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Endowing titanium surfaces with multifunctional properties can reduce implant-related infections and enhance osseointegration. In this study, titanium dioxide nanotubes with strontium doping (STN) were first created on the titanium surface using anodic oxidation and hydrothermal synthesis techniques. Next, casein phosphopeptide (CCP) and an antimicrobial peptide (HHC36) were loaded into the STN with the aid of vacuum physical adsorption (STN-CP-H), giving the titanium surface a dual function of "antimicrobial-osteogenic". The surface of STN-CP-H has a suitable roughness and good hydrophilicity, which is conducive to osteoblasts. STN-CP-H had a 99 % antibacterial rate against S. aureus and E. coli and effectively prevented the growth of bacterial biofilm. Meanwhile, the antibacterial mechanism of STN-CP-H was initially explored with the help of transcriptome sequencing technology. STN-CP-H could greatly increase osteoblast adhesion, proliferation, and expression of osteogenic markers (alkaline phosphatase, runt-related transcription) when CCP and Sr worked together synergistically. In vivo, the STN-CP-H coating could effectively promote new osteogenesis around titanium implant bone and had no toxic effects on heart, liver, spleen, lung and kidney tissues. A potential anti-infection bone healing material, STN-CP-H bifunctional coating developed in this work efficiently inhibited bacterial infection of titanium implants and encouraged early osseointegration., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Inc. All rights reserved.)

Despite fluoride's widespread use in preventing dental caries, it remains a significant oral disease with some drawbacks. Consequently, new preventative agents have emerged that can function independently of fluoride. Our aim is to demonstrate the efficacy of newly developed varnishes, 3% Rennou (theobromine calcium and phosphate) and 1% Rennou, in remineralizing initial caries. In our experiment, 40 human enamel samples were randomly allocated into four groups of 10 samples each as: Group 1 (G1): 5% NaF (Colgate PreviDent®), Group 2 (G2): 1% Rennou®, Group 3 (G3): 3% Rennou® and Group 4 (G4): Casein Phosphopeptide-Amorphous Calcium Phosphate + Fluoride (MI Varnish TM GC). To produce an artificial carious lesion in the enamel, the samples were kept in a demineralizing solution for 72 hours. Samples underwent pH cycling for 6 days in order to induce remineralization. The means of the three measurements were compared, and the percentage of Surface Microhardness Recovery in (SMHR%) was calculated. Scanning Electron Microscopy (SEM) was used for qualitative assessment of surface changes. G1 had the highest SMHR% value, followed by G3, G2 and G4. The One-way ANOVA (Analysis of Variance) showed significant differences in the SMHR% values among the groups after six days of cycling ( p < 0.001). In pairwise comparisons, groups did not show differences in means of SMHR% except for G1 and G4 ( p = 0.006). In the SEM Images, after treatment within the NaF group, many flaky sediments were found on the enamel surface. Similarly, the maximum mineral gain was seen in the NaF and Rennou groups. SEM images of both varnish surfaces revealed a uniform layer interspersed with shapeless precipitates. All varnishes treated artificial enamel lesions to varying degrees. However, both concentrations of Rennou showed remineralization potential comparable to 5% NaF in acceptable statistical measurements. Thus, it could be used as a potentially effective preventive measure for pediatric patients., Competing Interests: The authors declare no conflict of interest., (©2024 The Author(s). Published by MRE Press.)

Ulcerative colitis (UC) stands as a chronic inflammatory intestinal disease. This study aimed to explore a sustained-release strategy to alleviate DSS-induced colitis in mice using polyelectrolyte complexes (PECs) encapsulating an alkaloid, isoliensinine (ISO). The drug delivery platform, termed "Saloplastics (SAL)", was prepared by fabrication of PECs through the solid-liquid phase separation of sodium caseinate (SC) and ε-polylysine (EPL), followed by centrifugation to yield compact structures. Coarse-grained molecular dynamics simulations demonstrated that SAL had a nanorod-like structure formation between EPL and SC, which implied that the self-assembly of SAL is driven by hydrophobic aggregation and strong electrostatic attractions. A comprehensive evaluation of SAL was conducted, including characterizations of its physicochemical and biological properties. The results showed SAL had thermal plasticization properties and excellent swelling capacity as well as susceptibility to hydrolysis by pH and proteinase in simulated gastric fluid. Moreover, SAL displayed a porous morphology with high surface area for immobilizing organic molecules. ISO@SAL, formulated by ISO encapsulated in SAL, not only demonstrated high potency in alleviating DSS-induced colitis in mice, but also increased the dosing intervals from one day to three days. In conclusion, SAL is a biocompatible sustained-release oral drug delivery platform for gastrointestinal applications., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

This experiment was carried out to investigate the effect of bioactive peptides derived from casein on performance, nutrient digestibility, enzyme activity and intestinal microbial population in broiler chickens. In this study, 350 1-day-old male Ross 308 broiler chicks were distributed among 35 pens in a completely randomized design with seven treatments, five replicates and 10 chicks in each replicate. The experimental treatments included: basal diet without any additives (control), basal diet + Avilamycin antibiotic, basal diet + 200, 400, 600, 800 and 1000 mg of peptides per kg of diet. Results showed no significant effects of the experimental treatments on weight gain and feed conversion ratio during the starter period, but there was a significant improvement in weight gain in grower, finisher and whole periods in chicks fed with diet containing 1000 mg/kg peptides (p < 0.05). Adding peptides improved intestinal morphology (in duodenum and ileum) (p < 0.05). Supplementation of casein peptides significantly reduced thiobarbituric acid reactive substances concentration in breast and thigh meat. The activity of amylase, lipase and protease enzymes improved in treatments containing 800 and 1000 mg peptides in comparison to the control treatment. The use of casein-peptides increased intestinal Lactobacillus and decreased Coliform populations., (© 2024 Wiley‐VCH GmbH. Published by John Wiley & Sons Ltd.)

Objective: This study aimed to assess the remineralization efficacy of casein phosphopeptide-amorphous calcium phosphate (CPP-ACP), silver diamine fluoride/potassium iodide (SDF/KI), and sodium fluoride with functionalized tricalcium phosphate (NaF/fTCP) on artificial early enamel lesion using laser fluorescence and micro-CT analysis., Methodology: On extracted impacted third molars, artificial enamel lesions were prepared. Twenty-eight specimens were randomly assigned to four groups (n = 7 per group): a control group (artificial saliva), CPP-ACP (GC Tooth Mousse), SDF/KI (Riva Star), and NaF/fTCP (Clinpro White varnish). Following the manufacturer's instructions, the remineralization agents were applied to demineralized surfaces. Laser fluorescence and micro-CT were used to evaluate the remineralization efficacy of the agents and analyzes were performed during four stages: before demineralization, after demineralization, 1st day of remineralization and 30th day of remineralization. Shapiro-Wilk test, repeated measures two-way ANOVA, and Spearman correlation tests were used for statistical analysis. A significant level of p < 0.05 was established., Results: SDF/KI significantly reduced the lesion area and lesion volume on the demineralized enamel surface after 30 days of remineralization. In the T3 period, SDF/KI increased the mineral density statistically significantly compared to the T1 period. The laser fluorescence values for all three remineralizing agents exhibited a linear decrease. A significant correlation between the fluorescence values and the mineral density was found (p = 0.01)., Conclusion: All three investigated agents were showed positive remineralization efficacy on artificial enamel lesion. However, SDF/KI, containing silver diamine fluoride and potassium iodide exhibited superior than other agents in promoting remineralization., Clinical Significance: Although all three remineralization agents showed positive remineralization efficacy on artificial enamel lesions, SDF had higher remineralization performance over the other two agents. SDF has potential to prevent progression of demineralization in treating children with high caries risk in the long-term., (© 2024 The Author(s). Journal of Esthetic and Restorative Dentistry published by Wiley Periodicals LLC.)

Cordyceps militaris, a medicinal fungus, has gained considerable attention owing to its potential health benefits, notably the production of bioactive compounds such as cordycepin. Cordycepin possesses significant antifungal, antibacterial, and antiviral properties. The present study focused on optimizing the fermentation conditions for C. militaris to boost the production of mycelia and cordycepin, alongside investigating its antifungal properties using in silico and in vitro approaches. The optimal conditions, yielding the highest cordycepin and mycelial biomass, were a temperature of 20°C and a pH range of 4-6, with glucose and sucrose as carbon sources and yeast extract and casein hydrolysate as nitrogen sources. Under these conditions, cordycepin production peaked at low pH (600-1000 mg/L) and with carbon and maltose (400-500 mg/L). The low temperature favored cordycepin production (400 mg/L), whereas casein hydrolysate as a nitrogen source boosted cordycepin yield (600 mg/L). The docking analysis indicated that cordycepin had the highest binding affinity for the tubulin beta chain 2 (-10.4 kcal/mol) compared to the fungicide tebuconazole (-7.9 kcal/mol for both targets). The in silico results were corroborated by in vitro studies, where the mycelial extract of C. militaris inhibited approximately 75% of fungal growth at a concentration of 6000 ppm. These findings suggest that optimizing fermentation conditions significantly enhances cordycepin production, and cordycepin shows antifungal solid activity, making it a promising agent for biocontrol in agriculture., (© 2024 Wiley‐VCH GmbH.)

Objective: To investigate the anticaries effects of graphene oxide (GO) and graphene quantum dots (GQDs) combined with casein phosphopeptide-amorphous calcium phosphate (CPP-ACP) on enamel in a biofilm-challenged environment., Material and Methods: GO and GQDs were synthesised using citric acid. The antibiofilm and biofilm inhibition effects for Streptococcus mutans were evaluated by scanning electron microscopy (SEM), confocal laser scanning microscopy (CLSM), and colony-forming units (CFU). Remineralisation ability was determined by assessing mineral loss, calcium-to-phosphorus ratio, and surface morphology. To create a biofilm-challenged environment, enamel blocks were immersed in S. mutans to create the lesion and then subjected to artificial saliva/biofilm cycling for 7 days. Anticaries effects of GO, GQDs, GQDs@CPP-ACP, GO@CPP-ACP, and CPP-ACP were determined by broth pH and mineral changes after 7-day pH cycling. Biocompatibility was tested using a Cell Counting Kit-8 (CCK8) assay for human gingival fibroblasts (HGF-1)., Results: GQDs and GO presented significant antibiofilm and biofilm inhibition effects compared to the CPP-ACP and control groups (P < 0.05). The enamel covered by GQDs and GO showed better crystal structure formation and less mineral loss (P < 0.05) than that covered by CPP-ACP alone. After 7 days in the biofilm-challenged environment, the GO@CPP-ACP group showed less lesion depth than the CPP-ACP and control groups (P < 0.05). GO and GQDs showed good biocompatibility compared to the control group by CCK8 (P > 0.05) within 3 days., Conclusion: GO and GQDs could improve the anti-caries effects of CPP-ACP, and CPP-ACP agents with GO or GQDs could be a potential option for enamel lesion management., Clinical Significance: GO and GQDs have demonstrated the potential to significantly enhance the anticaries effects of CPP-ACP. Incorporating these nanomaterials into CPP-ACP formulations could provide innovative and effective options for the management of enamel lesions, offering improved preventive and therapeutic strategies in dental care., Competing Interests: Declaration of competing interest All authors declared that they have no conflicts of interest., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Objectives: This study aimed to investigate if CPP-ACP / infiltrating resin was superior in treating enamel demineralization during orthodontic therapy compared with fluoride varnish, in order to provide early-intervention implications for dental professionals., Materials and Methods: In the in-vitro study, premolars were grouped into four: remineralization with fluoride varnish / CPP-ACP, sealing with infiltrating resin, and negative control. Experimental demineralization of enamel surfaces was analyzed using techniques of QLF, SEM, EDS and micro-hardness testing. An in-vivo intervention study was conducted on patients randomly assigned into three groups. At the baseline and every-3-month follow-up, QLF parameters were compared temporally and parallelly to yield potential implications for promotion in clinical practice., Results: The in-vitro study performed on 48 experimental tooth surfaces demonstrated that sealing with infiltrating resin reduced enamel surface porosity and increased surface micro-hardness significantly. In the in-vivo intervention study on 163 tooth surfaces, it was suggested that for those who meet the criteria of -10 < ΔF < -6 and - 1000 < ΔQ < -20 at the baseline, all these treatment methods could achieve acceptable outcomes; with the rising of absolute values of ΔF and ΔQ, sealing with infiltrating resin showed more evident advantages., Conclusion: For enamel demineralization during orthodontic therapy, all the treatment methods involved in this study showed acceptable effectiveness but had respective characteristics in treatment effects. QLF parameters could be used as indicators for clinical early-intervention strategy with regards to this clinical issue., Clinical Relevance: With QLF parameters, clinical early-intervention strategy for enamel demineralization during orthodontic therapy could be optimized., (© 2024. The Author(s).)

Background: Acute myeloid leukemia (AML) is a hematological neoplasm of rapid and progressive onset, and is the most common form of leukemia in adults. Chemoresistance to conventional treatments such as cytarabine (Ara-C) and daunorubicin is a main cause of relapse, recurrence, metastasis, and high mortality in AML patients. It is known that sodium caseinate (SC), a salt derived from casein, a milk protein, inhibits growth and induces apoptosis in acute myeloid leukemia cells but not in normal hematopoietic cells. However, it is unknown whether SC retains its antileukemic effect in cytarabine-resistant AML cell lines., Objective: To evaluate the antineoplastic effect of SC in cytarabine-resistant leukemia models., Methods: The SC inhibits the growth and induces apoptosis in parental WEHI-3 AML cells. Here, we generated two cytarabine-resistant sublines, WEHI-CR25 and WEHI-CR50, which exhibit 6- and 16-fold increased resistance to cytarabine, respectively, compared to the parental WEHI-3 cells. Thus, these sublines mimic a chemoresistant model., Results: We demonstrate that WEHI-CR25 and WEHI-CR50 cells retain sensitivity to SC, similar to parental WEHI-3 cells. This sensitivity results in inhibited cell proliferation, induced apoptosis, and increased expression of ENT1 and dCK, molecules involved in the entry and metabolism of Ara-C, while decreasing MDR1 expression. Additionally, we observed that SC prolonged the survival of WEHI-CR50 tumor-bearing mice, despite their resistance to Ara-C., Conclusion: This is the first evidence that SC, a milk protein, may inhibit proliferation and induce apoptosis in cytarabine-resistant cells.

Hypertension is defined as a systolic blood pressure (SBP) of over 140 mmHg or diastolic blood pressure (DBP) of over 90 mmHg. Hypertension is widely known to be a factor affecting human health, so its prevention is considered important. We investigated the effect of casein-derived tripeptide Met-Lys-Pro (MKP) on blood pressure in a randomized, placebo-controlled, parallel-group study. Participants were healthy adults with SBP between 120 and 139 mmHg, and/or DBP between 80 and 89 mmHg. A total of 121 participants were randomly assigned to the MKP group or placebo group. Participants received either a test powder containing 100 μg of MKP or a placebo powder without MKP for 12 weeks. As a result, SBP and DBP were significantly lower in the MKP group than in the placebo group. No adverse events associated with the MKP intake were observed. This study showed that MKP has a beneficial effect on lowering blood pressure in healthy adults with high-normal and elevated blood pressure and can be safely used for continuous intake.

Glycomacropeptide (GMP) has a unique amino acid profile which may make less satiating than other dietary proteins. This study assessed the feasibility and likely acceptability of a leucine-enriched GMP drink and determined appetite response in older adults (OA). Thirteen OA (11f; 70 ± 4 years) were recruited for sensory assessments of a leucine-enriched GMP drink when mixed with water and with fruit smoothie, compared with whey protein isolate (WHEY). Participants also partook in a single focus group exploring acceptability to protein and supplementation. Separately, a counterbalanced, double-blind study with twelve OA (8f; 69 ± 3 years) was conducted to determine appetite and gut hormone responses. Fasting subjective appetite was recorded using visual analogue scales and a fasted venous blood sample was collected (to measures acyl-ghrelin, PYY, GLP-1, and CCK) before participants consumed either: GMP protein (27g + 3g leucine, 350 mL water), WHEY (30g, 350 mL water), or water. Participants rested for 240 min, with appetite measures and blood sampling throughout. An ad libitum pasta-based meal was then consumed. Sensory testing revealed low pleasantness rating for GMP in water vs. WHEY (16 ± 14 vs 31 ± 24, p = 0.016). GMP addition to smoothie reduced pleasantness (26 ± 21 vs. 61 ± 29, p = 0.009) and worsened the aroma (46 ± 15 vs. 69 ± 28, p = 0.014). The focus group revealed uncertainty of protein needs and a scepticism of supplements, with preference for food. Gut hormone response did not differ between GMP and WHEY (nAUC for all gut hormones p > 0.05). There was no difference between conditions for lunch ad libitum intake (549 ± 171 kcal, 512 ± 238 kcal, 460 ± 199 kcal for GMP, WHEY, and water, p = 0.175), or for subjective appetite response. Leucine-enriched GMP was not less satiating than WHEY, and low palatability and scepticism of supplements question the likely acceptability of GMP supplementation. Providing trusted nutritional advice and food enrichment/fortification may be preferred strategies for increasing protein intake in OA., Competing Interests: Declaration of competing interest This work was supported by a Pump-Priming Award from the Food 4 Years Network. Milk Specialties Global provided a non-monetary contribution in the form of test drinks. L.O. is an employee of Milk Specialties Global. L.O. contributed to the conception of the research question, but did not contribute to data collection, analyses, or interpretation. All other authors declare no conflicts of interest., (Copyright © 2024. Published by Elsevier Ltd.)

The steady increase in the prevalence of inflammatory bowel disease (IBD) is regarded as a worldwide health issue. Gut microorganisms could modulate host immune and metabolic status and are associated with health effects. Probiotics, Lactobacillus rhamnosus GG (LGG), are beneficial microorganisms that ameliorate disease and exert advantageous effects on intestinal homeostasis. However, the viability of probiotics will suffer from various risk factors in the digestive tract. In this view, we developed a probiotic coating with nanocomposite using tannic acid (TA) and casein phosphopeptide (CPP) through layer-by-layer technology to overcome the challenges after oral administration. LGG showed an improved survival rate in simulated gastrointestinal conditions after coated. The coating (LGG/TA-Mg 2+ /CPP) had potent reactive oxygen species (ROS) scavenging ability and improved the survival rate of colorectal epithelial cells after H 2 O 2 stimulation. In DSS-induced colitis, administration of LGG/TA-Mg 2+ /CPP ameliorated intestinal inflammation and reduced the disruption of barrier function. Furthermore, LGG/TA-Mg 2+ /CPP increased the abundance and diversity of the gut microbiota. In the mouse model of DSS colitis, LGG/TA-Mg 2+ /CPP can better activate the EGFR/AKT signaling pathway, thereby protecting the epithelial barrier function of the colon epithelium. In conclusion, the probiotic coating with nanocomposite may become a delivery platform for probiotics applied to IBD., Competing Interests: Declaration of Competing Interest The authors have declared no conflict of interest., (Copyright © 2024 The Authors. Published by Elsevier Masson SAS.. All rights reserved.)

Dental Implants are expected to possess both excellent osteointegration and antibacterial activity because poor osseointegration and infection are two major causes of titanium implant failure. In this study, we constructed layer-by-layer self-assembly films consisting of anionic casein phosphopeptides-amorphous calcium phosphate (CPP-ACP) and cationic poly (L-lysine) (PLL) on sandblasted and acid etched (SLA) titanium surfaces and evaluated their osseointegration and antibacterial performance in vitro and in vivo. The surface properties were examined, including microstructure, elemental composition, wettability, and Ca 2+ ion release. The impact the surfaces had on the adhesion, proliferation and differentiation abilities of MC3T3-E1 cells were investigated, as well as the material's antibacterial performance after exposure to the oral microorganisms such as Porphyromonas gingivalis (P. g) and Actinobacillus actinomycetemcomitans (A. a). For the in vivo studies, SLA and Ti (PLL/CA-3.0) 10 implants were inserted into the extraction socket immediately after extracting the rabbit mandibular anterior teeth with or without exposure to mixed bacteria solution (P. g & A. a). Three rabbits in each group were sacrificed to collect samples at 2, 4, and 6 weeks of post-implantation, respectively. Radiographic and histomorphometry examinations were performed to evaluate the implant osseointegration. The modified titanium surfaces were successfully prepared and appeared as a compact nano-structure with high hydrophilicity. In particular, the Ti (PLL/CA-3.0) 10 surface was able to continuously release Ca 2+ ions. From the in vitro and in vivo studies, the modified titanium surfaces expressed enhanced osteogenic and antibacterial properties. Hence, the PLL/CPP-ACP multilayer coating on titanium surfaces was constructed via a layer-by-layer self-assembly technology, possibly improving the biofunctionalization of Ti-based dental implants., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Functional drugs nano delivery systems manufactured from natural active products are promising for the field of biomedicines. In this study, an anti-ulcerative colitis (UC) curcumin loaded biopolymeric nanocomposite (CZNH) was fabricated and investigated. CZNH nanocomposite was obtained using the anti-solvent precipitation method, wherein curcumin-loaded zein colloidal particles served as the core, while sodium casein (NaCas) and hyaluronic acid (HA) formed the outermost layer of CZNH nanocomposite. Fourier transform infrared (FT-IR) spectrum and transmission electron microscopy (TEM) findings demonstrated that CZNH nanocomposite was a double-layer spherical micelle (250 nm) resulting from the hydrogen bond interactions and electrostatic adsorptions between zein, NaCas, and HA. Furthermore, CZNH nanocomposite exhibited prominent resuspension and storage stability in aqueous solution, which can be stored at 4 °C for approximately 30 days. In vivo anti-UC studies showed that CZNH nanocomposite could effectively alleviate UC symptoms via mediating inflammatory factors [tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), and IL-6], myeloperoxidase (MPO), and oxidative stress factor [malondialdehyde (MDA), superoxide dismutase (SOD), and glutathione peroxidase (GSH-Px)]. This study suggested that the CZNH nanocomposite showed great promise as an efficient curcumin nanocarrier for UC therapy., Competing Interests: Declaration of competing interest The authors declare no competing financial interest., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Background: White-spot lesions are considered an initial carious stage characterized by an outer enamel layer with significantly reduced mineralization. This study was conducted to assess the combined effect of Biomin F toothpaste and Diode laser on remineralization of white spot lesions., Materials and Methods: An invitro study conducted on a total of 30 premolars divided into three groups; Group A (Biomin F Tooth paste), Group B (Biomin F with laser application for 30 sec), Group C (Negative control). The three groups were submitted to three stages; stage 1:Baseline,stage 2:After demineralization ,and stage 3:After remineralization. In each stage, elemental analysis(calcium, phosphorus, and fluoride)was measured quantitatively using Energy Dispersive X-ray (EDX) analysis and qualitatively by micrographs using scanning electron microscope. The data were tested to find significant difference between mineral changes during stages by using (ANOVA) test and Bonferroni test., Results: Calcium, phosphorus and fluoride ions decreased in all groups after demineralization. In stage 3, after application of remineralizing agents, Calcium ions increased significantly in groups A and B where p<.05. As regards to the phosphorus ions, a significant increase was observed in all groups with group A showed the highest gain as phosphorus level percentage change (%mass) was 56.52±18.02 . Fluoride ions increased significantly in groups A and B (p<0.05) but decreased significantly in group C. There was no statistical significant difference between group A and B (p ≥.05) in calcium, phosphorus, and fluoride level after remineralization., Conclusion: Within the limitation of the present study, we concluded that Biomin F toothpaste is promising in the repairing of white spot lesions on the surface of the demineralized enamel. Diode laser did not affect the remineralizing ability of Biomin F toothpaste., (© 2024. The Author(s).)

Calcium is the most abundant mineral in the human body and is involved in critical physiological and cellular processes. It is essential for the development, maintenance, and integrity of bone tissue throughout life. Identifying new natural food-grade chelating agents to improve calcium uptake is of increasing interest. Casein phosphopeptides (CPPs), highly phosphorylated peptides obtained after enzymatic hydrolysis of caseins, represent promising calcium-chelating candidates. The aim of this study was to investigate, using cell culture models, the ability of a digested milk matrix enriched in CPPs to regulate calcium transport through the intestinal barrier and elucidate the involved mechanisms. To this end, a CPP-preparation underwent in vitro static digestion and was subsequently incubated with an intestinal barrier model to monitor calcium uptake and transport. Our results demonstrated that the digested CPP preparation enhanced the trans -epithelial calcium transport via paracellular pathways and that CPPs, identified by peptidomics, crossed the intestinal barrier in the same time.

Cancer is one of the leading causes of death worldwide, with over 10 million fatalities annually. While tumors can be surgically removed and treated with chemotherapy, radiotherapy, immunotherapy, hormonal therapy, or combined therapies, current treatments often result in toxic side effects in normal tissue. Therefore, researchers are actively seeking ways to selectively eliminate cancerous cells, minimizing the toxic side effects in normal tissue. Caseins and its derivatives have shown promising anti-cancer potential, demonstrating antitumor and cytotoxic effects on cells from various tumor types without causing harm to normal cells. Collectively, these data reveals advancements in the study of caseins and their derivative peptides, particularly providing a comprehensive understanding of the molecular mechanism of action in cancer therapy. These mechanisms occur through various signaling pathways, including (i) the increase of interferon-associated STAT1 signaling, (ii) the suppression of stemness-related markers such as CD44, (iii) the attenuation of the STAT3/HIF1-α signaling, (iv) the down-expression of uPAR and PAI-1, (v) the loss of mitochondrial membrane potential and reduced intracellular ATP production, (vi) the increase of caspase-3 activity, and (vii) the suppression of TLR4/NF-кB signaling. Therefore, we conclude that casein could be an effective adjuvant for cancer treatment., (© 2024. The Author(s).)

Milk varieties and specific proteins exhibit anxiolytic-like actions in mice and rats exposed to several tests, the most prominent being the elevated plus-maze. Administrations of α s1 -casein, its 91-100 (α-casozepine), 91-97, 91-93, and 91-92 fragments, the 60-69 fragment of β-casein, lactoferrin, β-lactotensin, wheylin-1, wheylin-2, and α-lactalbumin have been reported to increase open arm exploration relative to enclosed arm exploration. Anxiolytic-like actions have also been described for 91-93 and 91-92 fragments of α s1 -casein, wheylin-1, α-lactalbumin, and lactoferrin in the open-field. Some effects appear to be mediated by the GABA A receptor complex, since antagonists mitigated the anxiolytic-like actions of α s1 -casein, the 91-92 fragment of α s1 -casein, and wheylin-1. Other neurotransmitters purported to affect such behaviors include 5HT, dopamine, and neurotensin. Further research is needed to identify their neuropharmacological actions., Competing Interests: Declaration of competing interest The author reports no conflict of interest for this review., (Copyright © 2024 Elsevier Inc. All rights reserved.)

Human milk reduces risk for necrotizing enterocolitis in preterm infants. Necrotizing enterocolitis occurs in the ileocecal region where thousands of milk protein-derived peptides have been released from digestion. Digestion-released peptides may exert bioactivity, such as antimicrobial and immunomodulatory activities, in the gut. In this study, we applied mass spectrometry-based peptidomics to characterize peptides present in colostrum before and after in vitro digestion. Sequence-based computational modeling was applied to predict peptides with antimicrobial activity. We identified more peptides in undigested samples, yet the abundances were much higher in the digested samples. Heatmapping demonstrated highly different peptide profiles between undigested and digested samples. Four peptides (α S1 -casein [157-163], α S1 -casein [157-165], β-casein [153-159] and plasminogen [591-597]) were selected, synthesized and tested against common pathogenic bacteria associated with necrotizing enterocolitis. All four exhibited bacteriostatic, though not bactericidal, activities against Klebsiella aerogenes , Citrobacter freundii and Serratia marcescens , but not Escherichia coli .

Special attention is given to cow's milk and its variants, with ongoing discussions about health-related impacts primarily focusing on the A1 variant in contrast to the A2 variant. The difference between these variants lies in a single amino acid alteration at position 67 of β-casein. This alteration is presumed to make the A1 variant more susceptible to enzymatic breakdown during milk digestion, leading to an increased release of the peptide β-casomorphin-7 (BCM-7). BCM-7 is hypothesized to interact with µ-opioid receptors on immune cells in humans. Although BCM-7 has demonstrated both immunosuppressive and inflammatory effects, its direct impact on the immune system remains unclear. Thus, we examined the influence of A1 and A2 milk on Concanavalin A (ConA)-stimulated human peripheral blood mononuclear cells (PBMCs), as well as the effect of experimentally digested A1 and A2 milk, containing different amounts of free BCM-7 from β-casein cleavage. Additionally, we evaluated the effects of pure BCM-7 on the proliferation of ConA-stimulated PBMCs and purified CD4 + T cells. Milk fundamentally inhibited PBMC proliferation, independent of the β-casein variant. In contrast, experimentally digested milk of both variants and pure BCM-7 showed no influence on the proliferation of PBMCs or isolated CD4 + T cells. Our results indicate that milk exerts an anti-inflammatory effect on PBMCs, regardless of the A1 or A2 β-casein variant, which is nullified after in vitro digestion. Consequently, we deem BCM-7 unsuitable as a biomarker for food-induced inflammation.

Background: Yogurt, due to its rich casein phosphopeptide (CPP) content could promote remineralization of enamel. Contrary to the age-old usage of animal milk based yogurt, vegan dairy products are gaining momentum due to multiple reasons. Considering this change, the aim of the present study was to assess the in vitro effect of animal and plant-based yogurt extracts on demineralization of enamel., Methods: Enamel windows were prepared on the crowns of sixty premolar teeth using nail paint. Teeth were divided into four groups of fifteen each and were treated with distilled water, demineralizing agent, solution with a mixture of demineralizing agent and yogurt supernatants, respectively for 96 hrs. Quantitative analysis was done using EDXRF (baseline and postexperimental calcium and phosphorus content). Additionally, confocal microscopic analysis was performed to assess the extent of demineralization., Results: The animal-based yogurt (Group III) had the highest post experimental calcium value (mean±SD=81.15±5.02) and positive percentage of change in calcium level (15%; P=0.007) among the groups. This was followed by plant-based yogurt (Group IV) (mean calcium= 76.18±5.12; positive percentage change =8.11%; P=0.003)., Conclusions: Animal based yogurt could provide higher protection against enamel demineralization when compared to plant-based yogurt.

Dietary factors, particularly proteins, have been extensively explored to combat cognitive impairment. We have previously reported that dietary fish (Alaska Pollock) protein (APP) is more effective than casein (CAS) or fish oil in maintaining short-term memory in senescence-accelerated mice prone 10 (SAMP10). To examine the specificity of the protective effect of APP intakes against short-term memory decline, we assessed the impact of various dietary animal proteins, including APP, CAS, chicken breast protein (CP), and whey protein (WP), against age-related cognitive function in SAMP10 mice. After feeding the experimental diets for 5 months, memory was assessed using the Y-maze. The APP group exhibited a significant increase in spontaneous alternation behavior as an indicator of working memory when group compared with groups fed with other protein source. Additionally, the APP group displayed significantly higher neurofilament heavy chain positivity than the CAS and CP groups, as evidenced immunohistochemical analysis. Gut microbiota analysis indicated that dietary APP significantly enhanced the relative abundance of Lactobacillus, which positively correlated with spontaneous alternation behavior. Collectively, these findings suggest that dietary APP is more effective than CAS, CP, or WP in preventing age-related short-term memory decline and morphological abnormalities in the hippocampal axons of SAMP10 mice. Moreover, APP-mediated improvements in cognitive deficits may be associated with changes in microbiota diversity. PRACTICAL APPLICATION: This research suggests that dietary fish protein from Alaska Pollock may be more efficient in prevention short-term memory decline in mice, compared to other animal proteins. This finding has practical implications for nutritional optimization, developing the new health food products, and elucidating the relationship between the impact of specific proteins on gut microbiota and prevention of age-related cognitive decline., (© 2024 Institute of Food Technologists.)

Design: An in vitro study to determine the immediate and sustained effect of fluoride varnish and its combination with fluoride toothpastes in preventing the development of root caries., Case Selection: Human root dentine samples (150) were randomly divided into five experimental protocols of 30 specimens each: 1) fluoride varnish (22,600 ppm fluoride and 1-5% CPP-ACP); 2) fluoride varnish followed by Paste One (1100 ppm sodium fluoride and CPP-ACP); 3) fluoride varnish followed by Paste Plus (900 ppm sodium fluoride and CPP-ACP); 4) fluoride varnish followed by Paste One and Paste Plus; and 5) no treatment (control). A layer of varnish was applied to specimens except the control group and was left in situ for 18 h. The varnish layer was removed, and the various toothpaste treatments were initiated. Half of the specimens in each group were assigned to a short-term incubation model in which they were immediately subjected to a 7-day cariogenic challenge consisting of a combination of human saliva and artificial saliva containing 2% sucrose. The other half of the specimens in each group were assigned to the long-term incubation model in which the experimental protocol was continued for 8 weeks before initiating the seven-day cariogenic challenge. The protocols were evaluated by assessing dentine porosity (rhodamine intensity), mineral density, biofilm biomass, and viability assays., Data Analysis: Confocal laser scanning microscopy was used to determine dentine porosity and Levene's test was used to verify the assumption of equality of variances and normal distribution of errors before two-way ANOVA and the Games-Howell test were carried out at a significance level of 0.05 for both incubation models. Microcomputed tomography was used to determine mineral density with statistical analysis involving Levene's test, two-way ANOVA and Tukey's test at a significance level of 0.05 for both incubation models. Biomass was evaluated using a biofilm biomass assay with analysis of optical density data using Levene's test, ANOVA and Scheffe's test at a significance level of 0.05., Results: For both the short- and long-term incubation models, all the experimental regimes resulted in a statistically significant decrease in dentine porosity and an increase in mineral density when compared to the control group. Fluoride varnish followed by both pastes and fluoride varnish followed by Paste One resulted in a statistically significant decrease in dentine porosity for some depths in both models when compared to fluoride varnish alone. Changes in dentine porosity and mineral density were observed within groups over time. All the experimental regimes demonstrated anti-biofilm effects. Immediate and sustained anti-caries effects were observed for all preventive protocols, with the combination of fluoride varnish and Paste One resulting in superior additional anti-caries effects., Conclusions: The authors concluded that all protocols demonstrated immediate and sustained anti-caries effects against the development of root caries despite variations in effects over time. The combination of fluoride varnish and Paste One resulted in additional anti-caries effects that were consistently superior, with no additional effects being observed when Paste Plus was added in combination. The authors suggest that, within the study's limitations, topical fluoride varnish seems to have a protective effect on root surfaces for up to eight weeks and that fluoride varnish should be considered as an important adjunct strategy in the prevention of root caries in older adults., (© 2024. The Author(s).)

Soft assembly of peptide and curcumin (Cur) molecules enables functional integration by finding dynamic equilibrium states through non-covalent interactions. Herein, we developed two soft assembly systems, curcumin-egg white peptides (Cur-EWP) aggregations (AGs) and Cur-EWP-casein-quaternary chitosan (Cur-EWP-CA-QC) nanoparticles (NPs) to comparatively investigate their therapeutic effects on ulcerative colitis in mice and elucidate their underlying mechanism. Results revealed that Cur-EWP AGs, despite gastrointestinal tract instability, exhibited a propensity for swift accumulation within the colorectal region, enriching mucus-associated and short-chain fatty acid (SCAF)-producing bacteria, restoring the intestinal barrier damage. Whereas, Cur-EWP-CA-QC NPs, benefiting from their remarkable stability and exceptional mucosal adsorption properties, not only enhanced permeability of Cur and EWP in the small intestine to activate the immune response and boost tight junction protein expression but also, in their unabsorbed state, regulated the intestinal flora, exerting potent anti-inflammatory activity. Soft assembly of peptides and hydrophobic nutraceuticals could synergize biological activities to modulate chronic diseases., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Aim: Different remineralizing pretreatments Casein phosphopeptide-amorphous calcium phosphate fluoride (CPP-ACPF), tricalcium phosphate fluoride (TCP-F), self-assembling peptide (SAP) P11-4 and 10 % Nanohydroxyapatite (nHA) gel activation via invisible infrared light on the dentin microhardness (MH) and micro shear bond strength (µSBS) of composite restoration., Methods: Seventy-five human molar teeth were collected and the dentinal surface of all the samples was exposed to different demineralizing solutions. (n = 15) Group 1 (demineralized dentin), Group 2 (CPP ACP), Group 3 (TCP-F), Group 4 (SAP P11-4), Group 5 (nHA gel activation via invisible infrared light). MH assessment was performed using Vickers hardness. Each group of 10 samples was subjected to composite restoration buildup and µSBS were tested. The debonded samples were then observed under a stereo-microscope for failure analysis. ANOVA was conducted, along with Tukey's post hoc analysis, to examine the µSBS of composite and MH of the remineralized surface., Results: nHA gel activation via invisible infrared light pretreated specimens showed the maximum outcomes of surface hardness (331.2 ± 77.3) and bond strength (10.38 ± 2.77). However, Group 4 (SAP P11-4) (148.3 ± 29.2) remineralized dentin displayed minimum scores of MH and µSBS (5.88 ± 1.01)., Conclusion: Remineralizing pretreatment nHA gel activation via invisible infrared light and casein phosphopeptide-amorphous calcium phosphate fluoride seem to improve the dentin MH and µSBS of the composite restoration., (Copyright © 2024. Published by Elsevier B.V.)

This study aimed to evaluate in vitro the effect protocols and anticaries agents containing casein amorphous calcium fluoride phosphopeptide-phosphate (CPP-ACPF, MI Paste Plus), sodium trimetaphosphate (TMP) and fluoride (F), in remineralization of caries lesions. Bovine enamel blocks with initial caries lesions were divided into groups (n = 12): 1) Toothpaste without F-TMP-MI Plus (Placebo); 2) Toothpaste 1100 ppm F (1100F), 3) 1100F + MI Paste Plus (1100F-MI Paste Plus), 4) Toothpaste with 1100F + Neutral gel with 4,500 ppm F + 5%TMP (1100F + Gel TMP) and 5) Toothpaste with 1100F + Neutral gel with 9,000 ppm F (1100F + Gel F). For the 4 and 5 groups the gel was applied only once for 1 minute, initially to the study. For the 3 group, after treatment with 1100F, MI Paste Plus was applied 2x/day for 3 minute. After pH cycling, the percentage of surface hardness recovery (%SHR); integrated loss of subsurface hardness (ΔKHN); profile and depth of the subsuperficial lesion (PLM); concentrations of F, calcium (Ca) and phosphorus (P) in enamel was determined. The data were analyzed by ANOVA (1-criterion) and Student-Newman-Keuls test (p < 0.001). Treatment with 1100F alone led to ~ 28% higher remineralization when compared to treatment with 1100F associated with MI Paste Plus (p < 0.001). The 1100F and 1100F + Gel F groups showed similar values for %SHR (p = 0.150). 1100F + Gel TMP treatment also remineralized the enamel surface by ~ 30% and 20% when compared to the 1100F + Gel F and 1100F groups (p < 0.001). The lower lesion depth (ΔKHN) was observed for the 1100F + Gel TMP group (p < 0.001), where it was 54% and 44% lower in comparison to the 1100F and 1100F + Gel F groups (p < 0.001). Polarized light microscopy photomicrographs showed subsurface lesions in all groups, but these lesions were present to a lower extent in the 1100F + Gel TMP group (p < 0.001). Treatment with 1100F + Gel TMP promoted an increase in the concentration of Ca in the enamel by ~ 57% and ~ 26% when compared to the 1100F and 1100F + MI Paste Plus groups (p < 0.001), respectively. There were no significant differences between the 1100F, 1100F + MI Paste Plus and 1100F + Gel F groups (p > 0.001). Similar values of P in the enamel were observed in the 1100F, 1100F + MI Paste Plus and 1100F + Gel F groups (p > 0.001), except for the 1100F + Gel TMP group, which presented a high concentration (p < 0.001). We conclude that the 1100F+TMP gel treatment/protocol led to a significant increased remineralization when compared to the other treatments/protocols and may be a promising strategy for patients with early caries lesions.

The aim of this study was to assess the addition of 2% sodium caseinate in a commercial egg yolk-based medium in frozen ovine semen. Eight Dorper males were used for the study. The ejaculate was divided into two portions and frozen without (G1) or with the addition of 2% sodium caseinate (G2). Kinetic parameters were evaluated using CASA (computer-assisted sperm analysis), and membrane and acrosome integrity as well as oxidative stress were assessed using flow cytometry. After thawing, a thermoresistance test was conducted at time points T0 and T90. For the fertility test, 100 ewes were inseminated with semen from two rams selected based on in vitro parameters, one with good post-thaw quality (+70% total motility) and the other with low post-thaw quality (-55% total motility). For the fertility test, the females were divided into 4 groups for insemination: low-quality ram without caseinate (GBS = 25) and with caseinate (GBC = 25), and high-quality ram without caseinate (GAS = 25) and with caseinate (GAC = 25). Regarding the results of sperm kinetics, there was a statistically significant difference in the parameters of average path velocity (VAP) and curvilinear velocity (VCL) between the group frozen with BotuBov and the group with added caseinate. At time point T90, straight-line velocity maintained a trend (p < .06), with BotuBov® (BB group) being superior to caseinate this time, and in the linearity parameter, caseinate was superior to BotuBov®. Flow cytometry analysis showed no difference between any of the evaluated tests. In the fertility test, there was no statistically significant difference in the pregnancy rate between the BotuBOV® group (23%, 11/48) and the sodium caseinate group (BC group) (33%, 17/52), and no differences were observed in the male versus diluent interaction (p = .70). In conclusion, sodium caseinate supplementation did not influence sperm kinetic parameters and the fertility of sheep., (© 2024 Wiley‐VCH GmbH. Published by John Wiley & Sons Ltd.)

Developing novel antimicrobial wound dressings that have the potential to address the challenges associated with chronic wounds is highly imperative in providing effective infection control and wound healing support. Biocompatible electrospun nanofibers with their high porosity and surface area enabling efficient drug loading and delivery have been investigated in this regard as viable candidates for chronic wound care. Here, we design Casein/Polyvinyl alcohol (CAN/PVA) nanofibers reinforced with silver nanoparticles (Ag NPs) by the electrospinning technique to develop diabetic wound healing scaffolds. The prepared samples were characterized using spectroscopic and electron microscopic techniques. The biocompatibility of the polymer samples were assessed using 3 T3 fibroblast cell lines and the maximum cell viability was found to 95 % at a concentration of 50 μg/mL for the prepared nanofibers. Scratch assay tests were also performed to analyze the wound healing activity of the nanofibers wherein they demonstrated increased migration and proliferation of fibroblast 3 T3 cells. Moreover, these nanofibers also exhibit antibacterial efficiency against Gram-negative bacteria, Escherichia coli (E.coli). Therefore, the antimicrobial nature of the electrospun nanofibers coupled with their moisture absorption properties and wound healing ability render them as effective materials for wound dressing applications., Competing Interests: Declaration of competing interest We wish to confirm that there are no known conflicts of interest associated with this publication and there has been no significant financial support for this work that could have influenced its outcome., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Sperm adhering to glass slides is one of the main problems during fish sperm motility analyses with CASA systems. To mitigate this, albumin is the supplement added most frequently to activating solutions. However, there is no data on the use of supplements other than albumin (in various concentrations) in analyses of European whitefish (Coregonus lavaretus) sperm motility. This issue was investigated in the presented research using three anti-adhesive supplements (albumin, casein, Pluronic F-127) that were added to Billard solution (BS: 20 mM Tris, 1 mM CaCl 2 , 154 mM NaCl, 30 mM glycine at pH 9.0) at different concentrations (0.0; 0.1; 0.2; 0.5; 1.0; 2.0%). It was noted that the addition of the lowest concentration (0.1%) of albumin, casein, or the pluronic to BS had a significant effect on the motility and kinetic parameters of whitefish sperm compared to pure BS. BS supplemented with 0.2-0.5% albumin was the most appropriate variant used for whitefish sperm motility activation in the present experiment. BS supplemented with the pluronic at 1.0-2.0% concentrations resulted in significantly higher values of almost all CASA parameters compared to casein at the same concentrations. Moreover, CASA parameters determined in this variant of the pluronic (1.0-2.0%) were similar to those when BS was supplemented with the same albumin concentrations. This indicated that instead of albumin, the pluronic at higher concentrations in BS might be used to analyze whitefish sperm motility., Competing Interests: Declaration of Competing Interest The authors declare that they have no conflict of interest., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Sorafenib, a multikinase inhibitor is used to treat hepatocellular and renal carcinoma. However, a low solubility impedes its bioavailability and thus, effectiveness. This study aims to enhance its effectiveness by using novel camel milk casein nanoparticles as a delivery system. This study evaluates the cytotoxicity of sorafenib encapsulated in camel milk casein nanoparticles against human hepatocarcinoma cells (HepG2 cells) in vitro. Optimal drug loaded nanoparticles were stable for 1 month, had encapsulation efficiency of 96%, exhibited a particle size of 230 nm, zeta potential of -14.4 and poly disparity index of 0.261. Treatment with it led to cell morphology and DNA fragmentation as a characteristic of apoptosis. Flow cytometry showed G1 phase arrest of cell cycle and 26% increased apoptotic cells population upon treatment as compared to control. Sorafenib-loaded casein nanoparticles showed 6-fold increased ROS production in HepG2 cells as compared to 4-fold increase shown by the free drug. Gene and protein expression studies done by qPCR and western blotting depicted upregulation of tumor suppressor gene p53, pro-apoptotic Bax, and caspase-3 along with downregulated anti-apoptotic Bcl-2 gene and protein expression which further emphasized death by apoptosis. It is concluded regarding the feasibility of these casein nanoparticles as a delivery system with enhanced therapeutic outcomes against hepatocellular carcinoma cells., (© 2024 Wiley-VCH GmbH.)

Objectives: White spot lesions are more susceptible to staining agents due to their porous structure. This study examines the impact of white spot lesion treatments on discoloration caused by pediatric supplements., Method and Materials: Three treatments (fluoride, casein phosphopeptide-amorphous calcium phosphate [CPP-ACP], resin infiltration), a control, and their respective syrup-based subgroups (iron and black elderberry syrups) were established, each with eight teeth. Artificial white spot lesions were induced, and weekly applications of fluoride varnish, daily applications of CPP-ACP paste, or a single resin infiltration procedure were performed on the white spot lesions within the treatment groups over 4 weeks. Simultaneously, samples were exposed daily to iron or black elderberry syrups. Spectrophotometer measurements were taken at baseline, after demineralization (T0), and after 1 (T1), 2 (T2), and 4 weeks (T4). ΔE00 values were calculated. Statistical analysis was conducted using a three-way mixed-design ANOVA, with the significance level set at P = .05., Results: At T4, ΔE00 values from all groups exceeded the clinical acceptability limit of 1.8. At T2 and T4, the ΔE00 values obtained from the black elderberry syrup subgroups were significantly higher (P < .001). At T4, the highest ΔE00 values were seen in the CPP-ACP groups (P < .001). The lowest ΔE00 values at T2 and T4 were observed in the resin infiltration groups (P < .05)., Conclusions: Supplements containing ferrous sulfate and black elderberry extract caused color changes in white spot lesions that exceeded the clinical acceptability limit. Resin infiltration of white spot lesions provides advantages over remineralization treatments, particularly in minimizing discoloration induced by pediatric supplements.

The aim of this study was to investigate the immunomodulatory effects of A2 β-casein (β-CN) in cyclophosphamide-induced immunosuppressed BALB/c mice. Experiments conducted in vitro revealed that A2 β-CN digestive products have potent immunostimulatory activities. Animal studies demonstrated that A2 β-CN improved the immunological organ index reduction trend caused by cyclophosphamide, reduced the pathological damage to the spleen tissue in immunosuppressed mice, increased the release of IL-17A, IgG, and IgA, and reduced the production of IL-4. By regulating the relative abundance of advantageous bacteria like Oscillospira , Lactobacillus , and Bifidobacteria and harmful bacteria like Coprococcus and Desulfovibrionaceae , A2 β-CN improved gut microbiota disorders in immunosuppressed mice. Moreover, A2 β-CN promoted the production of short-chain fatty acids and increased the diversity of the gut microbiota. Therefore, ingestion of A2 β-CN is beneficial to the host's immune system and gut health. These findings provide insights for the future application of A2 β-CN-related dairy products.

Soy protein has shown remarkable effectiveness in reducing fat mass compared with other protein sources, and exercise has the potential to further enhance this fat loss effect. Previous studies have demonstrated that soy protein intake leads to decreased fatty acid synthesis, which contributes to its fat-loss properties. However, the exact mechanism by which these lipids are consumed remains unclear. To investigate this, we conducted a comprehensive study using C57/BL6 male mice, comparing the effects of soy and casein proteins with and without exercise (Casein-Sed, Casein-Ex, Soy-Sed, and Soy-Ex groups) under high- and low-protein conditions (14% or 40% protein). Our findings revealed that combining soy protein intake with exercise significantly reduced epididymal white adipose tissue (eWAT) weight, particularly in the high-protein diet group. Further analysis revealed that exercise increased the expression of lipid oxidation-regulatory proteins, including mitochondrial oxidative phosphorylation protein (OXPHOS) complexes, in the plantaris muscle regardless of the protein source. Although soy protein intake did not directly affect muscle mitochondrial protein expression, the activity of OXPHOS complex I was additively enhanced by exercise and soy protein under the 40% protein condition. Notably, complex I activity inversely correlated with eWAT weight in the soy protein diet group. These results highlight the potential link between improved complex I activity induced by soy protein and fat mass reduction, which emphasizes the promising benefits of combining soy protein with exercise in promoting fat loss. NEW & NOTEWORTHY The findings revealed that soy protein intake combined with exercise resulted in reduced adipose tissue weight compared with that obtained with casein protein intake. Furthermore, the joint impact of exercise and soy protein consumption resulted in enhanced activity of oxidative phosphorylation protein (OXPHOS) complex I in fast-twitch muscles, which appears to be associated with fat mass reduction. These findings elucidate the potential additive effects of soy protein and exercise on body weight management.

Objective: To assess the in-vitro effect of single applications of CPP-ACP pastes and different fluoridated solutions on the prevention of dental caries around orthodontic brackets., Material and Methods: Tooth/bracket sets (n=65) were immersed in artificial saliva (1h at 37ºC) and randomly subjected to single applications (100µL; 1min) of casein phosphopeptide-amorphous calcium phosphate (CPP-ACP emulsion), CPP-ACP with fluoride (CPP-ACPF emulsion), solutions of titanium tetrafluoride (TiF4) or sodium fluoride (NaF), or no treatment (CG). Multispecies biofilm (5 x 105 CFU/mL) was formed in the presence of 2% sucrose. After 24 h, the pH and the concentration of total soluble fluoride (TSF) were analyzed by culture medium. The presence of active white spot lesions (WSL) evaluated by macroscopic examination and the percent surface mineral loss (%SML) were analyzed. Also, the topography of enamel was detected by analysis of scanning electron microscopy (SEM). The data was assessed by chi-square, Kruskal-Wallis, and Mann-Whitney tests (p < 0.05)., Results: Fluoride-containing compounds led to a smaller pH reduction than did CPP-ACP and CG (p<0.05). There was difference in TSF between the groups (p<0.05), denoted as TiF4> NaF > CPP-ACPF > CPP-ACP > CG. Regarding the presence of WSL and %SML, the NaF group obtained lower values (p<0.05), while TiF4 and CPP-ACPF were similar (p>0.05). SEM demonstrated that fluoride-free groups had a larger surface dissolution., Conclusion: Fluoridated groups including solutions and CPP-ACPF were more effective than CPP-ACP in reducing enamel demineralization around orthodontic brackets after a single application.

In this study, the nano-assemblies of bovine casein hydrolyzed peptides (HP) modified by fatty acids with various alkyl chain lengths (C8, C10, C12 and C14) were synthesized. The physicochemical properties of HP-C 8 -HP-C 14 nano-assemblies were characterized using spectra, laser particle size analyzer, contact angle meter, scanning electron microscope (SEM) and cryo-transmission electron microscope (Cryo-TEM). HP-C 8 and HP-C 10 self-assembled into a hollow cube cage with an average size of ~500 nm, and the assembly of HP-C 12 showed a flower-shaped morphology with more dispersed behavior, and droplet size was observed as ~20 nm. The in vitro cytotoxicity against human breast cancer cells MCF-7 was tested using CCK-8 assay and flow cytometry analysis. HP-C 12 showed the highest cytotoxicity for MCF-7 cells with an inhibition rate of 66.03 % ± 0.35 % with an IC 50 value of 7.4 μM among HP-C n . HP-C 8 , HP-C 10 and HP-C 12 significantly affected on the migration, invasion and apoptosis of MCF-7 cells. The apoptosis mechanism may depend on the upregulation of anti-apoptotic protein Bcl-2 as well as pro-apoptotic proteins Bax and caspase-8. The dead MCF-7 cells were analyzed with UHPLC-MS/MS using untargeted metabolomics, revealing key metabolic pathways., Competing Interests: Declaration of competing interest The authors declare no conflict of interest., (Copyright © 2023. Published by Elsevier B.V.)

Scope: Non-alcoholic fatty liver disease (NAFLD) is a sexually dimorphic disease influenced by dietary factors. Here, the metabolic and hepatic effects of dietary amino acid (AA) source is assessed in Western diet (WD)-induced NAFLD in male and female mice., Methods and Results: The AA source is either casein or a free AA mixture mimicking the composition of casein. As expected, males fed a casein-based WD display glucose intolerance, fasting hyperglycemia, and insulin-resistance and develop NAFLD associated with changes in hepatic gene expression and microbiota dysbiosis. In contrast, males fed the AA-based WD show no steatosis, a similar gene expression profile as males fed a control diet, and a distinct microbiota composition compared to males fed a casein-based WD. Females are protected against WD-induced liver damage, hepatic gene expression, and gut microbiota changes regardless of the AA source., Conclusions: Free dietary AA intake prevents the unhealthy metabolic outcomes of a WD preferentially in male mice., (© 2023 The Authors. Molecular Nutrition & Food Research published by Wiley-VCH GmbH.)

Objective: This study aims to assess the effects of the most recent remineralizing agents, casein phosphopeptide-amorphous calcium phosphate (CPP-ACP), CO2 laser irradiation + topical fluoride (CO2 laser + TF), and Nanosilver fluoride - M. oleifera (NSF-MOLE), on the shear bond strength (SBS) and bond failure between resin composite and remineralized caries affected dentin (CAD)., Materials and Methods: Fifty human molars with occlusal caries reaching approximately halfway through the dentin were immersed in a 4% thymol solution. The infected dentin was removed using an excavator and the CAD surface was exposed. The sample was allocated into five groups (n=10) based on the remineralizing agent applied. Group 1: no remineralizing agent, group 2: CPP-ACP, group 3: Riva Star, group 4: NSF:MOLE, and group 5: (CO2 laser + TF). The shear bond testing procedure was conducted utilizing a universal testing machine and a stereo-microscope was used to study the failure pattern. The researchers utilized a one-way analysis of variance. The Tukey post hoc test was conducted for multiple comparison tests., Results: Group 4 (NSF-MOLE) (13.77±1.94 MPa) treated testers recognized the highest bond values of tooth color restoration to the CAD surface. Nonetheless, group 1 test samples with no mineralization unveiled the minimum outcome of bond integrity (9.12±1.14 MPa). Intergroup comparison exploration showed that group 2 (CPP-ACP), group 4 (NSF-MOLE) (13.77±1.94 MPa), and group 5 (CO2 laser + TF) established comparable values of SBS. Furthermore, group 3 (Riva Star) displayed better SBS than group 1 but lower than group 2, group 4, and group 5., Conclusions: Remineralization of CAD using modern regimes (CPP-ACP, NSF-MOLE, and CO2 laser + TF has the potential to be used to enhance the bond strength of CAD to composite restoration.

Background: White spot lesions (WSLs) appear as white, opaque areas on the teeth and are caused by a demineralization of the enamel., Objective: The purpose of this systematic review and meta-analysis was to determine whether casein phosphopeptide-amorphous calcium phosphate (CPP-ACP) has beneficial effects on WSLs and provides remineralizing benefits that are superior to those of placebo or no treatment., Methods: Following a thorough analysis of the literature across major online sources, 103 papers - 54 of which underwent in-depth evaluation - were found. We used inclusion and exclusion criteria to choose research articles that were appropriate for our review., Results: 10 papers met this review's requirements and were subsequently included. As compared to the controls, CPP-ACP demonstrated increased efficacy in treatment of WSLs in terms of remineralisation and other parameters which included microhardness tests and enamel surface roughness. The forest plots present the results of meta-analyses comparing the effect of CPP-ACP versus control/other agents on white spot lesions (WSLs). In terms of odds ratios (OR), CPP-ACP showed a small increase in the odds of WSLs compared to control/other agents. In-vitro studies showed no significant difference in relative risk (RR) between CPP-ACP and control/other agents. Randomized control trials indicated a small increase in the risk of WSLs with CPP-ACP treatment, but the overall effect was not statistically significant., Conclusion: There was evidence to support the use of CPP-ACP over other agents for WSL prevention or treatment, but further research is needed to confirm these findings.

The health benefits of soy foods are attributed to the high-quality protein and the bioactive compounds such as isoflavones. We previously reported that feeding obese (fa/fa) Zucker rats soy protein concentrates (SPCs) with low isoflavone (LIF) and high isoflavone (HIF) for 9 weeks significantly reduced liver steatosis compared to a casein control (C) diet. The current study extended the dietary treatments to 18 weeks to investigate the long-term effect of LIF and HIF SPC diets. 6-week-old male lean (L, n  = 21) and obese (O, n  = 21) Zucker rats were fed a casein C diet, LIF and HIF SPC diets for 18 weeks and body weight (BW) was recorded twice weekly. Rats were killed after 18 weeks to measure liver steatosis and serum aspartate aminotransferase and alanine aminotransferase. Obese rats had significantly greater final BW, liver weight, liver weight as the percentage of BW, and steatosis score compared to lean rats in all three dietary groups. The obese high-isoflavones (OHIF) group had significantly higher BW compared to obese control (OC) group ( P  < .0001) and obese low-isoflavones (OLIF) group ( P  = .01). OC group had significantly greater liver weight, liver weight as the percentage of BW, and liver steatosis score compared to OLIF ( P  = .0077, P  < .0001 and P  < .0001, respectively) and OHIF ( P  = .0094, P  < .0001, and P  < .0001, respectively) groups. Taken together, long-term feeding of SPC diets protected against liver steatosis regardless of isoflavone levels.

Objectives: The effectiveness of three different groups of polyethylene glycol (PEG)-based gels containing powders on dentin hypersensitivity (DH) treatment were assessed and compared with Actimins® as commercial reference group., Methods: Hydroxyapatite nanorods (nHA) and sol-gel-derived 45S5 bioglass (SGD 45S5) powders were synthesized through hydrothermal and sol-gel methods, respectively. First, 25 demineralized dentin disks were divided into five groups. Then, the prepared gels based on 45S5 bioglass with and without hydrolyzed casein (HC) as experimental, nHA gel and Actimins® as positive and commercial reference groups were applied twice a day on disks by a micro applicator. To mimic the oral environment, treated disks were immersed in artificial saliva in a water bath at 37 °C for 7 days. However, in the negative control group, no agent was applied on the samples. FE-SEM, EDS, AFM, and XRD were performed to assess tubule occlusion. One-way ANOVA test was used for statistical analysis and p*<0.05 was set as the significance level., Results: The nHA with an average aspect ratio of 2.77 and the SGD 45S5 powders with a polygonal morphology and the average size of 48.64±11.38 µm were synthesized. After treatment, tubule occlusion in HC-SGD 45S5 and nHA gels were shown to be higher than other groups. The root mean square roughness (R rms ) of the above-mentioned gels showed to be 121.54±9.25 nm, and 312.6 ± 9 nm, respectively., Conclusion: The nHA containing group exhibited the highest tubule occlusion efficiency (i.e., tubule diameter of 0.92±0.32 µm) with a superior mineral precipitation. HC as a novel material demonstrates to be potentially beneficial in DH treatment., Clinical Significance: DH as a common issue may be reduced or eliminated by occlusion of patent dentinal tubules. There are various types of desensitizing agents capable of controlling the DH by the occlusion of patent dentinal tubules. The desensitizing gels developed in this study showed to be promising for clinical and home-use applications., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023. Published by Elsevier Ltd.)

Single amino acid (AA) supplementations in foods are increasing, however their potential nutritional and physiological impacts are not fully understood. This study examined the effects of L-lysine (Lys) supplementation on protein quality of diets, serum AA concentrations and associations between the ratio of supplemental Lys to dietary protein (X) with body weight gain (BWG) in Sprague-Dawley male rats. Rats were fed one of 10 diets containing either 7% or 20% casein and supplemented with 0% (Control), 1.5%, 3%, 6% Lys or 6% Lys + 3% L-arginine (Arg) (8 rats/diet group) for 1 week. Lys supplementation reduced the protein quality of the casein-based diets (p < 0.01). BWG was reduced by supplemental Lys when X > 0.18. Free Lys supplementation dose-dependently increased serum Lys levels (p < 0.01), while increased protein-bound Lys (1.4% vs 0.52%) had little effect on serum Lys (p > 0.05). In the 7% casein diets, ≥ 1.5% supplemental Lys reduced serum alanine, asparagine, glycine, isoleucine, leucine, serine, tyrosine, valine, carnitine, ornithine, and increased urea. Supplementation of ≥ 3% Lys additionally reduced tryptophan and increased histidine, methionine and α-aminoadipic acid (α-AAA) compared to the Control (p < 0.05). In the 20% casein diets, addition of ≥ 1.5% Lys reduced serum asparagine and threonine, and ≥ 3% Lys reduced leucine, proline, tryptophan, valine, and ornithine, and 6% Lys reduced carnitine, and increased histidine, methionine, and α-AAA. Overall, this study showed that free Lys supplementation in a Lys-sufficient diet reduced the protein quality of the diets and modified the serum concentrations of many amino acids. Excess free Lys intake adversely affected growth and utilization of nutrients due to AA imbalance or antagonism. Overall lower protein intake increases susceptibility to the adverse effects of Lys supplementation., (© 2023. Crown.)

Milk casein is a rich source of antimicrobial peptides (AMPs) and the most common way to produce AMPs is enzymatic hydrolysis in vitro. In this study, active casein antimicrobial peptide (CAMPs) mixtures were generated by optimized proteolytic cleavage of milk casein. These natural-safe CAMPs mixtures exhibited high activity in the inhibition of Streptococcus mutans and Porphyromonas gingivalis. Morphological characterization suggested the pathogenic bacteria presented incomplete or irregular collapsed membrane surface after the treatment with active CAMPs mixtures. The CAMPs inhibition activity was also effective in the attachment and development of microbial biofilm. Potential CAMPs sequences were unambiguously determined by unbiased proteomic analysis and 301 potential CAMPs were obtained. The activity of 4 novel CAMPs was successfully confirmed by using synthetic standards. This study provides a promising milk CAMPs resource for the development of safe agents in oral bacteria inhibition and functional foods., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

According to the amyloid hypothesis, in the early phases of Alzheimer's disease (AD), small soluble prefibrillar aggregates of the amyloid β-peptide (Aβ) interact with neuronal membranes, causing neural impairment. Such highly reactive and toxic species form spontaneously and transiently in the amyloid building pathway. A therapeutic strategy consists of the recruitment of these intermediates, thus preventing aberrant interaction with membrane components (lipids and receptors), which in turn may trigger a cascade of cellular disequilibria. Milk α s1 -Casein is an intrinsically disordered protein that is able to inhibit Aβ amyloid aggregation in vitro , by sequestering transient species. In order to test α s1 -Casein as an inhibitor for the treatment of AD, it needs to be delivered in the place of action. Here, we demonstrate the use of large unilamellar vesicles (LUVs) as suitable nanocarriers for α s1 -Casein. Proteo-LUVs were prepared and characterized by different biophysical techniques, such as multiangle light scattering, atomic force imaging, and small-angle X-ray scattering; α s1 -Casein loading was quantified by a fluorescence assay. We demonstrated on a C. elegans AD model the effectiveness of the proposed delivery strategy in vivo . Proteo-LUVs allow efficient administration of the protein, exerting a positive functional readout at very low doses while avoiding the intrinsic toxicity of α s1 -Casein. Proteo-LUVs of α s1 -Casein represent an effective proof of concept for the exploitation of partially disordered proteins as a therapeutic strategy in mild AD conditions.

Purpose: This study aimed to evaluate and compare the protective effect of fluoride varnish (Enamelast™, Ultradent Inc., Cologne, Germany), casein phosphopeptide-amorphous calcium phosphate fluoride/CPP-ACPF (MI Paste Plus, GC Corp., Tokyo, Japan) and self-assembling P 11-4 peptide (Curodont™ Protect, Credentis AG, Windisch, Switzerland), against acidic erosion of primary teeth., Methods: Forty primary anterior teeth were randomly assigned to four groups (n = 10): group 1: control, group 2: fluoride varnish, group 3: CPP-ACPF and group 4: self-assembling P 11-4 peptide. After applying remineralising agents, except for the control group, all specimens underwent an erosive challenge of carbonated soft drink and artificial saliva for 15 cycles of 6 s each at 6-h intervals for a day. Groups were compared in terms of surface microhardness, roughness readings, and surface scanning with an extra-oral scanner (D800; 3Shape A/S) before and after the erosive process., Results: All experimental groups showed superior results than the control group regarding microhardness, surface roughness, and substance loss. The fluoride varnish group showed significantly favourable results in microhardness change. There was no significant difference between the experimental groups regarding surface roughness and enamel loss measurements., Conclusion: 5% NaF fluoride varnish, CPP-ACPF and self-assembling P 11-4 peptide protect the enamel of primary teeth against erosion compared to artificial saliva alone., (© 2023. The Author(s), under exclusive licence to European Academy of Paediatric Dentistry.)

Irritable bowel syndrome (IBS) is a common gastrointestinal disorder that affects 10-15% of the global population and presents symptoms such as abdominal discomfort, bloating and altered bowel habits. IBS is believed to be influenced by gut microbiota alterations and low-grade inflammation. Bovine kappa-casein glycomacropeptide (GMP), a bioactive dairy-derived peptide, possesses anti-adhesive, prebiotic and immunomodulatory properties that could potentially benefit IBS patients. This pilot study investigated the effects of daily supplementation with 30 g of GMP for three weeks on gut health in five people with IBS. We assessed alterations in gut microbiota composition, fecal and blood inflammatory makers, and gut-related symptoms before, during and after the GMP feeding period. The results revealed no changes in fecal microbiota, subtle effects on systemic and intestinal immune makers, and no changes in gut-related symptoms during and after the GMP supplementation. Further research is needed to assess the potential benefits of GMP in IBS patients, including the examination of dosage and form of GMP supplementation.

Preliminary findings from multiple studies indicate that dietary intake of soy-derived isoflavones exert beneficial effects on the skin including defense against oxidant damage, stimulation of collagen synthesis, and increased hydration. This study aims to investigate how oral supplementation of a soy protein isolate with added isoflavones (SPII) affects components of photoaging such as facial wrinkles and dyspigmentation, and skin biophysical measures such as skin hydration and sebum excretion in postmenopausal women. This 6-month prospective, randomized double-blind controlled study was conducted on 44 postmenopausal women with Fitzpatrick skin types I, II, and III who were randomized to receive either casein protein or SPII. A high-resolution facial photography system was used to measure wrinkle severity and pigmentation at 0, 8, 16, and 24 weeks. Skin biophysical measurements included skin hydration and sebum production. The average wrinkle severity was decreased in the SPII intervention group at week 16 and week 24 by 5.9% and 7.1%, respectively, compared to the baseline. Compared to the casein group, average wrinkle severity was significantly decreased at week 16 ( p < 0.05) and week 24 ( p < 0.0001). Facial pigment intensity was decreased by -2.5% ( p < 0.05) at week 24, whereas there was no significant change in the casein group. Compared to baseline, skin hydration in the SPII group was significantly increased by 39% and 68% on the left and right cheeks ( p < 0.05), respectively, at 24 weeks. There were no significant differences in sebum production. Dietary soy protein supplementation with isoflavones may improve skin photoaging, including wrinkles and dyspigmentation, and increase skin hydration in postmenopausal women with Fitzpatrick skin types I, II, and III.

The purpose of this systematic review and meta-analysis of in vitro studies was to evaluate the effect of the 2780 nm Er,Cr:YSGG laser combined with casein phosphopeptide-amorphous calcium phosphate (CPP-ACP) for enamel remineralisation. The electronic PubMed, Cochrane Library, Web of Science, and EMBASE databases were searched, with no language or date restrictions, up to January 2023. Two reviewers independently performed research information extraction and quality assessment. Continuous variables were analysed by standard mean difference (SMD) with a 95% confidence interval (CI). The statistical analyses were conducted using Review Manager (Version 5.4; Rev Man) and Cochrane Collaboration (2020). Finally, four trials were included for meta-analysis. According to the comprehensive results, the effect of the Er,Cr:YSGG laser combined with CPP-ACP on enamel remineralisation was significantly better than that of CPP-ACP alone: surface microhardness (SMD =  - 1.83, 95% CI: [- 2.98, - 0.69], P = 0.002); lesion depth (SMD = 6.63, 95% CI: [4.98, 8.28], P < 0.001). Under the limitations of this meta-analysis, the results show that the Er,Cr:YSGG laser combined with CPP-ACP has a better effect on enamel remineralisation than CPP-ACP alone. The combination of the Er,Cr:YSGG laser and CPP-ACP may be a feasible method to prevent and treat enamel demineralisation., (© 2023. The Author(s).)

Purpose: When blended, animal and plant proteins can complement each other in terms of amino acid composition and release time. In this study, we investigated whether the blended protein diet has a better feeding effect than the single protein diet, and to reveal the differences in growth and intestinal microbiota composition caused by the blended protein diet., Methods: Forty Sprague Dawley (SD) rats received diets with different protein sources, including casein (C), whey protein (WP), black soybean protein (BSP), and black soybean-whey blended protein (BS-WP), for eight weeks. To investigate the effects of blended protein supplement on gut microbiota and metabolites, we performed a high throughput 16S rDNA sequencing and fecal metabolomics profiling. In addition, we determined growth and serum biochemical indices, and conducted intestinal morphology analyses., Results: Compared to those in the BSP and WP groups, the daily body weight gain and feed conversion efficiency increased in the BS-WP group. Serum biochemical indices indicated that the protein utilization efficiency of the WP and BS-WP groups was relatively high, and the BS-WP blended protein diet improved the protein adoption rate. The BS-WP blended protein diet also improved intestinal tissue morphology and promoted intestinal villi development compared to the single protein diets. Furthermore, dietary protein altered the composition of gut microbiota, the gut microbial diversity of rats fed with the BS-WP diet was significantly (P < 0.05) higher than that of the other groups. The difference in dietary protein corresponded with an alteration of fecal amino acids and their metabolites, and tryptophan and tyrosine metabolism were the key mechanisms leading to the changes in fecal microbial composition., Conclusion: Dietary protein sources played an important role in the growth and development of rats by influencing intestinal metabolism and microbial composition. The BS-WP blended protein diet was more conducive to nutrient absorption than the single protein diet. Furthermore, blended protein increased the diversity of intestinal microbes and aided the establishment of intestinal barrier function., (© 2023. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany.)

Parkinson's disease (PD) is the fastest-growing neurodegenerative disease, with pathogenic causes elusive and short of effective treatment options. Investigations have found that dairy products positively correlate with the onset of PD, but the mechanisms remain unexplored. As casein is an antigenic component in dairy products, this study assessed if casein could exacerbate PD-related symptoms by stimulating intestinal inflammation and unbalanced intestinal flora and be a risk factor for PD. Using a convalescent PD mouse model induced by 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP), the results showed casein reduced motor coordination, caused gastrointestinal dysfunction, reduced dopamine content, and induced intestinal inflammation. Meanwhile, casein disturbed gut microbiota homeostasis by increasing the Firmicutes/Bacteroidetes ratio, decreasing α-diversity, and caused abnormal alterations in fecal metabolites. However, these adverse effects of casein attenuated much when it had hydrolyzed by acid or when antibiotics inhibited the intestinal microbiota of the mice. Therefore, our results suggested that casein could reactivate dopaminergic nerve injury and intestinal inflammation and exacerbate intestinal flora disorder and its metabolites in convalescent PD mice. These damaging effects might be related to disordered protein digestion and gut microbiota in these mice. These findings will provide new insights into the impact of milk/dairy products on PD progression and supply information on dietary options for PD patients., (Copyright © 2023 IBRO. Published by Elsevier Ltd. All rights reserved.)

Background: This study aimed to assess the impact of theobromine and casein phospho-peptides/amorphous calcium phosphate with fluoride (CPP-ACPF) on the resin-dentine bond strength, microhardness, and dentine morphology., Methods: A total of 18 sound human molars for micro-tensile bond strength (µTBS), 20 sound human premolars for microhardness, and 30 premolars for Scanning electron microscopy/energy-dispersive X-ray spectroscopy (SEM/EDX) were used. Based on the pre-treatment used, teeth were split into six groups; sound dentine, demineralized dentine, and demineralized dentine treated with theobromine (Sigma Aldrich) and MI paste plus (GC International, USA) for two-time intervals; 5 min and 1 month. The bonded teeth were sectioned to produce 1 mm 2 resin-dentine sticks which were evaluated for µTBS using a universal testing device (Instron 3365, USA). The dentine microhardness was tested by using the Vickers microhardness tester (Nexus 4000 TM, Netherlands). The pre-treated dentine surface was examined using SEM/EDX (Neoscope JCM-6000 plus Joel benchtop SEM, Japan). µTBS results were analysed with two-way ANOVA. Microhardness and EDX results were analysed with two-way mixed model ANOVA. The significance level was set at (p ≤ 0.05)., Results: While both remineralizing materials at the two-time intervals demonstrated µTBS comparable to sound dentine (46.38 ± 12.18), the demineralized group demonstrated statistically the lowest µTBS (p < 0.001). Whether used for 5 min or 1 month, theobromine significantly increased the microhardness (50.18 ± 3.43) and (54.12 ± 2.66) respectively (p < 0.001), whereas MI paste only increased the hardness (51.12 ± 1.45) after 1 month (p < 0.001)., Conclusions: The pre-treatment of demineralized dentine with theobromine for 5 min or 1 month could enhance its bond strength and microhardness while for MI paste plus, only 1-month application was efficient to ensure remineralization., (© 2023. The Author(s).)