RESUMO
OBJECTIVE: Carcinoembryonic antigen-related cell adhesion molecule 1 (CEACAM1) is a member of the carcinoembryonic antigen family. Although its expression has been found in chronic oral inflammatory epithelium, this study aimed to know whether CEACAM1 in oral keratinocytes participates in host immune response against Candida albicans . METHODOLOGY: We investigated CEACAM1 expression in oral keratinocytes induced by C. albicans as well as by Candida cell wall component ß-glucan particles (ß-GPs). Furthermore, the effects of CEACAM1 on ß-GPs-induced heme oxygenase-1 (HO-1) expression and its related signals were examined. RESULTS: Fluorescence staining showed CEACAM1 expression in oral keratinocytes (RT7) cells, whereas quantitative reverse transcription (RT)-PCR indicated that both live and heat-killed C. albicans increased CEACAM1 mRNA expression in RT7 cells. Examinations using quantitative RT-PCR and western blotting indicated that CEACAM1 expression was also increased by ß-GPs derived from C. albicans . Specific siRNA for CEACAM1 decreased HO-1 expression induced by ß-GPs from C. albicans as well as the budding yeast microorganism Saccharomyces cerevisiae . Moreover, knockdown of CEACAM1 decreased ß-GPs-induced ROS activity in the early phase and translocation of Nrf2 into the nucleus. CONCLUSION: CEACAM1 in oral keratinocytes may have a critical role in regulation of HO-1 for host immune defense during Candida infection.
Assuntos
Heme Oxigenase-1 , beta-Glucanas , Heme Oxigenase-1/metabolismo , Heme Oxigenase-1/farmacologia , beta-Glucanas/farmacologia , beta-Glucanas/metabolismo , Antígeno Carcinoembrionário/metabolismo , Antígeno Carcinoembrionário/farmacologia , Molécula 1 de Adesão Celular/metabolismo , Glucanos/metabolismo , Glucanos/farmacologia , Candida , Queratinócitos , Candida albicans/fisiologiaRESUMO
OBJECTIVE: Although oral fibroblasts are thought to have the potential to enhance host defenses against Candida albicans , it is unknown whether they are able to recognize Candida cell components to increase the expression of antifungal peptides, such as defensin factors, against Candida infection. METHODOLOGY: We performed expression profiles of defensin genes induced by heat-killed C. albicans in oral immortalized fibroblasts (GT1) using cDNA microarray analysis. From those results, quantitative RT-PCR was used to examine the effects of Candida ß-glucan-containing particles (ß-GPs) on ß-Defensin 118 (DEFB 118) expression in oral mucosal cells. Furthermore, the antifungal activities of recombinant DEFB 118 against C. albicans and C. glabrata were investigated using fungicidal assays. RESULTS: Microarray analysis showed that DEFB118, ß-Defensin 129 (DEFB129), and α-Defensin 1 (DEFA1) genes were induced by heat-killed C. albicans and that their mRNA expressions were also significantly increased by live as well as heat-killed C. albicans . Next, we focused on DEFB118, and found that GT1, primary fibroblasts, and RT7 (oral immortalized keratinocytes) constitutively expressed DEFB118 mRNA expression in RT-PCR. Furthermore, C. albicans ß-GPs significantly increased the expression of DEFB118 mRNA in GT1 and primary fibroblasts. Although DEFB118 mRNA expression in RT7 was significantly induced by both live and heat-killed C. albicans, C. albicans ß-GPs failed to have an effect on that expression. Finally, recombinant DEFB118 significantly decreased the survival of both strains of C. albicans in a dose-dependent manner, whereas no effects were seen for both C. glabrata strains. CONCLUSION: DEFB118, induced by C. albicans ß-GPs from oral fibroblasts, may play an important role in oral immune responses against C. albicans infection.
Assuntos
beta-Defensinas , beta-Glucanas , Antifúngicos/farmacologia , Candida albicans , Fibroblastos , Glucanos/metabolismo , RNA Mensageiro/metabolismo , beta-Defensinas/genética , beta-Defensinas/metabolismo , beta-Defensinas/farmacologia , beta-Glucanas/metabolismo , beta-Glucanas/farmacologiaRESUMO
Abstract Objective Carcinoembryonic antigen-related cell adhesion molecule 1 (CEACAM1) is a member of the carcinoembryonic antigen family. Although its expression has been found in chronic oral inflammatory epithelium, this study aimed to know whether CEACAM1 in oral keratinocytes participates in host immune response against Candida albicans . Methodology We investigated CEACAM1 expression in oral keratinocytes induced by C. albicans as well as by Candida cell wall component β-glucan particles (β-GPs). Furthermore, the effects of CEACAM1 on β-GPs-induced heme oxygenase-1 (HO-1) expression and its related signals were examined. Results Fluorescence staining showed CEACAM1 expression in oral keratinocytes (RT7) cells, whereas quantitative reverse transcription (RT)-PCR indicated that both live and heat-killed C. albicans increased CEACAM1 mRNA expression in RT7 cells. Examinations using quantitative RT-PCR and western blotting indicated that CEACAM1 expression was also increased by β-GPs derived from C. albicans . Specific siRNA for CEACAM1 decreased HO-1 expression induced by β-GPs from C. albicans as well as the budding yeast microorganism Saccharomyces cerevisiae . Moreover, knockdown of CEACAM1 decreased β-GPs-induced ROS activity in the early phase and translocation of Nrf2 into the nucleus. Conclusion CEACAM1 in oral keratinocytes may have a critical role in regulation of HO-1 for host immune defense during Candida infection.
RESUMO
Abstract Objective: Although oral fibroblasts are thought to have the potential to enhance host defenses against Candida albicans , it is unknown whether they are able to recognize Candida cell components to increase the expression of antifungal peptides, such as defensin factors, against Candida infection. Methodology: We performed expression profiles of defensin genes induced by heat-killed C. albicans in oral immortalized fibroblasts (GT1) using cDNA microarray analysis. From those results, quantitative RT-PCR was used to examine the effects of Candida β-glucan-containing particles (β-GPs) on β-Defensin 118 (DEFB 118) expression in oral mucosal cells. Furthermore, the antifungal activities of recombinant DEFB 118 against C. albicans and C. glabrata were investigated using fungicidal assays. Results: Microarray analysis showed that DEFB118, β-Defensin 129 (DEFB129), and α-Defensin 1 (DEFA1) genes were induced by heat-killed C. albicans and that their mRNA expressions were also significantly increased by live as well as heat-killed C. albicans . Next, we focused on DEFB118, and found that GT1, primary fibroblasts, and RT7 (oral immortalized keratinocytes) constitutively expressed DEFB118 mRNA expression in RT-PCR. Furthermore, C. albicans β-GPs significantly increased the expression of DEFB118 mRNA in GT1 and primary fibroblasts. Although DEFB118 mRNA expression in RT7 was significantly induced by both live and heat-killed C. albicans, C. albicans β-GPs failed to have an effect on that expression. Finally, recombinant DEFB118 significantly decreased the survival of both strains of C. albicans in a dose-dependent manner, whereas no effects were seen for both C. glabrata strains. Conclusion: DEFB118, induced by C. albicans β-GPs from oral fibroblasts, may play an important role in oral immune responses against C. albicans infection.
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OBJECTIVE: This study aimed to clarify the association between oral human cytomegalovirus (HCMV) and periodontitis in Japanese adults. METHODOLOGY: In total, 190 patients (75 men and 115 women; mean age, 70.2 years) who visited Hiroshima University Hospital between March 2018 and May 2020 were included. Oral rinse samples were taken to examine the presence of HCMV DNA using real-time polymerase chain reaction (PCR). P. gingivalis was detected by semi-quantitative PCR analysis. RESULTS: HCMV DNA was present in nine of 190 patients (4.7%). There were significant associations between HCMV presence and the presence of ≥4-mm-deep periodontal pockets with bleeding on probing (BOP) (P<0.01) and ≥6-mm-deep periodontal pockets with BOP (P=0.01). However, no significant relationship was observed between HCMV presence and periodontal epithelial surface area scores. Logistic regression analysis revealed that the presence of ≥4-mm-deep periodontal pockets with BOP was significantly associated with HCMV (odds ratio, 14.4; P=0.01). Propensity score matching was performed between patients presenting ≥4-mm-deep periodontal pockets with BOP (i.e., active periodontitis) and patients without ≥4-mm-deep periodontal pockets with BOP; 62 matched pairs were generated. Patients who had ≥4-mm-deep periodontal pockets with BOP showed a higher rate of HCMV presence (9.7%) than those who lacked ≥4-mm-deep periodontal pockets with BOP (0.0%). There was a significant relationship between HCMV presence and ≥4-mm-deep periodontal pockets with BOP (P=0.03). A significant relationship was found between HCMV/P. gingivalis DNA presence and ≥4-mm-deep periodontal pockets with BOP (P=0.03). CONCLUSIONS: Coinfection of oral HCMV and P. gingivalis was significantly associated with active periodontitis. Moreover, interactions between oral HCMV and P. gingivalis may be related to the severity of periodontal disease.
Assuntos
Infecções por Bacteroidaceae/epidemiologia , Infecções por Citomegalovirus/epidemiologia , Periodontite , Idoso , Coinfecção , Estudos Transversais , Citomegalovirus , Feminino , Humanos , Japão/epidemiologia , Masculino , Bolsa Periodontal/microbiologia , Bolsa Periodontal/virologia , Periodontite/epidemiologia , Periodontite/microbiologia , Periodontite/virologia , Porphyromonas gingivalis , PrevalênciaRESUMO
Abstract Objective This study aimed to clarify the association between oral human cytomegalovirus (HCMV) and periodontitis in Japanese adults. Methodology In total, 190 patients (75 men and 115 women; mean age, 70.2 years) who visited Hiroshima University Hospital between March 2018 and May 2020 were included. Oral rinse samples were taken to examine the presence of HCMV DNA using real-time polymerase chain reaction (PCR). P. gingivalis was detected by semi-quantitative PCR analysis. Results HCMV DNA was present in nine of 190 patients (4.7%). There were significant associations between HCMV presence and the presence of ≥4-mm-deep periodontal pockets with bleeding on probing (BOP) (P<0.01) and ≥6-mm-deep periodontal pockets with BOP (P=0.01). However, no significant relationship was observed between HCMV presence and periodontal epithelial surface area scores. Logistic regression analysis revealed that the presence of ≥4-mm-deep periodontal pockets with BOP was significantly associated with HCMV (odds ratio, 14.4; P=0.01). Propensity score matching was performed between patients presenting ≥4-mm-deep periodontal pockets with BOP (i.e., active periodontitis) and patients without ≥4-mm-deep periodontal pockets with BOP; 62 matched pairs were generated. Patients who had ≥4-mm-deep periodontal pockets with BOP showed a higher rate of HCMV presence (9.7%) than those who lacked ≥4-mm-deep periodontal pockets with BOP (0.0%). There was a significant relationship between HCMV presence and ≥4-mm-deep periodontal pockets with BOP (P=0.03). A significant relationship was found between HCMV/P. gingivalis DNA presence and ≥4-mm-deep periodontal pockets with BOP (P=0.03). Conclusions Coinfection of oral HCMV and P. gingivalis was significantly associated with active periodontitis. Moreover, interactions between oral HCMV and P. gingivalis may be related to the severity of periodontal disease.
Assuntos
Humanos , Masculino , Feminino , Idoso , Periodontite/microbiologia , Periodontite/epidemiologia , Periodontite/virologia , Infecções por Bacteroidaceae/epidemiologia , Infecções por Citomegalovirus/epidemiologia , Bolsa Periodontal/microbiologia , Bolsa Periodontal/virologia , Prevalência , Estudos Transversais , Porphyromonas gingivalis , Citomegalovirus , Coinfecção , Japão/epidemiologiaRESUMO
OBJECTIVE: The objective of this study was to clarify differences in bacterial accumulation between gastrointestinal cancer patients who underwent severely invasive surgery and those who underwent minimally invasive surgery. MATERIAL AND METHODS: We performed a preliminary investigation of gastrointestinal cancer patients who were treated at the Department of Surgery, Takarazuka Municipal Hospital, from 2015 to 2017 (n=71; 42 laparoscopic surgery, 29 open surgery) to determine changes in bacterial numbers at different sites of the oral cavity (tongue dorsum, gingiva of upper anterior teeth, palatoglossal arch), as well as mouth dryness and tongue coating indices. Specifically, patients received professional tooth cleaning (PTC), scaling, tongue cleaning, and self-care instruction regarding tooth brushing from a dental hygienist a day before the operation. Professional oral health care was also performed by a dental hygienist two and seven days after surgery. Oral bacteria numbers were determined using a bacterial counter with a dielectrophoretic impedance measurement method. RESULTS: The number of bacteria at all three examined sites were significantly higher in the open surgery group when compared to the laparoscopic surgery group on the second postoperative day. Relevantly, bacterial count in samples from the gingiva of the upper anterior teeth remained greater seven days after the operation in patients who underwent open surgery. Furthermore, the dry mouth index level was higher in the open surgery group when compared to the laparoscopic surgery group on postoperative days 2 and 7. CONCLUSIONS: Even with regular oral health care, bacterial numbers remained high in the upper incisor tooth gingiva in gastrointestinal cancer patients who received open surgery. Additional procedures are likely needed to effectively reduce the number of bacteria in the gingival area associated with the upper anterior teeth.
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Neoplasias Gastrointestinais/microbiologia , Neoplasias Gastrointestinais/cirurgia , Boca/microbiologia , Saúde Bucal , Assistência Perioperatória , Adulto , Idoso , Idoso de 80 Anos ou mais , Carga Bacteriana , Temperatura Corporal , Proteína C-Reativa/análise , Feminino , Neoplasias Gastrointestinais/patologia , Humanos , Laparoscopia , Contagem de Leucócitos , Masculino , Pessoa de Meia-Idade , Estadiamento de Neoplasias , Período Pós-Operatório , Período Pré-Operatório , Estatísticas não Paramétricas , Fatores de TempoRESUMO
Abstract Objective The objective of this study was to clarify differences in bacterial accumulation between gastrointestinal cancer patients who underwent severely invasive surgery and those who underwent minimally invasive surgery. Material and Methods We performed a preliminary investigation of gastrointestinal cancer patients who were treated at the Department of Surgery, Takarazuka Municipal Hospital, from 2015 to 2017 (n=71; 42 laparoscopic surgery, 29 open surgery) to determine changes in bacterial numbers at different sites of the oral cavity (tongue dorsum, gingiva of upper anterior teeth, palatoglossal arch), as well as mouth dryness and tongue coating indices. Specifically, patients received professional tooth cleaning (PTC), scaling, tongue cleaning, and self-care instruction regarding tooth brushing from a dental hygienist a day before the operation. Professional oral health care was also performed by a dental hygienist two and seven days after surgery. Oral bacteria numbers were determined using a bacterial counter with a dielectrophoretic impedance measurement method. Results The number of bacteria at all three examined sites were significantly higher in the open surgery group when compared to the laparoscopic surgery group on the second postoperative day. Relevantly, bacterial count in samples from the gingiva of the upper anterior teeth remained greater seven days after the operation in patients who underwent open surgery. Furthermore, the dry mouth index level was higher in the open surgery group when compared to the laparoscopic surgery group on postoperative days 2 and 7. Conclusions Even with regular oral health care, bacterial numbers remained high in the upper incisor tooth gingiva in gastrointestinal cancer patients who received open surgery. Additional procedures are likely needed to effectively reduce the number of bacteria in the gingival area associated with the upper anterior teeth.
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Humanos , Masculino , Feminino , Adulto , Idoso , Idoso de 80 Anos ou mais , Saúde Bucal , Assistência Perioperatória , Neoplasias Gastrointestinais/cirurgia , Neoplasias Gastrointestinais/microbiologia , Boca/microbiologia , Período Pós-Operatório , Fatores de Tempo , Temperatura Corporal , Proteína C-Reativa/análise , Laparoscopia , Estatísticas não Paramétricas , Período Pré-Operatório , Carga Bacteriana , Neoplasias Gastrointestinais/patologia , Contagem de Leucócitos , Pessoa de Meia-Idade , Estadiamento de NeoplasiasRESUMO
OBJECTIVE: The objective of this study was to clarify differences regarding HPV16 infection and gene amplification between the oral cavity and oropharynx in healthy individuals. MATERIAL AND METHODS: The subjects were 94 healthy asymptomatic individuals (41 males, 53 females; mean age 58.6 years, range 16-97 years) who visited the Department of Oral and Maxillofacial Reconstructive Surgery of the Hiroshima University Hospital from 2014 to 2015. Oral epithelial cells were collected from oral rinse and pharynx gargle samples and placed in saline. The human endogenous retrovirus gene ERV3-1 was used as a reference to estimate the number of human cells in each sample. DNA samples were extracted from approximately 10,000 human cells and tested for HPV16 DNA by PCR using a type-specific primer. Similarly, we analyzed the HPV16 viral copy number in HPV16-positive cases using real-time PCR to examine genomic amplification. RESULTS: The percentage of HPV16-positive cases was higher in the gargle (28.7%) as compared to the rinse (16.0%) samples. In the oral rinse samples, males (26.8%) showed a significantly higher rate of HPV16 than females (7.5%) (P=0.021). Importantly, in older subjects (aged 60-89 years), gargle samples showed a significantly higher rate of HPV16 (33.3%) than oral rinse samples (13.7%) (P=0.034). The average number of viral copies was approximately 8 times higher in the gargle than in the oral rinse samples (0.16±0.27 vs. 1.35±1.26 copy numbers per cell), a significant difference (P<0.001). CONCLUSION: Our findings suggest that the oropharynx is more susceptible to HPV16 infection as compared to the oral cavity, while HPV16 gene amplification is also more commonly found in the oropharynx.
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Amplificação de Genes/fisiologia , Papillomavirus Humano 16/genética , Boca/virologia , Orofaringe/virologia , Infecções por Papillomavirus/epidemiologia , Infecções por Papillomavirus/virologia , Adolescente , Adulto , Fatores Etários , Idoso , Idoso de 80 Anos ou mais , Contagem de Células , Variações do Número de Cópias de DNA , DNA Viral , Feminino , Humanos , Japão/epidemiologia , Masculino , Pessoa de Meia-Idade , Prevalência , Reação em Cadeia da Polimerase em Tempo Real , Fatores de Risco , Fatores de Tempo , Adulto JovemRESUMO
ABSTRACT Objective The objective of this study was to clarify differences regarding HPV16 infection and gene amplification between the oral cavity and oropharynx in healthy individuals. Material and Methods The subjects were 94 healthy asymptomatic individuals (41 males, 53 females; mean age 58.6 years, range 16-97 years) who visited the Department of Oral and Maxillofacial Reconstructive Surgery of the Hiroshima University Hospital from 2014 to 2015. Oral epithelial cells were collected from oral rinse and pharynx gargle samples and placed in saline. The human endogenous retrovirus gene ERV3-1 was used as a reference to estimate the number of human cells in each sample. DNA samples were extracted from approximately 10,000 human cells and tested for HPV16 DNA by PCR using a type-specific primer. Similarly, we analyzed the HPV16 viral copy number in HPV16-positive cases using real-time PCR to examine genomic amplification. Results The percentage of HPV16-positive cases was higher in the gargle (28.7%) as compared to the rinse (16.0%) samples. In the oral rinse samples, males (26.8%) showed a significantly higher rate of HPV16 than females (7.5%) (P=0.021). Importantly, in older subjects (aged 60-89 years), gargle samples showed a significantly higher rate of HPV16 (33.3%) than oral rinse samples (13.7%) (P=0.034). The average number of viral copies was approximately 8 times higher in the gargle than in the oral rinse samples (0.16±0.27 vs. 1.35±1.26 copy numbers per cell), a significant difference (P<0.001). Conclusion Our findings suggest that the oropharynx is more susceptible to HPV16 infection as compared to the oral cavity, while HPV16 gene amplification is also more commonly found in the oropharynx.