Objective: To investigate the inhibitory effect of neoeriocitrin (Neo) on ligature-induced experimental periodontitis in mice and evaluate its biosafety, providing experimental evidence for novel candidate drugs in periodontitis treatment. Methods: This study has been approved by the Animal Welfare and Ethical Safety Committee. A periodontitis model was established in C57BL/6J mice using silk ligation. The mice were divided into control, periodontitis model, and three Neo treatment groups (5, 10, 20 mg/kg Neo). After 2 weeks of intervention, alveolar bone resorption was analyzed by micro-computed tomography (micro-CT); periodontal tissue pathological changes were observed via hematoxylin-eosin (HE) and Masson staining; osteoclasts were counted using tartrate-resistant acid phosphatase staining; serum inflammatory factor levels [tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), interleukin-1β (IL-1β), interleukin-10 (IL-10)] were detected by enzyme-linked immunosorbent assay; gut microbiota changes were analyzed by 16S rRNA sequencing; and biosafety was assessed through HE staining of the heart, liver, spleen, lung, kidney, and colon, as well as zonula occludens-1 (ZO-1) immunohistochemical staining of the colon. Results: Compared with the periodontitis model group, the 10 mg/kg Neo treatment group showed significantly alleviated alveolar bone resorption, manifested as reduced cementoenamel junction to alveolar bone crest distance, increased bone mineral density, bone volume fraction, and trabecular thickness. Additionally, reduced inflammatory cell infiltration, improved collagen fiber arrangement, and a significantly decreased number of osteoclasts were observed in periodontal tissues. Serum levels of pro-inflammatory factors TNF-α and IL-6 were also reduced. Furthermore, 10 mg/kg Neo intervention modulated the gut microbiota structure without causing significant multi-organ toxicity or impairing intestinal barrier function. Conclusion Neo can effectively mitigate the progression of experimental periodontitis by inhibiting alveolar bone resorption, reducing periodontal tissue inflammation, modulating systemic immunity, and improving gut microbiota. With good biosafety, Neo is a promising candidate drug for the treatment of periodontitis.

Periodontitis leads to progressive destruction of periodontal supporting tissues, and conventional anti-infective therapies often fail to simultaneously achieve infection control and tissue regeneration. Due to the increasing threat of antibiotic resistance, the development of novel bioactive molecules with both anti-infective and pro-regenerative properties has become a focus of research. Antimicrobial peptides (AMPs), characterized by multi-target antimicrobial mechanisms and immunomodulatory functions, have attracted considerable attention in periodontal tissue regeneration. Current evidence indicates that AMPs not only inhibit periodontal pathogens and biofilm formation, but they also regulate macrophage polarization, suppress inflammatory cytokine release, and activate signaling pathways, such as Wnt/β-catenin and MAPK/ERK. These effects promote osteogenic differentiation of periodontal ligament stem cells and bone marrow mesenchymal stem cells, while enhancing angiogenesis through VEGF induction, thereby establishing a favorable immune microenvironment for periodontal regeneration. Despite these promising findings, the in vivo stability, dosage safety, and long-term mechanisms of AMPs remain to be fully elucidated. Future studies should focus on clarifying the molecular mechanisms by which AMPs regulate the periodontal immune microenvironment and angiogenesis-osteogenesis coupled regeneration. In addition, integrating structural optimization, advanced delivery systems, and artificial intelligence-assisted peptide discovery may accelerate the development and clinical translation of novel antimicrobial peptides.

This study aims to comprehensively analyze the material basis of toad visceral oil(hereafter referred to as toad oil), and explore the pharmacological effect of toad oil on atopic dermatitis(AD). Ultra-high performance liquid chromatography-linear ion trap/orbitrap high-resolution mass spectrometry(UHPLC-LTQ-Orbitrap-MS) and gas chromatography-mass spectrometry(GC-MS) were employed to comprehensively identify the chemical components in toad oil. The animal model of AD was prepared by the hapten stimulation method. The modeled animals were respectively administrated with positive drug(0.1% hydrocortisone butyrate cream) and low-and high-doses(1%, 10%) of toad oil by gavage. The effect of toad oil on AD was evaluated with the AD score, ear swelling rate, spleen index, and pathological section results as indicators. A total of 99 components were identified by UHPLC-LTQ-Orbitrap-MS, including 14 bufadienolides, 7 fatty acids, 6 alkaloids, 10 ketones, 18 amides, and other compounds. After methylation of toad oil samples, a total of 20 compounds were identified by GC-MS. Compared with the model group, the low-and high-dose toad oil groups showed declined AD score, ear swelling rate, and spleen index, alleviated skin lesions, and reduced infiltrating mast cells. This study comprehensively analyzes the chemical composition and clarifies the material basis of toad oil. Meanwhile, this study proves that toad oil has a good therapeutic effect on AD and is a reserve resource of traditional Chinese medicine for external use in the treatment of AD.
Animals ; Dermatitis, Atopic/immunology* ; Disease Models, Animal ; Mice ; Male ; Gas Chromatography-Mass Spectrometry ; Humans ; Bufonidae ; Oils/administration & dosage* ; Chromatography, High Pressure Liquid ; Female ; Mice, Inbred BALB C

Objective:To investigate the feasibility of areal bone mineral density (aBMD) quantification and the diagnostic performance for abnormal bone mass using the coronal spectral localizer radiography (SLR) from photon-counting detector CT (PCD-CT).Methods:This cross-sectional study prospectively enrolled 67 participants who underwent both dual-energy X-ray absorptiometry (DXA) and dual-source PCD-CT examinations within 7 days at Ruijin Hospital, Shanghai Jiao Tong University School of Medicine between April 2024 and September 2025. The aBMD values of L1-L4 derived from DXA and calculated based on PCD-CT SLR were obtained (aBMD DXA and aBMD SLR, respectively). Participants were categorized into four groups based on body mass index (BMI): underweight (BMI<18.5 kg/m2, n=3), normal weight (18.5 kg/m2≤BMI<24 kg/m2, n=33), overweight (24 kg/m2≤BMI<28 kg/m2, n=22), and obese (BMI≥28 kg/m2, n=9) groups. Using aBMD DXA as the golden reference, T-scores were calculated, and participants were classified into normal bone mass (T-score≥-1.0, n=42), osteopenia (-2.50.05). Across the normal bone mass, osteopenia, and osteoporosis groups, significant differences were found in aBMD DXA, aBMD SLR, and AE ( P<0.001), but not in RAE ( P=0.426). Using aBMD SLR-based T-score classification, 38, 16, and 13 participants were classifying as having normal bone mass, osteopenia, and osteoporosis. For diagnosing osteoporosis, aBMD SLR achieved an accuracy of 94.0% (63/67), sensitivity of 90.9% (10/11), and specificity of 94.6% (53/56). For detecting abnormal bone mass, the accuracy, sensitivity, and specificity were 94.0% (63/67), 100% (25/25), and 90.5% (38/42), respectively. Conclusions:Quantitative bone density assessment based on PCD-CT SLR is feasible and accurate, unaffected by body habitus or bone mass status, and demonstrates high diagnostic performance for osteoporosis and abnormal bone mass.

Objective:To study the effects of obesity on alveolar bone loss and gut microbiota in mice with periodontitis.Methods:Twenty-four seven-week-old female C57BL/6J mice were randomly divided into four groups based on table of random numbers ( n=6 in each group): normal-fat diet group (NFD group), high-fat diet group (HFD group), normal-fat diet and periodontitis group (NFD_PD group) and high-fat diet and periodontitis group (HFD_PD group). NFD and HFD groups were fed with normal or high-fat diet for twelve weeks respectively; NFD_PD and HFD_PD groups were induced to periodontitis by ligating the bilateral maxillary second molars with 5-0 silk thread at the fourth week after feeding with normal or high-fat diet respectively. The body weight was measured weekly. The mice were euthanized for collecting the samples at the end of the 12th week. Liver, kidneys, perirenal and retroperitoneal fat were weighed. Serum was collected to detect the level of serum lipids and inflammatory factors. The right maxilla bones were scanned by micro-CT. HE staining was performed to observe the periodontal tissue. The cecum contents were collected for gut microbiota 16S rRNA gene sequencing. Spearman correlation analysis was performed to analyze the correlation between the abundance of gut microbiota and serum inflammatory level and CT value. Results:After 12 weeks of high-fat diet fed, the body weight of HFD group [(26.52±1.96) g] was significantly higher than that of NFD group [(20.95±0.63) g] ( t=6.63, P<0.001). The body weight of HFD_PD group [(23.82±1.12) g] was significantly higher than that of NFD_PD group [(20.73±0.47) g] ( t=6.23, P=0.001). The serum levels of total cholesterol, triglyceride and low density lipoprotein in HFD group and HFD_PD group were significantly higher than those in NFD group and NFD_PD group ( P<0.01). The distance from the cemento-enamel junction to the alveolar bone crest (CEJ-ABC) on the mesial site of maxillary second molar in HFD_PD group [(647.46±47.46) μm] was significantly higher than that in NFD_PD group [(440.48±68.08) μm] ( t=5.58, P<0.001). HE staining showed that the maxillary second molar attachment loss, collagen fiber destruction and inflammatory cell infiltration were more significant serious in HFD_PD group compared with NFD_PD group. The levels of interleukin (IL)-1β, IL-6 and monocyte chemotactic protein-1 (MCP-1) of serum in HFD_PD group [(17.11±1.92), (31.61±3.20) and (204.42±35.96) ng/L, respectively] were significantly higher than those in NFD_PD group [(10.44±1.65), (19.96±2.09) and (147.36±10.76) ng/L, respectively] ( P<0.001, P<0.001, P=0.004). The 16S rRNA gene analysis revealed that the Bacteroides/Firmicutes ratio in HFD_PD group (4.00±3.30) was significantly higher than that in NFD_PD group (0.62±0.19) ( t=2.50, P=0.030). The abundance of Oscillospira in HFD_PD group [(12.25±0.05) %] was significantly higher than that in NFD_PD group [(2.80±0.01) %] ( t=4.64, P<0.001). The abundance of Parabacteroides in HFD_PD group [(0.25±0.27)% ] was significantly lower than that in NFD_PD group [(2.04±0.02)%] ( t=2.32, P=0.043). The β-diversity analysis of gut microbiota based on Bray-Curtis distance showed that samples of HFD_PD group and NFD_PD group were obviously grouped. Correlation analysis showed that the abundance of Oscillospira was positively correlated with IL-1β, IL-6, MCP-1 concentration and CEJ-ABC value in serum significantly ( r values were 0.80, 0.79, 0.80, 0.89, P<0.05). The abundance of Parabacteroides was negatively correlated with IL-1β, IL-6 concentration and CEJ-ABC value in serum significantly ( r values were -0.71, -0.71, -0.86, -0.95, P<0.05). Conclusions:Obesity promotes alveolar bone resorption in periodontitis mice and changes the gut microbiota. Oscillospira and Parabacteroides may play a key role.

Objective: To examine the effect of intracellular vesicles (IVs) and extracellular vesicles (EVs) that originated from periodontal ligament stem cells (PDLSCs) on the osteogenic differentiation of PDLSCs within a lipopolysaccharide (LPS)-simulated inflammatory microenvironment, and to provide new insights for the application of IVs in the repair and regeneration of periodontal tissue in periodontitis. Methods: Ethical approval was obtained from the institution. Human-origin PDLSCs were extracted, and the IVs and EVs from PDLSCs at the 3rd-6th passages were gathered and identified using transmission electron microscopy, nano flow cytometry (Nano FCM) analysis, and Western Blot. The 3rd-6th generations of PDLSCs were categorized into the following groups: Control group, LPS group, LPS + 100 μg/mL EVs group (LPS+EVs group), and LPS + 100 μg/mL IVs group (LPS+IVs group). The effects of the IVs and EVs on the anti-inflammatory and osteogenic differentiation of PDLSCs in an inflammatory microenvironment were assessed by using a Cell Counting Kit-8 (CCK-8), enzyme-linked immunosorbent assay (ELISA), quantitative real-time polymerase chain reaction (qRT-PCR), Western Blot, alkaline phosphatase (ALP) staining, and alizarin red staining (ARS). Results: Under transmission electron microscopy, the IVs and EVs derived from PDLSCs displayed a double-layer membrane structure. NanoFCM analysis revealed that the average diameters of the IVs and EVs were 79.6 nm and 82.1 nm, respectively. Western Blot analysis indicated that the surface proteins CD9, CD63, and CD81 of the IVs and EVs were positively expressed, while calnexin was negatively expressed, indicating that IVs and EVs were successfully obtained. Compared with the Control group, the proliferation of PDLSCs in the LPS group was reduced, while the levels of inflammatory cytokine interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) in the cell supernatant were increased, the mRNA expressions of osteogenic differentiation-related genes, including osteoblast-related genes runt-related transcription factor 2 (RUNX2), alkaline phosphatase (ALP), osteocalcin (OCN) of PDLSCs were reduced, the protein expressions of RUNX2 and osteopontin (OPN) were also decreased (P<0.05); compared with the LPS group, the proliferation of PDLSCs in the LPS+EVs group and LPS+IVs group were significantly increased, while the levels of IL-6, TNF-α were significantly reduced, and the mRNA expressions of RUNX2, ALP, OCN were significantly increased, the protein expressions of RUNX2 and OPN were also significantly increased (P<0.05). Further, in the inflammatory microenvironment, Compared with EVs, IVs more significantly promote the proliferation of PDLSCs, inhibit TNF-α expression, enhance the expression of RUNX2 mRNA, upregulate the expression of RUNX2 and OPN proteins, increase ALP activity, and promote the formation of mineralized nodules (P<0.05). Conclusion IVs and EVs derived from PDLSCs can boost the proliferation of PDLSCs in an inflammatory microenvironment, inhibit the expression of inflammatory factors, and advance the osteogenic differentiation of PDLSCs. The anti-inflammatory and osteogenic effects of IVs are superior to those of EVs.

Periodontal disease is highly prevalent, exerting detrimental effects on oral health and posing serious threats to systemic health. Over the past three decades, research exploring the impact of periodontal disease on systemic diseases has rapidly advanced. Nevertheless, numerous challenges and unanswered questions remain, necessitating further investigation. Therefore, this article first elucidates the association between periodontal disease and systemic diseases. Then, the key evidence supporting their relationship is graded according to the Oxford Centre for Evidence-Based Medicine levels of evidence criteria. Specifically, periodontal disease emerges as an independent risk factor for diabetes mellitus (level A evidence) and cardiovascular diseases (level B evidence). As such, it represents a potential risk factor for rheumatoid arthritis (level B evidence), chronic obstructive pulmonary disease (level B evidence), and inflammatory bowel disease (level B evidence). Furthermore, periodontal disease is closely linked to adverse pregnancy outcomes. Second, this article delineates the plausible mechanisms through which periodontal disease influences systemic diseases, explicitly showing that the foundational elements underlying their connection are bacteria and inflammation. The circulation pathway and saliva pathway specifically mediate this connection. Finally, in light of the current ambiguities surrounding the relationships between periodontal disease and certain systemic diseases, as well as the insufficient depth of mechanism research, this article outlines several considerations for future clinical research and animal experiment designs. Implementing large-sample, multi-center, high-quality clinical studies, utilizing multi-omics analyses for more in-depth exploration of mechanisms, and actively promoting clinical translational research are recommended. This article aims to advance the field of periodontal medicine, while simultaneously offering evidence-based insights to inform the implementation of public health policies.

Objective: To investigate the role of butyric acid-producing bacteria in long bone homeostasis in mice with periodontitis under a high-fat/high-sugar diet and to provide new insights for the prevention and treatment of periodontitis and related bone metabolic diseases. Methods: This study has been approved by the Animal Welfare and Ethics Committee of the Experimental Animal Center. Initially, 14 mice were randomly divided into the CON group (the control group) and the LIG group (the periodontitis group). Mice in the LIG group had experimental periodontitis induced by ligating the second maxillary molars bilaterally and were fed a high-fat and high-sugar diet. After 8 weeks, samples were collected. Micro-computed tomography (Micro-CT) was used to analyze alveolar bone resorption and various parameters of the proximal tibia trabecular bone, including bone mineral density (BMD), bone volume per tissue volume (BV/TV), trabecular number (Tb.N), trabecular thickness (Tb.Th), and trabecular separation (Tb.Sp). After decalcification, hematoxylin and eosin (HE) staining was performed on maxillary bone sections to assess periodontal tissue inflammation and connective tissue destruction. Quantitative real-time polymerase chain reaction (qRT-PCR) was used to detect related genes in the distal femur and proximal tibia bone tissues, including osteocalcin (OCN), osteogenic transcription factor (Osterix), osteoprotegerin (OPG), tartrate resistant acid phosphatase (TRAP), osteoclast-associated receptor (OSCAR), receptor activator of nuclear factor kappa-B (RANK), and receptor activator of nuclear factor kappa-B ligand (RANK-L). Subsequently, the other 28 mice were randomly divided into the CON group (the control group), LIG group (the periodontitis group), CON + butyric acid-producing bacteria (BP) group, and LIG + BP group. The breeding, sampling, and sample detection methods remained the same. Finally, the other 28 mice were randomly divided into the CON group (the control group), LIG group (the periodontitis group), CON + sodium butyrate (SB) group, and LIG + SB group. The breeding, sampling, and sample detection methods remained the same. Results: ①Periodontitis modeling was successful. Compared with the CON group, the LIG group exhibited significant alveolar bone resorption of the maxillary second molar, aggravated periodontal tissue inflammation, and connective tissue destruction. ②Periodontitis exacerbated long bone resorption in mice fed a high-fat high-sugar diet. Compared with the CON group, the LIG group had significantly lower BMD, BV/TV, Tb.N, and Tb.Th (P<0.05), and significantly higher Tb.Sp (P<0.05). HE staining of the proximal tibia showed that the trabeculae in the LIG group were sparse and disordered, with some areas showing fractures or dissolution. The expression of osteoblast markers (OCN, Osterix, OPG) was significantly lower in the LIG group (P<0.05), while the expression of the osteoclast marker TRAP showed an increasing trend (P>0.05). The ratio of RANK-L/OPG was significantly higher in the LIG group compared with the CON group (P<0.05). ③ Supplementation with butyric acid-producing bacteria alleviates periodontitis-induced disruption of long bone homeostasis in mice fed a high-fat/high-sugar diet. Compared with the LIG group, BMD and Tb.Th were significantly higher in the LIG + BP group. HE staining of the proximal tibia showed that bone resorption was mitigated in the LIG + BP group compared with the LIG group. The expression of OCN and Osterix was significantly higher in the LIG + BP group, while the expression of osteoclast-specific genes (OSCAR, RANK, RANK-L) was significantly lower (P<0.05). ④ Supplementation with butyrate alleviates periodontitis-induced disruption of long bone homeostasis in mice fed a high-fat/high-sugar diet. Compared with the LIG group, BV/TV and Tb.N were significantly higher in the LIG + SB group, and Tb.Sp was significantly lower (P<0.05). HE staining of the proximal tibia showed that bone resorption was mitigated in the LIG + SB group compared with the LIG group. The expression of Osterix, OPG, OSCAR, TRAP, and RANK was significantly lower in the LIG + SB group compared with the LIG group (P<0.05). Conclusion Periodontitis disrupts the long bone homeostasis of mice fed a high-fat high-sugar diet, aggravating long bone resorption. Supplementation with butyric acid-producing bacteria or butyrate can effectively alleviate the disruption of long bone homeostasis caused by periodontitis.

Periodontitis and rheumatoid arthritis (RA) are chronic inflammatory diseases that share similar inflammatory mechanisms and characteristics. Programmed cell death (PCD) has recently garnered attention for its crucial role in regulating inflammation and maintaining tissue homeostasis, as well as for its potential to link these two diseases. The various forms of PCD--including apoptosis, pyroptosis, and necroptosis--are closely controlled by signaling pathways such as Toll-like receptor 4 (TLR4) /NF-κB and MAPK. These pathways determine cell fate and influence inflammatory responses, tissue destruction, and repair, and they both play important roles in the pathogenesis of RA and periodontitis. In periodontitis, periodontal pathogens such as Porphyromonas gingivalis (P. gingivalis) and its virulence factors, including lipopolysaccharide (LPS), induce pyroptosis and necroptosis in immune cells such as macrophages via the TLR4/NF-κB pathway, which leads to an excessive release of pro-inflammatory cytokines such as interleukin (IL)-1β and tumor necrosis factor (TNF)-α. Concurrently, these pathogens inhibit the normal apoptotic process of immune cells, such as neutrophils, prolonging their survival, exacerbating immune imbalance, and aggravating periodontal tissue destruction. Similarly, in RA synovial tissue, fibroblast-like synoviocytes (FLS) acquire apoptosis resistance through signaling pathways such as the Bcl-2 family, JAK/STAT, and NF-κB, allowing for the consistent proliferation and secretion of matrix metalloproteinases and pro-inflammatory cytokines. Meanwhile, the continuous activation of pyroptotic pathways in neutrophils and macrophages results in the sustained release of IL-1β, further exacerbating synovial inflammation and bone destruction. Notably, dysregulated PCD fosters inter-organ crosstalk through shared inflammatory mediators and metabolic networks. Damage-associated molecular patterns (DAMPs) and cytokines that originate from periodontal lesions can spread systemically, influencing cell death processes in synovial and immune cells, thereby aggravating joint inflammation and bone erosion. By contrast, systemic inflammation in RA can upregulate osteoclastic activity or interfere with the normal apoptosis of periodontal cells via TNF-α and IL-6, ultimately intensifying periodontal immune imbalance. This review highlights the pivotal bridging role of PCD in the pathogenesis of both periodontitis and RA, providing a reference for therapeutic strategies that target cell death pathways to manage and potentially mitigate these diseases.

Periodontitis is a chronic inflammatory disease of the periodontal supporting tissues caused by plaque microorganisms, whereas inflammatory bowel disease (IBD) is a chronic inflammatory disease characterized by gastrointestinal tract damage. Studies have revealed a close association between periodontitis and IBD, and gut microbiota has been shown to play an important role in the development of IBD. When the gut microbiota is disturbed, it leads to intestinal barrier disruption, triggers immune-inflammatory responses, and influences IBD progression. There are significant differences between the salivary microbiota of periodontitis patients and healthy individuals, and periodontal pathogens can enter the intestinal tract with saliva and participate in the development of IBD by influencing the interactions between gut microbiota composition, immune responses, metabolite production, and intestinal barrier function. Current gut microbiota-targeted intervention strategies, such as fecal microbiota transplantation (FMT) and probiotic supplementation, have shown potential therapeutic value in the treatment of periodontitis. These approaches may exert synergistic effects on both periodontitis and IBD through microbiota modulation. This review summarizes research progress on the relationship between periodontitis and IBD to provide a foundation for the prevention and treatment of these two diseases.