Avaliação da 7-hidroxicumarina como agente terapêutico em modelo de periodontite experimental induzida por lipopolissacarídeo

Abstract

Periodontitis is characterized by a multifactorial etiology, wherein the immune response to microbial challenges triggers inflammation of the gingival tissues. This response leads to the activation of osteoclasts and subsequent reabsorption of the alveolar bone, ultimately resulting in the loss of dental elements. This phenomenon highlights the crucial importance of effective inflammatory control in managing periodontitis. Within this context, bioactive compounds extracted from medicinal plants, such as 7-hydroxycoumarin (7HC), represent a promising alternative. This endeavor commenced with an evaluation of the current state of knowledge, achieved through a technological prospecting analysis utilizing the Orbit patent database to explore the biotechnological potential of 7HC. The technological prospecting was structured into two distinct phases. The initial phase involved an evaluation of the biotechnological potential of 7-hydroxycoumarin (7HC) in a general context, utilizing the following search syntax: (umbelliferone)/ti/ab/clms/keyw) OR (7hydroxycoumarin)/ti/ab/clms/keyw), where "ti" denotes Title, "ab" refers to Abstract, and "clms" indicates Claim. The subsequent phase focused on assessing the biotechnological potential of 7HC, specifically within the field of dentistry. This evaluation was conducted using the search syntax: (umbelliferone)/ti/ab/clms/keyw AND (A61Q-011/00)/IPC/CPC), with the classification A61Q-011/00 being associated with products utilized for oral care. The conducted technological assessment demonstrated that 7HC, also referred to as umbelliferone, exhibits antiinflammatory, antioxidant, and antimicrobial properties, in addition to stimulating collagen synthesis. These attributes may play a significant role in the clinical treatment of periodontal disease. Consequently, the objective of this study was to evaluate the therapeutic efficacy of 7HC at concentrations of 50 mg/kg and 100 mg/kg in an animal model of periodontal disease induced by lipopolysaccharide (LPS). Additionally, a comparative study was conducted to assess the induction of periodontal disease in two mouse strains, Swiss and C57BL/6, through the administration of lipopolysaccharide (LPS) in the gingival tissue. The mice were categorized into groups based on the type of treatment received. Periodontal disease induction was achieved by administering 1 μl of LPS (20 μg/μl) into the gingival tissue on alternate days. 7HC was administered daily at concentrations of 50 mg/kg and 100 mg/kg from day 14 to day 28 of the study. Gingival tissue samples were collected to assess the expression of key biomarkers associated with active periodontal disease, utilizing both ELISA and qRT-PCR methodologies. Furthermore, the maxillae were excised to evaluate bone resorption, using staining with 1% methylene blue. The findings demonstrated that 7HC, at both concentrations examined, effectively inhibited the expression of pro-inflammatory cytokines, including IL-1β, IL6, MMP-9, and TIMP-1. Additionally, at a dosage of 100 mg/kg, 7HC stimulated the expression of anti-inflammatory cytokines, namely IL-10 and TGF-β, which contributed to a reduction in alveolar bone loss. This study presents 7HC as a promising bioactive compound for use in periodontal therapy. The comparative analysis of the Swiss and C57BL/6 mouse strains indicates that periodontitis in the Swiss strain commences at an earlier stage, specifically on the seventh day postinduction, and progresses with greater severity compared to the C57BL/6 strain. This finding suggests that, depending on the research objectives, utilizing the Swiss strain may yield advantages in terms of reducing both time requirements and operational costs.