Modern Approaches to Acellular Therapy in Bone and Dental Regeneration.

An approach called cell-free therapy has rapidly developed in regenerative medicine over the past decade. Understanding the molecular mechanisms and signaling pathways involved in the internal potential of tissue repair inspires the development of new strategies aimed at controlling and enhancing these processes during regeneration. The use of stem cell mobilization, or homing for regeneration based on endogenous healing mechanisms, prompted a new concept in regenerative medicine: endogenous regenerative medicine. The application of cell-free therapeutic agents leading to the recruitment/homing of endogenous stem cells has advantages in overcoming the limitations and risks associated with cell therapy. In this review, we discuss the potential of cell-free products such as the decellularized extracellular matrix, growth factors, extracellular vesicles and miRNAs in endogenous bone and dental regeneration.

First Experimental Study of the Influence of Extracellular Vesicles Derived from Multipotent Stromal Cells on Osseointegration of Dental Implants.

Herein, the aim was to study the state of the bone tissue adjacent to dental implants after the use of extracellular vesicles derived from multipotent stromal cells (MSC EVs) of bone marrow origin in the experiment. In compliance with the rules of asepsis and antiseptics under general intravenous anesthesia with propofol, the screw dental implants were installed in the proximal condyles of the tibia of outbred rabbits without and with preliminary introduction of 19.2 µg MSC EVs into each bone tissue defect. In 3, 7, and 10 days after the operation, the density of bone tissue adjacent to different parts of the implant using an X-ray unit with densitometer was measured. In addition, the histological examinations of the bone site with the hole from the removed device and the soft tissues from the surface of the proximal tibial condyle in the area of intra-bone implants were made. It was found out that 3 days after implantation with the use of MSC EVs, the bone density was statistically significantly higher by 47.2% than after the same implantation, but without the injection of MSC EVs. It is possible that as a result of the immunomodulatory action of MSC EVs, the activity of inflammation decreases, and, respectively, the degree of vasodilation in bones and leukocyte infiltration of the soft tissues are lower, in comparison with the surgery performed in the control group. The bone fragments formed during implantation are mainly consolidated with each other and with the regenerating bone. Day 10 demonstrated that all animals with the use of MSC EVs had almost complete fusion of the screw device with the bone tissue, whereas after the operation without the application of MSC EVs, the heterogeneous histologic pattern was observed: From almost complete osseointegration of the implant to the absolute absence of contact between the foreign body and the new formed bone. Therefore, the use of MSC EVs during the introduction of dental implants into the proximal condyle of the tibia of rabbits contributes to an increase of the bone tissue density near the device after 3 days and to the achievement of consistently successful osseointegration of implants 10 days after the surgery.

Materials and Methods for All-Ceramic Dental Restorations Using Computer-Aided Design (CAD) and Computer-Aided Manufacturing (CAM) Technologies-A Brief Review.

The materials used in dentistry for the fabrication of all-ceramic restorations have undergone great and rapid developments over the last two decades. Among the most common ceramic materials in dentistry are those based on zirconium and lithium disilicate. Due to the properties of these materials, they are in great demand in the field of dental restoration production. Thus, dental restorations that will use those materials are commonly machined in CAD/CAM systems, which offer the possibility of manufacturing all-ceramic dental restorations in a very short period of time. This article reviews the modern materials in the field of all-ceramic dental restorations, their manufacturing processes, as well as what determines which ceramic materials are used for the production of CAD/CAM blanks and their production technology.

3Y-TZP/Ta Biocermet as a Dental Material: An Analysis of the In Vitro Adherence of Streptococcus Oralis Biofilm and an In Vivo Pilot Study in Dogs.

The surface adhesion of bacterial cells and the in vivo biocompatibility of a new ceramic-metal composite made of zirconium dioxide and tantalum were evaluated. Within the framework of an in vitro study using the crystal violet staining and colony counting methods, a relatively similar adhesion of Streptococcus oralis to the 3Y-TZP/Ta biocermet (roughness Ra = 0.12 ± 0.04 µm) and Ti-Al6-V4 titanium alloy (Ra = 0.04 ± 0.01 µm) was found. In addition, in an in vivo preliminary study focused on the histological analysis of a series of rods implanted in the jaws of beagle dogs for a six-month period, the absence of any fibrous tissue or inflammatory reaction at the interface between the implanted 3Y-TZP/Ta biocermets and the new bone was found. Thus, it can be concluded that the developed ceramic-metal biocomposite may be a promising new material for use in dentistry.

Evaluation of serum and urine biomarkers for severe COVID-19.

Introduction: The new coronavirus disease, COVID-19, poses complex challenges exacerbated by several factors, with respiratory tissue lesions being notably significant among them. Consequently, there is a pressing need to identify informative biological markers that can indicate the severity of the disease. Several studies have highlighted the involvement of proteins such as APOA1, XPNPEP2, ORP150, CUBN, HCII, and CREB3L3 in these respiratory tissue lesions. However, there is a lack of information regarding antibodies to these proteins in the human body, which could potentially serve as valuable diagnostic markers for COVID-19. Simultaneously, it is relevant to select biological fluids that can be obtained without invasive procedures. Urine is one such fluid, but its effect on clinical laboratory analysis is not yet fully understood due to lack of study on its composition. Methods: Methods used in this study are as follows: total serum protein analysis; ELISA on moderate and severe COVID-19 patients' serum and urine; bioinformatic methods: ROC analysis, PCA, SVM. Results and discussion: The levels of antiAPOA1, antiXPNPEP2, antiORP150, antiCUBN, antiHCII, and antiCREB3L3 exhibit gradual fluctuations ranging from moderate to severe in both the serum and urine of COVID-19 patients. However, the diagnostic value of individual anti-protein antibodies is low, in both blood serum and urine. On the contrary, joint detection of these antibodies in patients' serum significantly increases the diagnostic value as demonstrated by the results of principal component analysis (PCA) and support vector machine (SVM). The non-linear regression model achieved an accuracy of 0.833. Furthermore, PCA aided in identifying serum protein markers that have the greatest impact on patient group discrimination. The study revealed that serum serves as a superior analyte for describing protein quantification due to its consistent composition and lack of organic salts and drug residues, which can otherwise affect protein stability.

Microbial Signatures in COVID-19: Distinguishing Mild and Severe Disease via Gut Microbiota.

The COVID-19 pandemic, caused by the SARS-CoV-2 virus, has significantly impacted global healthcare, underscoring the importance of exploring the virus's effects on infected individuals beyond treatments and vaccines. Notably, recent findings suggest that SARS-CoV-2 can infect the gut, thereby altering the gut microbiota. This study aimed to analyze the gut microbiota composition differences between COVID-19 patients experiencing mild and severe symptoms. We conducted 16S rRNA metagenomic sequencing on fecal samples from 49 mild and 43 severe COVID-19 cases upon hospital admission. Our analysis identified a differential abundance of specific bacterial species associated with the severity of the disease. Severely affected patients showed an association with Enterococcus faecium, Akkermansia muciniphila, and others, while milder cases were linked to Faecalibacterium prausnitzii, Alistipes putredinis, Blautia faecis, and additional species. Furthermore, a network analysis using SPIEC-EASI indicated keystone taxa and highlighted structural differences in bacterial connectivity, with a notable disruption in the severe group. Our study highlights the diverse impacts of SARS-CoV-2 on the gut microbiome among both mild and severe COVID-19 patients, showcasing a spectrum of microbial responses to the virus. Importantly, these findings align, to some extent, with observations from other studies on COVID-19 gut microbiomes, despite variations in methodologies. The findings from this study, based on retrospective data, establish a foundation for future prospective research to confirm the role of the gut microbiome as a predictive biomarker for the severity of COVID-19.

Decellularized Matrix Induced Spontaneous Odontogenic and Osteogenic Differentiation in Periodontal Cells.

The regeneration of periodontal tissues is a decisive factor in the treatment of periodontitis. Currently, to achieve complete periodontal regeneration, many studies have evaluated the effectiveness of decellularized tissue-engineered constructs on periodontal regeneration. We studied the possibilities of osteogenic and odontogenic differentiation of periodontal progenitor and stem cells (SCs) of the periosteum and periodontal ligament, in decellularized tooth matrix (dTM) and periodontal ligament (dPDL), in 2D and 3D culture. The cell culture of periodontal cells without decellularized matrices was used as control. On the 14th day of cultivation of PDLSCs, PSCs, and PDLSCs + PSCs on dTM and/or dPDL scaffolds in 2D conditions, in all scaffold variants, a dense monolayer of spindle-shaped cells was intensely stained for markers of osteogenic differentiation, such as osteopontin and osteocalcin. Periodontal cells in the collagen I hydrogel (3D-dimensional culture) were more diverse in shape and, in combination of dTM and dPDL, in addition to osteogenic expression, expressed dentin sialophosphoprotein, an odontogenic differentiation marker. Thus, collagen I hydrogel contributed to the formation of conditions similar to those in vivo, and the combination of dTM with dPDL apparently formed a microenvironment that promoted osteogenic and odontogenic differentiation of periodontal cells.

Influence of Extracellular Matrix Components on the Differentiation of Periodontal Ligament Stem Cells in Collagen I Hydrogel.

Regeneration of periodontal tissues requires an integrated approach to the restoration of the periodontal ligament, cementum, and alveolar bone surrounding the teeth. Current strategies in endogenous regenerative dentistry widely use biomaterials, in particular the decellularized extracellular matrix (dECM), to facilitate the recruitment of populations of resident cells into damaged tissues and stimulate their proliferation and differentiation. The purpose of our study was to evaluate the effect of the exogenous components of the extracellular matrix (hyaluronic acid, laminin, fibronectin) on the differentiation of periodontal ligament stem cells (PDLSCs) cultured with dECM (combinations of decellularized tooth matrices and periodontal ligament) in a 3D collagen I hydrogel. The immunohistochemical expression of various markers in PDLSCs was assessed quantitatively and semi-quantitatively on paraffin sections. The results showed that PDLSCs cultured under these conditions for 14 days exhibited phenotypic characteristics consistent with osteoblast-like and odontoblast-like cells. This potential has been demonstrated by the expression of osteogenic differentiation markers (OC, OPN, ALP) and odontogenic markers (DSPP). This phenomenon corresponds to the in vivo state of the periodontal ligament, in which cells at the interface between bone and cementum tend to differentiate into osteoblasts or cementoblasts. The addition of fibronectin to the dECM most effectively induces the differentiation of PDLSCs into osteoblast-like and odontoblast-like cells under 3D culture conditions. Therefore, this bioengineered construct has a high potential for future use in periodontal tissue regeneration.

Experimental and Statistical Analysis of the Effect of Heat Treatment on Surface Roughness and Mechanical Properties of Thin-Walled Samples Obtained by Selective Laser Melting from the Material AlSi10Mg.

Statistical analysis of mechanical properties of thin-walled samples (~500 microns) obtained by selective laser melting from AlSi10Mg material and subjected to heat treatment for 1 h at temperatures from 260 °C to 440 °C (step of aging temperature change 30 °C) has shown that the maximum strain hardening in the stretching diagram section from yield strength to tensile strength is achieved at the heat treatment temperature equal to 290 °C. At carrying out of correlation analysis, a statistically significant positive correlation between deformation corresponding to yield strength and the sum of heights of the largest protrusions and depths of the largest depressions of the surface roughness profile within the basic length of the sample (Rz) and the full height of the surface roughness profile (Rmax) was established. It was found that the reason for the correlation is the presence of cohesive states between the extreme values of the surface roughness profile that persist along the entire length of the specimen.

Dental Implants: Modern Materials and Methods of Their Surface Modification.

The development of dental implantology is based on the detailed study of the interaction of implants with the surrounding tissues and methods of osteogenesis stimulation around implants, which has been confirmed by the increasing number of scientific publications presenting the results of studies related to both the influence of the chemical composition of dental implant material as well as the method of its surface modification on the key operational characteristics of implants. The main materials for dental implant manufacturing are Ti and its alloys, stainless steels, Zr alloys (including ceramics based on ZrO2), and Ta and its alloys, as well as other materials (ceramics based on Al2O3, Si3N4, etc.). The review presents alloy systems recommended for use in clinical practice and describes their physical-mechanical and biochemical properties. However, when getting into the body, the implants are subjected to various kinds of mechanical influences, which are aggravated by the action of an aggressive biological environment (electrolyte with a lot of Cl- and H+); it can lead to the loss of osteointegration and to the appearance of the symptoms of the general intoxication of the organism because of the metal ions released from the implant surface into the biological tissues of the organism. Since the osteointegration and biocompatibility of implants depend primarily on the properties of their surface layer (it is the implant surface that makes contact with the tissues of the body), the surface modification of dental implants plays an important role, and all methods of surface modification can be divided into mechanical, physical, chemical, and biochemical methods (according to the main effect on the surface). This review discusses several techniques for modifying dental implant surfaces and provides evidence for their usefulness.

Investigation of Tribological Characteristics of PEO Coatings Formed on Ti6Al4V Titanium Alloy in Electrolytes with Graphene Oxide Additives.

Coatings with a thickness from ~40 to ~50 µm on Ti6Al4V titanium alloys were formed by plasma electrolytic oxidation (PEO) in a silicate-hypophosphite electrolyte with the addition of graphene oxide. The PEO treatment was carried out in the anode-cathode mode (50 Hz) at a ratio of anode and cathode currents of 1:1; their sum density was 20 A/dm2, and the treatment's duration was 30 min. The effect of the graphene oxide's concentration in the electrolyte on the thickness, roughness, hardness, surface morphology, structure, composition, and tribological characteristics of the PEO coatings was studied. Wear experiments, under dry conditions, were carried out in a ball-on-disk tribotester with an applied load of 5 N, a sliding speed of 0.1 m·s-1, and a sliding distance of 1000 m. According to the obtained results, the addition of graphene oxide (GO) into the base silicate-hypophosphite electrolyte leads to a slight decrease in the coefficient of friction (from 0.73 to 0.69) and a reduction in the wear rate by more than 1.5 times (from 8.04 to 5.2 mm3/N·m), with an increase in the GO's concentration from 0 to 0.5 kg/m3, respectively. This occurs due to the formation of a GO-containing lubricating tribolayer upon contact with the coating of the counter-body in the friction pair. Delamination of the coatings during wear occurs due to contact fatigue; with an increase in the concentration of GO in the electrolyte from 0 to 0.5 kg/m3, this process slows down by more than four times.

An In Vivo Rat Study of Bioresorbable Mg-2Zn-2Ga Alloy Implants.

In the present study, pins made from the novel Mg-2Zn-2Ga alloy were installed within the femoral bones of six Wistar rats. The level of bioresorption was assessed after 1, 3, and 6 months by radiography, histology, SEM, and EDX. Significant bioresorption was evident after 3 months, and complete dissolution of the pins occurred at 6 months after the installation. No pronounced gas cavities could be found at the pin installation sites throughout the postoperative period. The animals' blood parameters showed no signs of inflammation or toxication. These findings are sufficiently encouraging to motivate further research to broaden the experimental coverage to increase the number of observed animals and to conduct tests involving other, larger animals.

Adsorption Ability of Graphene Aerogel and Reduced Graphene Aerogel toward 2,4-D Herbicide and Salicylic Acid.

Within this work, new aerogels based on graphene oxide are proposed to adsorb salicylic acid (SA) and herbicide 2,4-Dichlorophenoxyacetic acid (2,4-D) from aqueous media. Graphene oxide aerogel (GOA) and reduced graphene oxide aerogel (rGOA) were obtained by freeze-drying processes and then studied by Raman spectroscopy, Fourier-transform infrared spectroscopy (FT-IR), and Brunauer-Emmett-Teller (BET) analysis. The influence of contact time and the concentration of the adsorbates were also assessed. It was found that equilibrium for high adsorption is reached in 150 min. In a single system, the pseudo-first-order, pseudo-second-order kinetic models, Intraparticle diffusion, and Elovich models were used to discuss the detail of the aerogel adsorbing pollutant. Moreover, the Langmuir, Freundlich, and Temkin adsorption models were applied to describe the equilibrium isotherms and calculate the isotherm constants.

Bone Remodeling Interaction with Magnesium Alloy Implants Studied by SEM and EDX.

The development direction of bioresorbable fixing structures is currently very relevant because it corresponds to the priority areas in worldwide biotechnology development. Magnesium (Mg)-based alloys are gaining high levels of attention due to their promising potential use as the basis for fixating structures. These alloys can be an alternative to non-degradable metal implants in orthopedics, maxillofacial surgery, neurosurgery, and veterinary medicine. In our study, we formulated a Mg-2Zn-2Ga alloy, prepared pins, and analyzed their biodegradation level based on SEM (scanning electron microscopy) and EDX (energy-dispersive X-ray analysis) after carrying out an experimental study on rats. We assessed the resorption parameters 1, 3, and 6 months after surgery. In general, the biodegradation process was characterized by the systematic development of newly formed bone tissue. Our results showed that Mg-2Zn-2Ga magnesium alloys are suitable for clinical applications.

Investigation of MAO Coatings Characteristics on Titanium Products Obtained by EBM Method Using Additive Manufacturing.

Coatings with a thickness from 27 to 62 µm on electron beam melted Ti-6Al-4V have been formed by micro-arc oxidation (MAO) in a silicate-hypophosphite electrolyte. MAO tests in the anode-cathode mode (50 Hz) with an anode-to-cathode current ratio of 1:1 and sum current densities 10 and 20 A/dm2 were carried out. The duration of the MAO treatment was 30 and 60 min. The effect of the processing parameters on the structural properties of the MAO treated coatings was studied. The current density and treatment time significantly affect the coating thickness and surface roughness. The values of these characteristics increase as the current density increases. The effect of thermal cycling tests on surface morphology, thickness and roughness, and elemental and phase composition of MAO coatings was analyzed. After 50 cycles of thermal cycling from +200 °C to -50 °C, no cracking or delamination of coatings was observed. Coatings formed in 30 min at a current density of 20 A/dm2 turned out to be the best in terms of such indicators as surface morphology, thickness, and roughness.

Evaluation of Mechanical and Electrical Performance of Aging Resistance ZTA Composites Reinforced with Graphene Oxide Consolidated by SPS.

This paper presents a study of Al2O3-ZrO2 (ZTA) nanocomposites with different contents of reduced graphene oxide (rGO). The influence of the rGO content on the physico-mechanical properties of the oxide composite was revealed. Graphene oxide was obtained using a modified Hummers method. Well-dispersed ZTA-GO nanopowders were produced using the colloidal processing method. Using spark plasma sintering technology (SPS), theoretically dense composites were obtained, which also reduced GO during SPS. The microstructure, phase composition, and physico-mechanical properties of the sintered composites were studied. The sintered ZTA composite with an in situ reduced graphene content of 0.28 wt.% after the characterization showed improved mechanical properties: bending strength was 876 ± 43 MPa, fracture toughness-6.8 ± 0.3 MPa·m1/2 and hardness-17.6 ± 0.3 GPa. Microstructure studies showed a uniform zirconia distribution in the ZTA ceramics. The study of the electrical conductivity of reduced graphene oxide-containing composites showed electrical conductivity above the percolation threshold with a small content of graphene oxide (0.28 wt.%). This electrical conductivity makes it possible to produce sintered ceramics by electrical discharge machining (EDM), which significantly reduces the cost of manufacturing complex-shaped products. Besides improved mechanical properties and EDM machinability, 0.28 wt.% rGO composites demonstrated high resistance to hydrothermal degradation.

Prevalence and Risk of Dental Erosion in Patients with Gastroesophageal Reflux Disease: A Meta-Analysis.

AIM: The present paper aims to systematize data concerning the prevalence and risk of dental erosion (DE) in adult patients with gastroesophageal reflux disease (GERD) compared to controls. MATERIALS AND METHODS: Core electronic databases, i.e., MEDLINE/PubMed, EMBASE, Cochrane, Google Scholar, and the Russian Science Citation Index (RSCI), were searched for studies assessing the prevalence and risk of DE in adult GERD patients with publication dates ranging from 1 January 1985 to 20 January 2022. Publications with detailed descriptive statistics (the total sample size of patients with GERD, the total sample size of controls (if available), the number of patients with DE in the sample of GERD patients, the number of patients with DE in the controls (if available)) were selected for the final analysis. RESULTS: The final analysis included 28 studies involving 4379 people (2309 GERD patients and 2070 control subjects). The pooled prevalence of DE was 51.524% (95 CI: 39.742-63.221) in GERD patients and 21.351% (95 CI: 9.234-36.807) in controls. An association was found between the presence of DE and GERD using the random-effects model (OR 5.000, 95% CI: 2.995-8.345; I2 = 79.78%) compared with controls. When analyzing studies that only used validated instrumental methods for diagnosing GERD, alongside validated DE criteria (studies that did not specify the methodologies used were excluded), a significant association between the presence of DE and GERD was revealed (OR 5.586, 95% CI: 2.311-13.503; I2 = 85.14%). CONCLUSION: The meta-analysis demonstrated that DE is quite often associated with GERD and is observed in about half of patients with this extremely common disease of the upper gastrointestinal tract.

Diagnostics of non-carious lesions of dental hard tissues with the methods of optical spectroscopy and radiography / Diagnóstico de lesões não cariosas de tecidos duros dentais com os métodos de espectroscopia óptica e radiografia

Objective: This work aims to use new spectroscopic and radiographic methods to study the dental hard tissue erosion and abfraction, as these lesions are actually quite difficult to be diagnosed in clinical practice. Material and Methods: This in vivo study was conducted on 60 patients with early erosion and 60 patients with abfraction at the cervical area by means of laser-induced fluorescence (LIF) and multilayer spiral computed tomography (MSCT). Results: In comparative dental hard tissues studies LIF spectra from intact and affected areas, it was noted multidirectional fluorescence intensity dependence between areas affected by abfraction and intact areas. MSCT technique allowed assessing the degree of damaged areas, especially at deeper stages. Conclusion: Although LIF and MSCT methods presented limitations, it was shown their effectiveness in the diagnosis of abfraction and erosion by detecting changes in the morphological structure as well as in the chemical and mineral composition of affected dental hard tissues. So LIF and MSCT methods can be successfully used in order to develop new medical devices which will provide most accurate clinical diagnose of different stages of dental erosion and abfraction.(AU)

Objetivo: Este trabalho tem como objetivo utilizar novos métodos espectroscópicos e radiográficos para estudar a erosão e a abração do tecido duro dentário, pois essas lesões são realmente bastante difíceis de serem diagnosticadas na prática clínica. Material e Métodos: Este estudo in vivo foi realizado em 120 pacientes com erosão e abração precoces na área cervical por meio de fluorescência induzida por laser (LIF) e tomografia computadorizada em espiral multicamada (MSCT). Resultados: Em estudos comparativos espectros de LIF, de tecidos duros dentais intactos e afetados, observouse dependência da intensidade da fluorescência multidirecional entre as áreas afetadas pela abração e as áreas intactas. A técnica MSCT permitiu avaliar o grau de áreas danificadas, principalmente em estágios mais profundos. Conclusão: Embora os métodos LIF e MSCT tenham apresentado limitações, foi demonstrada sua eficácia no diagnóstico de abração e erosão, detectando alterações na estrutura morfológica e na composição química e mineral dos tecidos duros afetados. Portanto, os métodos LIF e MSCT podem ser utilizados com sucesso, a fim de desenvolver novos dispositivos médicos que fornecerão um diagnóstico clínico mais preciso de diferentes estágios de erosão e abração dentária.(AU)