Lysosomal dysfunction-derived autophagy impairment of gingival epithelial cells in diabetes-associated periodontitis with altered protein acetylation.

Diabetes-associated periodontitis (DP) presents severe inflammation and resistance to periodontal conventional treatment, presenting a significant challenge in clinical management. In this study, we investigated the underlying mechanism driving the hyperinflammatory response in gingival epithelial cells (GECs) of DP patients. Our findings indicate that lysosomal dysfunction under high glucose conditions leads to the blockage of autophagy flux, exacerbating inflammatory response in GECs. Single-cell RNA sequencing and immunohistochemistry analyses of clinical gingival epithelia revealed dysregulation in the lysosome pathway characterized by reduced levels of lysosome-associated membrane glycoprotein 2 (LAMP2) and V-type proton ATPase 16 kDa proteolipid subunit c (ATP6V0C) in subjects with DP. In vitro stimulation of human gingival epithelial cells (HGECs) with a hyperglycemic microenvironment showed elevated release of proinflammatory cytokines, compromised lysosomal acidity and blocked autophagy. Moreover, HGECs with deficiency in ATP6V0C demonstrated impaired autophagy and heightened inflammatory response, mirroring the effects of high glucose stimulation. Proteomic analysis of acetylation modifications identified altered acetylation levels in 28 autophagy-lysosome pathway-related proteins and 37 sites in HGECs subjected to high glucose stimulation or siATP6V0C. Overall, our finding highlights the pivotal role of lysosome impairment in autophagy obstruction in DP and suggests a potential impact of altered acetylation of relevant proteins on the interplay between lysosome dysfunction and autophagy blockage. These insights may pave the way for the development of effective therapeutic strategies against DP.

Diabetic Macrophage Exosomal miR-381-3p Inhibits Epithelial Cell Autophagy Via NR5A2.

PURPOSE: To explore the mechanism underlying autophagy disruption in gingival epithelial cells (GECs) in diabetic individuals. METHODS AND MATERIALS: Bone marrow-derived macrophages (BMDMs) and GECs were extracted from C57/bl and db/db mice, the exosomes (Exo) were isolated from BMDMs. qRT‒PCR and Western blotting were performed to analyse gene expression. The AnimalTFDB database was used to identify relevant transcription factors, and miRNA sequencing was utilised to identify relevant miRNAs with the aid of the TargetScan/miRDB/miRWalk databases. A dual-luciferase assay was conducted to verify intermolecular targeting relationships. RESULTS: Similar to BMDMs, BMDM-derived Exos disrupted autophagy and exerted proinflammatory effects in GEC cocultures, and ATG7 may play a vital role. AnimalTFDB database analysis and dual-luciferase assays indicated that NR5A2 is the most relevant transcription factor that regulates Atg7 expression. SiRNA-NR5A2 transfection blocked autophagy in GECs and exacerbated inflammation, whereas NR5A2 upregulation restored ATG7 expression and ameliorated ExoDM-mediated inflammation. MiRNA sequencing, with TargetScan/miRDB/miRWalk analyses and dual-luciferase assays, confirmed that miR-381-3p is the most relevant miRNA that targets NR5A2. MiR-381-3p mimic transfection blocked autophagy in GECs and exacerbated inflammation, while miR-381-3p inhibitor transfection restored ATG7 expression and attenuated ExoDM-mediated inflammation. CONCLUSION: BMDM-derived Exos, which carry miR-381-3p, inhibit NR5A2 and disrupt autophagy in GECs, increasing periodontal inflammation in diabetes.

Multisensory Preclinical Training Strategy of Periodontal Scaling for Undergraduates.

BACKGROUND: Invisibility of subgingival scaling is the most important negative factor affecting the performance of periodontal treatment. A multisensory teaching strategy is used in the preclinical training of undergraduates in order to increase the haptic-auditory-visual feedback, aiming to overcome the invisibility and achieve minimal postoperative complications, improving patients' treatment experience. METHODS: One hundred undergraduate dental students in grade 5 were divided into a multisensory teaching strategy group (MTS: n = 50) and a conventional training pattern group (CTP: n = 50). All participants attended a lecture on using an ultrasonic subgingival scaler (USS) and Gracey curettes (GRA), followed by a 3-week training programme. Students in the MTS group were trained in a haptic/auditory-visual feedback manner, whereas students in the CTP group were trained conventionally. After the training phase, paired students in the 2 different groups performed subgingival scaling in paired patients with equivalent teeth of periodontitis using USS and GRA. Objective and subjective postoperative evaluations were recorded. Probing depth (PD) and gingival index (GI) were evaluated before and 4 weeks after scaling by the same periodontal specialist. RESULTS: MTS significantly reduced treatment time and ameliorated postoperative complications (gingival injury, haemorrhage and root surface roughness; P < .05). Postoperative sensitivity was reduced in the MTS group from day 1 to day 7 (D1-D5: P < .001; D6: P = .002; D7: P = .003), whereas postoperative pain was reduced on day 1 (P = .006), compared with that in the CTP group. The PD reduction was not significant between the groups (MTS: 3.17 ± 0.95 mm vs CTP: 3.07 ± 0.97 mm, P > .05), whereas the GI change showed a significant difference between the groups (MTS: 1.71 ± 0.41 vs CTP: 1.67 ± 0.41, P < .05). CONCLUSIONS: Multisensory teaching strategies in the preclinical periodontal training of undergraduates can reduce postoperative complications (gingival injury, postoperative haemorrhage, and root surface roughness) and provide a better treatment experience (decreased treatment time and postoperative pain and sensitivity) for patients.