and unscheduled DNA synthesis (UDS) of cultured human gingival keratinocytes (GK). Arecoline and AN extract suppressed the growth of GK over 5 days of incubation in a dose-dependent fashion. At concentrations of 100, 200 and 400 μg/ml, AN extract suppressed the growth of GK by 31%, 46% and 90%, respectively. The IPB extracts exerted less inhibitory effect on the growth of GK. IPB extract (200–400 μg/ml) decreased cell numbers by 20–40% over 5 days of incubation. Moreover, at a concentration of 0.1, 0.2 and 0.4 mM, arecoline suppressed cell growth by 44%, 77% and 96%, respectively. However, only AN extract induced TDS and UDS in cultured GK within 6 h of exposure. Induction of UDS by AN extract was concomitant with the presence of apparent intracellular vaculoization. Arecoline was also toxic to GK, but did not induce intracellular vacuolization. At a concentration range of 200–1600 μ/ml, AN extract induced TDS by 2.1- to 6.5-fold. Furthermore, at a concentration of 400–1600 μ/ml, AN extract elevated the UDS by 2.4- to 5.5-fold more than that of untreated control. On the contrary, IPB extract (200–1600 μ/ml) and arecoline (0.2–1.6 mM) inhibited the TDS and UDS of GK to a different extent. Simultaneous exposure of confluent GK to AN extract, IPB extract and arecoline for 1 to 5 days led to different degrees of cytotoxicity that was dose-and time-dependent. These results indicate that AN, IPB and arecoline take part in the pathogenesis of BQ chewing-related oral mucosal lesions, possibly through both genotoxic and non-genotoxic mechanisms.     

Observations on the effect of areca nut extracts on oral fibroblast proliferation

The effects of aqueous extracts of raw, baked and boiled areca nuts were tested on cultured human buccal mucosa fibroblasts. Cells were exposed to extract concentrations of 0, 50, 100, 150, 300 and 500 μg/ml. The arecoline and arecaidine content was determined in the extracts with HPLC and raw nut contained 5.5% in m. baked nut 6.6% mm and boiled nut 7.1% m/m. Extract concentrations of 50 to 150 μg/ml inhibited cell growth in a concentration-dependent manner but did not lead to total cell death during a 7 day period. However, total cell death did occur with concentrations of 300 and 500 μg/ml. It is concluded that areca nut extract is toxic to cultured fibroblasts and inhibits their proliferation in a concentration-dependent manner.     

Arecoline and oral keratinocytes may affect the collagen metabolism of fibroblasts

Background:  The characteristic of oral submucous fibrosis (OSF) is related with the disturbance of synthesis and degradation of extracellular matrix. Arecoline, the areca nut (betel nut) component of betel quid, plays a major role in pathogenesis of OSF. But the exact mechanism how arecoline influences the collagen metabolism is unclear.
Methods:  Oral keratinocytes and fibroblasts were cocultured and keratinocytes were pre-treated by arecoline. Fibroblasts alone, fibroblasts stimulated by arecoline, fibroblasts cocultured with keratinocytes and fibroblasts cocultured with keratinocyte pre-treated by arecoline were included as the four groups in the present study. The concentration of collagen, the content and activity of matrix metalloproteinase (MMP) and the concentration of tissue inhibitor of metalloproteinase (TIMP) were assessed.
Results:  The collagen production of fibroblasts decreased when cocultured with keratinocytes; when cocultured with keratinocytes pre-treated by arecoline, fibroblasts produced more soluble collagen than non-pretreated coculture group. MMP-9 was produced only in coculture groups. There was no significant difference in the two coculture groups. The activation ratio of pro-MMP-2 in arecoline pre-treated keratinocytes-fibroblasts coculture group was significantly higher than that of non-coculture groups, but no significant difference existed in the two coculture groups. TIMP-1 produced by arecoline pre-treated keratinocytes-fibroblasts coculture group was significantly higher than those by the other three groups.
Conclusion:  TIMP-1 and the interaction of oral keratinocytes and fibroblasts play important role in pathogenesis of OSF.     

槟榔碱对HaCaT细胞增殖活性及S100A7 mRNA表达的影响

目的：通过检测角质形成细胞株HaCaT在不同浓度的槟榔碱刺激下,其生长增殖活性的变化,以及槟榔碱浓度升高对HaCaT细胞S100A7 mRNA表达的影响,进一步探讨口腔粘膜下纤维性变（oral submucous fibrosis,OSF）的发病机制。方法：应用不同浓度槟榔碱处理HaCaT细胞24h,采用MTT法检测HaCaT细胞增殖活性的改变;采用逆转录聚合酶链式反应（RT-PCR）方法检测HaCaT在槟榔碱浓度升高刺激后细胞中S100A7 mRNA表达的变化。结果：高浓度（80～320mg/L）的槟榔碱抑制HaCaT细胞的增殖（P〈0.05）,80、160和320mg/L的槟榔碱相对于未加入槟榔碱的对照组分别使细胞数量下降到89%、84%和61%,对细胞增殖的抑制分别为11%、16%和39%;而在相对较低的槟榔碱浓度（0～40mg/L）下未见对HaCaT细胞的增殖有明显抑制或促进作用。高浓度槟榔碱处理后的细胞可以见到明显的形态改变。浓度为40～80mg/L的槟榔碱轻度上调HaCaT细胞中S100A7 mRNA的表达（P〈0.05）。结论：较低浓度的槟榔碱对于HaCaT细胞的生长没有明显的抑制与促进作用,高浓度的槟榔碱能显著抑制HaCaT细胞的生长。槟榔碱对HaCaT细胞具有细胞毒性。槟榔碱在一定浓度下可以诱导HaCaT细胞S100A7 mRNA的表达增加。S100A7基因及其编码蛋白可能在口腔粘膜下纤维性变的发病中起了重要作用。     

槟榔碱对人牙周膜成纤维细胞凋亡的影响

目的 研究槟榔碱(Arecoline)对体外培养的人牙周膜成纤维细胞(human periodontal ligament fibro-blast,hPDLFs)中凋亡及相关蛋白p-JNK、p-p53、Bcl-2表达的影响.方法 采用组织块培养法培养原代hPDLFs,并传代纯化后用于实验.采用浓度为0μg/mL、20μg/mL、40μg/mL、80μg/mL的槟榔碱处理牙周膜成纤维细胞12 h,通过MTT法检测细胞增殖,流式细胞仪检测细胞凋亡,蛋白免疫印记法检测槟榔碱对hPDLFs中p-JNK、p-p53、Bcl-2表达的情况.结果 槟榔碱作用于hPDLFs后,细胞增殖受到抑制,细胞凋亡率随着药物浓度增加而增加,p-JNK、p-p53蛋白表达逐渐增强,Bcl-2蛋白表达逐渐降低,呈现浓度依赖性.结论 提示槟榔碱可通过激活JNK和p53的磷酸化水平导致hPDLFs的凋亡,对牙周组织的再生有抑制作用.     

Effects of extracts from Bacteroides gingivalis, Bacteroides intermedius, and Bacteroides asaccharolyticus on the growth of fibroblast lines obtained from healthy and inflamed human gingiva

The effect of sonicates from Bacteroides gingivalis, Bacteroides intermedius , and Bacteroides asaccharolyticus on the uptake of (³H)-thymidine by different gingival fibroblast cell lines was studied. Extracts from B. intermedius stimulated thymidine uptake by as much as 150% over that of controls. This effect varied according to both the fibroblast line and the concentration of the bacterial extract. B. asaccharolyticus extracts inhibited the thymidine uptake of all fibroblast cell lines at high concentrations, but in low doses the thymidine uptake was stimulated in some fibroblast lines by up to 200%. B. gingivalis extract caused a strong, dose-dependent inhibition of thymidine uptake in all of the fibroblast lines tested. Low concentrations of B. gingivalis extract (10 μg protein/ml) inhibited DNA-synthesis by 12-60%, and high concentrations (100 μg protein/ml) by 70-90%. The results suggest that the response of periodontal fibroblasts to oral Bacteroides varies considerably depending on the Bacteroides species, as well as on the individual characteristics of the cell lines being tested.     

电磁辐射和不良习惯对牙周健康影响的研究

目的：探讨电磁辐射和不良习惯对牙周健康的影响,为今后开展牙周病预防和治疗提供参考.方法：对335名接受口腔健康检查的人员,依据是否使用电脑,按性别、年龄进行配对分组,采用现场检查和填写调查表的形式,对社区牙周指数（CPI）均值进行统计分析.结果：有不良习惯人员的CPI均值明显高于无不良习惯者,同时有咀嚼槟榔和吸烟习惯人员的CPI均值明显高于只有吸烟或只有咀嚼槟榔者;男性CPI均值明显高于女性;年龄大的CPI均值明显高于年龄小者;使用电脑人员CPI均值与不使用者无差异,但使用液晶（LCD）屏电脑人员的CPI均值明显低于使用阴极射线管（CRT）屏电脑或CRT和LCD混用者.结论：应选择低辐射种类电脑,特别须戒除吸烟或咀嚼槟榔的不良习惯,以保护牙周组织不受破坏.     

Areca nut extract-treated gingival fibroblasts modulate the invasiveness of polymorphonuclear leukocytes via the production of MMP-2

Background:  Areca nut chewing is associated with an increase in the incidence of oral neoplastic or inflammatory diseases. Aberrations in matrix metalloprotease (MMP) expression are associated with the pathogenesis of oral diseases. This study investigated the potential effects of areca nut extract (ANE) on human gingival fibroblasts and the consequential impacts on inflammatory pathogenesis.
Methods:  Analyses of senescence marker, cell viability, changes of the cell cycle, and cell granularity in gingival fibroblasts together with an assessment of the invasiveness of polymorphonuclear (PMN) leukocytes after treatment with the supernatant of ANE-treated gingival fibroblasts were performed to characterize the phenotypic impacts. Western blotting and gelatin zymography were used to assay the expression and activity of MMP-2.
Results:  Chronic subtoxic (<10 μg/ml) ANE treatment resulted in premature growth arrest, appearance of senescence-associated β-galactosidase activity and various other senescence-associated phenotypes in gingival fibroblasts. Gingival fibroblasts established from older individuals had a higher propensity to become ANE-induced senescent gingival fibroblasts. An activation of MMP-2 was identified in senescent cells. PMN leukocytes treated with the supernatant of ANE-induced senescent cells exhibited a significant increase in invasiveness, which was abrogated by both a MMP-2 blocker and a MMP-2 nullifying antibody.
Conclusions:  This study provides evidence whereby MMP-2 secreted from ANE-induced senescent gingival fibroblasts would facilitate the invasiveness of PMN leukocytes, which could be associated with the oral inflammatory process in areca chewers.     

Stimulation of human buccal mucosa fibroblasts in vitro by betel-nut alkaloids

Oral submucous fibrosis (OSF) is characterized by excessive collagen production by mucosal fibroblasts and is associated with the habitual chewing of betel-nuts (Areca catechu); nut extracts stimulate fibroblast activity in vitro. The metabolism of arecoline, the major alkaloid in the nut, by human buccal mucosa fibroblasts in vitro was investigated; alkaloid metabolites extracted from culture media were analysed by gas chromatography and thin-layer chromatography. [3H]-arecoline was metabolized predominantly to [3H]-arecaidine and this was accompanied by a concentration-dependent stimulation of collagen synthesis and cell proliferation. Arecaidine was a more potent stimulator than arecoline. The rate of hydrolysis of a series of synthetic arecaidine esters (methyl, ethyl, butyl, propyl and pentyl) by fibroblasts was closely correlated with the extent of stimulation of collagen synthesis. Thus fibroblasts are responsive to the major metabolite of arecoline and hydrolysis of the ester group may be necessary for this action. Exposure of buccal mucosa fibroblasts to these alkaloids in vivo may contribute to the accumulation of collagen in OSF.     

Inhibitory effects of areca nut extracts on phagocytosis of Actinobacillus actinomycetemcomitans ATCC 33384 by neutrophils

BACKGROUND: Areca quid chewers have a higher prevalence of periodontal disease than non-chewers. Little is known about the influence of areca quid on the immune system. This study was to determine the possible effects of the areca nut on phagocytic activity of human neutrophils. METHODS: Aqueous extracts of ripe areca nut without husk (rANE), fresh and tender areca nut with husk (tANE), a major alkaloid (arecoline), and a phenolic component ([+]-catechin) of areca nut were examined for their effects on cellular viability using trypan blue exclusion assay. The possible effects on the phagocytic activity of neutrophils against a periodontal pathogen, Actinobacillus actinomycetemcomitans ATCC 33384, were determined using flow cytometry and confocal laser scanning microscopy. RESULTS: At the concentrations tested, rANE, tANE, arecoline, and (+)-catechin did not significantly affect viability of neutrophils. However, rANE, tANE, arecoline, and (+)-catechin inhibited the phagocytic activity of neutrophils in a dose-dependent manner. Approximately 50% of the relative phagocytic activity of neutrophils was affected when 50 microg/ml of rANE, 400 microg/ml of tANE, 20,000 microg/ml of arecoline, or 2,500 microg/ml of (+)- catechin was used. Decreased levels of internalized fluorescent bacteria were also demonstrated. However, arecoline or (+)-catechin alone could not be used to explain the inhibitory effects observed for rANE and tANE. CONCLUSIONS: Components of areca nut reduced the uptake of A. actinomycetemcomitans ATCC 33384 by human neutrophils. The inhibition of areca nut on phagocytosis of neutrophils may be one possible mechanism by which the areca nut compromises the periodontal health of areca quid chewers.     

槟榔碱对颊黏膜成纤维细胞增殖及迁移的影响

目的通过观察槟榔碱诱发人颊黏膜成纤维细胞增殖、迁移能力以及微丝形态的改变,探讨槟榔碱在口腔黏膜下纤维性变(OSF)的致病途径。方法使用甲噻唑四唑氮(MTT)比色法、划痕法、激光共聚焦扫描显微镜检测不同浓度槟榔碱(5、10、20、40、80μg·mL^-1)对成纤维细胞增殖、迁移、微丝形态的影响。结果5、10、20μg·mL^-1的槟榔碱可轻微提高颊黏膜成纤维细胞增殖迁移能力及微丝聚合(P<0.05);40、80μg·mL^-1的槟榔碱抑制颊黏膜成纤维细胞增殖、迁移能力及微丝解聚(P<0.05)。结论槟榔碱改变颊黏膜成纤维细胞增殖、迁移能力及微丝形态分布,可能在口腔黏膜下纤维性变病理过程中起着重要的作用。