Elucidating the Role of STAT3 in Epithelial-microbiome Interaction During Periodontitis and Peri-implantitis

Summary

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.

Conditions

• Periodontitis
• Peri-implantitis

Sponsors & Collaborators

• Universidad Complutense de Madrid

  collaborator OTHER

• University of Chile

  lead OTHER

Eligibility

Min Age

18 Years

Sex

ALL

Healthy Volunteers

Yes

Timeline & Regulatory

Start

2023-06-01

Primary Completion

2025-01-10

Completion

2025-01-30

Countries

• Chile

Study Locations