Elucidating the Role of STAT3 in Epithelial-microbiome Interaction During Periodontitis and Peri-implantitis
试验速览
- 阶段
- 不适用
- 状态
- 已完成
- 发起方
- 入组人数
- 43
- 试验地点
- 1
- 主要终点
- Percentage of epithelial cells that present activated STAT3 protein
研究概览
简要总结
Periodontitis and peri-implantitis are highly prevalent immuno-inflammatory pathologies, characterized by a series of intracellular signals, which, as a consequence of the interaction of the gingival epithelium with the dysbiotic microbiome, lead to an exacerbated immuno-inflammatory response, ultimately resulting in tissue loss around teeth and implants. STAT3 is a transcription factor that plays a key role in inflammatory diseases, which when phosphorylated (pSTAT3) promotes the transcription of genes with immuno-defensive functions. STAT3 phosphorylation has only been reported during experimental periodontitis, but our preliminary results reveal a higher number of epithelial cells with the presence of pSTAT3 in subjects with periodontitis. Other studies have reported greater STAT3 activation in oral keratinocyte cells stimulated with LPS from P. gingivalis, which and F. nucleatum are abundant bacterial species in both pathologies. On the other hand, inhibition of the STAT3 pathway is a therapeutic target in multiple inflammatory diseases, where inhibitors such as C188-9 have shown a direct effect on STAT3 signaling. A better understanding of this signaling pathway in periodontitis and peri-implantitis could allow for future complementary therapies for these diseases.
Based on the above, the hypothesis of this project is:
"During periodontitis and peri-implantitis, there is greater activation of STAT3, particularly at the level of the gingival epithelium, inducing an increase in immune-defensive function in response to microbial dysbiosis and specific constituents of the microbiota associated with these pathologies compared to gingival health."
General objective: To evaluate STAT3 activation and its immune-defensive function during gingival health, periodontitis, and peri-implantitis, specifically at the level of the gingival epithelium, in response to microbial dysbiosis and specific bacteria associated with these periodontal conditions.
Objective 1: To characterize STAT3 activation and immune-defensive function in gingival tissue during gingival health, periodontitis, and peri-implantitis. The investigators will obtain gingival tissue samples from the three conditions through a clinical study. These samples will be processed by Western blot and immunofluorescence to determine pSTAT3. In addition, RT-qPCR will determine the expression of genes related to its immune-defensive function.
Objective 2: To determine STAT3 activation and its immune-defensive function in oral keratinocytes in response to the subgingival biofilm associated with gingival health, periodontitis, and peri-implantitis: Subgingival microbiota samples will be obtained from the three conditions, and the microbiome related to each condition will be characterized by massive sequencing of the 16S rDNA gene. Furthermore, through an in vitro study, oral keratinocyte cells will be stimulated with the obtained samples, and the expression of genes related to their immune-defense function will be determined by Western blot, pSTAT3, and RT-qPCR. The effect of STAT3 inhibition using C188-9 will be subsequently evaluated.
Objective 3: Determine STAT3 activation and its immune-defense function in oral keratinocytes in response to specific bacteria associated with gingival health, periodontitis, and peri-implantitis. Through an in vitro study, oral keratinocyte cells will be infected with reference strains of S. sanguinis, P. gingivalis, and F. nucleatum, and the expression of genes related to their immune-defense function will be determined by Western blot, pSTAT3, and RT-qPCR.
The investigators hope to achieve greater activation of STAT3 and increased expression of genes related to the immune-defensive response of the gingival epithelium during periodontitis and peri-implantitis. The investigators also hope to determine the effect of STAT3 inhibition, expecting a reduction in the expression of genes related to this response.
详细描述
Aim: To characterize STAT3 activation and immune defense function in gingival tissue during gingival health, periodontitis, and peri-implantitis.
Subject Selection:
This study will comply with the Declaration of Helsinki (October 2013) and Chilean legal and regulatory requirements. The study will be conducted at the Dental Clinic of the Faculty of Dentistry of the University of Chile.
Subjects attending the Dental Clinic of the Faculty of Dentistry of the University of Chile and diagnosed with gingival health, periodontitis, and peri-implantitis, according to the classification of periodontal and peri-implant diseases, will be recruited and examined.
All subjects must sign an informed consent form previously validated by an ethics committee and meet the following inclusion and exclusion criteria:
研究设计
- 研究类型
- Observational
- 观察模型
- Cohort
- 时间视角
- Cross Sectional
入排标准
- 年龄范围
- 18 Years 至 —（Adult, Older Adult）
- 性别
- All
- 接受健康志愿者
- 是
入选标准
- •Patients in the gingival health group:
- •Meet the case definition of gingival health (Chapple et al. 2018).
- •≥ 18 years of age.
- •Willingness to donate gingival tissue and collect microbiological samples for the study.
- •Patients in the periodontitis group:
- •Meet the case definition of periodontitis (Papapanou et al. 2018).
- •≥ 18 years of age.
- •Willingness to donate gingival tissue and collect microbiological samples for the study.
- •Patients in the peri-implantitis group:
- •Meet the case definition of periodontitis and peri-implantitis (Berglundh et al., 2018).
- •≥ 18 years of age.
- •Willingness to donate gingival tissue and collect a microbiological sample for the study.
- •Exclusion Criteria
- •Having used systemic antibiotics (intravenous, intramuscular, or oral) in the last six months.
- •Having used corticosteroids or other immunosuppressants (e.g., cyclosporine), cytokine therapy, methotrexate, or any other immunosuppressive chemical agent, or high-dose commercially available probiotics (≥ 108 colony-forming units per day) in tablet, capsule, or powder form within the last three months. The use of fermented foods/liquids, e.g., milk, yogurt, etc., does not apply to the exclusion of the individual.
- •Being pregnant or breastfeeding.
- •Subjects who cannot give consent.
排除标准
- 未提供
结局指标
主要结局
Percentage of epithelial cells that present activated STAT3 protein
时间窗: Baseline
Measured as a percentage of epithelial cells that present the activated protein from 0 to 100, where 0 means no epithelial cells activate the protein, and 100% means all cells present it.
次要结局
未报告次要终点