KL-Biome (Postbiotic Formulation of Lactiplantibacillus plantarum KM2) Improves Dexamethasone-Induced Muscle Atrophy in Mice.

Sarcopenia refers to an age-related decrease in muscle mass and strength. The gut-muscle axis has been proposed as a promising target to alleviate muscle atrophy. The effect of KL-Biome-a postbiotic preparation comprising heat-killed Lactiplantibacillus plantarum KM-2, its metabolites, and an excipient (soybean powder)-on muscle atrophy was evaluated using dexamethasone (DEX)-induced atrophic C2C12 myoblasts and C57BL/6J mice. KL-Biome significantly downregulated the expression of genes (Atrogin-1 and MuRF1) associated with skeletal muscle degradation but increased the anabolic phosphorylation of FoxO3a, Akt, and mTOR in C2C12 cells. Oral administration of KL-Biome (900 mg/kg) for 8 weeks significantly improved muscle mass, muscle function, and serum lactate dehydrogenase levels in DEX-treated mice. KL-Biome administration increased gut microbiome diversity and reversed DEX-mediated gut microbiota alterations. Furthermore, it significantly increased the relative abundances of the genera Subdologranulum, Alistipes, and Faecalibacterium prausnitzii, which are substantially involved in short-chain fatty acid production. These findings suggest that KL-Biome exerts beneficial effects on muscle atrophy by regulating gut microbiota.

The interaction of milk sphingomyelin and proteins on stability and microstructure of dairy emulsions.

The interaction between dairy proteins [micellar casein (MC) vs. whey protein isolate (WPI)] and phospholipids [PL; soy phosphatidylcholine (PC) vs. milk sphingomyelin (SM)] in an oil-in-water emulsion system was investigated. Sole PC-stabilized emulsion (1%, wt/vol) showed a significantly larger mean particle diameter (6.5 µm) than SM-stabilized emulsions (3.8 µm). The mean particle diameters of emulsions prepared by the combination of protein (1%, wt/vol) and PL (1%, wt/vol) did not significantly differ from the emulsions prepared with a single emulsifier (MC, WPI, and SM). Emulsion instability differed significantly among samples by a centrifugation-mediated accelerated stability test. Emulsion instability increased in the order of MC+SM < MC+PC, WPI+SM < WPI+PC < MC < SM < WPI < PC. Protein surface load determined by aqueous phase depletion was significantly decreased only in WPI+PC emulsion, whereas no significant difference was found between the MC+SM and WPI+SM emulsions. Topographic and phase images of emulsion surface by atomic force microscopy showed surface layers prepared by protein+PL combinations were composites with different mechanical properties, and PL formed a more compact domain than proteins. A smoother phase image was observed in MC+PL combinations than in WPI+PL counterparts. Based on the microstructure analysis using confocal laser scanning microscopy, combination and MC+SM formed a uniform and thick surface coating of fat droplets. More PC aggregates were observed in the emulsions containing PC (sole PC, MC+PC, and WPI+PC) compared with their SM counterparts. Based on these results, the appropriate selection of the PL matrix is important to modulate the emulsion stability of dairy emulsion products.

Changes in Particle Size, Sedimentation, and Protein Microstructure of Ultra-High-Temperature Skim Milk Considering Plasmin Concentration and Storage Temperature.

Age gelation is a major quality defect in ultra-high-temperature (UHT) pasteurized milk during extended storage. Changes in plasmin (PL)-induced sedimentation were investigated during storage (23 °C and 37 °C, four weeks) of UHT skim milk treated with PL (2.5, 10, and 15 U/L). The increase in particle size and broadening of the particle size distribution of samples during storage were dependent on the PL concentration, storage period, and storage temperature. Sediment analysis indicated that elevated storage temperature accelerated protein sedimentation. The initial PL concentration was positively correlated with the amount of protein sediment in samples stored at 23 °C for four weeks (r = 0.615; p < 0.01), whereas this correlation was negative in samples stored at 37 °C for the same time (r = -0.358; p < 0.01) due to extensive proteolysis. SDS-PAGE revealed that whey proteins remained soluble over storage at 23 °C for four weeks, but they mostly disappeared from the soluble phase of PL-added samples after two weeks' storage at 37 °C. Transmission electron micrographs of PL-containing UHT skim milk during storage at different temperatures supported the trend of sediment analysis well. Based on the Fourier transform infrared spectra of UHT skim milk stored at 23 °C for three weeks, PL-induced particle size enlargement was due to protein aggregation and the formation of intermolecular ß-sheet structures, which contributed to casein destabilization, leading to sediment formation.

Effects of Probiotic Culture Supernatant on Cariogenic Biofilm Formation and RANKL-Induced Osteoclastogenesis in RAW 264.7 Macrophages.

Postbiotics are a promising functional ingredient that can overcome the limitations of viability and storage stability that challenge the production of probiotics. To evaluate the effects of postbiotics on oral health, eight spent culture supernatants (SCSs) of probiotics were prepared, and the effects of SCSs on Streptococcus mutans-induced cariogenic biofilm formation and the receptor activator of the nuclear factor κB ligand (RANKL)-induced osteoclastogenesis were evaluated in RAW 264.7 macrophages. SCS of Lactobacillus salivarius MG4265 reduced S. mutans-induced biofilm formation by 73% and significantly inhibited tartrate-resistant acid phosphatase (TRAP) activity, which is a biomarker of mature osteoclasts in RAW 264.7 macrophages. The suppression of RANKL-induced activation of mitogen activated the protein kinases (c-Jun N-terminal kinase, extracellular signal-regulated kinase, and p38) and nuclear factor κB pathways, as well as the upregulation of heme oxygenase-1 expression. The suppression of RANK-L-induced activation of mitogen also inhibited the expression of transcriptional factors (c-fos and nuclear factor of activated T cells cytoplasmic 1) and, subsequently, osteoclastogenesis-related gene expression (tartrate-resistant acid phosphatase-positive (TRAP), cathepsin K, and matrix metalloproteinase-9).Therefore, SCS of L. salivarius MG4265 has great potential as a multifunctional oral health ingredient that inhibits biofilm formation and suppresses the alveolar bone loss that is associated with periodontitis.

The Effect of Chrysin-Loaded Phytosomes on Insulin Resistance and Blood Sugar Control in Type 2 Diabetic db/db Mice.

Although a variety of beneficial health effects of natural flavonoids, including chrysin, has been suggested, poor solubility and bioavailability limit their practical use. As a promising delivery system, chrysin-loaded phytosomes (CPs) were prepared using egg phospholipid (EPL) at a 1:3 molar ratio and its antidiabetic effects were assessed in db/db diabetic mice. Male C57BLKS/J-db/db mice were fed a normal diet (control), chrysin diet (100 mg chrysin/kg), CP diet (100 mg chrysin equivalent/kg), metformin diet (200 mg/kg) or EPL diet (vehicle, the same amount of EPL used for CP preparation) for 9 weeks. Administration of CP significantly decreased fasting blood glucose and insulin levels in db/db mice compared with the control. An oral glucose tolerance test and homeostatic model assessment for insulin resistance were significantly improved in the CP group (p < 0.05). CP treatment suppressed gluconeogenesis via downregulation of phosphoenolpyruvate carboxykinase while it promoted glucose uptake in the skeletal muscle and liver of db/db mice (p < 0.05). The CP-mediated improved glucose utilization in the muscle was confirmed by upregulation of glucose transporter type 4, hexokinase2 and peroxisome proliferator-activated receptor γ during treatment (p < 0.05). The CP-induced promotion of GLUT4 plasma translocation was confirmed in the skeletal muscle of db/db mice (p < 0.05). Based on the results, CP showed greater antidiabetic performance compared to the control by ameliorating insulin resistance in db/db mice and phytosome can be used as an effective antidiabetic agent.

Enzyme-assisted extraction of cactus bioactive molecules under high hydrostatic pressure.

BACKGROUND: To improve the extraction and recovery of bioactive materials from cactus, the present study investigated the effect of polysaccharide-degrading enzymes [Rapidase-Viscozyme mixture, 1/3 (v/v)] treatment under high hydrostatic pressure (HHP). RESULTS: The dry weight of the extract increased with the use of increasing pressure regardless of enzyme treatment. However, the polyphenol content showed a tendency to decrease with the increase in pressure in the cactus extract with or without enzyme treatment. The enzyme-assisted extraction resulted in an increase of dry weight and polyphenol content in the cactus extract. The total sugar and reducing sugar contents of the cactus extract increased with increasing pressure in enzyme-assisted extraction. The uronic acid content of the cactus extract showed a pattern similar to that of the reducing sugars. The enzyme-assisted extraction also increased the contents of taxifolin, quercetin and isorhametin. The cactus extract obtained through enzyme-assisted extraction showed intense scavenging activity of both DPPH and ABTS radicals. The crude polysaccharides isolated from the extract (51.2% at 1000 µg mL⁻¹ for HHP extraction at 300 MPa) had higher anti-complementary activity than the others except for lipopolysaccharide (60.00% at 1000 µg mL⁻¹). HHP extraction and enzyme-assisted extraction using HHP showed an increase of anti-complementary activity compared with the heat and enzyme controls, respectively. CONCLUSION: Overall, the use of HHP in enzyme-assisted extraction resulted in more efficient extraction than the use of enzyme treatment alone.

Inhibition of Pectobacterium carotovorum-mediated potato soft rot by carboxymethyl cellulose-based antibacterial edible coating containing green tea extract.

This study was conducted to propose a new strategy for preventing Pectobacterium carotovorum-mediated potato soft rot through the development of a carboxymethyl cellulose (CMC)-based antibacterial coating incorporated with green tea extract (GTE). GTE/CMC films resulted in increased water vapor permeability due to the incorporation of polar groups in GTE. In the antibacterial test against P. carotovorum, the MBC value of GTE was 2 mg/mL. The time-kill assay demonstrated that GTE/CMC (2 × MBC) completely eradicated bacteria within 0.5 h (~6.4 log CFU/mL reduction). The potential of GTE/CMC to prevent potato soft rot was evaluated by monitoring the potato appearance, maceration area, and texture properties. The GTE/CMC-coated potatoes exhibited significantly reduced maceration area and remained firm for 3 days. Moreover, there was no change in the antimicrobial efficacy for 8 weeks. The developed GTE/CMC could be used as a biological coating system for postharvest storage and soft rot prevention. Supplementary Information: The online version contains supplementary material available at 10.1007/s10068-024-01548-6.

Lactiplantibacillus plantarum LM1001 Improves Digestibility of Branched-Chain Amino Acids in Whey Proteins and Promotes Myogenesis in C2C12 Myotubes.

Lactiplantibacillus plantarum is a valuable potential probiotic species with various proven health-beneficial effects. L. plantarum LM1001 strain was selected among ten strains of L. plantarum based on proteolytic activity on whey proteins. L. plantarum LM1001 produced higher concentrations of total free amino acids and branched-chain amino acids (Ile, Leu, and Val) than other L. plantarum strains. Treatment of C2C12 myotubes with whey protein culture supernatant (1%, 2% and 3%, v/v) using L. plantarum LM1001 significantly increased the expression of myogenic regulatory factors, such as Myf-5, MyoD, and myogenin, reflecting the promotion of myotubes formation (p<0.05). L. plantarum LM1001 displayed ß-galactosidase activity but did not produce harmful ß-glucuronidase. Thus, the intake of whey protein together with L. plantarum LM1001 has the potential to aid protein digestion and utilization.

Postbiotic potential of Bacillus velezensis KMU01 cell-free supernatant for the alleviation of obesity in mice.

Attention toward the preventive effects of postbiotics on metabolic diseases has increased because of greater stability and safety over probiotics. However, studies regarding the bioactive effects of postbiotics, especially from probiotic Bacillus strains, are relatively limited. The anti-obesity effects of the cell-free culture supernatant of Bacillus velezensis KMU01 (CFS-B.vele) were evaluated using high-fat-diet (HFD)-induced mice. HFD-induced mice (n = 8 per group) received equal volumes of (1) CFS-B.vele (114 mg/kg) in PBS, (2) Xenical in PBS, or (3) PBS alone by oral gavage daily for 13 weeks. The results demonstrated that CFS-B.vele changed the gut microbiota and showed anti-obesity effects in HFD-induced obese mice. The elevated Firmicutes/Bacteroidota ratio induced by HFD was decreased in the CFS-B.vele group compared to the other groups (p < 0.05). The CFS-B.vele intervention led to the enrichment of SCFA-producers, such as Roseburia and Eubacterium, in the cecum, suggesting their potential involvement in the amelioration of obesity. Due to these changes, the various obesity-related biomarkers (body weight, fat in tissue, white adipose tissue weight and size, serum LDL-cholesterol level, hepatic lipid accumulation, and adipogenesis/lipogenesis-related gene/protein expression) were improved. Our findings suggest that CFS-B.vele has potential as a novel anti-obesity agent through modulation of the gut microbiota.

Saponins from soy bean and mung bean inhibit the antigen specific activation of helper T cells by blocking cell cycle progression.

Treatment of helper T (Th) cells with saponins from soy bean and mung bean prevented their activation by inhibiting cell proliferation and cytokine secretion. However, the saponins did not affect the expression of major histocompatibility complex class II (A(b)) and co-stimulatory molecule (CD86) on professional antigen-presenting cells. Instead, the saponins directly inhibited Th cell proliferation by blocking the G(1) to S phase cell cycle transition. Moreover, blocking of the cell cycle by the saponins was achieved by decreased expression of cyclin D1 and cyclin E, and constitutive expression of p27(KIP1). Saponins also increased stability of p27(KIP1) in Th cells after antigenic stimulation.

Effect of phospholipid matrix on emulsion stability, microstructure, proteolysis, and in vitro digestibility in model infant formula emulsion.

The effects of adding different phospholipid (PL) matrices [milk sphingomyelin (SM) vs soy phosphatidylcholine (PC)] on emulsion stability, microstructure, and in vitro simulated lipid digestion were examined using a Model Infant Formula Emulsion (MIFE). The emulsion stability of MIFE increased significantly with PL addition (0.1 and 0.2 %). Compared to sole MIFE or MIFE + PC, the incorporation of SM resulted in increased emulsion stability (p < 0.05) and a greater amount of free fatty acid release (p < 0.05) during in vitro simulated digestion. This was mainly due to the reduction of intensive droplet aggregation, thus providing a large surface area and improved digestibility. This is further experimentally supported by the evolution of particle size distribution, zeta-potential, and microstructure analysis using confocal laser scanning microscopy. The incorporation of SM in the emulsion formation significantly delayed digestion of ß-lactoglobulin during in vitro digestion. Lipid digestibility in MIFE was altered depending on the type of PL matrix, and SM displayed a superior effect to PC. Thus, the creation of a novel emulsion interface by the appropriate selection of emulsifiers can be used to improve lipid digestion in infants and obtain desirable nutritional consequences.

Synergistic antimicrobial effect and mode of action of palmarosa oil-loaded nanoemulsion and citric acid against Pectobacterium carotovorum.

The synergistic antimicrobial activity of palmarosa oil (Cymbopogon martini, PO)-loaded nanoemulsion (PO-NE) and citric acid (CA) against Pectobacterium, the major pathogen for soft-rot disease, was evaluated. The combination of PO-NE and CA (PO-NE + CA) significantly improved the storage stability of PO-NE at 30 °C. Compared to the anti-Pectobacterium activity of alone, PO-NE + CA reduced the minimum inhibitory concentration (MIC) by 1/4 and 1/2, respectively. Bactericidal efficacy of PO-NE + CA against P. carotovorum PCC3 was similar of PO-NE alone in the MIC in time-kill kinetic assay. PO-NE treatment mainly influenced membrane integrity, while CA treatment strongly stimulated intracellular ATP depletion. This synergistic combination effectively reduced the use of PO-NE, imparting a strong flavor note without sacrificing the antimicrobial efficacy against Pectobacterium. Supplementary Information: The online version contains supplementary material available at 10.1007/s10068-022-01217-6.

Effects of Lactobacillus reuteri MG5346 on Receptor Activator of Nuclear Factor-Kappa B Ligand (RANKL)-Induced Osteoclastogenesis and Ligature-Induced Experimental Periodontitis Rats.

Effects of culture supernatants of Lactobacillus reuteri MG5346 (CS-MG5346) on receptor activator of nuclear factor-kappa B ligand (RANKL)-induced osteoclastogenesis were examined. CS-MG5346 treatment up to 400 µg/mL significantly reduced tartrate-resistant acid-phosphatase (TRAP) activity, the phenotype biomarker of osteoclast, without affecting cell viability. CS-MG5346 inhibited the expression of osteoclast specific transcriptional factors (c-fos and nuclear factor-activated T cells c1) and their target genes (TRAP, cathepsin, and matrix metallo-proteinase-9) in a dose-dependent manner (p<0.05). The administration of L. reuteri MG5346 (2×108 CFU/day) for 8 wks significantly improved furcation involvement, but no difference was observed in alveolar bone loss in ligature-induced experimental periodontitis rats. The elevated RANKL/ osteoprotegerin ratio, the biomarker of periodontitis, was significantly lowered in the gingival tissue by administration of L. reuteri MG5346 (p<0.05). L. reuteri MG5346 showed excellent stability in simulated stomach and intestinal fluids and did not have antibiotic resistance. Based on the results, L. reuteri MG5346 has the potential to be a promising probiotic strain for oral health.

Erratum to: Effect of Modified Casein to Whey Protein Ratio on Dispersion Stability, Protein Quality and Body Composition in Rats.

[This corrects the article DOI: 10.5851/kosfa.2021.e42.].

Roles of Milk Fat Globule Membrane on Fat Digestion and Infant Nutrition.

Milk fats are present as globules emulsified in the aqueous phase of milk and stabilized by a delicate membrane architecture called milk fat globule membrane (MFGM). The unique structure and composition of the MFGM play an important role in fat digestion and the metabolic programming of neonates. The objective of this review is to compare the structure, composition, and physicochemical characteristics of fat globules in human milk, bovine milk, and infant formula. It provides an overview of the fat digestion process and enzymes in healthy infants, and describes the possible roles of the MFGM in association with factors affecting fat digestion. Lastly, the health benefits of the MFGM on infant nutrition and future perspectives are discussed with a focus on brain development, metabolic response, and gut health.

Effects of Lactobacillus curvatus MG5246 on inflammatory markers in Porphyromonas gingivalis lipopolysaccharide-sensitized human gingival fibroblasts and periodontitis rat model.

This study investigated the effects of Lactobacillus curvatus MG5246 on periodontitis inflammation. Cell-free supernatants (CFS) prepared from L. curvatus MG5246 decreased prostaglandin E2 production and cyclooxygenase-2 gene expression by 60% and 78% in Porphyromonas gingivalis-lipopolysaccharide stimulated human gingival fibroblasts at 400 µg/mL. Gene expressions of tumor necrosis factor-α, interleukin-6, matrix metalloproteinases, and chemokines were significantly downregulated by CFS treatment (p < 0.05). L. curvatus MG5246 (2 × 108 CFU/day, 8 weeks) administration significantly improved alveolar bone loss in the ligature-induced periodontitis rat model. Elevated mRNA expression of the receptor activator of nuclear factor-κB ligand/osteoprotegerin ratio in the gingival tissue was significantly decreased by L. curvatus MG5246 administration (p < 0.05). Moreover, L. curvatus MG5246 showed sufficient tolerance in simulated gastrointestinal conditions (gastric tolerance: 89.48%, intestinal tolerance: 98.62%) and did not show antibiotic resistance and hemolytic activity. Therefore, L. curvatus MG5246 has the potential as novel oral probiotics.

Ecklonia cava Extract Exerts Anti-Inflammatory Effect in Human Gingival Fibroblasts and Chronic Periodontitis Animal Model by Suppression of Pro-Inflammatory Cytokines and Chemokines.

Periodontitis is one of the most common chronic inflammatory diseases. The anti-inflammatory effect of the extract from brown algae Ecklonia cava was analyzed in lipopolysaccharide (LPS)-stimulated human gingival fibroblasts (HGF-1), the most abundant cells in gingival tissue. The gene expressions of cyclooxygenase-2 and interleukin-6 were decreased by 78 and 50%, respectively, at 100 µg/mL Ecklonia cava extract (ECE) treatment. The gene expressions of matrix metalloproteases (MMP-2 and MMP-8) and chemokines (macrophage inflammatory protein 1-alpha and stromal cell-derived factor 1) were also significantly down-regulated by ECE treatment (p < 0.05). The increased reactive oxygen species (ROS) production in HGF-1 cells by LPS stimulation was decreased by 30% at 100 µg/mL ECE treatment. The mitogen-activated protein kinase pathway and the nuclear factor-kappa B (NF-κB) signal activated by ROS were suppressed by ECE in a dose-dependent manner. ECE treatment (400 mg/kg, 8 weeks) significantly improved alveolar bone resorption in the ligature-induced chronic periodontitis rat model. ECE supplementation also lowered elevated mRNA expression of the receptor activator of nuclear factor-kappa B (RANKL)/osteoprotegerin (OPG) in the gingival tissue (p < 0.05). Therefore, ECE mitigated gingival tissue destruction and bone resorption associated with chronic periodontitis condition.

Effect of Modified Casein to Whey Protein Ratio on Dispersion Stability, Protein Quality and Body Composition in Rats.

The present study was designed to investigate the effects of protein formula with different casein (C) to whey protein (W) ratios on dispersion stability, protein quality and body composition in rats. Modification of the casein to whey protein (CW) ratio affected the extent of protein aggregation, and heated CW-2:8 showed a significantly increased larger particle (>100 µm) size distribution. The largest protein aggregates were formed by whey protein self-aggregation. There were no significant differences in protein aggregation when the CW ratios changed from 10:0 to 5:5. Based on the protein quality assessment (CW-10:0, CW-8:2, CW-5:5, and CW-2:8) for four weeks, CW-10:0 showed a significantly higher feed intake (p<0.05), but the high proportion of whey protein in the diet (CW-5:5 and CW-2:8) increased the feed efficiency ratio, protein efficiency ratio, and net protein ratio compared to other groups. Similarly, CW-2:8 showed greater true digestibility compared to other groups. No significant differences in fat mass and lean mass analyzed by dual-energy x-ray absorptiometry were observed. A significant difference was found in the bone mineral density between the CW-10:0 and CW-2:8 groups (p<0.05), but no difference was observed among the other groups. Based on the results, CW-5:5 improved protein quality without causing protein instability problems in the dispersion.

Effect of Homogenization Pressure on Plasmin Activity and Mechanical Stress-Induced Fat Aggregation of Commercially Sterilized Ultra High Temperature Milk during Storage.

Commercially sterilized ultra high temperature (UHT) milk was manufactured at different homogenization pressures (20, 25, and 30 MPa), and changes in fat particle size, mechanical stress-induced fat aggregation, plasmin activity, and lipid oxidation were monitored during ambient storage of the UHT milk for up to 16 wk. The particle sizes of milk fat globules were significantly decreased as homogenization pressure increased from 20 to 30 MPa (p<0.05). The presence of mechanical stress-induced fat aggregates in milk produced at 20 MPa was significantly higher than for UHT milk produced at either 25 or 30 MPa. This difference was maintained all throughout the storage. There were no significant differences in plasmin activity, trichloroacetic acid (12%, w/v) soluble peptides, and the extent of lipid oxidation. Based on these results, an increase of homogenization pressure from 20 (the typical homogenization pressure employed in the Korea dairy industry) to 25-30 MPa significantly decreased mechanical stress-induced fat aggregation without affecting susceptibility to lipid oxidation during storage.

Anti-Inflammatory Effect of Ecklonia cava Extract on Porphyromonas gingivalis Lipopolysaccharide-Stimulated Macrophages and a Periodontitis Rat Model.

Ecklonia cava, an edible marine brown alga (Laminariaceae), is a rich source of phlorotannins. This study aimed to investigate the anti-inflammatory effect of Ecklonia cava ethanol extract (ECE, dieckol 10.6%, w/w) on Porphyromonas gingivalis lipopolysaccharide-stimulated inflammation in RAW 264.7 cells and in ligature-induced periodontitis in rats. The levels of nitric oxide (NO) and prostaglandin E2 were decreased by more than half on treatment with 100 µg/mL ECE. Downregulated tumor necrosis factor-α, interleukin (IL)-1ß, and IL-6 gene expression confirmed the anti-inflammatory properties of ECE. ECE treatment upregulated heme oxygenase-1 (HO-1) expression by 6.3-fold and increased HO-1/nuclear factor erythroid 2-related factor 2 (Nrf-2) signaling decreased nuclear factor-κB (NF-κB) translocation. ECE administration (400 mg/kg) significantly reduced gingival index, restricted tooth mobility, and prevented alveolar bone loss (p < 0.05). These beneficial effects were due to decreased inflammatory cell infiltration, IL-1ß production, and matrix metalloproteinase expression in gingival tissues. The ratio of receptor activator of nuclear factor-κB ligand (RANKL)/osteoprotegerin, a biomarker of periodontitis and osteolysis, was significantly decreased by ECE administration (p < 0.05). Thus, ECE has potential therapeutic effects for the alleviation of periodontal disease.