Osteoblast Attachment Research Articles

Influence of Sandblasted and Acid-etched Commercially Pure Grade II Titanium Surface on Human Osteosarcoma Osteoblast Cell Proliferation and Attachment: A Pilot Study

Influence of Sandblasted and Acid-etched Commercially Pure Grade II Titanium Surface on Human Osteosarcoma Osteoblast Cell Proliferation and Attachment: A Pilot Study

Study of the influence of Zr on the mechanical properties and functional response of Ti-Nb-Ta-Zr-O alloy for orthopedic applications

In Ti-Nb-Ta-Zr based β-titanium alloys intended for orthopedic applications, Zr does not affect the stability of low modulus β-phase, unlike Nb and Ta. The present study attempts to investigate the influence of Zr on the overall mechanical and functional responses of a Ti-Nb-Ta-Zr-O alloy in contrast to a new Ti-Nb-Ta-O alloy. In each material, different crystallographic textures were produced by varying the processing route. While both alloys were found to show low elastic modulus values due to their β-only microstructures, Ti-Nb-Ta-O alloy had lower elastic modulus because of its favorable crystallographic orientation caused by absence of Zr. The tensile strength values were remarkably high for both due to the presence of interstitial oxygen. The hardening effect of Zr was also evident from the higher strength of Ti-Nb-Ta-Zr-O as compared to Ti-Nb-Ta-O alloy. Although the corrosion resistance and in vitro biological behavior of the two alloys were satisfactory, the Ti-Nb-Ta-Zr-O alloy showed lower corrosion rate and improved osteoblast attachment than the Ti-Nb-Ta-O alloy. Thus, whereas the two alloys show promising performance in terms of their mechanical and functional response, presence of Zr marginally improves the performance in the Ti-Nb-Ta-Zr-O for orthopedic applications.

A high current anodization to fabricate a nano-porous structure on the surface of Ti-based implants.

In this study, an oxide layer on Ti-based implants is fabricated by using a high current anodization (HCA) technique in the nitrate electrolyte. This layer is composed of micro-pits and nano-porous arrays in the honeycomb structure. The results show that both the roughness and the layer thickness are related to the reaction time, whereas the size of nano-pores has little to do with the anodization duration. Compared to the nano-tubular arrays constructed by the conventional anodization, this nano-porous layer shows significantly improved mechanical stability. Furthermore, the in vitro assay of osteoblasts shows that cells behaviors on this surface can be modulated by the topology of this special layer. A suitable hierarchical structure composed of micro-pits and nano-porous structure can significantly stimulate osteoblasts attachment, activity, spreading and ALP function. Therefore, this hierarchical surface layer may provide a promising approach, which endows the Ti-based implants with better stability and osseointegration.

Compressive performance of an arbitrary stiffness matched anatomical Ti64 implant manufactured using Direct Metal Laser Sintering

The reduction of stress shielding following Segmental Bone Defect (SBD) repair requires stiffness matching strategies. Accordingly, this work introduces a Ti6Al4V (Ti64) SBD tibial implant that mimics the segmented bone anatomy using a digital bio-model derived from X-Ray μCT Scan data. The implant features a sheathed periodic unit cell design that can perform slightly lower than the segmented bone being replaced for potential stiffness matching. Finite Element Analysis (FEA) was carried out for the selection of unit cell and to predict the implant performance. The results were then compared to compression test data from a Ti64 Grade 23 implant manufactured using Direct Metal Laser Sintering (DMLS) to assess predictability. The outcome of this research shows an anatomical stiffness matched design that maybe suitable for SBD repair of a tibial segment that can be manufactured using DMLS. The developed implant exhibits Young's Modulus (E) of 12.03, 11.94 and 14.58 GPa using Maxwell's criterion, FEA and experimental (highest) methodologies respectively. This is slightly lower than the segmented bone that exhibited 18.01 GPa (ETibia) to allow for stiffness matching following a period of osseointegration depending on ‘critical size’. Furthermore, the surface roughness of the implant was found to be favourable for osteoblasts attachment.

Autologous platelet-rich plasma application for the optimization of bone tissue reparative regeneration in osteopenia

Objective: analysis of scientific literature on the effectiveness of using Plateled-Rich Plasma to optimize the treatment of compression fractures of vertebral bodies of thoracic and lumbar localization against the background of postmenopausal and senile osteoporosis. All the methods of using Platelet-Rich Plasma, described in the literature, can be divided into several main groups. Platelet-Rich Plasma can be mixed with bone material; is applied to the lesion site before application of bone material; is applied over the bone material; used as a biological membrane. Platelet-Rich Plasma contain fibrillar and cellular components and is able to act as a carrier of cells which are important for the regeneration of soft and bony tissues. It can strengthen the attachment of human osteoblasts to membranes and, as a consequence, induce osteogenesis. Many authors are the supporters of the stimulating effect of Platelet-Rich Plasma on reparative osteogenesis. After injection of Platelet-Rich Plasma in the case of complicated fracture the following was observed: odema in the damaged tissues is less pronounced, there are less signs of acute and chronic inflammation, the phase of alteration is changing more quickly by regenerative-reparative processes. The natural clot contains fibrin, fibronectin and vitronectin, they are also called adhesion molecules necessary for cell migration, osteoconduction, epithelialization and osseointegration. Biological properties of Platelet-Rich Plasma are due not only to the presence of growth factors (TGF-β and PDGF), but without them it immediately induces the formation of a fibrin clot, followed by stimulation of cell adhesion and collagen synthesis. Conclusion. Based on the published literature, a high potential of plasma therapy by Platelet-Rich Plasma for bone tissue regeneration is demonstrated. No adequate clinical studies on the effectiveness of Platelet-Rich Plasma therapy that would meet all modern criteria have yet been conducted. Therapy of Platelet-Rich Plasma is a promising branch in medicine and deserves further research in terms of new effective and safe method of application in medical practice.

Cigarette Smoke and E-Cigarette Vapor Dysregulate Osteoblast Interaction With Titanium Dental Implant Surface.

The purpose of this study was to determine the possible deleterious effects of e-cigarette vapor on osteoblast interaction with dental implant material. Osteoblasts were cultured onto Ti6Al4V titanium implant disks and were then exposed or not to whole cigarette smoke (CS), as well as to nicotine-rich (NR) or nicotine-free (NF) e-vapor for 15 or 30 minutes once a day for 1, 2, or 3 days, after which time various analyses were performed. Osteoblast growth on the titanium implant disks was found to be significantly ( P < .001) reduced following exposure to CS and to the NR and NF e-vapors. Osteoblast attachment to the dental implant material was also dysregulated by CS and the NR and NF e-vapors through a decreased production of adhesion proteins such as F-actin. The effects of CS and e-cigarette vapor on osteoblast growth and attachment were confirmed by reduced alkaline phosphatase (ALP) activity and tissue mineralization. The adverse effects of CS and the NR and NF e-vapors on osteoblast interaction with dental implant material also involved the caspase-3 pathway, as the caspase-3 protein level increased following exposure of the osteoblasts to CS or e-vapor. It should be noted that the adverse effects of CS on osteoblast growth, attachment, ALP, and mineralized degradation were greater than those of the NR and NF e-vapors, although the latter did downregulate osteoblast interaction with the dental implant material. Overall results suggest the need to consider e-cigarettes as a possible contributor to dental implant failure and/or complications.

An Innovative Approach to Manganese-Substituted Hydroxyapatite Coating on Zinc Oxide⁻Coated 316L SS for Implant Application.

In this paper, the synthesis of porous manganese substituted hydroxyapatite (Mn-HAp) coating on zinc oxide (ZnO) coated stainless steel (316L SS) using the electrodeposition technique is reported. The structural, functional, morphological, and elemental analyses are characterized by various analytical techniques including X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FT-IR), field emission scanning electron microscopy (FE-SEM), transmission electron microscopy (TEM), and X-ray photoelectron spectroscopy (XPS). Results of electrochemical techniques such as cyclic polarization and impedance show that the Mn-HAp coating on ZnO coated 316L SS has the highest corrosion resistance in simulated body fluid (SBF) solution. Moreover, dissolution of metal ions was extremely reduced, as evaluated by inductively coupled plasma-atomic emission spectroscopy (ICP-AES). The adhesion and hardness of Mn-HAp/ZnO bilayer coatings have superior mechanical properties over individual coatings. Further, the biocompatibility of in vitro osteoblast attachment, cell viability, and live/dead assessment also confirmed the suitability of Mn-HAp/ZnO bilayer coating on 316L SS for orthopedic applications.

Antibacterial Performance of TiCaPCON Films Incorporated with Ag, Pt, and Zn: Bactericidal Ions Versus Surface Microgalvanic Interactions.

It is very important to prevent bacterial colonization at the early postoperative stages. There are four major strategies and their corresponding types of antibacterial surfaces specifically designed to fight infection: bactericide release, anti-adhesion, pH-sensitive, and contact-killing. Herein, we aimed at determining the antibacterial efficiency of different types of bactericidal ions and revealing the possible contribution of surface microgalvanic effects arising from a potential difference on heterogeneous surfaces. We considered five types of TiCaPCON films, with Ag, Zn, Pt, Ag + Zn, and Pt + Zn nanoparticles (NPs) on their surface. The Ag-modified film demonstrated a pronounced antibacterial effect at a very low Ag ion concentration of 0.11 ppb in physiological solution that was achieved already after 3 h of immersion in Escherichia coli ( E. coli) bacterial culture. The Zn-containing sample also showed a noticeable antibacterial effect against E. coli and Staphylococcus aureus ( S. aureus) strains, wherein the concentration of Zn ions was 2 orders of magnitude higher (15 ppb) compared with the Ag ions. The presence of Ag NPs accelerated the leaching of Zn ion out of the TiCaPCON-Ag-Zn film, but no synergistic effect of the simultaneous presence of the two bactericidal components was observed. After the incubation of the samples with Ag, Zn, and Ag + Zn NPs in E. coli and S. aureus suspensions for 24 and 8 h, respectively, all bacterial cells were completely inactivated. The Pt-containing film showed a very low Pt ion release, and therefore the contribution of this type of ions to the total bactericidal effect could be neglected. The results of the electrochemical studies and Kelvin probe force microscopy indicated that microgalvanic couples were formed between the Pt NPs and the TiCaPCON film, but no noticeable antibacterial effect against either E. coli or S. aureus strains was observed. All ion-modified samples provided good osteoblastic cell attachment, spreading, and proliferation and therefore were concluded to be nontoxic for cells. In addition, the TiCaPCON films with Ag, Pt, and Zn NPs on their surface demonstrated good osteoconductive characteristics.

Surface property alterations and osteoblast attachment to contaminated titanium surfaces after different surface treatments: An in vitro study.

Studies have reported a high prevalence of peri-implantitis. The etiology of peri-implantitis remains unclear and no available treatments result in total resolution of established peri-implantitis. To investigate the factors that interfere with osteoblast adhesion to contaminated titanium surfaces after different surface treatments. Grade 4 titanium discs were randomly divided into 5 groups and each group was divided into 2 subgroups, with one contaminated with Aggregatibacter actinomycetemcomitans (A. actinomycetemcomitans), and the other contaminated with Porphyromonas gingivalis (P. gingivalis). Group 1 did not receive bacterial inoculation or surface debridement and served as a control. Group 2 received A. actinomycetemcomitans or P. gingivalis inoculation, separately. Group 3 received bacterial inoculation and titanium curette debridement, followed by normal saline irrigation. Group 4 received bacterial inoculation, curette debridement, normal saline irrigation, and ultrasonication. Group 5 received bacterial inoculation, curette debridement, normal saline irrigation, and placement in 0.12% chlorhexidine. After various surface treatments, the surface roughness and hydrophilicity of the titanium surface were measured, the number of adhered osteoblast cells was calculated, and the amount of residual lipopolysaccharide (LPS) was quantified. A. actinomycetemcomitans and P. gingivalis biofilms noticeably reduced surface hydrophilicity. Groups 3-5 showed decreased hydrophilicity and fewer adhered osteoblast cells compared with the control group. Although ultrasonication was more effective in removing LPS than curette debridement and chlorhexidine, cell adhesion was not as high as with clean titanium discs. The non-surgical treatment used in this study was not effective in removing LPS from titanium surfaces and increasing osteoblast adhesion. A more effective method to remove LPS completely is required to enhance the treatment outcome of peri-implantitis.

Enhancement of calcification by osteoblasts cultured on hydroxyapatite surfaces with adsorbed inorganic polyphosphate.

Inorganic polyphosphate has been expected to accelerate bone regeneration. However, there are limited evidences to prove that polyphosphate adsorbed on the surface of a hydroxyapatite plate enhances calcification of cultured osteoblasts. In this study, we examined the effect of polyphosphate adsorbed onto the surface of a hydroxyapatite plate on the attachment, proliferation, differentiation, and calcification of osteoblasts. After hydroxyapatite plates were soaked in solutions of polyphosphate, the plate surfaces were analyzed by scanning electron microscopy and toluidine blue staining to confirm adsorption of polyphosphate. The hydroxyapatite plates were further subjected to the measurements of surface roughness, water contact angle, and the binding capacity of calcium ions. Cell culture experiments were carried out using MC3T3-E1 pre-osteoblastic cells. It was found that soaking a hydroxyapatite plate in a polyphosphate solution gave rise to an increase in surface roughness and reduction in water contact angle in a concentration-dependent manner, suggesting the adsorption of polyphosphate onto the surface of a hydroxyapatite plate. It was further observed that surface-adsorbed polyphosphate exhibited an inhibitory effect on cell adhesion and proliferation. In contrast, cell differentiation was promoted on hydroxyapatite plates with adsorbed polyphosphate, when assessed from expression of differentiation marker genes including alkaline phosphatase, osteopontin, and osteocalcin. In addition, calcification of the culture was enhanced on hydroxyapatite plates with relatively low density of adsorbed polyphosphate. Our results as a whole provided an evidence to show that there is a narrow window with regard to the surface density of adsorbed polyphosphate for the enhancement of osteoblast calcification.

Improving osteoblasts cells proliferation via femtosecond laser surface modification of 3D-printed poly-ε-caprolactone scaffolds for bone tissue engineering applications

Synthetic polymer biomaterials incorporating cells are a promising technique for treatment of orthopedic injuries. To enhance the integration of biomaterials into the human body, additional functionalization of the scaffold surface should be carried out that would assist one in mimicking the natural cellular environment. In this study, we examined poly-e-caprolactone (PCL) fiber matrices in view of optimizing the porous properties of the constructs. Altering the porosity of a PCL scaffold is expected to improve the material’s biocompatibility, thus influencing its osteoconductivity and osteointegration. We produced 3D poly-e-caprolactone (PCL) matrices by a fused deposition modeling method for bone and cartilage tissue engineering and performed femtosecond (fs) laser modification experiments to improve the surface properties of the PCL construct. Femtosecond laser processing is one of the useful tools for creating a vast diversity of surface patterns with reproducibility and precision. The processed surface of the PCL matrix was examined to follow the effect of the laser parameters, namely the laser pulse energy and repetition rate and the number (N) of applied pulses. The modified zones were characterized by scanning electron microscopy (SEM), confocal microscopy, X-ray computed tomography and contact angle measurements. The results obtained demonstrated changes in the morphology of the processed surface. A decrease in the water contact angle was also seen after fs laser processing of fiber meshes. Our work demonstrated that a precise control of material surface properties could be achieved by applying a different number of laser pulses at various laser fluence values. We concluded that the structural features of the matrix remain unaffected and can be successfully modified through laser postmodification. The cells tests indicated that the micro-modifications created induced MG63 and MC3T3 osteoblast cellular orientation. The analysis of the MG63 and MC3T3 osteoblast attachment suggested regulation of cells volume migration.

All-in-one trifunctional strategy: A cell adhesive, bacteriostatic and bactericidal coating for titanium implants.

Strategies to inhibit initial bacterial adhesion are extremely important to prevent infection on biomaterial surfaces. However, the simultaneous attraction of desired eukaryotic cells remains a challenge for successful biomaterial-host tissue integration. Here we describe a method for the development of a trifunctional coating that repels contaminating bacteria, kills those that adhere, and promotes osteoblast adhesion. To this end, titanium surfaces were functionalized by electrodeposition of an antifouling polyethylene glycol (PEG) layer and subsequent binding of a peptidic platform with cell-adhesive and bactericidal properties. The physicochemical characterization of the samples via SEM, contact angle, FTIR and XPS analysis verified the successful binding of the PEG layer and the biomolecules, without altering the morphology and topography of the samples. PEG coatings inhibited protein adsorption and osteoblast-like (SaOS-2) attachment; however, the presence of cell adhesive domains rescued osteoblast adhesion, yielding higher values of cell attachment and spreading compared to controls (p < 0.05). Finally, the antibacterial potential of the coating was measured by live/dead assays and SEM using S. sanguinis as a model of early colonizer in oral biofilms. The presence of PEG layers significantly reduced bacterial attachment on the surfaces (p < 0.05). This antibacterial potential was further increased by the bactericidal peptide, yielding values of bacterial adhesion below 0.2% (p < 0.05). The balance between the risk of infection and the optimal osteointegration of a biomaterial is often described as "the race for the surface", in which contaminating bacteria and host tissue cells compete to colonize the implant. In the present work, we have developed a multifunctional coating for a titanium surface that promotes the attachment and spreading of osteoblasts, while very efficiently inhibits bacterial colonization, thus holding promise for application in bone replacing applications.

The Role of Membrane Curvature in Nanoscale Topography-Induced Intracellular Signaling.

Over the past decade, there has been growing interest in developing biosensors and devices with nanoscale and vertical topography. Vertical nanostructures induce spontaneous cell engulfment, which enhances the cell-probe coupling efficiency and the sensitivity of biosensors. Although local membranes in contact with the nanostructures are found to be fully fluidic for lipid and membrane protein diffusions, cells appear to actively sense and respond to the surface topography presented by vertical nanostructures. For future development of biodevices, it is important to understand how cells interact with these nanostructures and how their presence modulates cellular function and activities. How cells recognize nanoscale surface topography has been an area of active research for two decades before the recent biosensor works. Extensive studies show that surface topographies in the range of tens to hundreds of nanometers can significantly affect cell functions, behaviors, and ultimately the cell fate. For example, titanium implants having rough surfaces are better for osteoblast attachment and host-implant integration than those with smooth surfaces. At the cellular level, nanoscale surface topography has been shown by a large number of studies to modulate cell attachment, activity, and differentiation. However, a mechanistic understanding of how cells interact and respond to nanoscale topographic features is still lacking. In this Account, we focus on some recent studies that support a new mechanism that local membrane curvature induced by nanoscale topography directly acts as a biochemical signal to induce intracellular signaling, which we refer to as the curvature hypothesis. The curvature hypothesis proposes that some intracellular proteins can recognize membrane curvatures of a certain range at the cell-to-material interface. These proteins then recruit and activate downstream components to modulate cell signaling and behavior. We discuss current technologies allowing the visualization of membrane deformation at the cell membrane-to-substrate interface with nanometer precision and demonstrate that vertical nanostructures induce local curvatures on the plasma membrane. These local curvatures enhance the process of clathrin-mediated endocytosis and affect actin dynamics. We also present evidence that vertical nanostructures can induce significant deformation of the nuclear membrane, which can affect chromatin distribution and gene expression. Finally, we provide a brief perspective on the curvature hypothesis and the challenges and opportunities for the design of nanotopography for manipulating cell behavior.

Titanium-based implant comprising a porous microstructure assembled with nanoleaves and controllable silicon-ion release for enhanced osseointegration.

Osseointegration is crucial for early fixation as well as for long-term implantation success, hence numerous efforts have been made to tune the surface topography or chemical composition of biomedical implants to improve osseointegration. In this work, various nanostructures, including nanoflocs, nanobundles, nanorods, and nanoleaves, were introduced to the surface of silicon (Si)-incorporated microporous structure to form Si-incorporated micro-nano hierarchical structures on titanium (Ti)-based implants. The osseointegration of the implants were systemically assessed in vivo and in vitro. The in vitro evaluations showed that the nanostructures promoted the protein adsorption, thus modulating the early cellular responses, including the attachment and spreading of osteoblasts and human endothelial cells (HUVECs), and subsequent cell proliferation and differentiation. Furthermore, compared with the single microporous structure, the nanostructures located over the microporous structure protected the Si ions from quick release and allowed the long-term sustained Si-ions release, which further contributed to the proliferation and differentiation of osteoblasts and vascular endothelial growth factor (VEGF) secretion as well as the tube formation of HUVECs. Collectively, the favorable nano-surface structures, especially the nanoleaves structure, and the constant Si-ion release together led to robust osteogenic and angiogenic activities. More importantly, in vivo micro-CT evaluation and histological observations further verified that the Si-incorporated micro-nano hierarchical implant with nanoleaves structure could efficiently promote new bone formation, thus indicating it was an attractive candidate as a next-generation bone-implant material.

Immobilisation of hydroxyapatite-collagen on polydopamine grafted stainless steel 316L: Coating adhesion and in vitro cells evaluation.

The utilisation of hydroxyapatite and collagen as bioactive coating materials could enhance cells attachment, proliferation and osseointegration. However, most methods to form crystal hydroxyapatite coating do not allow the incorporation of polymer/organic compound due to production phase of high sintering temperature. In this study, a polydopamine film was used as an intermediate layer to immobilise hydroxyapatite-collagen without the introduction of high sintering temperature. The surface roughness, coating adhesion, bioactivity and osteoblast attachment on the hydroxyapatite-collagen coating were assessed as these properties remains unknown on the polydopamine grafted film. The coating was developed by grafting stainless steel 316L disks with a polydopamine film. Collagen type I fibres were then immobilised on the grafted film, followed by the biomineralisation of hydroxyapatite. The surface roughness and coating adhesion analyses were later performed by using AFM instrument. An Alamar Blue assay was used to determine the cytotoxicity of the coating, while an alkaline phosphatase activity test was conducted to evaluate the osteogenic differentiation of human fetal osteoblasts on the coating. Finally, the morphology of cells attachment on the coating was visualised under FESEM. The highest RMS roughness and coating adhesion were observed on the hydroxyapatite-collagen coating (hydroxyapatite-coll-dopa). The hydroxyapatite-coll-dopa coating was non-toxic to the osteoblast cells with greater cells proliferation, greater level of alkaline phosphate production and more cells attachment. These results indicate that the immobilisation of hydroxyapatite and collagen using an intermediate polydopamine is identical to enhance coating adhesion, osteoblast cells attachment, proliferation and differentiation, and thus could be implemented as a coating material on orthopaedic and dental implants.

A new insight into in vitro behaviour of poly(\u03b5-caprolactone)/bioactive glass composites in biologically related fluids

In the present work, the role of content, size and chemical composition of gel-derived bioactive glass particles from the SiO2–CaO–P2O5 system in modulating the in vitro bioactivity, osteoinductive properties and long-term (up to 15 months) degradation behaviour of poly(ε-caprolactone)-based composite films was investigated. Bioactivity was assessed in simulated body fluid (SBF) and HEPES-free Dulbecco’s modified Eagle medium (DMEM) supplemented with 10% foetal bovine serum (FBS), while hydrolytic degradation tests were performed in phosphate buffer saline. Obtained composite films showed excellent calcium phosphate (CaP) layer forming ability in both SBF and DMEM-10% FBS. However, kinetics of bioactivity process strongly depended on the type of medium used. The layer of amino acids and proteins, derived from cell culture medium, on the surfaces of composites created barrier that inhibited release of the ions on the one hand, while increasing nucleation density of calcium phosphates, affecting the morphology of formed CaP layers on the other. The presence of bioactive glass fillers was shown to impart osteoinductive properties to obtained films, supporting osteoblast attachment and proliferation, as well as stimulating cell differentiation and also matrix mineralization process in vitro. We showed that kinetics of bioactivity process and also osteoinductive properties of composite films could be easily modulated with the use of different contents and chemical compositions of fillers. The results showed that modification of PCL matrix with bioactive glass particles accelerated its degradation. We proved that the degradation rate of composites could be controlled and optimized for bone regeneration, in particular by using bioactive fillers causing different calcium phosphate layer forming ability on the surfaces of composites, depending on particle size and chemical composition. We have presented new opportunities to design and obtain multifunctional composites with tunable degradation and bioactivity kinetics, as well as biological properties that can meet complex requirements of bone tissue engineering.

In situ silica nanoparticles-reinforced biodegradable poly(citrate-siloxane) hybrid elastomers with multifunctional properties for simultaneous bioimaging and bone tissue regeneration

Most of current biomedical implants do not possess multifunctional properties including biomimetic elastomeric behavior, enhancing cellular activity, real-time imaging tracking ability. In this study, through an in situ nanoparticles formation process, the silica nanoparticles (SNs) reinforced poly(citrate-siloxane) hybrid elastomers (PCS-SN) with multifunctional properties were developed. SNs with uniform size and spherical morphology were distributed in PCS polymer matrix homogeneously. The structure, elastomeric mechanical properties, biodegradation, photoluminescent ability and osteoblasts biocompatibility of PCS-SN could be well controlled. The addition of SNs significantly enhanced the elastomeric mechanical properties and photoluminescent capacity of PCS. PCS-SN-2.5% exhibited significantly low in vivo inflammatory response and long-term fluorescent imaging performance in vivo. The attachment and proliferation of osteoblasts (MC3T3-E1) was also significantly improved, demonstrating the excellent osteogenic cellular biocompatibility of PCS-SN-2.5%. The novel hybrid structure and multifunctional properties make PCS-SN-2.5% hybrid elastomers promising for simultaneous bone tissue regeneration and bioimaging applications.

Suspended graphene oxide nanoparticle for accelerated multilayer osteoblast attachment.

Mimicking bone tissues having layered structures is still a significant challenge because of the lack of technologies to assemble osteoblast cell types into bone structures. One of the promising and attractive materials in biomedical and different engineering fields is graphene and graphene-based nanostructures such as graphene oxide (GO) because of their unique properties. In most studies, GO was synthesized using chemical vapor deposition method, and was coated on the substrate. In this study, we proposed a simple technique for assembly of cells that facilitates the construction of osteoblast-like structures using suspended GO synthesized by graphite powder, H2 SO4 , and KMnO4 .Toxicity effects of GO on human mesenchymal stem cells (hMSCs) derived from bone marrow were analyzed. In addition to normal MSCs, toxicity effects of GO on human cancer cell line saos-2 as an abnormal cell line that possess several osteoblastic features, was examined. The attachment and expression of osteoblast cells genes were evaluated after differentiation of MSCs to osteoblast cells in presence of suspended GO by scanning electron microscopy and real time PCR. We found that the toxicity effects of GO are dose dependent and in oseogenic medium containing suspended GO the expression level of osteoblast genes osteopontin and osteocalcin and cell adhesion markers connexin were higher than control group. Interestingly, through this method GO was found to induce multilayer osteoblast cell morphology and enhance the number of cell layer. We expect that the presented method would become a highly useful approach for bone tissue engineering. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 106A: 293-303, 2018.

Synthesis and Characterization of an Injectable and Hydrophilous Expandable Bone Cement Based on Poly(methyl methacrylate).

Poly(methyl methacrylate) (PMMA), the most common bone cement, has been used as a graft substitute in orthopedic surgeries such as vertebroplasty. However, an undesirable minor crack in the bone-cement interface provoked by shrinkage during polymerization and high elastic modulus of conventional PMMA bone cement dramatically increases the risk of vertebral body refracture postsurgery. Thus, herein, a hydrophilous expandable bone cement was synthesized based on a PMMA commercial cement (Mendec Spine Resin), acrylic acid (AA), and styrene (St). The two synthesized cements (PMMA-PAA, PMMA-PAA-PSt) showed excellent volumetric swelling in vitro and cohesive bone-cement contact in rabbit femur cavity defect. The elastic modulus and compressive strength of the new cements were lower than PMMA. Furthermore, the in vitro analysis indicated that the new cements had lower cytotoxicity than PMMA, including superior proliferation and lower apoptotic rates of Sprague-Dawly rat-derived osteoblasts. Western blotting for protein expression and RT-PCR analysis of osteogenesis-specific genes were conducted on SD rat-derived osteoblasts from both PMMA and new cements films; the results showed that new cements enhanced the expression of osteogenesis-specific genes. Scanning electron microscopy demonstrated improved morphology and attachment of osteoblast on new cement discs compared to the PMMA discs. Additionally, the histological morphologies of the bone-cement interface from the rabbit medial femoral condyle cavity defect model revealed direct and cohesive contact with the bone in the new cement groups in contrast to a minor crack in the PMMA cement group. The sign of a new bone growing into the cement has been found in the new cements after 12 weeks, thereby indicating the osteogenic capacity in vivo. In conclusion, the synthesized hydrophilous expandable bone cements based on PMMA and poly(acrylic acid) (PAA) are promising candidates for vertebroplasty.

Effects of an Erbium:Yttrium‐Aluminum‐Garnet Laser and Ultrasonic Scaler on Titanium Dioxide‐Coated Titanium Surfaces Contaminated With Subgingival Plaque: An In Vitro Study to Assess Post‐Treatment Biocompatibility With Osteogenic Cells

Effects of conventional ultrasonic scaler versus an erbium:yttrium-aluminum-garnet (Er:YAG) laser on titanium surfaces contaminated with subgingival plaque from patients with peri-implantitis are evaluated in terms of: 1) plaque and biocorroded titanium oxide coating removal; 2) surface change induction; and 3) residual biocompatibility toward osteoblasts. Subgingival plaque-coated titanium disks with a moderately rough surface were fixed with ethanol and treated with an ultrasonic scaler (metal tip) or Er:YAG laser (20.3 or 38.2 J/cm2) in non-contact mode. Fluorescent detection of residual plaque was performed. Disk surface morphology was evaluated by scanning electron microscopy. Viability, attachment, proliferation, and differentiation of Saos-2 osteoblasts on new and treated disks were assayed by propidium iodide/DNA stain assay and confocal microscopic analysis of cytoskeleton, Ki67, expression of osteopontin and alkaline phosphatase, and formation of mineralized nodules. Both methods resulted in effective debridement of treated surfaces, the plaque area being reduced to 11.7% with the ultrasonic scaler and ≤0.03% with the Er:YAG laser (38.2 J/cm2). Ultrasound-treated disks showed marked surface changes, incomplete removal of the titanium dioxide (TiO2) layer, and scanty plaque aggregates, whereas the Er:YAG laser (38.2 J/cm2) completely stripped away the plaque and TiO2 layer, leaving a micropitted surface. Both treatments maintained a good biocompatibility of surfaces to Saos-2 osteoblasts. Air-water cooling kept disk temperature below the critical threshold of 47°C. This study shows that an ultrasonic scaler with metal tip is less efficient than high-energy Er:YAG irradiation to remove the plaque and TiO2 layer on anodized disks, although both procedures appear capable of restoring an adequate osseoconductivity of treated surfaces.