Adhesive Index Research Articles

АДГЕЗИВНАЯ АКТИВНОСТЬ МИКРООРГАНИЗМОВ К СТОМАТОЛОГИЧЕСКИМ МАТЕРИАЛАМ ДЛЯ 3D ПЕЧАТИ И ВАКУУМНОЙ ФОРМОВКИ

Relevance: The adhesive activity of the oral microbiome to new generation materials is the subject of study by the global medical community, and the issue of choosing structural materials is still being discussed in the scientific community and is of a controversial nature. Objective: To study the adhesion of representatives of the oral microbiome to samples of modern dental materials under experimental conditions (in vitro). Materials and methods: Samples of photopolymer resins for 3D printing: Dental Crown LCD, Dental Sand A1-A2, Dental Clear Pro, (n=30), (1-3 groups), thermoforming plates for vacuum forming: EV Gasket Bleaching 080, Bioplast multicolor 3.0*123 mm, Bioplast color 3.0*125 mm (n=30), (groups 4-6); clinical isolates of the microbiome as test strains: Staphylococcus aureus, Streptococcus oralis, Neisseria subflava, Actinomyces oris, Candida albicans (n=5). Research methods: experimental, microbiological, cultural, analytical and statistical. Results: The degree of adhesion of representatives of the oral microbiome to the studied structural materials is in the range from 0 to 4.91 (Ia) and depends on both the species composition of the microbiome and the physico-chemical properties of the materials. The high adhesion index for all microbiota groups was noted for vacuum forming materials (4.91), and the lowest was Dental Crown LCD (0), which makes it possible to recommend it for practical use. Conclusions: The choice of structural material should be made, including taking into account the adhesive activity of the oral microbiome in order to reduce the risk of complications. It is recommended to give preference to materials with a low adhesive activity index.

The influence of professional oral hygiene on the indicators of local immunity

Professional oral hygiene is an important part of measures aimed at preventing and maintaining the patient’s health. Local immune response is the first barrier of immune protection capable of preventing infection diseases. The aim of the study - the influence of professional hygiene on the indicators of local immunity has been studied. The study involved two groups of patients: the first group included 19 individuals with chronic caries of varying depths (class I), and the second group comprised 15 individuals with moderate periodontal disease. To evaluate nonspecific oral protection, the index of natural colonization of the buccal epithelium and the activity of microbial adsorption by oral mucosal cells were assessed. Specific immunity was evaluated by measuring the concentration of immunoglobulins in unstimulated saliva. All indicators were recorded both before and after professional oral hygiene procedures. When studying the index of natural colonization of the buccal epithelium, it was found that professional cleaning in both groups led to normalization of the spectrum of bacteria adhered to the epithelium: there was a decrease in the amount of gram-negative microflora, atypical for this biotope, while maintaining a high adhesion index of gram-positive cocci. In persons with caries, before sanation, “unsatisfactory” resistance of oral mucosa was revealed in a third of the examined, after sanation, the number of such patients decreased by 3 times, with “satisfactory” resistance increased by 2 times. In the group with periodontal disease, all patients exhibited “unsatisfactory” mucosal resistance before the hygiene procedures. After sanitation, 20 % “unsatisfactory” resistance persisted. In the majority of patients (80 %), the resistance of the oral mucosa improved to “satisfactory”. The concentration of all classes of immunoglobulins in both groups before sanitation was significantly below the norm, after the measures taken, the indicators increased significantly and reached the level typical for healthy individuals. Carrying out professional cleaning of the teeth leads to the normalization of local immune status even without additional medical immunocorrection.

Ride Test on Vehicles Travelling Over Speed Bumps: Simulation with CarSim Software

This study explores the effects of different speed bump geometries—flat-topped, sinusoidal, and parabolic—on vehicle dynamics and ride comfort using CarSim simulations. The analysis focuses on key parameters such as vertical forces on the suspension, vertical acceleration, and the wheel surface adhesion index. The results show that flat-topped bumps generate the highest vertical forces, reaching peaks of up to 6,000 N on the front suspension, leading to increased discomfort. Sinusoidal bumps, in contrast, generate smoother transitions, with vertical forces peaking at approximately 3,500 N, improving ride comfort. At vehicle speeds of 30 km/h, the vertical forces on the suspension increase significantly, with flat-topped bumps reducing the wheel surface adhesion index to as low as 0.6, indicating a higher risk of wheel slip and compromised vehicle stability. In contrast, sinusoidal bumps maintain a more favorable adhesion index of 0.85 at similar speeds. These reductions in adhesion elevate the risk of loss of control, especially at higher speeds. The findings suggest that adaptive suspension systems, capable of adjusting damping and stiffness based on the bump geometry and vehicle speed, would enhance ride quality and stability. Additionally, smoother bump designs, such as sinusoidal profiles, are recommended to reduce the impact on vehicle dynamics, particularly in urban environments. These insights contribute to improving both vehicle design and road safety, ensuring safer and more comfortable driving experiences.

Evaluation of Mechanical and Elemental Properties of Bioceramic-Coated Orthodontic Brackets and Enamel Surface.

The aim is to coat orthodontic brackets with two different bioactive materials and to compare the mechanical and morphological properties of coated brackets and tooth surfaces. A total of 120 stainless steel brackets were divided equally into three groups, that is, the uncoated brackets and nanohydroxyapatite (nHA)-coated, and nanobioactive glass (nBG)-coated brackets using a spin coater machine. The brackets were bonded on the enamel surface and underwent remineralization/demineralization cycles for days 1, 7, 14, and 30. At each time interval, the bond strength of the brackets was assessed using mechanical loading. An optical and scanning electron microscope (SEM) were used for surface evaluation, and the adhesive remanent index (ARI) values were obtained and quantified. One-way analysis of variance using Tukey's test was used to compare the differences among the groups. A uniform distribution of nanoparticles occurred on the surfaces of brackets. The shear bond strength (SBS) showed no significant differences in any tested groups on days 1, 7, and 14. However, control and nBG showed a significant difference from nHA at day 30. On days 7, 14, and 30, the nHA group showed the highest SBS values among the groups. For ARI, most samples showed an adhesive nature of failure at the enamel-brackets interface. The images confirmed the presence of coated particles on brackets and remnants of adhesives after SBS. This study confirmed that the nHA- and nBG-coated brackets have a high potential for application in orthodontics regarding structural and mechanical properties.

Improving the physicochemical and sensory properties and volatile profile of goat ricotta cream with Limosilactobacillus mucosae CNPC007 supplementation

This study assessed the effects of the autochthonous strain Limosilactobacillus mucosae CNPC007 on the techno-functional characteristics, physicochemical properties, volatile profile, and sensory aspects of goat ricotta cream during refrigerated storage. Three cheese formulations were tested: CRC (without probiotic), RCLM (with L. mucosae CNPC007), and RCLA (with commercial probiotic Lactobacillus acidophilus La-5). L. mucosae CNPC007 influenced color, adhesiveness, lactose content, acidity, and proteolysis depth index in goat ricotta cream. RCLA and RCLM maintained a probiotic viable cell count >6 log CFU g−1. RCLM had the highest proteolysis depth index and free amino acid release. Seventeen distinct volatile compounds were identified in the formulations. Probiotic supplementation did not affect global acceptance or purchase intention. RCLA and RCLM were preferred over CRC in the ranking test and considered ideal for goat aroma in the JAR test. L. mucosae CNPC007 supplementation has the potential to improve the nutritional, functional, aromatic, and sensory aspects of goat ricotta cream.

Interfacial adhesion features of high-viscosity asphalt and aggregate under water conditioning: Micro perspective and molecular simulation

Interface properties between asphalt and aggregate intimately affect the service performance and long-term durability of asphalt pavement. This study seeks to explore the interfacial adhesion features of high-viscosity asphalt (HVA)-aggregate under water conditioning to provide the understanding for the moisture damage of HVA-aggregate interface and asphalt pavement. To this end, the HVA was initially suffered from different durations of water conditioning, and atomic force microscopy (AFM) test was employed to gather the microscopic phase structure and adhesion force of water-conditioned HVA and aggregate, to analyze the adhesion variations of HVA-aggregate interface. Further, molecular dynamics method was used to simulate the changing process and influencing mechanism of adhesion effect at the HVA-aggregate interface under water immersion. The findings indicated that with the increasing water immersion, Catana phase of HVAs vanished, and concave and convex structures appeared on the surface, along with pit groove dispersion phase structures. Longer immersion time resulted in augmented surface roughness of HVA and a more dispersed height distribution in the area domain, most notably during the initial stages of water immersion. Aging increased sensitivity to immersion duration in terms of surface roughness, mechanical properties and adhesion ability of HVAs. Water immersion had a diminishing effect on the mechanical properties of HVA at different aging degrees, although it increased the micro-mechanical modulus of HVAs and impaired the interface adhesion with aggregates. Under water immersion, the phase structure and mechanical properties of HVAs and the interface adhesion indexes all distinguished a prominent negative Pearson correlation (correlation coefficient |R| > 0.7000), and these factors were critical for HVA-aggregate interface adhesion, as alterations in surface phase structure and micro-mechanical traits of HVAs triggered variations in the HVA-aggregate interface adhesion. In addition, water content in the interface layer, simulation temperature and mineral anisotropy significantly affected the interfacial adhesion of HVA and aggregate in water environment. The introduction of water and the rise of temperature considerably weakened the interfacial adhesion effect of asphalt-aggregate system, and the interface adhesion failure occurred once the water layer thickness surpassed 5 Å. Comparatively, the HVA-calcite system demonstrated better interfacial adhesion competence than the HVA-α-quartz system, and the HVA-calcite {1 0 4} system showed the optimal interfacial adhesion.

Comparative Evaluation of Shear Bond Strength, Enamel Microfracture, and Adhesive Remnant Index of APC Flash-Free System Versus Traditional Bonding: An In-Vitro Study.

In orthodontics, shear bond strength plays an important role because it provides a good bond between the brackets and tooth surface; it avoids fracture of the tooth surface and prevents debonding of brackets from the tooth surface. All of these allow sufficient treatment time. Many factors, including the adhesive, its thickness, its strength, the bonding procedure, the clinician's ability, the base design, the geometry of the bracket, the material, and the kind of bracket all contribute to the shear bond strength. Brackets joined using conventional adhesive and adhesive pre-coated (APC) flash-free glue were the subjects of this comparison and evaluation research, which aimed to measure shear bond strength, enamel microfracture, and adhesive residual index. 60 recently removed premolars from humans were used in this investigation. Before mounting on the acrylic block, the teeth were meticulously cleaned and preserved in artificial saliva. Two groups were formed from the collected premolars the control group and the experimental group. For the control group, we used American Orthodontics (AO) Master/Mini Master series brackets glued with resin composite kits. To make sure the adhesive was uniformly thick, we flashed extra adhesive around the brackets. In the meantime, samples were bonded using 3M Unitek APC flash-free technology in the experimental group. The research indicated that there was a statistically significant difference between the two groups to the adhesive remnant index (ARI) and mean shear bond strength. The shear bond strength of the experimental group averaged 10.96 megapascals (MPa), whereas the control group's was 5.70 MPa. The control group's ARI score was 2.97, whereas the experimental group's score was 2.4. There was no statistically significant change seen in enamel microfracture. A more robust shear bond may be possible using APC flash-free brackets. Compared to conventional bonding techniques and brackets, APC flash-free brackets have a lower adhesive residual index. The APC flash-free bracket technology also causes more enamel microfracture than conventional bonding and bracketing methods.

Preparation and performance evaluation of a novel orthodontic adhesive incorporating composite dimethylaminohexadecyl methacrylate-Polycaprolactone fibers.

This study addressed enamel demineralization, a common complication in fixed orthodontic treatment, by evaluating a novel orthodontic adhesive with DMAHDM-PCL composite fibers. These fibers, produced through electrospinning, were incorporated into orthodontic adhesive to create experimental formulations at different concentrations and a control group. The study assessed antimicrobial properties, biosafety, and mechanical characteristics. New orthodontic adhesive exhibited significant bacteriostatic effects, reducing bacterial biofilm activity and concentrations. Incorporating 1% and 3% DMAHDM-PCL did not affect cytocompatibility. Animal tests confirmed no inflammatory irritation. Shear bond strength and adhesive residual index results indicated that antimicrobial fibers didn't impact bonding ability. In conclusion, orthodontic adhesives with 3% DMAHDM-PCL fibers are potential antimicrobial bonding materials, offering a comprehensive solution to enamel demineralization in orthodontic patients.

Количественная оценка антиадгезивного и антибактериального эффектов биологически активной добавки Уронекст® в отношении уропатогенов: микробиологическое исследование.

Introduction. Antibacterial drugs are the basis of pharmacotherapy of lower urinary tract infections (UTI), and there is a growing interest in alternative nonantibacterial methods of their treatment and prevention. Purpose of the study. Quantitative evaluation of the anti-adhesive and antibacterial effects of the dietary supplement «Uronext®» containing D-mannose, vitamin D3 and cranberry extract against uropathogens isolated from patients with recurrent UTI. Materials and Methods. 124 strains of uropathogens isolated from the urine of women with recurrent UTI were studied. At the first stage of the study we determined the sensitivity of microorganisms to Uronext dietary supplement. In its presence, uropathogens were the material for the second stage of the study, during which the minimum inhibitory concentration of «Uronext®», its effect on the adhesion index (AI) of uropathogens, as well as the degree and duration of antimicrobial activity were evaluated. Results. The direct antibacterial effect of «Uronext®» against 53.7% of Gram-negative and 51.7% – of Gram-positive microorganisms strains was revealed. A pronounced anti-adhesive effect of the investigated product was noted, especially against Gram-negative bacteria. The anti-adhesive effect of «Uronext®» was the greatest in the first 4 hours and lasted for the whole 24-hour observation period. The minimum inhibitory concentration of «Uronext®» for Gram-negative bacteria was 8 times lower than for Gram-positive bacteria. Antibacterial activity of «Uronext®» was manifested already 1 hour after the product was added to the cell culture, reached the highest value in 2 hours and remained at the maximum level during 5 hours of observation. The mentioned tendency was characteristic for all investigated microorganisms. Conclusion. The conducted study allowed to establish and quantitatively evaluate the expression of anti-adhesive and antibacterial action of the product «Uronext®» against the main uropathogens. The presence of these effects determines the pathogenetic validity of the prescription of the dietary supplement «Uronext®» to patients with recurrent UTI.

Effect of unilateral surface compression technique and heat treatment on industrial application performance of poplar

The utilization of unilateral surface compression technique offers significant advantages such as reduced volume loss of wood and energy consumption, making it highly promising for widespread applications. This work focused on fast-growing poplar (Ctrl) as research subject. The unilateral surface compression technique was employed to prepare compressed wood (USW), while superheated steam was utilized to suppress rebound effects (resulting in the formation of US＆HW). The results showed that the combination of densification and heat treatment technology significantly enhanced the application performance of wood. Compared to Ctrl, the densified area exhibited a significant reduction in the volume of cell cavities, with a stepped distribution evident on the VDP. Furthermore, heat treatment resulted in a decrease in the relative amount of hydroxyl in the wood, as well as a slight increase in crystallinity. This effectively reduced deformation in dense wood, resulting in a 20.02% reduction in its recovery rate, without causing excessive damage to the VDP. Additionally, Heat treatment resulted in a darker wood color, reduced glossiness, increased roughness, and further increase the water contact angle of the compressed wood. The mechanical performance of US＆HW achieved an excellent grade, with a score of 48, while all six processing performance grades of US＆HW exceeded Grade 3, surpassing those of both USW and Ctrl. The finishing performance of the US＆HW was good, although slightly lower than that of USW in terms of ball impact performance. And the adhesion, hardness and abrasion resistance indexes of the paint film meet the requirements of excellent products in the standard.

Обеспечение безопасного контроля радиационно-защитных свойств спецодежды пожарных

The radiation-protective clothes production to equip firefighters guarding nuclear power plants has been established in Russia. Special protective clothes were designed at the Institute for Nuclear Research of the Russian Academy of Sciences under the supervision of Boris Benetsky (1933–2015). According to the state standard GOST R 53264—2019, the radiation-protective properties of special protective clothes are tested with gamma and beta rays in accordance with the methodology developed at the Institute for Nuclear Research of the Russian Academy of Sciences, which ensures the radiation safety of the test. It is required to check the decontamination coefficient; however, this testing method is still hazardous due to the use of open-type radiation-hazardous substances. To ensure the safety of the test, the simulation method has been proposed based on the substitution of the hazardous substance with a safe dust-like pollutant, whereas the decontamination is to be assessed by the adhesion index. The adhesive tape method to assess the contamination of the surface is used. Two casts are taken from two sections of the tested surface before and after the treatment with a detergent. The images of the casts are converted into digital form via a scanner and then computer-processed by the specially designed GLASS application. The contamination of the casts depends on the amount of dust stuck. Upon the determination of the average contamination of each cast, data is obtained to calculate the adhesion index. The result of the test of the same material by simulation method differs from the standard method by 20 %.

Orthodontic adhesive loaded with different proportions of ZrO2 silver-doped nanoparticles: An in vitro μTBS, SEM, EDX, FTIR, and antimicrobial analysis.

Zirconium dioxide silver-doped nanoparticles (ZrO2AgDNPs) impacts the adhesive material in terms of its physical characteristics, antimicrobial properties, degree of conversion (DC), and micro-tensile bond strength (μTBS) of orthodontic brackets to the enamel surface. A comprehensive methodological analysis utilizing a range of analytical techniques, including scanning electron microscopy coupled with energy-dispersive x-ray spectroscopy (EDX), Fourier-transform infrared (FTIR) spectroscopy, DC analysis, and μTBS testing. A light-curable orthodontic adhesive, specifically Transbond XT, was combined with ZrO2AgDNPs at 2.5% and 5%. As a control, an adhesive with no incorporation of ZrO2AgDNPs was also prepared. The tooth samples were divided into three groups based on the weightage of NPs: group 1: 0% ZrO2AgDNPs (control), group 2: 2.5 wt% ZrO2AgDNPs, and group 3: 5 wt% ZrO2AgDNPs. EDX graph demonstrated silver (Ag), Zirconium (Zr), and Oxygen (O2), The antibacterial efficacy of adhesives with different concentrations of NPs (0%, 2.5%, and 5%) was assessed using the pour plate method. The FTIR spectra were analyzed to identify peaks at 1607 cm-1 corresponding to aromatic CC bonds and the peaks at 1638 cm-1 indicating the presence of aliphatic CC bonds. The μTBS was assessed using universal testing machine (UTM) and bond failure of orthodontic brackets was seen using adhesive remanent index (ARI) analysis. Kruskal-Wallis test assessed the disparities in survival rates of Streptococcus mutans. Analysis of variance (ANOVA) and post hoc Tukey multiple comparisons test calculated μTBS values. The lowest μTBS was observed in group 1 adhesive loaded with 0% ZrO2AgDNPs (21.25 ± 1.22 MPa). Whereas, the highest μTBS was found in group 3 (26.19 ± 1.07 MPa) adhesive loaded with 5% ZrO2AgDNPs. ZrO2AgDNPs in orthodontic adhesive improved μTBS and has acceptable antibacterial activity against S mutans. ZrO2AgDNPs at 5% by weight can be used in orthodontic adhesive alternative to the conventional method of orthodontic adhesive for bracket bonding. RESEARCH HIGHLIGHTS: The highest μTBS was found in orthodontic adhesive loaded with 5% ZrO2AgDNPs. ARI analysis indicates that the majority of the failures fell between 0 and 1 among all investigated groups. The colony-forming unit count of S. mutans was significantly less in orthodontic adhesive loaded with nanoparticles compared with control. The 0% ZrO2AgDNPs adhesive showed the highest DC.

Investigation of adhesive shear strength of universal adhesives for fixing ceramic braces

Relevance. At the present stage, the choice of high-quality adhesive material is an important task of dentistry. The choice of the ideal adhesive material remains an urgent task to this day.The aim of the study was to study in laboratory conditions the adhesive shear strength for fixing ceramic braces of a universal adhesive system containing 3D-SR monomer (Universal Bond II (Tokuyama Dental)) and adhesive cement ESTECEM II PLUS (Tokuyama Dental) before and after thermal cycling, with and without the use of varnish.Materials and methods. For the preparation of samples, removed teeth without carious lesions and large fillings were used. Samples of 60 teeth were divided into 2 groups (n=30) with and without varnish. In each group, the adhesive shear strength was studied before and after thermal cycling (n=15). The following materials were used: two-component self-curing dental adhesive system Universal Bond II (Tokuyama Dental), varnish SHIELD FORCE PLUS (Tokuyama Dental), adhesive fixing cement double curing ESTECEM II PLUS (Tokuyama Dental). Tests of the adhesive shear strength of the prepared samples were carried out on a universal testing machine «SYNTHEZ 5» at a traverse speed of 5 mm/min according to the methodology of GOST R 56924-2016 (р. 7.15).Results. The adhesive strength index of the control group (before thermal cycling) was 4.33±1.28 MPa, in the main subgroup (after thermal cycling) 3.48±0.81 MPa. The presence of varnish on the enamel surface did not affect the adhesive strength of the bonding of the bracket system and tooth tissue and amounted to 4.50±0.25 MPa and 3.14±0.82 MPa, respectively (p= 0.0005).Conclusions. The presence of varnish on the enamel surface did not affect the adhesive strength of the connection of the bracket systems with tooth tissues in the control (before thermal cycling) and the main (after thermal cycling) subgroups (p≤ 0.05). The adhesive strength index in the control subgroups (before thermal cycling) and the main ones (after thermal cycling) of both groups differed slightly. Thermal cycling reduces the adhesive strength of braces with tooth tissues when using universal adhesive Universal Bond II (Tokuyama Dental) (p≤ 0.05).

Novel Root Conditioner Apple Cider Vinegar versus Citric Acid on Teeth with Periodontal Disease: A Pilot In-vitro SEM Analysis

Introduction: Root biomodification in periodontal therapy aims at removing the smear layer and enhances the clot stability necessary for periodontal regeneration. Citric Acid (CA), EDTA, fibronectin has been used routinely. But if used injudiciously acidic erosion of the periodontal tissues might occur. Apple Cider Vinegar (ACV) an organic acetic has been reported to possess antimicrobial, anti-inflammatory and antioxidant effects. Aim: To assess the effect of smear layer removal and fibrin clot adhesion of ACV in comparison to CA on periodontally affected teeth. Materials and Methods: This pilot study was conducted at Sri Venkateswara Dental College and Hospital, Chennai, India, for three months, starting in January 2024 and ending in March 2024. 8 Teeth extracted due to periodontal reasons were cleaned, scaled and root planed. From the extracted teeth 16 dentin block samples were obtained, of which eight underwent CA testing (Control group) and eight underwent ACV testing (test group). The SEM analysis was done to assess the efficiency of smear layer removal and fibrin clot adhesion. Results: Under qualitative Scanning Electron Microscope (SEM) analysis, though ACV (50%) and CA (50%) group exhibited significant level of smear layer removal, complete dentinal tubule opening (RMSI score 2) was observed only in ACV category. In regard to Blood Element Adhesion Index (BEAI) both the groups demonstrated moderate fibrin clot adhesion (BEA score 2) in 50% of their samples. Considering both the parameters, root conditioning effects of ACV was better than CA. Conclusion: Teeth treated with ACV had a comparable effect as the CA in terms of root biomodification. This novel agent with several medicinal effects can be considered for future studies to determine its regenerative effectiveness.

IN VITRO ASSESSMENT OF PROBIOTIC SURVIVAL IN SIMULATED GASTROINTESTINAL CONDITIONS AND ADHESIVE PROPERTIES

The study focused on evaluating the functional properties of four lactic acid bacteria (LAB) strains – Lactobacillus fermentum 30, Lactobacillus cellobiosus 36, Lacticaseibacillus paracasei 36/1, and Lac-ticaseibacillus paracasei 30/1 – and their associations, aiming to assess acid resistance, bile tolerance, enzyme stability, and adhesion capabilities using in vitro methods. The survival rates of these LAB strains and their associations were investigated under simulated gastric juice (SGJ) and simulated intestinal juice (SIJ). In SGJ, the association L. fermentum 30 + L. cellobiosus 36 showed higher survival rates compared to individual strains, with approximately 75% viability after 2 hours incubation. Similarly, the associa-tion L. paracasei 30/1 + L. paracasei 36/1 exhibited enhanced survival in SGJ, maintaining about 80% viability. In SIJ, both associations demonstrated improved survival compared to individual strains, with L. fermentum 30 + L. cellobiosus 36 and L. paracasei 30/1 + L. paracasei 36/1 maintaining approxi-mately 70% and 75% viability, respectively. The adhesion capabilities of these LAB strains and associa-tions were evaluated using human erythrocytes. All strains displayed high adhesive activity, particularly notable in L. paracasei 36/1 + L. paracasei 30/1, which demonstrated a significantly higher adhesion index compared to other strains. These findings highlight the robust survival and adhesive properties of the LAB strains and their associations under simulated gastrointestinal conditions. Further research is warranted to explore their potential applications in promoting gut health and combating gastrointestinal disorders.

A Phase 2, Open-Label Study to Assess the Pharmacokinetics, Pharmacodynamics and Safety of Orally Administered Epeleuton in Patients with Sickle Cell Disease

Background. Sickle cell disease (SCD) is a rare hereditary multi-system disorder that is caused by a single mutation in the beta allele of the hemoglobin gene, leading to the production of sickle hemoglobin (HbS). HbS polymerizes when deoxygenated, resulting in red blood cell (RBC) sickling and membrane damage, ultimately leading to a cascade of events which include hemolysis, chronic anemia, inflammation, and vaso-occlusion. Despite recent progress, there is a need for additional disease-modifying therapeutic options. Epeleuton is 15-hydroxy eicosapentaenoic acid (15(S)-HEPE) ethyl ester, a novel synthetic second generation w-3 fatty acid which targets key drivers of hemolysis and multicellular adhesion. In preclinical studies in Townes humanized SCD mice and SCD patient RBCs, epeleuton acts as a disease-modifying agent by simultaneously decreasing (a) systemic and local inflammatory activation, (b) endothelial adhesion, (c) organ damage and (d) intravascular hemolysis and sickling. Epeleuton has been administered to 343 subjects in clinical trials, with a favorable safety and tolerability profile. Objective To evaluate the pharmacokinetics, pharmacodynamics and safety of orally administered epeleuton in patients with SCD. Study Design and Methods. This open-label, phase 2, multicenter study (NCT05861453) will enroll approximately 30 adult patients with a confirmed diagnosis of SCD (HbSS and HbSß0-thalassemia). Participants must have hemoglobin levels of 5.0-9.5g/dL at screening, 2-10 vaso-occlusive crises (VOCs) in the past 12 months, and a high adhesion index at screening defined as RBC laminin adhesion > 400 cells. Concomitant treatment with a stable dose of hydroxyurea is permitted during the study. Key exclusion criteria include receiving an RBC transfusion within three months of enrolment, receiving voxelotor, crizanlizumab or l-glutamine within six weeks of enrolment and, prior hematopoietic stem cell transplant. The trial has a 28-day screening period, followed by a 16-week treatment phase and a 30-day post-treatment follow-up period. Study participants will attend the clinic on 6 occasions at screening, baseline, weeks 4, 8 and 16 (end-of-treatment) and the week 20 post-treatment follow-up visit. Enrolled patients will receive epeleuton 2g BID (4g daily) for 16 weeks. Study endpoints include change from baseline in hemoglobin levels, hemolytic markers, assessments of RBC health, soluble adhesion molecules, RBC adhesion, whole blood adhesion and rates of VOC. Conclusion: Treatments that can simultaneously target hemolysis and cellular adhesion may have an important disease-modifying utility for patients with SCD. The results of this study are anticipated to confirm the potential of epeleuton to address multiple aspects of SCD and to guide the design and conduct of future studies in patients with SCD.

Healthcare Utilization and Cost of Care Analysis of Patients with Sickle Cell Disease with High Vs. Low Biomarkers: A Retrospective Chart Review

Sickle cell disease (SCD) is a red blood cell (RBC) disorder that is associated with frequent and unpredictable vaso-occlusive episodes (VOEs) including pulmonary hypertension, acute chest syndrome, chronic anemia, priapism, splenic and renal dysfunction, and stoke. The total cost to manage a SCD patient with a life expectancy of 64 years is between $1.3-$2.1M. Within the United States alone, up to 67% of adults have ≥3 VOEs per yearthat accounts for most of the healthcare utilization, including 78% of emergency room (ER) visits and 95% of hospitalizations. Despite the same single gene mutation of beta-globin, the frequency of VOEs amongst patients is highly variable. The pathophysiology of VOEs involves a complex interplay between circulating blood cells and elevated plasma factors that promote abnormal interactions to the vascular endothelium. Nearly a half century ago studies demonstrated RBC adhesion strongly correlates with disease severity however only recently standardized testing has become available to assess adhesion clinically. More specifically, we developed a suite of RBC health biomarkers and deployed our flow adhesion bioassays in our landmark, observational study (ELIPSIS). We established that flow adhesion of whole blood to vascular cell adhesion molecule-1 (FA -WB-VCAM) at baseline stratified SCD patients with severe disease phenotypes and validated the clinical predictive value of FA-WB-VCAM two years post-ELIPSIS. The objective of this study was to determine whether baseline FA-WB-VCAM biomarker levels could predict patients more likely to receive direct healthcare contact (ER visits/hospital admissions) to manage SCD complications. This study was a retrospective assessment of clinical FA-WB-VCAM in individuals with SCD during steady state between January 2020 through March 2023. Flow adhesion assays were performed using pulsatile, shear flow (1.67Hz, 1.0 dyne/cm 2). SCD patients that received at least 3 steady state FA-WB-VCAM biomarker evaluations within 1-year were included in this analysis. Clinical site received an IRB waiver to obtain patient-reported ER visits and hospitalizations during relevant time periods. We calculated a composite adhesion index by averaging baseline FA-WB-VCAM biomarker evaluations and established SCD disease severity by quantifying the number of ER visits and hospital admissions one year after the most recent FA-WB-VCAM biomarker evaluation. The non-parametric Wilcoxon matched pairs signed rank test was used to test the statistical differences between groups. Data are presented as mean ± standard error of mean. A p-value < 0.05 was considered statistically significant. Out of a total of 3939 clinical tests performed, 366 samples were obtained for “baseline” indications. Of these, only 28 samples were analyzed from 9 unique patients who met the study criteria described above. The average flow adhesion of whole blood to VCAM-1 (FA-WB-VCAM) in SCD patients with high adhesion phenotypes (>400 cells/mm 2) was significantly higher than the low/moderate SCD cohort (mean high=603±78 cells/mm 2, mean low/moderate=343±28 cells/mm 2, p=0.02). More important, the high adhesion cohort had more ER visits (11 vs. 5) and hospitalizations (24 vs 9) and significantly higher healthcare costs ($35,871 vs. $13,563). This is the first healthcare cost analysis assessing the relationship between SCD patients stratified by their adhesive phenotype using the standardized, clinical adhesion biomarker FA-WB-VCAM, and healthcare utilization. The previous ELIPSIS study established that baseline FA-WB-VCAM stratifies patients based on their risk of developing self-reported VOCs during a 6-month longitudinal observation period and their risk of higher clinical disease severity in a 2-year follow-up study. Consistent with this data, the current study demonstrated that a cohort of SCD patients with elevated baseline adhesion phenotypes based on FA-WB-VCAM had a greater utilization of healthcare resources (ER visits and hospitalizations) and had a higher cost of care during the retrospective observation period. These data suggest that RBC health as indicated by “baseline” FA-WB-VCAM levels may predict lower health care utilization and costs, and that this biomarker may be a plausible surrogate endpoint to assess the pharmaco-economic impact of SCD-modifying therapies.

Assessment of Novel Erythroid Signaling Pathways Modulating Hydroxyurea-Mediated Reduction of Vaso-Occlusive Adhesive Interactions in Early Precursors of Sickle Red Blood Cells (RBCs)

Sickle cell disease (SCD) affects ~100,000 Americans and millions worldwide, with healthcare costs in the U.S. exceeding $1 billion annually to treat frequent and unpredictable vaso-occlusive episodes (VOEs). In addition to fetal hemoglobin induction, hydroxyurea (HU) reduces the frequency of VOEs by increasing nitric oxide (NO) bioavailability and decreasing adhesion receptor expression and RBC-endothelial interactions. Very late antigen-4 (VLA-4), the best characterized adhesion receptor in SCD, is highly expressed on circulating reticulocytes (immature RBCs) and white blood cells (WBCs) during VOEs and in SCD patients with severe disease phenotypes and, decreased in response to HU therapy. Like other integrins, VLA-4 is functionally regulated by cell signaling pathways to modulate activity and binding affinity to a wide variety of ligands that are elevated in the SCD micro-environment. There is highly intriguing evidence showing HU modulates adhesion by upregulating a NO/cGMP-dependent pathway to decrease adhesion receptor activity however specific mechanisms remain unclear. The objective of this study was to assess rapid cell signaling pathways involved in HU-mediated reduction in VLA-4 binding to endothelial vascular cell adhesion molecule-1 (VCAM-1). Whole blood (WB) was obtained from patients with SCD at benign clinic visits according to IRB-approved protocols. WB density-gradient centrifugation was used to isolate hematopoietic stem cells (HSCs) and WBCs from the buffy coat layer and, a CD71 monoclonal antibody (Ab) coupled with magnetic beads to isolate reticulocytes from the packed RBC layer. HSCs were matured to erythroblasts (EBs) in appropriate cell culture media and flow cytometric analyses assessed purity of blood samples using CD235a (exclusive WBC marker) and CD45 (exclusive RBC marker) Abs . Blood samples were pretreated with and without a therapeutically relevant hydroxyurea concentration (50mcM) followed by perfusion through micro-fluidic channels coated with VCAM-1 during pulsatile, shear flow (1.67Hz, 1.0 dyne/cm 2). Additionally, EBs were treated with HU combined and 1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1- one (ODQ; NO-dependent guanylate cyclase inhibitor). Adhered cells were quantified to establish an adhesion index. The non-parametric Wilcoxon matched pairs signed rank test was used to test the statistical differences between groups. Data are presented as mean ± standard error of mean. A p-value < 0.05 was considered statistically significant. There was a significant decrease in WB (p=0.01) and reticulocyte (p<0.0001) adhesion to VCAM-1 in HU-treated samples however, there was no effect on WBC (p>0.99) adhesion suggesting reticulocytes reduce the adhesive burden of blood cells in WB adhesion assays and modulation of VLA-4 binding differs in sickle RBCs when compared to WBCs. More recently, preliminary data from my lab demonstrated that EBs pretreated with HU reduces VLA-4 binding to VCAM-1 (from 126.5 ± 29.42 to 109.5 ± 29.09) by cGMP-dependent mechanisms (HU + ODQ; from 109.5 ± 29.09 to 120.2 ± 42.80). Laboratory-based static and flow adhesion assays have been used for decades to describe adhesion in SCD since earlier studies established that sickle RBCs are more adherent than non-sickle blood cells and, adhesion correlates with SCD severity. For example, the adhesion assay described above has been used to demonstrate that VLA-4 binding can stratify SCD patients based on disease severity and predict impending VOEs. Additionally, reticulocyte and HbF levels strongly correlate with VLA-4 binding to VCAM-1 and, VLA-4-mediated adhesion is increased during patient-reported VOEs and decreased in HU-treated SCD patients. This is the first SCD study demonstrating the role of HU in modulating VLA-4 binding to endothelial VCAM-1 through rapid erythroid signaling pathways. There is also compelling evidence involving downregulation of VLA-4 activity/binding affinity through NO/cGMP dependent mechanisms and enhancement of VLA-4 binding by kinase-dependent pathways. HU has also been shown to modulate adhesion by upregulating a NO/cGMP-dependent pathway to decrease adhesion receptor activity. Collectively, these data suggest that VLA-4 mechanisms may be targeted to therapeutically inhibit vaso-occlusive adhesive events that promote VOEs in SCD.

Smaart Criz: Sickle Cell Mechanisms of Activation, Adhesion, Rheology, and Thrombosis (SMAART) in Response to P-Selectin Inhibition

Sickle cell disease (SCD) pathophysiology involves uncontrolled inflammation, dysregulated coagulation, and adherence of erythrocytes, leukocytes, and platelets to endothelium, all contributing to disrupted rheology and ultimately vaso-occlusion. Crizanlizumab (Novartis), an FDA-approved humanized monoclonal antibody, binds to p-Selectin and blocks endothelial interactions with blood cells. The phase 2 SUSTAIN trial demonstrated a reduction in the median rate of pain crisis, as well as prolonged time to first and second crisis ( Ataga 2017). Not all participants benefited, however, nor have all treated patients achieved pain reduction in the post-marketing period. Paradoxical infusion-related pain has also been reported ( Kanter 2022). Baseline differences between responders, non-responders, and those suffering infusion-related pain remain uncharacterized. In January 2023, Novartis issued a preliminary update on the ongoing global crizanlizumab phase 3 STAND trial which indicated no statistically significant pain reduction with crizanlizumab therapy compared to placebo. A better understanding of patient suitability for the various disease modifying therapies for SCD would however be beneficial. Our center is a pilot site for a lab conducting an assay measuring flow adhesion of whole blood to P-selectin (FA-WB-Psel) utilizing a protein coated microfluidic chamber by Functional Fluidics. Previously published data identified a steady state reference range for FA-WB-Psel in non-SCD as well as SCD patients ( Pittman 2021). The SMAART Criz study aims to characterize downstream biological effects of crizanlizumab (FA-WB-Psel) and identify biomarkers with the potential to predict treatment response, with the goal of facilitating more individualized prescribing. This is an interim analysis of an investigator-initiated, prospective, IRB-approved, 18-week longitudinal study at the Children's Hospital of Michigan. Patients desiring crizanlizumab treatment were offered enrollment. Following informed consent, samples were obtained for FA-WB-Psel before and after each crizanlizumab dose. Flow adhesion of whole blood to vascular cell adhesion molecule-1 (FA-WB-VCAM) was assessed as a control substrate. Thromboelastography (TEG) and whole blood platelet aggregometry were performed to evaluate platelet function at baseline, week 6, and week 18. Patient reported response to treatment was used to measure response to therapy. As of August 2023, 7 patients have completed the study with a median age of 20 years (range 16-25 years; 4 female). Genotypes are HbSS (5), HbSC (1), and HbS-G-Philadelphia (1). Four of 7 patients self-reported clinical response to therapy (Table 1). Treatment was discontinued after 1st dose due to treatment-related acute pain in 2 participants, and after the 4th dose due to patient-reported worsening pain trend (Figure 1). Clinical responders had a median baseline FA-WB-Psel adhesion index of 86 cells/mm 2 (n=4, mean 106, range of 56-193). Clinical responders experienced a 65% reduction in FA-WB-Psel from baseline (mean 86) to the mean pre-infusion level calculated using visit 2 and 3 levels (p=0.05, later visits affected by dropout). Non-responders had a median baseline FA-WB-PSel of 49 cells/mm 2 (n=3, mean 46, range 13-75) with widely variable FA-WB-PSel responses (Fig. 1). The two patients with acute treatment-induced pain had the lowest baseline values at 13 and 49 cells/mm 2. No significant trends were observed in adhesion to a control substrate using FA-WB-VCAM. There was an overall decrease in both arachidonic acid (AA) and adenosine diphosphate (ADP) platelet aggregation over time (AA: 23.5 Ohms to 11.3 Ohms, p=0.254; ADP: 23.4 Ohms to 10.6 Ohms, p=0.036). Early indications from SMAART Criz are that administration of crizanlizumab decreases FA-WB-Psel immediately after infusion with continued decreases over time. Interim analysis suggests elevated baseline FA-WB-Psel may predict individuals with SCD likely to benefit from crizanlizumab, while normal FA-WB-Psel levels at baseline may predict non-response or even adverse reactions. This cohort showed an overall decrease in both AA and ADP aggregation, indicating reduced platelet aggregation following crizanlizumab treatment, a potential downstream biological change contributing to treatment effect (Table 1). Further investigation is warranted. Enrollment and analyses are ongoing.

Adhesion property of municipal solid waste incinerator bottom ash and limestone with asphalt based on surface energy theory

ABSTRACT The aim of this paper is to investigate the adhesion of municipal solid waste incinerator bottom ash (MSWIBA) and limestone to asphalt using the principles of surface energy theory. MSWIBA asphalt mixes are known for their satisfactory water stability, with adhesion being a crucial factor, as demonstrated in previous freeze-thaw splitting tests. Contact angles of ground MSWIBA and limestone were measured using a capillary rise test, eliminating the influence of material surface structure. Subsequently, adhesion work and spalling work were calculated to correlate with water-induced damage. The results showed that the sequence of adhesion work was fine MSWIBA > limestone > coarse MSWIBA, while the sequence of spalling work was coarse MSWIBA > limestone > fine MSWIBA. The adhesion of MSWIBA asphalt mixes was evaluated using the water boiling test image method to validate the findings of surface energy theory. In both asphalt tests, the spalling rate followed this sequence: coarse MSWIBA > limestone > fine MSWIBA. The Adhesion index ER ranked as follows: ER Fine MSWIBA > ER limestone > ER coarse MSWIBA. These results suggest that the water stability of MSWIBA asphalt mixes is influenced by the surface energy of the components. Consequently, indices derived from surface energy theory can be applied in formulating and predicting the properties of MSWIBA asphalt mixes.