Osteoblastic Cells Research Articles

Red light-emitting diode therapy minimizes the functional deleterious effects of the antiretroviral ritonavir on osteoblasts in vitro.

Long-term human immunodeficiency virus (HIV)-infected patients are considered at higher risk for osteoporosis. Among the various causes that lead these patients to lower bone health, there is the use of antiretroviral drugs (ARVs), especially protease inhibitors (PI), such as ritonavir (RTV). In this context, emerge the potential benefits of LED therapy, whose effects on bone cells are currently being extensively studied, showing a modulation in cell differentiation. However, it remains unclear if photobiostimulation might interfere with RTV effects on osteoblast differentiation. In the present study, we investigated the effects of red LED (625nm) irradiation (15 mW/cm2, 0.2J/cm2, and 8 mW/cm2, 0.12J/cm2) on osteoblast cell line MC3T3-E1 treated with RTV (2.5, 5, and 10μg/mL). Our results indicated that red LED irradiation was able to reverse, or at least minimize, the deleterious effects of RTV on the osteoblasts. Neither the ARV treatments 5 and 10μg/mL (104.4% and 95.01%) nor the LED protocols (100.3% and 105.7%) statistically altered cell viability, assessed by the MTT assay. Also, the alkaline phosphatase activity and mineralization showed a decrease in osteoblast activity followed by ARV exposure (39.3-73%), which was attenuated by LED in more than 70% with statistical significance (p < 0.05). In conclusion, photobiostimulation with red LED at 625nm was associated with improved beneficial biological effects as a potential inducer of osteogenic activity on RTV-affected cells. This is the first study that investigated the benefits of red LED irradiation over ARV-treated in vitro osteoblasts.

Targeting POLRMT by a first-in-class inhibitor IMT1 inhibits osteosarcoma cell growth in vitro and in vivo

Osteosarcoma (OS) is a highly aggressive form of bone cancer that predominantly affects adolescents and young adults. In this study, we have undertaken an investigation into the potential anti-OS cell activity of IMT1 (inhibitor of mitochondrial transcription 1), a first-in-class inhibitor of RNA polymerase mitochondrial (POLRMT). IMT1 exhibited a profound inhibitory effect on cell survival, proliferation, cell cycle progression, and migration in primary and immortalized OS cells. Furthermore, this POLRMT inhibitor elicited apoptosis in the OS cells, without, however, inducing cytotoxicity in human osteoblasts or osteoblastic cells. IMT1 disrupted mitochondrial functions in OS cells, resulting in mitochondrial depolarization, oxidative injury, lipid peroxidation, and ATP reduction in OS cells. Silencing POLRMT using targeted shRNA closely mimicked the actions of IMT1 and exerted potent anti-OS cell activity. Importantly, IMT1’s effectiveness was diminished in POLRMT-silenced OS cells. Subsequent investigations revealed that IMT1 suppressed the activation of the Akt-mammalian target of rapamycin (mTOR) cascade in OS cells. IMT1 treatment or POLRMT silencing in primary OS cells led to a significant reduction in Akt1-S6K-S6 phosphorylation. Conversely, it was enhanced upon POLRMT overexpression. The restoration of Akt-mTOR activation through the introduction of a constitutively active S473D mutant Akt1 (caAkt1) mitigated IMT1-induced cytotoxicity in OS cells. In vivo, oral administration of IMT1 robustly curtailed the growth of OS xenografts in nude mice. Furthermore, IMT1 suppressed POLRMT activity, impaired mitochondrial function, repressed Akt-mTOR activation, and induced apoptosis within xenograft tissues. Collectively, these findings underscore the potent growth-inhibitory effects attributed to IMT1 via targeted POLRMT inhibition. The utilization of this POLRMT inhibitor carries substantial therapeutic promise in the context of OS treatment.

MTRR gene rs1801394 polymorphism is associated with neonatal birth weight in pregnant women with fetal growth retardation

Aim: to assess associations between folate cycle gene polymorphism and neonatal birth weight in pregnant women with fetal growth retardation (FGR) and related functional effects in population of the Central Black Earth Region.Materials and Methods. 98 cases of women with FGR were enrolled to a retrospective molecular and genetic screening to assess prevalence 5 SNPs (single nucleotide polymorphisms) in genes involved in folic acid cycle and methionine metabolism (rs699517 TYMS, rs2790 TYMS, rs1979277 SHMT1, rs1805087 MTR, rs1801394 MTRR).Results. It was found out that allele A of the rs1801394 MTRR was associated with a lower neonatal birth weight (recessive model: β = –0.34 ± 0.13; p = 0.009). This polymorphic locus exerts crucial functional effects by determining the amino acid substitution in methionine synthase reductase (Ile22Met) localized in the region of modified histones, which mark enhancers and promoters in ectoderm, endoderm and mesoderm cell cultures, primary osteoblast cells, brain, fat nuclei, skeletal muscles, etc. In addition, rs1801394 MTRR is found DNA sites (motifs) responsible for sensitivity to transcription factors STAT and TBX5 being also related to MTRR gene mRNA expression level in subcutaneous and visceral adipose tissue, thyroid gland, fibroblast cell culture as well as various brain regions.Conclusion. Thus, the allele A of the rs1801394 polymorphism in MTRR gene is a risk factor for a lower neonatal birth weight.

The marine factor 3,5-dihydroxy-4-methoxybenzyl alcohol prevents TNF-α-mediated impairment of mineralization in mouse osteoblastic MC3T3-E1 cells: Impact of macrophage activation

The phenolic antioxidant 3,5-dihydroxy-4-methoxybenzyl alcohol (DHMBA), found in the Pacific oyster Crassostrea gigas, is a superior peroxyl radical scavenger compared to other materials, including Trolox. DHMBA may play an important role in the prevention of health disorders. This study elucidates whether DHMBA prevents the impairment of mineralization of mouse osteoblastic MC3T3-E1 cells under inflammatory conditions by using mouse macrophage RAW264.7 cells in vitro. Culturing with DHMBA (1–100 μM) did not affect the proliferation and death of MC3T3-E1 cells. DHMBA stimulated osteoblastic mineralization. DHMBA blocked the decrease in mineralization of MC3T3-E1 cells caused by culture with the inflammatory cytokine TNF-α. DHMBA inhibited the production of TNF-α by stimulation with lipopolysaccharide (LPS) in RAW264.7 cells. The growth of MC3T3-E1 cells was suppressed by coculture with macrophages under LPS stimulation through the crosstalk of both cells. Interestingly, the growth of MC3T3-E1 cells was suppressed by culturing with the conditioned medium obtained by culturing macrophages with LPS. The effect of the conditioned medium was blocked by the presence of DHMBA or Bay 11–7082, an inhibitor of the TNF-α pathway. The blocking effect of DHMBA was not further enhanced in the presence of Bay 11–7082. Mechanistically, DHMBA was found to decrease the levels of NF-κB p65 and the activity of NF-κB reporter expression in MC3T3-E1 cells. DHMBA was shown to prevent the impairment of osteoblastic mineralization via TNF-α signaling involved in macrophage activation in the bone marrow microenvironment. This study may provide a novel strategy for the therapy of osteoblastic impairment.

Osteoblast Attachment on Bioactive Glass Air Particle Abrasion-Induced Calcium Phosphate Coating.

Air particle abrasion (APA) using bioactive glass (BG) effectively decontaminates titanium (Ti) surface biofilms and the retained glass particles on the abraded surfaces impart potent antibacterial properties against various clinically significant pathogens. The objective of this study was to investigate the effect of BG APA and simulated body fluid (SBF) immersion of sandblasted and acid-etched (SA) Ti surfaces on osteoblast cell viability. Another goal was to study the antibacterial effect against Streptococcus mutans. Square-shaped 10 mm diameter Ti substrates (n = 136) were SA by grit blasting with aluminum oxide particles, then acid-etching in an HCl-H2SO4 mixture. The SA substrates (n = 68) were used as non-coated controls (NC-SA). The test group (n = 68) was further subjected to APA using experimental zinc-containing BG (Zn4) and then mineralized in SBF for 14 d (Zn4-CaP). Surface roughness, contact angle, and surface free energy (SFE) were calculated on test and control surfaces. In addition, the topography and chemistry of substrate surfaces were also characterized. Osteoblastic cell viability and focal adhesion were also evaluated and compared to glass slides as an additional control. The antibacterial effect of Zn4-CaP was also assessed against S. mutans. After immersion in SBF, a mineralized zinc-containing Ca-P coating was formed on the SA substrates. The Zn4-CaP coating resulted in a significantly lower Ra surface roughness value (2.565 μm; p < 0.001), higher wettability (13.35°; p < 0.001), and higher total SFE (71.13; p < 0.001) compared to 3.695 μm, 77.19° and 40.43 for the NC-SA, respectively. APA using Zn4 can produce a zinc-containing calcium phosphate coating that demonstrates osteoblast cell viability and focal adhesion comparable to that on NC-SA or glass slides. Nevertheless, the coating had no antibacterial effect against S. mutans.

Selenomethionine, a Trace Element, Increases Osteoblastic Activity of hFOB 1.19 Cells (an In Vitro Study).

Osteoporosis and resulting fractures affect a significant group of people in the world. It has been shown in many studies that selenium has positive effects on bone metabolism. Based on this information, the aim of this study is to investigate whether bone differentiation will start in a shorter time by applying selenomethionine (SeMet) to hFOB cells.First, hFOB 1.19 cells were cultured. Safe doses of SeMet were determined by MTT and LDH tests. Ossification levels were determined by alizarin red staining and measurement of alkaline phosphatase enzyme levels. The results were analyzed with statistical tests.It was observed that SeMet increased cell viability at concentrations of 10, 25, 50, 100, and 200µM in 24h. At these concentrations, cell viability increased above the control, the viabilities were as follows: 109.4%, 104.9%, 104.3%, 103.15%, and 100.27%. High doses of SeMet significantly reduce cell viability. According to Alizarin red staining, SeMet increases the amount of calcium deposits in hFOB cells in a dose-dependent manner. In the experimental groups, the highest ALP enzyme was determined in the 7-day SeMet application. The most effective dose was measured as 15µM.It was determined that SeMet, which is found as a trace element in living things in nature, increases the viability of hFOB cells, which are osteoblast cell precursors, and increases osteoblastic differentiation and osteoblastic activity in these cells. Our results are at a level that sheds light on an important problem in public health.

ED-71 ameliorates bone regeneration in type 2 diabetes by reducing ferroptosis in osteoblasts via the HIF1α pathway

Eldecalcitol (ED-71), a novel active form of vitamin D, shows potential in treating osteoporosis. However, its underlying mechanisms of action remain to be determined. This study aimed to investigate the effect of ED-71 on bone regeneration and to illustrate its mode of action. The in-vitro model was developed using rat primary osteoblasts cultured under high-glucose conditions, and these cells were treated with ED-71. Additionally, an in vivo model of cranial bone defects was established in type 2 diabetic rats, and ED-71 was administered by gavage. The results demonstrated that ED-71 prevented osteoblast cell death, enhanced rat primary osteoblasts' osteogenic capacity, and attenuated the overexpression of hypoxia-inducible factor 1α (HIF1α) induced by high glucose levels. Furthermore, ED-71 increased glutathione peroxidase 4 (GPX4) levels and inhibited ferroptosis in response to hyperglycemic stimulation. Notably, interference with the HIF1α activator and ferroptosis activator Erastin significantly reduced the therapeutic effects of edetate osteolysis. These findings were further tested in vivo experiments. These results suggest that ED-71 activates the HIF1α pathway in vivo and in vitro, effectively relieving the ferroptosis induced by high glucose. Significantly, ED-71 may improve osteogenic disorders caused by diabetes.

Ultrasound stimulated piezoelectric barium titanate and boron nitride nanotubes in nonconductive poly-ε-caprolactone nanofibrous scaffold for bone tissue engineering

Nanomaterials can provide unique solutions for the problems experienced in tissue engineering by improving a scaffold’s physico-bio-chemical properties. With its piezoelectric property, bone is an active tissue with easy adaptation and remodeling through complicated mechanisms of electromechanical operations. Although poly(ε-caprolactone) (PCL) is an excellent polymer for bone tissue engineering, it is lack of conductivity. In this study, piezoelectric barium titanates (BaTiO3) and boron nitride nanotubes (BNNTs) are used as ultrasound (US) stimulated piezoelectric components in PCL to mimic piezoelectric nature of bone tissue. Electric-responsive Human Osteoblast cells on the scaffolds were stimulated by applying low-frequency US during cell growth. Biocompatibility, cell adhesion, alkaline phosphatase activities and mineralization of osteoblast cells on piezo-composite scaffolds were investigated. BaTiO3 or BNNTs as reinforcement agents improved physical and mechanical properties of PCL scaffolds. In vitro studies show that the use of BaTiO3 or BNNTs as additives in non-conductive scaffolds significantly induces and increases the osteogenic activities even without US stimulation. Although BaTiO3 is one of the best piezoelectric materials, the improvement is more dramatic in the case of BNNTs with the increased mineralization, and excellent chemical and mechanical properties.

Osteogenic-angiogenic coupled response of cobalt-containing mesoporous bioactive glasses in vivo

The incorporation of cobalt ions into the composition of bioactive glasses has emerged as a strategy of interest for bone regeneration purposes. In the present work, we have designed a set of bioactive mesoporous glasses SiO2-CaO-P2O5-CoO (Co-MBGs) with different amounts of cobalt. The physicochemical changes introduced by the Co2+ ion, the in vitro effects of Co-MBGs on preosteoblasts and endothelial cells and their in vivo behaviour using them as bone grafts in a sheep model were studied. The results show that Co2+ ions neither destroy mesoporous ordering nor inhibit in vitro bioactive behaviour, exerting a dual role as network former and modifier for CoO concentrations above 3 % mol. On the other hand, the activity of Co-MBGs on MC3T3-E1 preosteoblasts and HUVEC vascular endothelial cells is dependent on the concentration of CoO present in the glass. For low Co-MBGs concentrations (1mg/ml) cell viability is not affected, while the expression of osteogenic (ALP, RUNX2 and OC) and angiogenic (VEGF) genes is stimulated. For Co-MBGs concentration of 5 mg/ml, cell viability decreases as a function of the CoO content. In vivo studies show that the incorporation of Co2+ ions to the MBGs improves the bone regeneration activity of these materials, despite the deleterious effect that this ion has on bone-forming cells for any of the Co-MBG compositions studied. This contradictory effect is explained by the marked increase in angiogenesis that takes place inside the bone defect, leading to an angiogenesis-osteogenesis coupling that compensates for the partial decrease in osteoblast cells. Statement of significanceThe development of new bone grafts implies to address the need for osteogenesis-angiogenesis coupling that allows bone regeneration with viable tissue in the long term. In this sense the incorporation of cobalt ions into the composition of bioactive glasses has emerged as a strategy of great interest in this field. Due to the potential cytotoxic effect of cobalt ions, there is an important controversy regarding the suitability of their incorporation in bone grafts. In this work, we address this controversy after the implantation of cobalt-doped mesoporous bioactive glasses in a sheep model. The incorporation of cobalt ions in bioactive glasses improves the bone regeneration ability of these bone grafts, due to enhancement of the angiogenesis-osteogenesis coupling.

Antibacterial effect of Ag-containing hydroxyapatite thin film fabricated by sputtering.

Infections related to joint prosthesis are still a major concern for orthopedic surgeons. Hydroxyapatite (HA) is a useful biocompatible material because of its good osteocompatibility. Antibacterial HA coatings have been fabricated with addition of antibacterial agents such as Ag to HA using the plasma spraying method. However, the plasma-sprayed HA coating suffers from fractures at large thicknesses. The sputter-coated HA thin film has a high density, and has been applied clinically for dental implants. However, there are no extensive studies on Ag-containing HA thin films. The aim of this study was to prepare an Ag-containing HA thin film by sputtering, and evaluate its antibacterial effect and cytotoxicity. The Ag-containing HA thin films were prepared by radio-frequency magnetron sputtering. The films were characterized using X-ray photoelectron spectroscopy, X-ray diffractometry, and scanning electron microscopy (SEM). The antibacterial activities and cytotoxicities of the films were also evaluated. The Ag/Ca molar ratio of the films increased with the Ag ratio in the target. The SEM observation of the hydrothermally treated films showed surfaces covered with globular particles. All Ag-containing HA films exhibited an antibacterial effect against E. Coli. The number of cells of the films decreased with the Ag ratio. The films subjected to the hydrothermal treatment exhibited a higher number of cells than the as-sputtered films. The Ag-containing HA thin film was effective in terms of antibacterial effect, and had a low cytotoxicity in the proliferation of osteoblast cells at a low Ag ratio in the target.

Surface-modified deproteinized human demineralized tooth matrix for bone regeneration: physicochemical characterization and osteoblast cell biocompatibility.

Tooth presents an intriguing option as a bone graft due to its compositional similarity to bone. However, the deproteinized human demineralized tooth matrix (dpDTM), developed to overcome the limited availability of autologous tooth grafts, has suboptimal pore size and surface roughness. This study aimed to fabricate a surface-modified dpDTM using acid etching and collagen coating, followed by in vitro evaluation of physicochemical and biological properties. The dpDTM was modified into two protocols: Acid-modified dpDTM (A-dpDTM) and collagen-modified dpDTM (C-dpDTM). Results demonstrated that A-dpDTM and C-dpDTM had increased pore sizes and rougher surfaces compared to dpDTM. Collagen immobilization was evidenced by nitrogen presence exclusively in C-dpDTM. All groups had a Ca/P molar ratio of 1.67 and hydroxyapatite as the sole constituent, with 65-67% crystallinity. Degradation rates significantly increased to 30% and 20% for C-dpDTM and A-dpDTM, respectively, compared to 10% for dpDTM after 120 days. Cumulative collagen release of C-dpDTM on Day 30 was 45.16 µg/ml. Osteoblasts attachment and proliferation were enhanced on all scaffolds, especially C-dpDTM, which displayed the highest proliferation and differentiation rates. In conclusion, surface modified of dpDTM, including A-dpDTM and C-dpDTM, significantly enhances bioactivity by altering surface properties and promoting osteoblast activity, thereby demonstrating promise for bone regeneration applications.

TRIAZINE-TRIONE BASED COMPOSITE MATERIALS FOR POTENTIAL BONE TISSUE ENGINEERING

Several synthetic polymers have been widely investigated for their use in bone tissue engineering applications, but the ideal material is yet to be engineered. Triazine-trione (TATO) based materials and their derivatives are novel in the field of biomedical engineering but have started to draw interest. Different designs of the TATO monomers and introduction of different chemical linkages and end-groups widens the scope of the materials due to a range of mechanical properties.The aim of our work is to investigate novel TATO based materials, with or without hydroxyapatite filler, for their potential in bone tissue engineering constructs. Initially the biocompatibility of the materials was tested, indirectly and directly, according to ISO standards. Following this the osteoconductive properties were investigated with primary osteoblasts and an osteoblastic cell line. Bone marrow derived mesenchymal stem cells were used to evaluate the osteogenic differentiation and consequently the materials potential in bone tissue engineering applications.

Effect of silver/zinc co-dopant on enhancing cytotoxicity and antibacterial property of spray pyrolyzed bioactive glasses

ABSTRACT Owing to the advancement of bioactive materials, bioactive glasses (BGs) are progressively being recognized as promising choices in fields of bone and tissue engineering with their superior properties such as bioactivity and biocompatibility. However, their applications are limited by the fact that the material itself does not possess antibacterial properties. To address this limitation, antibacterial BGs with silver-zinc co-dopant were proposed and successfully synthesized via the one-step spray pyrolysis technique in this work. The phase composition, chemical composition, surface morphology, and nanoscale structures of the obtained BG specimens were characterized using X-ray diffraction, energy dispersive spectroscopy, scanning electron microscopy, and transmission electron microscopy. Moreover, in-vitro bioactivity was assessed through immersion of the SBF solution, while the in-vitro antibacterial property was examined using Escherichia coli bacteria. Meanwhile, the viability of MC3T3-E1 osteoblast cells was evaluated using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Finally, the results showed positive effects of zinc addition, and the influence of silver and zinc dopants on the in-vitro bioactivity, antibacterial property, and cell viability were discussed.

Fabrication and biocompatibility evaluation of hydroxyapatite-polycaprolactone-gelatin composite nanofibers as a bone scaffold.

One approach to addressing bone defects involves the field of bone tissue engineering, with scaffolds playing an important role. The properties of the scaffold must be similar to those of natural bone, including pore size, porosity, interconnectivity, mechanical attributes, degradation rate, non-toxicity, non-immunogenicity, and biocompatibility. The primary goals of this study are as follows: first, to evaluate hydroxyapatite (HA)/polycaprolactone (PCL)/gelatin nanofiber scaffolds based on functional groups, fibre diameter, porosity, and degradation rate; second, to investigate the interaction between HA/PCL/gelatin scaffolds and osteoblast cells (specifically, the ATCC 7F2 cell line) using in vitro assays, including cell viability and adhesion levels. The fibre samples were fabricated using an electrospinning technique with a 15 kV voltage, a spinneret-collector distance of 10 cm, and a flow rate of 0.3 mL hour-1. The process was applied to five different HA/PCL/gelatin concentration ratios: 50 : 40 : 10; 50 : 30 : 20; 50 : 25 : 25; 50 : 20 : 30; 50 : 35 : 15 (in %wt). Fourier Transform Infrared (FTIR) spectrum analysis and tests revealed no differences in functional groups across the five compositions. The identified functional groups include PO4 3-, OH-, CO3 2- and C[double bond, length as m-dash]O stretching. Notably, an increase in PCL concentrations resulted in larger fiber diameters, ranging from 369-1403 nm with an average value of 929 ± 175 nm. The highest porosity percentage was (77.27 ± 11.57) %, and a sufficient degradation rate of up to 3.5 months facilitated the proliferation process of osteoblast cells. Tensile strength assessments revealed a significant increase in tensile strength with the addition of PCL, reaching a peak of 1.93 MPa. The MTT assay demonstrated a discernible increase in cell proliferation, as evidenced by increased cell viability percentages on days 1, 3, and 5. Concurrently, the fluorescence microscopy examination indicated an increase in cell numbers, which was especially noticeable on days 1 and 5. The SEM analysis confirmed the biocompatibility of the HA/PCL/gelatin nanofiber scaffold, as osteoblast cells attached and dispersed successfully five days after seeding. Based on these findings, the HA/PCL/gelatin nanofiber scaffold emerges as a very promising candidate for treating bone damage.

Evaluation of immunohistochemical expression of collagen type I in bone defect treated by local application of collagen and beta-tricalcium phosphate in rats.

Aims: This study aimed to estimate outcome of combined application of collagen/β-tricalcium phosphate (Coll /β- TCP) on bone defect healing by immuno-histochemical localization of collagen type I. Materials and Methods: A total of 20 adult male rats (albino rats) were used in this study. Four intra bony defects were created in both femurs of each rat, three defects were treated with Coll, β- TCP and combination of Coll /β- TCP while forth defect left to heal spontaneously as control. Scarif i cation of animals was done after 2- and 4-weeks healing periods (10 rats for each). Immunohistochemical localization of collagen type I (COL1) monoclonal antibody was done for all bone specimens. Results: Immunohistochemical results showed positive expression of (COL1) by bone cells osteoblast (OB), osteocyte (OC), osteoclast (OCL) and bone marrow stromal cells (BMSCs) at 2- and 4-week duration for all examined groups with variable rates of intensity. Mean values of immunoreactive score (IRS) decreased with time for bone cells and for BMSCs except for OC where values increased with time as the highest values were recorded in combination group at 4 weeks. Conclusions: Combined application of (Coll /β- TCP) was ef f ective in accelerating bone healing process through elevating the IRS of (COL1) noticed by osteo-blast, osteocyte and bone marrow stromal cells.

Static 3D Osteoblast Cell Culture on 3D Printed Titanium Scaffolds.

In vitro cell cultures are a very useful tool for the validation of biomaterial cytocompatibility, especially for bone tissue engineering scaffolds and bone implants. In this chapter, a protocol for a static three-dimensional osteoblast cell culture on titanium scaffolds and subsequent analysis of osteogenic capacity is presented. The protocol is explained for additively manufactured titanium scaffolds, but it can be extrapolated to other scaffolds with similar size and structure, while differing in composition or manufactured technology. Additionally, the protocol can be used for culture of other adherent cell types beyond osteoblast cells such as mesenchymal stem cells.

Nitriding Titanium by Plasma Ion Implantation: Surface Properties and Initial Osteoblast Cell Response

This in vitro study aimed to investigate the effect of plasma ion implantation nitriding on titanium's surface properties and initial cell response. Grade 4 titanium discs (12.7 × 2 mm) were blasted with aluminum oxide particles to create moderately rough surfaces. The experimental discs (TiN) were nitrided using the plasma ion implantation technique in a vacuum chamber. Surface characterization was performed using laser confocal microscopy, atomic force microscopy (AFM), and X-ray photoelectron spectroscopy (XPS). Surface wettability was assessed by measuring the contact angle of a sessile drop using a goniometer. Human osteoblast cells were seeded on the discs to evaluate cell attachment and proliferation at 1, 3, 5, and 7 days of culture using a tetrazolium compound assay. Alkaline phosphatase (ALP) activity was measured at day 7 to assess cell differentiation. Cell morphology was examined by scanning electron microscopy (SEM) and confocal laser scanning microscopy. The TiN group exhibited similar micro-roughness to the control group; however, it displayed a higher density of nanostructures, increased nitrogen content, and slightly improved wettability. Cell proliferation and ALP activity were similar between the groups after seven days of culture. In conclusion, plasma ion implantation nitriding enhances surface nanofeatures and wettability without compromising the biocompatibility of titanium, making it a promising surface modification technique for dental and orthopedic implants.

Examining the level of inflammatory cytokines TNF-α and IL-8 produced by osteoblasts differentiated from dental pulp stem cells.

The use of dental pulp stem cells (DPSCs) in clinical applications instead of bone marrow stem cells is a very promising method capable of significantly changing the future of medical treatment. If further studies prove that DPSCs and the cells differentiated from them do not stimulate the immune system, these cells can be used more reliably in treatment of autoimmune diseases. In this research, we examined the isolated DPSCs and differentiated osteoblasts from them in medium without inflammatory stimulants in terms of TLR3 and TLR4 gene expression and inflammatory cytokines, including TNF-α and IL-8 using qRT-PCR, and measured the concentration of inflammatory cytokines IL-8 and TNF-α produced by these two types of cells through ELISA. The obtained results showed that the expression level of inflammatory cytokines IL-8 and TNF-α in differentiated osteoblasts is significantly different as compared with DPSCs. However, no significant difference was observed in TLR-4 expression between two groups. An increase in TNF-α expression level was found to directly correlate with an increase in the expression of IL-8. The concentration of cytokine TNF-α in osteoblasts was significantly higher than that of IL-8 in DPSCs. In comparison to DPSCs, osteoblast cells first lead to inflammatory responses. These responses reduce overtime. However, DPSCs retain their immunomodulatory properties and do not show inflammatory responses.

Photodynamic Therapy With Resveratrol and an Nd:YAG Laser for Enterococcus faecalis Elimination.

Enterococcus faecalis is the leading cause of endodontic treatment failures. Despite various conventional disinfection approaches, microorganisms often persist in root canals. Photodynamic therapy (PDT) is an adjunct antimicrobial strategy employing a nontoxic photosensitizer (PS) and light source. This study evaluated the antimicrobial effect of PDT using an Nd:YAG laser and resveratrol (RSV) with or without pigment, and confirmed that RSV is nontoxic as a PS. We employed laser irradiation at a 3W output power, using RSV and red pigment as the PS, on an E. faecalis bacterial solution. Subsequently, colony-forming units were quantified. The impact of RSV on osteoblasts was measured using an MTT assay. E. faecalis counts declined after laser irradiation. The combined application of laser irradiation with RSV, red pigment, or both showed a reduction compared to no irradiation and control groups without RSV and red pigment. The 50% cytotoxic concentration against osteoblast cells from mice incubated with RSV for 48 h was 162 μM. The value with RSV and laser was 201 μM and that with RSV and red pigment was 199 μM. The value with RSV, laser and red pigment was 357 μM. The combination of Nd:YAG laser irradiation and RSV as the PS with pigment was efficacious for E. faecalis elimination without inducing any toxic effects on osteoblasts. This combination holds potential as a root canal irrigation strategy.

Comprehensive analysis of the clinical significance and molecular mechanism of T-box transcription factor 3 in osteosarcoma.

Background: T-box transcription factor 3 (TBX3) has been implicated in various malignant tumors, while its exact involvement in osteosarcoma (OS) remains unknown. Methods: Utilizing microarray data and bulk and single-cell RNA-seq data and qRT-PCR, we compared TBX3 mRNA expression levels in different stages of OS. Diagnostic ability testing and prognosis analysis were conducted to better understand the clinical importance of TBX3. Enrichment analysis was performed using gene groups with biological functions similar to TBX3 in different stages of OS to investigate the potential role of TBX3 in OS progression. In addition, we predicted medications targeted at TBX3 and identified downstream target genes to gain a comprehensive understanding of its therapeutic direction and regulatory mechanism. Results: TBX3 expression was highly upregulated in OS and was predominantly expressed in osteoblastic OS cells, with higher expression levels in metastatic tissues. TBX3 expression appeared somewhat suitable for discriminating between OS and normal samples, as well as different stages of OS. We found that TBX3 increased the malignant development of OS by altering cell cycle and cell adhesion molecules; exisulind and tacrolimus, which are targeted small-molecule medicines, were anticipated to counteract this dysregulation. The expression of CCNA2 could potentially be regulated by TBX3, contributing to OS advancement. Conclusion: TBX3 emerges as a potential biomarker for OS. In-depth research into its underlying molecular processes may offer new perspectives on treating OS.