口腔疾病防治 (Dec 2024)
Mitochondrial homeostasis imbalance in HGFs in periodontitis
Abstract
Objective To investigate the changes of mitochondrial homeostasis of human gingival fibroblasts (HGFs) in periodontitis, and to provide a basis for exploring the mechanism of periodontitis. Methods This study has been reviewed and approved by the Ethics Committee. Gingival tissue was collected from patients who underwent periodontal surgery at the Department of Periodontology, School of Stomatology, the Fourth Military Medical University, from June 1, 2023 to December 31, 2023. All of the subjects signed informed consent forms prior to surgery. Gingival connective tissues were collected from patients with chronic periodontitis (CP group) and healthy individuals (control group) undergoing flap surgery and crown lengthening surgery, respectively. There were 6 cases in each group. The primary HGFs obtained from healthy periodontal subjects were cultured and divided into the control group (cultured in complete medium for 24 h) and the Pg.LPS group (cultured in medium with 5μg/mL Pg.LPS for 24 h). The number, morphology, and structure of mitochondria in gingival connective tissue and HGFs were observed by transmission electron microscopy. The number, circumference, and surface area of mitochondria were quantitatively analyzed. MitoSOXTMRed, TMRM, and an ATP kit were used to determine the production levels of mitochondrial reactive oxygen species, mitochondrial membrane potential, and ATP in each group of HGFs. Results Transmission electron microscopy showed that the morphology and structure of mitochondria were abnormal in the gingival connective tissues of the periodontitis group and HGFs, which were stimulated by Pg.LPS. The mitochondrial ridges were broken or were not visible in these groups. The number of mitochondria decreased significantly, and the surface area and circumference of the mitochondria increased significantly (P < 0.05). In addition, after stimulation by Pg.LPS, the reactive oxygen species level in HGFs was significantly higher than that in the control group (P < 0.05). The mitochondrial membrane potential and the level of ATP production was significantly lower than that of the control group (P < 0.05). Conclusion The number, morphological structure, and function of mitochondria in HGFs changed significantly in periodontitis. The mitochondrial homeostasis is closely related to the occurrence and development of periodontitis.
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