Search Results (535)

Age-related macular degeneration (AMD) is a complex retinal disease influenced by genetic and metabolic factors. Genetic variants impact disease susceptibility, while alterations in amino acid and purine metabolism are involved in AMD development. This study aimed to examine the association between the COL2A1 rs1635529 polymorphism and AMD, as well as its relation to specific metabolites. The study comprised 919 participants: 261 with early AMD, 229 with exudative AMD, and 429 controls. DNA was extracted using the salting-out method, and genotyping was performed using real-time PCR. Metabolite levels were analysed with liquid chromatography–mass spectrometry. Statistical analysis was conducted using IBM SPSS Statistics 27.0. Logistic regression revealed that carriers of the GT + TT genotypes had a 1.63-fold higher risk of early AMD (p = 0.046). The T allele was also linked to a 1.67-fold elevated risk (p = 0.033). No significant associations were observed in exudative AMD. Furthermore, lower leucine levels were noted in exudative AMD patients, and inosine levels were reduced in GT genotype carriers within the early AMD group. The COL2A1 rs1635529 polymorphism showed a nominal association with early AMD, but not exudative AMD. Differences in leucine and inosine levels were observed, suggesting a potential link between genetic variation and metabolic alterations. These findings indicate possible involvement of collagen-related and metabolic pathways in early disease development; however, the results should be interpreted with caution and require validation in larger studies. Full article

This study aims to evaluate the therapeutic efficacy of emodin (EMO) in rheumatoid arthritis (RA) and to verify whether its underlying mechanism involves the blockade of pathological angiogenesis via the inhibition of the nuclear factor-kappa B (NF-κB)/hypoxia-inducible factor-1α (HIF-1α)/vascular endothelial growth factor (VEGF) signaling axis. Bovine type II collagen-induced arthritis (CIA) mouse models and lipopolysaccharide (LPS)-stimulated EA.hy926 endothelial cells were utilized in this study. The effects of EMO on joint pathological alterations, the expression of NF-κB/HIF-1α/VEGF axis proteins, inflammatory cytokines (tumor necrosis factor-alpha (TNF-α), interleukin-6 (IL-6), and interleukin-1 beta (IL-1β)), and angiogenic capacity were assessed using histopathological analysis, Western blotting, immunohistochemistry (IHC), immunofluorescence, and tube formation assays. Furthermore, small interfering RNA (siRNA) interference targeting key molecules was employed to validate the molecular mechanisms underlying the therapeutic effects of EMO. In the CIA model group, the ankle joints of mice exhibited pronounced inflammatory infiltration, synovial hyperplasia, and bone destruction. Compared with the model group, both the EMO and methotrexate (MTX) treatment groups demonstrated attenuated synovial hyperplasia and cartilage destruction, along with significantly downregulated expression levels of key NF-κB pathway proteins, HIF-1α, and VEGF in joint tissues (p < 0.001). In vitro experiments revealed that EMO treatment significantly reduced the LPS-induced secretion of pro-inflammatory cytokines (TNF-α, IL-6, and IL-1β) (p < 0.001), and decreased both the number and total length of tubular structures formed by endothelial cells compared to the control (p < 0.001). Notably, siRNA-mediated knockdown of p65 resulted in decreased intracellular protein levels of HIF-1α and VEGF, accompanied by a significant reduction in tube formation (p < 0.001). This study demonstrates that EMO alleviates pathological damage in RA by inhibiting the activation of the NF-κB signaling pathway, which subsequently downregulates pathological angiogenesis and inflammatory responses mediated by the HIF-1α/VEGF axis. These findings provide a robust experimental basis for the potential application of EMO as a therapeutic agent for RA. Full article

The in vivo effect of platelet-rich plasma (PRP) on supraspinatus tendon morphology and subacromial bursa cell gene expression in degenerative rotator cuff tears remains unclear. This randomized controlled trial evaluated the effect of preoperative leukocyte-poor PRP (LP-PRP) subacromial injection on supraspinatus tendon histology and subacromial bursa gene expression. Sixteen patients with full-thickness supraspinatus tears were randomized to receive an ultrasound-guided LP-PRP injection (n = 8) or no injection (n = 8) six weeks before arthroscopic repair. Tendon biopsies were assessed using the modified Movin score. Gene expression of collagen type I, II and III, metalloproteinase 3 and 13, and interleukin 1β and 6 genes from subacromial bursa cells was quantified using quantitative real-time PCR. The results of the two groups were compared to determine any statistically significant difference regarding all the examined parameters. The PRP group demonstrated a significantly lower total modified Movin score than controls (6.5 vs. 12.1, p = 0.002), with lower scores for fiber structure, fiber arrangement, nuclear rounding, inflammation and cell density (all p < 0.003), while angiogenesis did not differ (p = 0.149), indicating an architecture closer to that of normal tendon. No statistically significant differences in gene expression were observed (all p > 0.05), although collagen II and metalloproteinase 3 and 13 showed biologically relevant downregulation [fold change 0.23 (95%CI 0.05–1.09), 0.24 (95%CI 0.002–26.10), and 0.26 (95%CI 0.02–2.76), respectively]. The LP-PRP injection was associated with improved supraspinatus tendon histological characteristics and biologically relevant reductions in selected bursal genes, in the setting of supraspinatus tendon tear. Full article

Cartilage is an important yet vulnerable tissue with limited self-healing capacity, where damage often progresses to joint degeneration, which eventually leads to severe osteoarthritis (OA). Current tissue engineering strategies focus on biocompatible scaffolds for cartilage regeneration, particularly amnion (or amniotic membrane), emerging as a promising biomaterial due to its wide availability, low immunogenicity, and naturally derived microenvironment that is advantageous for cartilage regeneration. This systematic review aims to evaluate the existing evidence on the efficacy of amnion as a tissue scaffolding material for cartilage regeneration in both preclinical and clinical studies. Using terms such as “cartilage damage”, “cartilage injuries”, “amnion” and “amniotic membrane”, 19 relevant studies were identified across three major databases (PubMed, Scopus and Web of Science) until 25 December 2025. All preclinical and clinical studies that utilized amnion for cartilage repair or as cartilage tissue engineering scaffolding materials were included. Evidence quality was assessed using the OHAT and MINORS risk of bias tool. This study is prospectively registered in the PROSPERO database under the ID 1178444. The findings consistently indicate that amniotic scaffolds, regardless of processing methods or cell seeding, yield favorable outcomes without adverse effects across different species. In vitro analysis revealed that treatment groups with amnion show better cell attachment, viability, and proliferation, and higher content of cartilage-related markers expressed by the seeded cells, either chondrocyte, bone marrow-derived mesenchymal stem cells (MSCs), adipose tissue-derived MSCs, placenta-derived MSCs, umbilical cord-derived MSCs, amniotic MSCs or amniotic epithelial cells. In in vivo and ex vivo studies, amnion-treated groups demonstrated improved quality of the treated cartilage, with better integration, as indicated by higher histological scores and the presence of type II collagen (COL-II). There was an inconsistency in the reporting of cartilage defect dimensions in the in vivo models across the different studies. Nevertheless, the outcome measurements were consistently reported with histological analysis, with or without International Cartilage Repair Society (ICRS) scoring and immunohistochemistry (IHC) analysis, across the studies. Clinically, most subjects show improvement in the Knee Injury and Osteoarthritis Outcome Score (KOOS) Sports and Recreation score and KOOS Quality of Life score, as well as reduced Visual Analogue Scale (VAS) average and maximum pain scores. In conclusion, preclinical and clinical studies support amnion as an ideal scaffold material for cartilage tissue engineering and regeneration. Future research should focus on optimizing and standardizing amnion scaffold preparation at a production scale to facilitate the translation of these positive outcomes into clinical applications. This study is funded by the Ministry of Higher Education Malaysia via Prototype Research Grant Scheme (PRGS/1/2021/SKK01/UM/02/1) and UM International Collaboration Grant—2023 SATU Joint Research Scheme Program: ST007-2024. Full article

We evaluated human embryonic stem cell-derived mesenchymal stem cells (ES-MSCs) on collagen scaffolds for meniscus-like neotissue formation and ex vivo repair of human osteoarthritic (OA) meniscal defects. Collagen type I fibrous scaffolds were pneumatospun, and laminate scaffolds were fabricated from electrospun PLA/collagen; crosslinked; heparin conjugated; fibronectin coated; functionalized with TGFβ1, TGFβ3, or PDGFbb; seeded with ES-MSCs; and cultured for 4 weeks, followed by in vitro assessment or ex vivo implantation into 3.5 mm human meniscus defects for 5 weeks. Pneumatospinning generated highly porous scaffolds that supported uniform cell infiltration, while laminate scaffolds demonstrated interlocking fiber interfaces and enhanced mechanical properties. TGFβ1 and TGFβ3 immobilization enhanced scaffold bioactivity, defined as growth factor-mediated increases in meniscus-like matrix deposition, collagen fiber organization, and meniscogenic gene expression, by significantly increasing safranin O staining, collagen type II deposition, collagen fiber polarization, and ACAN expression. TGFβ3 additionally increased COL1A1 expression and pushout shear modulus; TGFβ1 increased peak pushout stress, indicating superior ex vivo mechanical integration. Laminate scaffolds resulted in extensive cell infiltration, robust neotissue formation (elastic modulus ~2.4 MPa), and improved ex vivo tissue integration when functionalized with TGFβ3. The data indicated that ES-MSC-seeded, heparin-conjugated, TGFβ-immobilized pneumatospun/electrospun collagen–PLA scaffolds support meniscogenic differentiation and biomechanical integration, with repair of focal meniscal defects and potential for partial meniscus replacement. Full article

Articular cartilage, a highly specialised and avascular tissue, exhibits limited regenerative potential following trauma or degenerative conditions such as osteoarthritis (OA). Conventional surgical interventions, including microfracture and autologous chondrocyte implantation (ACI), have shown limited long-term efficacy due to donor site morbidity and restricted cell proliferation. In this context, mesenchymal stromal cells (MSCs) have emerged as a promising alternative owing to their multipotency, self-renewal capacity, and low immunogenicity. While bone marrow (BM) remains the traditional source of MSCs, recent studies have reported that peripheral blood-derived mesenchymal stromal cells (PB-MSCs) may possess chondrogenic, osteogenic, and adipogenic potential comparable to that of BM-derived MSCs. PB-MSCs can be harvested through minimally invasive methods, thereby avoiding the complications associated with BM aspiration. Experimental evidence indicates that PB-MSCs exhibit strong cell viability, proliferative potential, and the ability to synthesise cartilage-specific extracellular matrix proteins, such as type II collagen and sulphated glycosaminoglycans, within three-dimensional scaffolds. Immunophenotypically, PB-MSCs express mesenchymal markers including CD29, CD44, CD90, and CD105 while lacking hematopoietic markers CD34 and CD45. Flow cytometry analyses reveal that CD105⁺ populations increase following cryopreservation, highlighting their clinical utility. In contrast to these experimentally defined PB-MSCs, the term peripheral blood stem cells (PBSCs) is used in clinical studies to describe heterogeneous, non-cultured peripheral blood-derived cell preparations, typically enriched in hematopoietic stem and progenitor cells following granulocyte colony-stimulating factor (G-CSF) mobilisation, without full mesenchymal characterisation. In vitro studies confirm successful tri-lineage differentiation, whereas in vivo investigations have demonstrated effective cartilage regeneration using PB-based clinical approaches, including postoperative intra-articular administration of hyaluronic acid (HA) combined with PBSCs, as well as implantation of PBSCs covered with a collagen membrane. Furthermore, advancements in biomaterial engineering, such as poly(ethylene glycol)–cysteine–arginine–glycine–aspartic acid (PEG-CRGD) hydrogels, have enhanced PB-MSC adhesion, proliferation, and chondrogenic differentiation while promoting immunomodulation through M2 macrophage polarisation. Despite these promising outcomes, the available evidence remains limited and heterogeneous, with substantial variability in cell definitions, experimental models, and clinical study designs, which currently constrains definitive conclusions regarding clinical efficacy. Future research should focus on optimising isolation protocols, understanding molecular pathways governing PB-MSC chondrogenesis, and standardising clinical applications. Overall, PB-MSCs represent a viable, less invasive, and translationally relevant cell source for cartilage regeneration and regenerative orthopaedic therapies Full article

Type II collagen (CII), the major structural protein in the cartilage extracellular matrix, is a promising biomaterial for scaffold design in cartilage tissue engineering. In this study, high-purity CII was successfully extracted from bovine cartilage, an abundant by-product of cattle slaughter, and its amino acid composition, triple-helical conformation, and thermal stability were verified. CII was subsequently combined with silk fibroin (SF) and chitosan (CS) to fabricate three-dimensional (3D) porous scaffolds via freeze-drying. The pore structure, porosity, swelling behavior, mechanical properties and in vitro degradation characteristics were systematically evaluated. Scaffolds with favorable structural integrity, mechanical performance, and degradation rates were further evaluated biologically using human primary chondrocytes. All CII-based composite scaffolds supported chondrocyte growth and promoted early extracellular matrix deposition. Notably, the scaffold with a CII:SF:CS ratio of 7:3:1 showed the highest GAG/DNA content, accompanied by upregulated gene expression related to the cartilage phenotype (COL2A1, ACAN, and SOX9) and reduced expression of the dedifferentiation marker COL1A1, indicating improved phenotype maintenance. Overall, within the tested range, CII70 (CII:SF:CS = 7:3:1) represents a practical compromise between scaffold stability and in vitro chondrocyte-related outcomes, providing a basis for selecting CII/SF/CS formulations for cartilage tissue engineering. Full article

Background: Joint inflammation is significantly connected with progressive joint deterioration, potentially increasing the incidence of persistent major clinical challenges and global disability. Nutrient-based preventive strategies have been explored to investigate the interventive efficacy of the proposed prescribed formula for joint inflammation. However, the synergistic ameliorative effects of the nutritional formula should be evaluated to investigate its impact on joint inflammation. Methods: A prescribed formula including turmeric (T), N-acetylglucosamine (G), enzymatically hydrolyzed bone powder (E), and undenatured type II collagen (U) was comprehensively evaluated for its synergistic effects on joint inflammation and the underlying mechanisms. A rat model established using the Hulth method was used to evaluate the interventive effects in vivo. Moreover, in vitro analysis using the murine chondrogenic cell line ATDC5 was performed to validate the intervention and its mechanism of action. Results: The prescribed formula was shown to synergistically reduce levels of inflammation-related cytokines, reduce oxidative stress, and enhance bone metabolism to promote joint regeneration. Micro-Computed Tomography (Micro-CT) analysis revealed restoration of joint architecture and ameliorated physiological status upon formula intervention. In vitro analysis further validated the synergistic alleviation of inflammation and oxidation, as well as reductions in MMP13 and CTX-1 levels, which implies that modulating bone metabolism alleviates the deterioration and inflammation of joint architecture. Conclusions: The synergistic formula in this study achieves synchronous modulation of several core pathological pathways, yielding synergistic modulation of joint inflammation. Nutrient-based interventions or preventive strategies show promising effects against joint inflammation and progressive mechanistic deterioration. Full article

Osteoarthritis (OA) in dogs is increasingly recognized as a condition with systemic inflammatory and metabolic components, potentially influenced by body condition and sex. This study aimed to characterize phenotypic differences in circulating inflammatory, metabolic, and biochemical biomarkers in dogs with OA according to body condition and sex. In this cross-sectional study, client-owned dogs were classified as healthy controls, thin dogs with OA (TOA), or obese dogs with OA (OOA). Circulating cytokines, adipokines, cartilage degradation markers, and routine biochemical parameters were measured in blood samples, including interleukin-1 beta, interleukin-4, interleukin-10, adiponectin, C-terminal telopeptide of type II collagen, and standard metabolic and hepatic markers. Data were analyzed using linear models fitted on log-transformed values, with group and sex as fixed effects, complemented by adjusted and sensitivity analyses. TOA dogs showed significantly higher interleukin-1 beta concentrations compared with controls (multiplicative effect 1.39, 95% confidence interval 1.05–1.82), indicating increased systemic inflammatory activity. In contrast, OOA dogs exhibited predominantly metabolic-associated alterations, including higher gamma-glutamyl transferase activity (multiplicative effect 1.22, 95% confidence interval 1.03–1.46) and higher cholesterol concentrations (multiplicative effect 1.22, 95% confidence interval 1.03–1.46). Several other biomarkers showed no clear group-related differences. Overall, these findings demonstrate that systemic biomarker profiles in canine OA vary primarily according to body condition, with secondary sex-related patterns, supporting the existence of biologically distinct OA phenotypes relevant for future diagnostic and therapeutic strategies. Full article

Background/Objectives: Obesity is characterized by chronic low-grade inflammation, which plays a central role in the development of its metabolic complications. The genetic factors influencing this inflammatory phenotype remain incompletely understood. This study aimed to analyze the associations of functional polymorphisms in genes involved in extracellular matrix remodeling (MMP2, MMP9, MMP12, COL1A1), metabolism (MTHFR, CYP3A5), and vascular regulation (NOS3, AGTR1) with plasma cytokine profiles and to identify inflammatory subphenotypes in patients with obesity. Methods: The study included 127 individuals, comprising 73 patients with excess body weight (body mass index, BMI ≥ 25 kg/m²) and 54 individuals with normal weight (BMI 18.5–24.9 kg/m²). Genotyping of selected polymorphisms was performed using real-time PCR. Plasma concentrations of 47 cytokines and chemokines were measured by multiplex immunoassay. Results: Nominally significant associations between genetic variants and cytokine levels were identified. Polymorphisms COL1A1 rs1107946 (CA genotype) and MMP9 rs17576 (AG genotype) were associated with a favorable inflammatory profile (decreased IL-6 and increased IL-10, respectively). In contrast, the AGTR1 rs5186 (AC genotipe) variant was associated with elevated TNF-α, IP-10/CXCL10, while the MTHFR rs1801131 (AC genotipe) variant was linked to increased MIP-1β/CCL4, both reflecting a pro-inflammatory shift. Complex, pleiotropic associations were observed for MMP2 rs243865 (elevated IL-7 and Fractalkine/CX3CL1) and NOS3 rs1799983 (elevated MCP-1/CCL2 and Eotaxin/CCL11). Cluster analysis revealed distinct patient subpopulations with differing inflammatory signatures. In one well-defined subgroup, an exploratory model (test R² = 0.537) identified IL-8, IL-15, and albumin as candidate biomarkers predictive of BMI. Conclusions: The study identifies candidate genetic polymorphisms and inflammatory biomarkers associated with distinct patterns of systemic inflammation in obesity. These hypothesis-generating findings underscore the phenotypic heterogeneity of obesity and provide a basis for further research into the stratification of patients by the risk of developing metabolic complications. Full article

Alveolar inflammation, elevated interleukin-17A (IL-17A), and fibrin deposition are common features in all forms of lung injury followed by fibrotic repair. Type II alveolar epithelial cell (AEC) viability, regulated by tumor suppressor protein p53 and changes in uPA-mediated fibrinolysis, has been linked to lung injury and pulmonary fibrosis (PF). Nevertheless, mechanistic details linking increased IL-17A with p53 and PAI-1 to lung injury and remodeling remain unclear. We found that IL-17A and its receptor (IL-17RA) are induced during various lung injuries. IL-17A augments IL-17RA, p53 and downstream PAI-1 with a concurrent decrease in uPA and its receptor (uPAR) in AECs. These changes promote AEC apoptosis, alveolar injury and PF. In addition, IL-17A causes a dose-dependent increase in IL-17RA and profibrogenic markers in lung fibroblasts (LFs), suggesting myofibroblast differentiation. We further found that inhibition of IL-17A by caveolin-1 scaffolding domain peptide (CSP) or its 7-mer deletion fragment (CSP7) inhibits AEC apoptosis, lung inflammation, and profibrogenic markers in LFs and PF. Further, treatment of mice with bleomycin-induced lung injury using CSP7, an anti-IL-17A antibody, or an IL-17RA blocking antibody attenuates total lung hydroxyproline and soluble collagen content, as well as levels of profibrogenic markers. These observations support the role of IL-17A/IL-17RA signaling in lung injury and post-injury remodeling. Full article

Canine osteoarthritis is a progressive degenerative joint condition characterized by inflammation, structural damage, pain, and impaired joint function. Osteoarthritis risk is influenced not only by well-established factors such as age, genetics, obesity and underlying arthropathies, but also by systemic conditions such as low-grade inflammation and sarcopenia. Current interventions are primarily focused on pain relief rather than mitigating physiological impairment, despite recent veterinary guidelines. In this study, several preclinical models were employed to identify postbiotic candidates with potential to promote canine joint health. An initial screening of nine heat-treated bacterial strains identified two Lacticaseibacillus rhamnosus strains, PRIOME^® JH and HT-PB01, which demonstrated significant anti-inflammatory activity on canine chondrocytes. Further characterization revealed that these two strains significantly increased type II collagen synthesis in canine chondrocytes and modulate immune responses in canine macrophages, enhancing Interleukin-10 secretion over Interleukin-12. Additionally, both strains showed potential in mitigating risk factors such as gut inflammation and permeability, fat accumulation and sarcopenia in Caenorhabditis elegans models. In conclusion, this study identified two promising postbiotic candidates, PRIOME^® JH and HT-PB01, with the potential to target key risk factors associated with osteoarthritis through multifactorial mechanisms. These findings emphasize conducting dog clinical trials to evaluate their relevance in osteoarthritis. Full article

The tendon-to-bone enthesis is a multiphasic structure with four structurally continuous and compositionally distinct regions: tendon, uncalcified fibrocartilage, calcified fibrocartilage and bone. Our study aimed to develop 3D scaffold-free in vitro spheroids and macro-tissues of the enthesis for applications as experimental tools to understand the development and repair of enthesis injury. This study hypothesises that integrating tendon and bone cell spheroids with bone marrow mesenchymal stem cell spheroids will facilitate the production of a fibrocartilaginous interface. 3D Spheroids: The biphasic (tendon–bone) and triphasic co-culture (tendon–stem cell–bone) of spheroids in growth media and chondrogenic media were investigated to establish fusion kinetics, and the cellular and ECM components produced via histology and immunohistochemistry. Complete fusion between spheroids occurred within 6-to-8 days in biphasic co-culture, and 15-to-20 days in triphasic co-culture. Compared to biphasic, the triphasic co-culture in chondrogenic media showed a continuous interface connecting the tendon and bone regions. The presence of collagen I, sulphated proteoglycans and collagen type II in the interface region of triphasic co-culture indicates fibrochondrogenic differentiation. 3D macro-tissues: The modular tissue engineering strategy was used in this study to produce enthesis macro-tissues using spheroids as building blocks. Spheroids were bio-assembled in the triphasic manner (12 tendon spheroids, 12 stem cell spheroids and 8 bone spheroids) in the custom-designed and 3D-printed temporary supports (Formlabs Clear Resin^®) using a customised spheroid bio-assembly system. The fusion of spheroids occurred by day 8 after bio-assembly, and they were removed from temporary supports and cultured in scaffold-free conditions. Although the bio-assembly methodology was successful in producing fused scaffold-free macro-tissues, the histological analysis revealed the presence of an extensive necrotic core due to the large-sized constructs. To conclude, the findings support the hypothesis that a triphasic co-culture has the potential to produce a structurally continuous fibrocartilaginous interface but requires further optimisation to produce macro-tissues with anatomical morphologies and reduced necrotic cores. Full article

Type II collagen is a primary fibrillar component of articular cartilage, and its early degradation is a key biomarker of joint-degenerative disorders such as osteoarthritis, rheumatoid arthritis, gout, etc. Reliable detection at low concentrations remains challenging due to limited assay accessibility, complex analytical procedures, and nonspecific responses in multicomponent biological matrices. This research reports the development of a Molecularly Imprinted Polymer (MIP)–based electrochemical sensor engineered for the selective recognition of type II collagen. A series of monomer formulations were evaluated, and the 1AAM:2VP composition produced a well-defined imprinted layer on screen-printed carbon electrodes, yielding the highest electrochemical sensitivity and linearity. The optimized sensor exhibited strong anodic and cathodic responses proportional to increasing collagen concentrations, with a calibration slope corresponding to an R² value of 0.9394. Minimal signal interference was observed, confirming high molecular selectivity. The limit of detection (LOD) was calculated to be approximately 0.065 µg/mL. These characteristics demonstrate that the proposed MIP sensor provides a low-cost, accessible, and highly selective analytical platform suitable for early-stage cartilage degeneration monitoring. Full article

Periodontitis is a chronic inflammatory condition characterized by dysregulated immune responses that promote alveolar bone destruction. Targeting inflammatory signaling pathways has therefore become an important area of investigation. This study investigated the anti-inflammatory and bone-protective effects of Magnolia kobus DC. extract (MKE) in a ligature-induced periodontitis rat model. Rats were assigned to five groups (n = 5 per group): non-ligature control, ligature control, doxycycline (20 mg/kg), MKE 100 mg/kg, and MKE 400 mg/kg, and treated orally for eight weeks. Periodontal damage and alveolar bone loss were assessed by micro-computed tomography (micro-CT), gingival index, and tooth mobility. Micro-CT analysis demonstrated a dose-dependent reduction in alveolar bone loss, as evidenced by a significant decrease in the cementoenamel junction–alveolar bone crest (CEJ–ABC) distance and reduced furcation involvement in MKE-treated groups compared with the ligature control group, while tooth mobility scores were significantly improved. Serum levels of receptor activator of nuclear factor kappa-B ligand, interleukin-1β, tumor necrosis factor-α, and cyclooxygenase-2 were significantly decreased, while nuclear factor kappa-B signaling was suppressed in gingival tissue. The extract also significantly reduced matrix metalloproteinases 3, 8, 9, and 13, and increased collagen type I and II expression. In summary, MKE exerted anti-inflammatory and bone-protective properties, effectively reducing alveolar bone loss and maintaining periodontal structure. These findings support MKE’s potential application as a natural anti-inflammatory and bone-protective agent and as a functional food ingredient for periodontitis prevention and treatment, meriting further clinical evaluation. Full article