Behavior Of Osteoblastic Cells Research Articles

Tailoring zirconia surface topography via femtosecond laser-induced nanoscale features: effects on osteoblast cells and antibacterial properties

The performance and long-term durability of dental implants hinge on the quality of bone integration and their resistance to bacteria. This research aims to introduce a surface modification strategy for zirconia implants utilizing femtosecond laser ablation techniques, exploring their impact on osteoblast cell behavior and bacterial performance, as well as the integral factors influencing the soft tissue quality surrounding dental implants. Ultrafast lasers were employed to craft nanoscale groove geometries on zirconia surfaces, with thorough analyses conducted using x-ray diffraction, scanning electron microscopy, atomic force microscopy, and water contact angle measurements. The study evaluated the response of human fetal osteoblastic cell lines to textured zirconia ceramics by assessing alkaline phosphatase activity, collagen I, and interleukin 1βsecretion over a 7 day period. Additionally, the antibacterial behavior of the textured surfaces was investigated usingFusobacterium nucleatum, a common culprit in infections associated with dental implants. Ciprofloxacin (CIP), a widely used antibacterial antibiotic, was loaded onto zirconia ceramic surfaces. The results of this study unveiled a substantial reduction in bacterial adhesion on textured zirconia surfaces. The fine biocompatibility of these surfaces was confirmed through the MTT assay and observations of cell morphology. Moreover, the human fetal osteoblastic cell line exhibited extensive spreading and secreted elevated levels of collagen I and interleukin 1βin the modified samples. Drug release evaluations demonstrated sustained CIP release through a diffusion mechanism, showcasing excellent antibacterial activity against pathogenic bacteria, includingStreptococcus mutans, Pseudomonas aeruginosa, andEscherichia coli.

Plasma Rich in Growth Factors in Bone Regeneration: The Proximity to the Clot as a Differential Factor in Osteoblast Cell Behaviour.

The osteogenic differentiation process, by which bone marrow mesenchymal stem cells and osteoprogenitors transform into osteoblasts, is regulated by several growth factors, cytokines, and hormones. Plasma Rich in Growth Factors (PRGF) is a blood-derived preparation consisting of a plethora of bioactive molecules, also susceptible to containing epigenetic factors such as ncRNAs and EVs, that stimulates tissue regeneration. The aim of this study was to investigate the effect of the PRGF clot formulation on osteogenic differentiation. Firstly, osteoblast cells were isolated and characterised. The proliferation of bone cells cultured onto PRGF clots or treated with PRGF supernatant was determined. Moreover, the gene expression of Runx2 (ID: 860), SP7 (ID: 121340), and ALPL (ID: 249) was analysed by one-step real-time quantitative polymerase chain reaction (RT-qPCR). Additionally, alkaline phosphatase (ALPL) activity determination was performed. The highest proliferative effect was achieved by the PRGF supernatant in all the study periods analysed. Concerning gene expression, the logRGE of Runx2 increased significantly in osteoblasts cultured with PRGF formulations compared with the control group, while that of SP7 increased significantly in osteoblasts grown on the PRGF clots. On the other hand, despite the fact that the PRGF supernatant induced ALPL up-regulation, significantly higher enzyme activity was detected for the PRGF clots in comparison with the supernatant formulation. According to our results, contact with the PRGF clot could promote a more advanced phase in the osteogenic process, associated to higher levels of ALPL activity. Furthermore, the PRGF clot releasate stimulated a higher proliferation rate in addition to reduced SP7 expression in the cells located at a distant ubication, leading to a less mature osteoblast stage. Thus, the spatial relationship between the PRGF clot and the osteoprogenitors cells could be a factor that influences regenerative outcomes.

Osteoblast cell behavior on polyetheretherketone dental implant surfaces treated with different grit size aluminum oxide particles: An in vitro analysis

The hydrophobic and bioinert nature of polyetheretherketone (PEEK) implants needs to be addressed for successful osseointegration. The purpose of this in vitro study was to evaluate the osteoblast cell behavior on PEEK implant surfaces treated with airborne-particle abrasion using different grit size aluminum oxide (Al2O3) particles. Disk-shaped specimens (n=96) were prepared from medical grade PEEK rods and were distributed into 4 groups (n=24) of untreated PEEK (PEEK 0), airborne-particle abrasion using 50-μm Al2O3 particles (PEEK 50), airborne-particle abrasion using 110-μm Al2O3 particles (PEEK 110), and airborne-particle abrasion using 150-μm Al2O3 particles (PEEK 150). The surface characteristics were assessed using water contact angle (WCA) measurements and scanning electron microscopy (SEM). MG-63 osteoblast cells were cultured, and the biocompatibility of PEEK was assessed using a CellTiter-blue cell viability assay and florescence staining at day 1, 3, and 7. The specimens were stained with Alizarin red to assess the osteoblast cell differentiation on day 10 and 14. The Levene test was used to test the homogeneity of variances. One-way and Welch ANOVA with post hoc corrections were used to assess the overall statistical significance of differences among the groups (α=.05). The lowest mean WCA was demonstrated in PEEK 150 (49.25 ±5.51) and the highest in PEEK 0 (89.14 ±4.24) (P<.001). SEM images of PEEK 150 illustrated a more complex structure with a large area of globular outcroppings throughout the surface. PEEK 150 showed the highest cell metabolic activity at each time point with florescence staining showing a substantial cell confluence at day 3 and 7. Although PEEK 150 did not show a significant increase in cell proliferation, the number of cells attached was significantly higher than other groups (P<.05). PEEK 110 and 150 also showed a substantial increase in the extent of mineralization. Airborne-particle abrasion using moderate Al2O3 grit size (110- or 150-μm) improved the hydrophilicity and osteoblast cell behavior on PEEK implants.

Proliferation and differential regulation of osteoblasts cultured on surface-phosphorylated cellulose nanofiber scaffolds

Phosphorus-containing polymers have received much attention for their excellent ability to regulate bone cell differentiation and calcification. Given the increasing concern about environmental issues, it is promising to utilize “green” biomaterials to construct novel cell culture scaffolds for bone tissue engineering. Herein, surface-phosphorylated cellulose nanofibers (P-CNFs) were fabricated as a novel green candidate for osteoblast culture. Compared with native CNF, P-CNFs possessed shorter fiber morphology with tunable phosphate group content (0–1.42 mmol/g). The zeta-potential values of CNFs were enhanced after phosphorylation, resulting in the formation of uniform and stable scaffolds. The cell culture behavior of mouse osteoblast (MC3T3-E1) cells showed a clear phosphate content-dependent cell proliferation. The osteoblast cells adhered well and proliferated efficiently on P-CNF0.78 and P-CNF1.05, with phosphate contents of 0.78 and 1.05 mmol/g, respectively, whereas the cells grown on native CNF substrate formed aggregates due to poor cell attachment and exhibited limited cell proliferation. In addition, the P-CNF substrates with optimal phosphate content provided a favorable cellular microenvironment and significantly promoted osteogenic differentiation and calcification, even in the absence of a differentiation inducer. The bio-based P-CNFs are expected to mimic the bone components and provide a means to regulate osteoblast proliferation and differentiation in bone tissue engineering.

Osteoblastic Cell Behavior and Gene Expression Related to Bone Metabolism on Different Titanium Surfaces.

The surface topography of titanium dental implants has a great influence on osseointegration. In this work, we try to determine the osteoblastic behavior and gene expression of cells with different titanium surfaces and relate them to the physicochemical properties of the surface. For this purpose, we have used commercial titanium discs of grade 3: as-received corresponds to machined titanium without any surface treatment (MA), chemically acid etched (AE), treated via sand blasting with Al2O3 particles (SB) and a sand-blasting treatment with acid etching (SB+AE). The surfaces have been observed using scanning electron microscopy (SEM) and the roughness, wettability and surface energy with dispersive and polar components have been characterized. Osteoblastic cultures were performed with SaOS-2 osteoblastic cells determining cell viability as well as alkaline phosphatase levels for 3 and 21 days, and osteoblastic gene expression was determined. The roughness values of the MA discs was 0.02 μm, which increases to 0.3 μm with acid attack and becomes the maximum for the sand-blasted samples, reaching values of 1.2 μm for SB and SB+AE. The hydrophilic behavior of the MA and AE samples with contact angles of 63° and 65° is superior to that of the rougher samples, being 75° for SB and 82° for SB+AE. In all cases, they show good hydrophilicity. GB and GB+AE surfaces present a higher polar component in the surface energy values, 11.96 and 13.18 mJ/m2, respectively, than AE and MA, 6.64 and 9.79 mJ/m2, respectively. The osteoblastic cell viability values at three days do not show statistically significant differences between the four surfaces. However, the viability of the SB and SB+AE surfaces at 21 days is much higher than that of the AE and MA samples. From the alkaline phosphatase studies, higher values were observed for those treated with sand blasting with and without acid etching compared to the other two surfaces, indicating a greater activity in osteoblastic differentiation. In all cases except in the Osterix (Ostx) -osteoblast-specific transcription factor-a decrease in gene expression is observed in relation to the MA samples (control). The most important increase was observed for the SB+AE condition. A decrease in the gene expression of Osteoprotegerine (OPG), Runt-related transcription factor 2 (Runx2), Receptor Activator of NF-κB Ligand (RANKL) and Alkaline Phosphatase (Alp) genes was observed in the AE surface.

Experimental analyses of surface treatment on titanium samples

The roughness of the dental implant surface will influence the behavior of osteoblastic cells regarding the spread, proliferation, differentiation and synthesis of proteins on the surface of titanium. Depending on its characteristics, roughness can be macro-, micro- and nano-roughness. Material and method. We achieved the obtaining of thin layers of biomaterials by the pulsed laser deposition method (PLD, Pulsed Laser Deposition) on the titanium samples and the deposition has been made by Spin coating method of a thin film based on hydroxyapatite, respectively hydroxyapatite-maleic copolymer, only on one side of the sample. Results. From a microscopic point of view, the qualities of bio-glass are remarkable. Thus, bio-glasses are amorphous materials and are successfully used as covering material for metallic and polymeric implants, to significantly increase their biocompatibility. Discussion. The biological properties of the materials for implants are of fundamental importance for the success of the treatments and they are reflected along with other characteristics of the implants, in the tissue reaction they cause under the conditions of their insertion in biological systems. Conclusion. Osseointegration could be impaired in people with diabetes, osteoporosis, with bone metabolism disorder where therapeutic success depends on the biomimetic and functional topography of the dental implant.

Effects of Different Titanium Surface Treatments on Adhesion, Proliferation and Differentiation of Bone Cells: An In Vitro Study.

The objective of this study was to evaluate the impacts of different sandblasting procedures in acid etching of Ti6Al4V surfaces on osteoblast cell behavior, regarding various physicochemical and topographical parameters. Furthermore, differences in osteoblast cell behavior between cpTi and Ti6Al4V SA surfaces were evaluated. Sandblasting and subsequent acid etching of cpTi and Ti6Al4V discs was performed with Al2O3 grains of different sizes and with varying blasting pressures. The micro- and nano-roughness of the experimental SA surfaces were analyzed via confocal, atomic force and scanning electron microscopy. Surface free energy and friction coefficients were determined. hFOB 1.19 cells were seeded to evaluate adhesion, proliferation and osteoblastic differentiation for up to 12 d via crystal violet assays, MTT assays, ALP activity assays and Alizarin Red staining assays. Differences in blasting procedures had significant impacts on surface macro- and micro-topography. The crystal violet assay revealed a significant inverse relationship between blasting grain size and hFOB cell growth after 7 days. This trend was also visible in the Alizarin Red assays staining after 12 d: there was significantly higher biomineralization visible in the group that was sandblasted with smaller grains (F180) when compared to standard-grain-size groups (F70). SA samples treated with reduced blasting pressure exhibited lower hFOB adhesion and growth capabilities at initial (2 h) and later time points for up to 7 days, when compared to the standard SA surface, even though micro-roughness and other relevant surface parameters were similar. Overall, etched-only surfaces consistently exhibited equivalent or higher adhesion, proliferation and differentiation capabilities when compared to all other sandblasted and etched surfaces. No differences were found between cpTi and Ti6Al4V SA surfaces. Subtle modifications in the blasting protocol for Ti6Al4V SA surfaces significantly affect the proliferative and differentiation behavior of human osteoblasts. Surface roughness parameters are not sufficient to predict osteoblast behavior on etched Ti6Al4V surfaces.

The sintering behavior, in vitro performance, and radiopacity of spark plasma sintered barium-containing calcium-phospho-alumino-borosilicate glass powders

The effects of substituting CaO with BaO on the crystallization and sintering behavior of bioactive glass powders in the SiO2–B2O3–Al2O3–CaO–Na2O–P2O5 system were observed during the Spark Plasma Sintering (SPS) process. The outcome of thermal analysis confirmed the surface crystallization mechanism for the glasses. It was revealed that the replacement of 3.3 mol.% BaO accelerated the crystallization kinetics of glass powder while replacing 9.0 mol.% BaO prevented crystallization. The X-ray diffraction results showed that upon increasing BaO replacement, pseudo-wollastonite phase was crystalized instead of wollastonite. Additionally, by replacing 3.3 and 9.0 mol.% of BaO, the beginning temperature of the sintering process was reduced from 450 °C to 440 and 425 °C, respectively. The relative densities of the sintered bodies containing 0.0% and 3.3 mol.% of BaO were 99.95 and 99.27, respectively. Substitution of 9.0 mol.% BaO resulted in a completely amorphous and full-dense sample (relative density of 100%). The in vitro apatite formation ability and osteoblastic cell behavior of the sintered bodies were evaluated. The substitution of BaO for CaO restricted the glass apatite formation ability; however, no cytotoxic effect was found. For all SPS-sintered glasses, an appropriate cell-surface interaction was observed, confirming good cell supporting behavior of produced bodies. Moreover, the radiopacity features were assessed and the higher radiopacity was achieved for Ba-containing bodies.

In Vitro Characterization of In Situ Alloyed Ti6Al4V(ELI)-3 at.% Cu Obtained by Laser Powder Bed Fusion.

The intensive cytotoxicity of pure copper is effectively kills bacteria, but it can compromise cellular behavior, so a rational balance must be found for Cu-loaded implants. In the present study, the individual and combined effect of surface composition and roughness on osteoblast cell behavior of in situ alloyed Ti6Al4V(ELI)-3 at.% Cu obtained by laser powder bed fusion was studied. Surface composition was studied using scanning electron microscopy, energy dispersive spectroscopy, and X-ray diffraction. Surface roughness measurements were carried out using confocal microscopy. In vitro osteoblast performance was evaluated by means of cell morphology observation of cell viability, proliferation, and mineralization. In vitro studies were performed at 1, 7, and 14 days of cell culture, except for cell mineralization at 28 days, on grounded and as-built (rough) samples with and without 3 at.% Cu. The addition of 3 at.% Cu did not show cell cytotoxicity but inhibited cell proliferation. Cell mineralization tends to be higher for samples with 3 at.% Cu content. Surface roughness inhibited cell proliferation too, but showed enhanced cell mineralization capacity and therefore, higher osteoblast performance, especially when as-built samples contained 3 at.% Cu. Cell proliferation was only observed on ground samples without Cu but showed the lowest cell mineralization.

Evaluation of osteoblastic cell behavior upon culture on titanium substrates photo-functionalized by vacuum ultra-violet treatment

Photo-functionalization of titanium orthopedic/prosthetic implants using ultraviolet illumination is known to improve osteogenesis. Therefore, in this study, we aimed to examine the influence of vacuum ultraviolet (VUV)-treated titanium surfaces on osteoblast cell adhesion, activity, and differentiation. Osteoblastic cells were cultured on titanium substrates treated with various VUV treatment conditions (0, 6.2, 18.7, and 37.4 J/cm2) and their behavior was evaluated. The results revealed that cell adhesion was increased whereas cell activity and differentiation ability were decreased upon cell culture on VUV-treated substrates. In particular, cell activity and differentiation ability were dramatically suppressed with 18.7 J/cm2 VUV irradiation. Within the limitations of this cell-based experiment, we clarified the VUV treatment conditions in which cell adhesion was improved but cell activity and differentiation ability were suppressed. These results indicate that VUV-treatment can be used to influence cell growth properties and can be used to accelerate or suppress cell differentiation on implant substrates.

A facile surface modification of poly(dimethylsiloxane) with amino acid conjugated self-assembled monolayers for enhanced osteoblast cell behavior

Polydimethylsiloxane (PDMS) is a biocompatible synthetic polymer and used in various applications due to its low toxicity and tunable surface properties. However, PDMS does not have any chemical cues for cell binding. Plasma treatment, protein coating or surface modification with various molecules have been used to improve its surface characteristics. Still, these techniques are either last for a very limited time or have very complicated experimental procedures. In the present study, simple and one-step surface modification of PDMS is successfully accomplished by the preparation of hydrophilic and hydrophobic amino acid conjugated self-assembled monolayers (SAMs) for enhanced interactions at the cell-substrate interface. Synthesis of histidine and leucine conjugated (3-aminopropyl)-triethoxysilane (His-APTES and Leu-APTES) were confirmed with proton nuclear magnetic resonance spectroscopy (1H NMR) and optimum conditions for the modification of PDMS with SAMs were investigated by X-ray photoelectron spectroscopy (XPS) analysis, combined with water contact angle (WCA) measurements. Results indicated that both SAMs enhanced cellular behavior in vitro. Furthermore, hydrophilic His-APTES modification provides a superior environment for the osteoblast maturation with higher alkaline phosphatase activity and mineralization. As histidine, leucine, and functional groups of these SAMs are naturally found in biological systems, modification of PDMS with them increases its cell-substrate surface biomimetic properties. This study establishes a successful modification of PDMS for in vitro cell studies, offering a biomimetic and easy procedure for potential applications in microfluidics, cell-based therapies, or drug investigations.

Osteoblastic Cell Behavior and Early Bacterial Adhesion on Macro-, Micro-, and Nanostructured Titanium Surfaces for Biomedical Implant Applications.

Surface treatments may significantly affect physical-chemical properties and surface biologic responses. This study aimed to investigate the influence of alterations in the physical-chemical properties of pure titanium with different surface topographies on biocompatibility and early microbiologic response. Titanium disks were exposed to five different surface treatments created through acid etching and anodizing methods. Surface morphology, 2D and 3D roughness, wettability, biocompatibility, and cell viability were evaluated. Osteoblast adhesion and bacterial adhesion tests were also executed. Data were statistically analyzed using analysis of variance followed by Tukey test, roughness (P < .05), and bacterial proliferation (P < .05). Five different surface morphologies were developed; double acid etching was shown to be significantly rougher than the others. The 2D roughness measurements were shown to be less consistent than the 3D measurements. All surfaces presented biocompatibility to allow cell behavior and differentiation. Osteoblasts presented better evolution in terms of adhesion and behavior in the nanomorphologies. High roughness significantly increased bacterial adhesion. Surface treatments may critically alter titanium properties and morphology. Therefore, roughness measurements with a wide area should be used in their evaluation. Nanotextured surfaces show a positive effect on bone cells and antibacterial response; their application is suggested when considering surface texturization for biomedical implants.

The Evaluation of Osteoblastic Cell Behavior on Treated Titanium Surface

Background: There are several potential advantages in optimizing the initial events of osseointegration in the benefit of clinical outcome. Objective: The objective of the present study was to evaluate the behavior of osteoblastic cells on surfaces treated by double acid etching using HNO3 and H2SO4. Methods: Commercially pure titanium (grade 4) discs measuring 6 mm in diameter and 2 mm in thickness were used. The discs were divided into two groups: machined group and double acid-etched discs (HNO3 and H2SO4). Surface characteristics were assessed using Scanning Electron Microscopy. Pre-osteoblastic MC3T3-E1 cells were used for cell culture on the tested surfaces to assess proliferation, viability (MTT), as well as secretion (ELISA) and cytoplasmic expression (Western blot) of type I collagen. Results: The data obtained were analyzed using t-test or two-way ANOVA followed by Bonferroni’s test at 95% significance. The titanium surfaces showed average roughness values for the machined and treated surfaces of 0.29 and 1.16, respectively (p&lt;0.05). An increase in cell proliferation was observed, which was corroborated by the viability assay. Both type I collagen secretion and intracellular expression were higher on the double acid-etched surface compared to the machine surfaces (p&lt;0.05). Conclusion: Implant surfaces treated by double acid etching positively affected the early events of the interaction between titanium and osteoblastic cells, suggesting optimization of osseintegration.

The Evaluation of Osteoblastic Cell Behavior on Treated Titanium Surface

Background: There are several potential advantages in optimizing the initial events of osseointegration in the benefit of clinical outcome. Objective: The objective of the present study was to evaluate the behavior of osteoblastic cells on surfaces treated by double acid etching using HNO3 and H2SO4. Methods: Commercially pure titanium (grade 4) discs measuring 6 mm in diameter and 2 mm in thickness were used. The discs were divided into two groups: machined group and double acid-etched discs (HNO3 and H2SO4). Surface characteristics were assessed using Scanning Electron Microscopy. Pre-osteoblastic MC3T3-E1 cells were used for cell culture on the tested surfaces to assess proliferation, viability (MTT), as well as secretion (ELISA) and cytoplasmic expression (Western blot) of type I collagen. Results: The data obtained were analyzed using t-test or two-way ANOVA followed by Bonferroni’s test at 95% significance. The titanium surfaces showed average roughness values for the machined and treated surfaces of 0.29 and 1.16, respectively (p&lt;0.05). An increase in cell proliferation was observed, which was corroborated by the viability assay. Both type I collagen secretion and intracellular expression were higher on the double acid-etched surface compared to the machine surfaces (p&lt;0.05). Conclusion: Implant surfaces treated by double acid etching positively affected the early events of the interaction between titanium and osteoblastic cells, suggesting optimization of osseintegration.

Morphologically controlled synthesis of (β- tricalcium phosphate) β-TCP particles from cuttlebone by sodium dodecyl sulfate (SDS) anionic surfactant: In vitro behavior of mouse osteoblast cells

β-TCP was successfully synthesized using cuttlebone as a raw material, which was considered to be a calcium source. The calcium source from cuttlebone and phosphoric acid were hydrothermally prepared for 2 h at ambient temperatures. The effect of the sodium dodecyl sulfate (SDS) concentration (ranging from 0 to 6 critical micelle concentrations (CMC)) was assessed. Fourier-transform infrared spectroscopy and X-ray diffraction were employed to investigate the structural properties of the β-TCP. The particles presented irregularly shaped morphologies with variations in size. Brunauer–Emmett–Teller and dynamic light scattering were employed to evaluate the surface properties. No significant change was observed with variations in the CMC. We carried out preliminary investigations of the cell cytotoxicity against mouse fibroblasts. None of the conditions presented cell cytotoxicity. We can conclude that β-TCP prepared from cuttlebone can be used as a medical material.

Influence of RGD Peptide on Morphology and Biocompatibility of 3D Scaffolds Based on PLA/Hydroxyapatite

AbstractThe developments of three‐dimensional and bioactive supports are essential for bone tissue engineering. Therefore, this study reports the morphology and biological behavior of osteoblast cells grown on bioactive 3D scaffolds, obtained by electro‐spinning and based on poly(lactic acid), PLA, and Hydroxyapatite, HAp, composites (PLA/HAp), which were superficially treated with RGD. Likewise, the characterization of the obtained 3D scaffolds is presented and discussed. The results indicate that it is possible to obtain structures with porosities higher than 90% with interconnected channels, which favor cell adhesion and growth. Also, the biological performance evaluation of the scaffolds indicates that surface treatment with RGD was effective and has a synergistic effect along with HAp. The cell adhesion was also observed by SEM and the images show cells extended and adhered to the scaffold's fibers along with neighboring cells. Also, the HAp had an influence on cell proliferation, which showed the maximum values with 2.5 and 5.0% wt of HAp. Therefore, 3D PLA/HAp scaffolds, obtained by electrospinning, treated with RGD represent promising material for use in bone tissue engineering.

Biocompatibility of Hydrophilicity Improved Apatite Cement

Hydrophilicity of apatite cement was increased after O3 gas treatment on apatite cement (AC) powder. It results on the improvement of the handling and mechanical properties of set AC. Behavior of osteoblastic cells to O3-treated set AC was evaluated including initial cell attachment, morphology of the attached cells and proliferation using rat bone marrow cell (RBM). Cells’ response to the set AC was the same regardless of O3 treatment. The cells well attached and spread with filopodial extensions even over the O3-treated set AC specimens. The rates of cell proliferation on set AC were also the same regardless of O3 treatment. The result indicated O3 treatment of AC powder would not affect to the osteoblast cell response of set AC.

Accelerated Bone Regeneration by Gold-Nanoparticle-Loaded Mesoporous Silica through Stimulating Immunomodulation.

Bone repair and regeneration are greatly influenced by the local immune microenvironment. In this regard, the immunomodulatory capability of biomaterials should be considered when evaluating their osteogenic effects. In this study, we investigated the modulatory effects of gold nanoparticle (AuNP)-loaded mesoporous silica nanoparticles (Au-MSNs) on macrophages and the subsequent effects on the behavior of osteoblastic lineage cells. The results demonstrate that Au-MSNs could generate a favorable immune microenvironment by stimulating an anti-inflammatory response and promoting the secretion of osteogenic cytokines by macrophages. As a result, there is an enhancement of osteogenic differentiation in preosteoblastic MC3T3 cells as assessed by the increased expression of osteogenic markers, alkaline phosphatase (ALP) production, and calcium deposition. The immunomodulatory effects and direct osteogenic stimulation by Au-MSNs synergistically increased the osteogenic differentiation capability of MC3T3 cells as a result of crosstalk between Au-MSN-conditioned macrophages and Au-MSN-treated osteoblasts in a coculture system. An in vivo study further revealed that Au-MSNs could accelerate new bone formation in a critical-sized cranial defect site in rats based on computed tomography analysis and histological examination. Together, this novel Au-MSNs could significantly promote osteogenic activity by modulating the immune microenvironment, showing its therapeutic potential for bone tissue repair and regeneration.

Human Osteoblast Cell Behaviour on Titanium Discs Treated with Argon Plasma.

(1) Background. Titanium is characterized by its biocompatibility and resistance to stress and fatigue. Treatment with argon plasma may favour growth of human osteoblasts with respect to cell adhesion and proliferation. The aim of this study was to analyse the behaviour of human osteoblasts (MG-63) on Grade IV and V titanium possessing a sand-blasted, acid-etched (SLA) surface. SLA is a widely used surface treatment to create micro- and macroretentions to enhance osteoconductive properties on the surface. (2) Methods. One group of each grade of titanium was decontaminated with argon plasma and compared. On each disc, 20 × 104 cells were cultivated for morphological analysis, study of cell viability (regarding a negative control [100% viability]) and mitochondrial energy balance. (3) Results. At 24 h titanium treated with SLA showed a higher percentage of cell viability (47.3 ± 8.1%) compared to titanium IV treated with argon plasma, which presented a percentage of 79.1 ± 1.1%. Grade V titanium treated with argon plasma presented a higher viability percentage 91.3 ± 3.0% whereas nontreated Grade V titanium presented 53.3 ± 4.0%. Cells cultivated on the surfaces with an argon-plasma treatment were enlarged in comparison to non-treated discs. The cells with smaller circularity with a greater spread and spindle shape were the ones cultivated on the Grade V titanium surface. Cells seeded on treated titanium IV and titanium V, treated or not, showed higher mitochondrial activity over nontreated titanium IV. (4) Conclusions. Cells cultivated on those Grade V titanium discs that were decontaminated with argon plasma presented higher levels of cell adhesion and proliferation, lower mitochondrial damage and a higher mean cell area compared to those not decontaminated with argon plasma.

Influence of bioactive glass-coating of zirconia implant surfaces on human osteoblast behavior in vitro

ObjectiveThe recently developed bioactive glass PC-XG3, which is suitable to coat zirconia implant surfaces with high adhesion strength may reduce the time of osseointegration and the marginal bone loss following implantation. The glass composition has been previously evaluated for cytotoxicity on fibroblast cells, and will now be used to evaluate the cell behavior of osteoblast cells. MethodsThree different surface morphologies were created with PC-XG3 on zirconia discs. A clinically tested zirconia implant surface as well as polished and machined zirconia served as a reference. Cell viability after 24 h, cell spreading after 30 min and 24 h and the respective morphology of human osteoblasts using scanning electron microscopy were evaluated. Additionally, the corrosive process of PC-XG3 in cell culture medium up to 7 d was measured. ResultsInitial cell behavior of human osteoblasts was not accelerated by the PC-XG3 surface when compared to zirconia. Additionally, it was found that a decreased surface roughness promoted initial cell spreading. Storage in cell culture medium resulted in the accumulation of C and N on the bioglass surface while Mg, Si, K and Ca were decreased and crack formation was observed. SignificanceSince initial spreading quality to a biomaterial is a crucial factor that will determine the subsequent cell function, proliferation, differentiation, and viability it can be assumed that a coating of zirconia implants with this bioactive glass will unlikely reduce osseointegration time.