Initial Adhesion Of Osteoblasts Research Articles

Construction of a Rough Surface with Submicron Ti2Cu Particle on Ti-Cu Alloy and Its Effect on the Antibacterial Properties and Cell Biocompatibility

Titanium-copper (Ti-Cu) alloy is an advanced antibacterial material with excellent mechanical properties, thermodynamic stability, corrosion resistance and biocompatibility. Sandblasting and acid-etching was applied to the Ti-3Cu alloy to construct a rough surface with Ti2Cu phase on the surface in order to improve the antibacterial properties and the osseointegration. The phase constitutes and the physical properties were characterized by X-ray diffraction (XRD), scanning electron microscopy (SEM) and confocal laser scanning microscope (CLSM), and the surface chemical properties were analyzed by X-ray photoelectron spectroscopy (XPS) and electrochemical testing. The antibacterial property was assessed by the plate-count method and the cell compatibility was evaluated by the CCK-8 test in order to reveal the effect of surface characteristics on the antibacterial ability and bioactivity. The results demonstrated a rough and lamellar surface structure with many submicron Ti2Cu particles on the surface of Ti-3Cu, which could enhance the antibacterial ability and promote the cell proliferation and the initial adhesion of osteoblasts. However, the surface treatment also reduced the corrosion resistance and accelerated the Cu ion release.

Single-cell adhesion of human osteoblasts on plasma-conditioned titanium implant surfaces in vitro

ObjectivesThis study aimed to demonstrate the effect of treating titanium-implant surfaces with plasma from two different sources on wettability and initial single-cell adhesion of human osteoblasts and to investigate whether aging affects treatment outcomes. MethodsTitanium disks with sandblasted and acid-etched (SLA) surfaces were treated with atmospheric pressure plasma (APP) and low-pressure plasma (LPP). For wetting behavior of the specimens after plasma treatment, the water contact angle was measured. The single-cell detachment force and amount of work of detachment of human osteoblasts were determined with single-cell force spectroscopy (SCFS). To evaluate the aging effect in APP-treated specimens, SCFS was conducted 10 and 60 min after treatment. ResultsSignificantly higher hydrophilicity was observed in the APP and LPP treatment groups than in the control group, but no significant difference was observed between the APP and LPP groups. No significant difference in cell-detachment force or work of detachment was observed, and there were no significant differences according to the conditioning mechanisms and storage time. SignificanceConditioning of the titanium surfaces with APP or LPP was not a significant influencing factor in the initial adhesion of the osteoblasts.

Poly-l-lysine/Sodium Alginate Coating Loading Nanosilver for Improving the Antibacterial Effect and Inducing Mineralization of Dental Implants.

In recent years, antibacterial surface modification of titanium (Ti) implants has been widely studied in preventing implant-associated infection for dental and orthopedic applications. The purpose of this study was to prepare a composite coating on a porous titanium surface for infection prevention and inducing mineralization, which was initialized by deposition of a poly-l-lysine (PLL)/sodium alginate(SA)/PLL self-assembled coating, followed by dopamine deposition, and finally in situ reduction of silver nanoparticles (AgNPs) by dopamine. The surface zeta potential, SEM, XPS, UV–vis, and water contact angle analyses demonstrate that each coating was successfully prepared after the respective steps and that the average sizes of AgNPs were 20–30 nm. The composite coating maintained Ag+ release for more than 27 days in PBS and induced mineralization when incubated in SBF. The antibacterial results showed that the composite coating inhibited/killed bacteria on the material surface and killed bacteria around them. In addition, although this coating inhibited the initial adhesion of osteoblasts, the mineralized surface greatly enhanced the cytocompatibility. Thus, we concluded that the composite coating could prevent bacterial infections and facilitate mineralization in vivo in the early postoperative period, and then, the mineralized surface could enhance the cytocompatibility.

The osteoinduction of RGD and Mg ion functionalized bioactive zirconia coating.

The objective of this study was to investigate the adhesion, proliferation and mineralization of osteoblasts on arginine-glycine-aspartic acid (RGD)- and magnesium ion (Mg+)-decorated zirconia coatings. The zirconia coatings were prepared via a plasma spray; RGD and Mg+ were immobilized via a silane-coupling agent and ion implantation, respectively. This study employed scanning electron microscopy (SEM) to observe the surface morphology of RGD- and Mg+-decorated zirconia coatings; surface roughness and wettability were also measured. The initial adhesion of osteoblasts was measured, and cell morphology and focal adhesion were observed. In addition, the expressions of the integrins a1, a2, a5, av, and ß1 were measured using RT-PCR. A cell count was conducted to measure proliferation. The expressions of ALP and OCN were detected based on a western blot analysis, and mineralized nodules were observed to visualize the mineralization of osteoblasts. A nanoscale surface structure could be found on the Mg+-decorated zirconia coating, and the RGD-decorated zirconia coating showed better wettability (p < 0.05). Cells on the RGD- and Mg+-decorated zirconia coating possessed better spreading properties than did cells on nondecorated surfaces, and more focal adhesion was observed. The higher expressions of the integrins a5, av and ß1 were found on the RGD-decorated zirconia coating (p < 0.05). The western blot results demonstrated that the introduction of Mg+ heightened the expressions of ALP and OCN. More and bigger mineralized nodules were observed on the Mg+- and RGD-decorated zirconia coating, which consisted of small mineralized nodules. RGD- and Mg+-functionalized zirconia coating facilitates the osteogenic reaction of osteoblasts. RGD improves the adhesion of osteoblasts, and Mg+ benefits the mineralization of osteoblasts. In addition, a synergistic effect was found between RGD and Mg+, allowing better performances with regard to adhesion, proliferation and mineralization when the two were used together rather than as separate decorations.

Surface Immobilization of TiO2 Nanotubes with Bone Morphogenetic Protein-2 Synergistically Enhances Initial Preosteoblast Adhesion and Osseointegration.

Although titanium (Ti) alloys have been widely used as implant materials, the bioinertness of pristine Ti impairs their bioactivity and early osseointegration. In the present work, we prepared TiO2 nanotubes (TNT) layer on the titanium (Ti) surface by anodic oxidation. The anodized surface was functionalized with human bone morphogenetic protein-2 coating to form the hBMP-2/TNT surface. The release behavior of hBMP-2 on the hBMP-2/TNT surface displayed a controlled and sustained pattern, compared to that on the hBMP-2/Ti surface, which showed a rapid release. In vitro cellular activity tests demonstrated that both TNT and hBMP-2/Ti surfaces, particularly the hBMP-2/TNT surface, enhanced adhesion, proliferation, and differentiation of osteoblast cells. Increased cell adhesion, improved cytoskeleton organization, and immunofluorescence staining of vinculin were observed on the modified surfaces. The TNT, hBMP-2/Ti, and hBMP-2/TNT surfaces, especially the hBMP-2/TNT surface, further displayed an upregulated gene expression of adhesion and osteogenic markers vinculin, collagen type 1, osteopontin, and osteocalcin, compared to the pristine Ti surface. In vivo experiments using a rat model demonstrated that the TNT and hBMP-2/Ti surfaces, in particular the hBMP-2/TNT surface, improved osseointegration and showed a superior bone bonding ability compared to Ti. Our study revealed a synergistic role played by TiO2 nanotubes nanotopography and hBMP-2 in promoting initial osteoblast adhesion, proliferation, differentiation, and osseointegration, thus suggesting a promising method for better modifying the implant surface.

Improved osteoblast adhesion and osseointegration on TiO2 nanotubes surface with hydroxyapatite coating.

To improve initial osteoblast adhesion and subsequent osseointegration, TiO2 nanotubes layer was constructed on the titanium (Ti) surface by anodic oxidation (AO), with an additional hydroxyapatite (HA) coating to form the AO/HA surface. Tests on in vitro cellular activity displayed that the AO surface, especially the AO/HA surface, promoted initial adhesion, proliferation and differentiation of osteoblast cells. The modified AO and AO/HA surfaces further presented an up-regulated gene expression of osteogenic and adhesion markers collagen type 1 (COL), osteopontin (OPN), osteocalcin (OCN) and vinculin. In addition, in vivo experiments with a rat model demonstrated that the AO surface, particularly the AO/HA surface, achieved earlier osseointegration and a superior bone bonding ability compared with Ti. Our study shed light on a synergistic role played by nanotopography and HA in promoting osteoblast adhesion, proliferation, differentiation and osseointegration, thus suggesting a promising method for better modifying the implant surface.

Initial osteoblast adhesion and subsequent differentiation on zirconia surfaces are regulated by integrins and heparin-sensitive molecule.

PurposeIt is well known that zirconia materials have good biocompatibility; however, little is known regarding the mechanism by which cells attach to these materials. The purpose of this study is to elucidate the mechanism of cell attachment.Materials and methodsIn this study, we examined the surface characteristics of ceria-stabilized zirconia/alumina nanocomposite (NANOZR), yttria-stabilized zirconia (Y-TZP) and commercially pure titanium (CpTi), and we evaluated the initial response of osteoblast-like cells to them with different inhibitors.ResultsUnder the same polishing treatment, the three materials, NANOZR, Y-TZP and CpTi, show similar surface wettability but different surface roughness. Osteoblasts could adhere to the surface of all three materials, and spindle shapes were clearer in serum-containing media compared to PBS and serum-free culture media, suggesting that serum-contained proteins are helpful for the initial cell adhesion and spreading. Cell adhesion and proliferation were disrupted in the presence of EDTA. RGD-peptide interfered with cell proliferation by affecting cell protrusion and stress fibers. Monoclonal antibody against non-RGD type integrin α2β1 enhanced proliferation in Y-TZP, CpTi and culture dish but not in NANOZR. Cell proliferation on NANOZR was specifically inhibited in the presence of heparin. Furthermore, under heparin administration, spindle shape formation was maintained but actin cytoskeleton was disrupted, resulting in loose cellular spreading.ConclusionThese results suggest that RGD type integrins and heparin-sensitive protein in coordination regulate cell morphology and proliferation on NANOZR, through the regulation of cell polarity and stress fiber formation, respectively.

Micro-Nanofibrillar Polycaprolactone Scaffolds as Translatable Osteoconductive Grafts for the Treatment of Musculoskeletal Defects without Infection.

The treatment of musculoskeletal defects is currently limited by the tissue-regenerative materials available to orthopedic surgeons: autologous bone grafts only have a finite amount of harvestable material within a given patient, while allografts are prone to severe immunological complications and host rejection. With this motivation, the production of poly(ε-caprolactone) (PCL) scaffolds as synthetic, biomimetic biomaterials was investigated, with a specific focus on potential orthopedic translation. PCL scaffolds were produced through three different fabrication techniques: electrospinning (ES), rotary jet spinning (RJS), and airbrush (AB). ES and RJS were observed to produce microfibrillar scaffolds, while all AB products were nanofibrous. Osteoblast viability, within the PCL scaffolds, and the osteogenic phenotype were assessed in vitro through a combination of adherence, metabolic activity, proliferation, gene expression, alkaline phosphatase bioactivity, and calcium deposition assays. While the polymeric scaffolds induced slight reductions in initial osteoblast adhesion and metabolic activity, seeded cells were able to proliferate and demonstrate the bone formation phenotype. AB products demonstrated reduced bacterial surface colonization when inoculated with both Gram-positive (Staphylococcus aureus) and Gram-negative (Pseudomonas aeruginosa) bacterial strains, in comparison to the microfibrous ES and RJS products, without any small-molecule antibiotics, antimicrobial peptides, or reactive nanomaterials included during scaffold synthesis.

Surface engineering of titanium implants with enzyme-triggered antibacterial properties and enhanced osseointegration in vivo.

Preventing bacterial infection and improving the osseointegration of titanium (Ti) and its alloys are both highly crucial factors for their long-term successful implantation in clinical applications. However, the straightforward applications of antibacterial surfaces on Ti-based materials remain limited due to their side effects on cytocompatibility. Herein, catechol-functionalized multilayer films composed of dopamine-modified hyaluronic acid (HA-c) and 3,4-dihydroxyhydrocinnamic acid-modified chitosan (Chi-c) were developed on Ti substrates modified with TiO2 nanotube arrays loaded with an antibacterial drug. The treated Ti substrate showed strong hydrophilicity, with a water contact angle of about 20°, and obviously inhibited early-stage bacterial adhesion. Moreover, this system displayed an enzyme-responsive release of antibacterial drug triggered by the hyaluronidase degradation of HA-c, which exhibited effective antibacterial ability and eliminated side effects caused by burst release of antibiotics. Meanwhile, the modified Ti substrates significantly promoted initial osteoblast adhesion through up-regulating the expression of adhesion-related genes, including integrin αv and β3. More importantly, this prepared coating with bacterial self-responsiveness improved osseointegration and prevented bacterial infection of Ti implants in vivo. Overall, our developed catechol-functionalized and bacterial self-responsive coating on Ti substrate has great significance in clinical applications of orthopedic and dental implants.

Biodegradable Ceramics Consisting of Hydroxyapatite for Orthopaedic Implants

This study aims to analyze hydroxyapatite (HAP) coatings enriched with Mg and Ti prepared by a magnetron sputtering technique on Ti6Al4V substrate. For preparation of the coatings, three magnetron targets (HAP, MgO and TiO2) were simultaneously co-worked. The concentration of Mg added was varied by modifying the power applied to the MgO target. In all coatings, the Ti concentration was maintained constant by keeping the same cathode power fed during the whole deposition. The influence of different Mg dopant contents on the formation of phase, microstructure and morphology of the obtained Ti-doped HAP coatings were characterized by Fourier transform infrared spectroscopy (FTIR) and scanning electron microscopy (SEM). Moreover, the effects of Mg addition upon corrosion, mechanical and biological properties were also investigated. Mg- and Ti-doped HAP coating obtained at low radio-frequency (RF) power fed to the MgO target provided material with high corrosion resistance compared to other coatings and bare alloy. A slight decrease in hardness of the coatings was found after the Mg addition, from 8.8 to 5.7 GPa. Also, the values of elastic modulus were decreased from 87 to 53 GPa, this being an advantage for biomedical applications. The coatings with low Mg concentration proved to have good deformation to yielding and higher plastic properties. Biological test results showed that the novel surfaces exhibited excellent properties for the adhesion and growth of bone cells. Moreover, early adherent vital cell numbers were significantly higher on both coatings compared to Ti6Al4V, suggesting that Mg ions may accelerate initial osteoblast adhesion and proliferation.

TiNxOy coatings facilitate the initial adhesion of osteoblasts to create a suitable environment for their proliferation and the recruitment of endothelial cells

Titanium-nitride-oxide coatings (TiNxOy) improve osseointegration of endosseous implants. The exact mechanisms by which these effects are mediated are poorly understood except for an increase of osteoblast proliferation while a high degree of differentiation is maintained. One hypothesis holds that TiNxOy facilitates the initial spreading and adhesion of the osteoblasts. The aim of this work was to investigate the molecular mechanisms of osteoblast adhesion on TiNxOy as compared to microrough titanium SLA. A global view of the osseointegrative process, that is, taking into account other cell groups, especially endothelial cells, is also presented. To this aim, gene expression and focal adhesion analysis, cocultures and wound assays were performed early after seeding, from 6 h to 3 days. We demonstrated that TiNxOy coatings enhance osteoblast adhesion and spreading when compared to the standard microrough titanium. The integrin β1, either in association with α1 or with α2 plays a central role in these mechanisms. TiNxOy coatings optimize the process of osseointegration by acting at several levels, especially by upregulating osteoblast adhesion and proliferation, but also by supporting neovascularization and the development of a suitable inflammatory environment.

Enhancing the regeneration of bone defects by alkalizing the peri-implant zone – anin vitroapproach

AbstractThe effects of alkaline pH on the initial adhesion of osteoblasts to titanium surfaces was analyzed by single cell force microscopy (SCFM). In the SCFM measurements, the same cells were used to compare their unspecific adhesion to uncoated titanium with their specific adhesion to collagen coated titanium. When the maximum detachment forces (MDFs) were compared at pH 7.4 and 8.0, only slight differences were found on pure titanium, while the MDFs were significantly increased at collagen coated surfaces at pH 8.0. Effects on the subsequent proliferation and gene expression were investigated in anin vitromodel system consisting of an alkalizing polyvinyl alcohol (PVA) matrix and a perforated titanium disc. The sodium hydroxide releasing matrix maintained the medium pH between pH 7.6 and pH 8.4 during the entire experiment. Under these conditions, cell counts were significantly increased with respect to the control system after 7 days in culture. These results were supported by gene expression analyses, which showed an upregulation of proliferation-controlling genes of the EGFR1 and PI3K/AKT pathways after 14 days in culture. The SCFM data were complemented by findings of an intensive regulation of genes known to be associated with focal adhesion such as Itga8 and Tnn.

Silk fibroin nanoparticles support in vitro sustained antibiotic release and osteogenesis on titanium surface

Increasing amounts of metal-based implants are used for orthopedic or dental surgeries throughout the world. Still several implant-related problems such as inflammation, loosening and bacterial infection are prevalent. These problems stem from the immediate microbial contamination and failure of initial osteoblast adhesion. Additionally, bacterial infections can cause serious and life-threatening conditions such as osteomyelitis. Here, antibiotic (gentamicin)-loaded silk protein fibroin (non-mulberry silkworm, Antheraea mylitta) nanoparticles are fabricated and deposited over the titanium surface to achieve sustained drug release in vitro and to alter the surface nano-roughness. Based on the altered surface topography, chemistry and antibacterial activity, we conclude that the nanoparticle-deposited surfaces are superior for osteoblast adhesion, proliferation and differentiation in comparison to bare Ti. This method can be utilized as a cost-effective approach in implant modification. From the Clinical EditorTitanium-based implants are commonly used in the field of orthopedics or dentistry. Surface modification of an implant is vital to ensure osseointegration. In this article, the author investigated the use of silk protein fibroins for metal surface modification and also for drug delivery against bacteria. The encouraging data should provide a new method in terms of nanotechnology in the respective clinical fields.

One-day treatment of small molecule 8-bromo-cyclic AMP analogue induces cell-based VEGF production for in vitro angiogenesis and osteoblastic differentiation.

Small molecule-based regenerative engineering is emerging as a promising strategy for regenerating bone tissue. Small molecule cAMP analogues have been proposed as novel biofactors for bone repair and regeneration and, while promising, the effect that these small molecules have on angiogenesis, a critical requirement for successful bone regeneration, is still unclear. Our previous research demonstrated that the small molecule cAMP analogue 8-bromoadenosine-3',5'-cyclic monophosphate (8-Br-cAMP) was able to promote initial osteoblast adhesion on a polymeric scaffold via cAMP signalling cascades. Here, we report that 8-Br-cAMP is capable of inducing in vitro cell-based VEGF production for angiogenesis promotion. We first demonstrated that treating osteoblast-like MC3T3-E1 cells with 8-Br-cAMP for 1 day significantly increased VEGF production and secretion. We then demonstrated that 8-Br-cAMP-induced cell-secreted VEGF is biologically active and may promote angiogenesis, as evidenced by increased human umbilical vein endothelial cells (HUVECs) migration and tubule formation. In addition, treatment of MC3T3-E1 cells with 8-Br-cAMP for as short as a single day resulted in enhanced ALP activity as well as matrix mineralization, demonstrating in vitro osteoblastic differentiation. A short-term 8-Br-cAMP treatment also addresses the concern of non-specific cytotoxicity, as our data indicate that a 1-day 8-Br-cAMP treatment scheme supports cellular proliferation of MC3T3-E1 cells as well as HUVECs. While the major concern associated with small molecule drugs is the risk of non-specific cytotoxicity, the short exposure treatment outlined in this paper provides a very promising strategy to mitigate the risk associated with small molecules. Copyright © 2013 John Wiley & Sons, Ltd.

Effects of the surface characteristics of nanoporous titanium oxide films on Ti-24Nb-4Zr-8Sn alloy on the initial adhesion of osteoblast-like MG-63 cells

The aim of the present study was to investigate the effects of the surface characteristics of nanoporous titanium oxide films, formed by anodization on Ti-24Nb-4Zr-8Sn (Ti2448) alloy, on the early adhesion of osteoblast-like MG-63 cells. Nanoporous titanium oxide films with two different pore sizes (30 and 90 nm) were formed by anodization in NH4F solution on Ti2448 alloy. The surface roughness of the nanoporous titanium oxide films was determined using a Surftest Formtracer and field emission scanning electron microscopy (FESEM). Cell viability was evaluated at different time points using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. To investigate the regulatory mechanisms involved in the focal adhesion of osteoblasts to Ti2448 alloy, we quantified the expression levels of integrin β1 and paxillin mRNAs on the nanoporous titanium oxide films during early osteoblast adhesion using real-time RT-PCR. Samples with a 30-nm nanoporous film exhibited a greater number of overlapping microporous structures with microprojections compared with the 90-nm nanoporous film samples. The MTT assay indicated that cell viability on the 30-nm nanoporous surface following 24 and 48 h of cell culture was higher than those observed on the unanodized control and 90-nm nanoporous surfaces. Integrin β1 mRNA expression levels on the 30-nm nanoporous surface following cell culture for 48 h were also significantly higher compared with those on the unanodized control and 90-nm nanoporous surfaces. The results demonstrated that a 30-nm nanoporous titanium oxide film on Ti2448 alloy may provide the optimum bioactive implant surface for the initial adhesion of osteoblasts.

Selectively Promoting or Preventing Osteoblast Growth on Titanium Functionalized with Polyelectrolyte Multilayers

AbstractTitanium plays an important role in medical applications, such as hip joint implants or fixation plates. These implants must perform differently depending on their clinical application. In particular, the osseointegrative properties required of the implant vary with clinical application. The present work is aimed at the functionalization of titanium surface using polyelectrolyte multilayers consisting of natural biopolymers and testing their cell adhesive properties with respect to the osseointegration capacity.Multilayered coatings were created from chitosan (Chi), hyaluronic acid (HA), and gelatine (Gel) through layer‐by‐layer deposition. Cell adhesion, proliferation, and viability were tested in vitro with the human osteoblast cell line CAL‐72 at timescales up to 7 d. Two multilayer coatings consisting of alternated chitosan/gelatin or chitosan/hyaluronic acid layers with the outmost layer of gelatin (Chi/Gel) or hyaluronic acid (Chi/HA), respectively, were tested. The experimental results showed that surfaces functionalized with Chi/Gel and Chi/HA multilayers demonstrated a good initial adhesion of osteoblasts. After 4 d culturing, osteoblast cells were almost completely detached from the substrates functionalized with Chi/HA multilayers. In contrast to Chi/HA, the proliferation of osteoblasts on substrates with Chi/Gel multilayer coatings was statistically significantly higher compared to the control titanium.We have shown that the growth of osteoblasts can be enhanced or completely prevented on a titanium surface functionalized with polyelectrolyte multilayers consisting of natural biopolymers, as desired. Both multilayer coatings, Chi/Gel and Chi/HA, have potential for applications in the field of titanium implants, where rapid osseointegration is essential, and/or where no ingrowth of the implant is desired, respectively.

Development and characterization of nanofibrous poly(lactic-co-glycolic acid)/biphasic calcium phosphate composite scaffolds for enhanced osteogenic differentiation

Poly(lactic-co-glycolic acid)(PLGA)/biphasic calcium phosphate (BCP) composite nanofibers with different BCP to PLGA ratios were fabricated using the electrospinning technique. The scanning electron microscopy (SEM) images showed a similar morphology and fibers in all groups. The incorporated BCP was dispersed homogenously throughout the nanofibers, and the surface roughness was affected by the input amount of BCP. The increase in amount of BCP incorporated was confirmed by several methods. BCP incorporation into the PLGA nanofibers did not affect the initial adhesion of osteoblasts and their adherent morphology. However, the proliferation of the cells cultured on the composite nanofibers for 10 days with larger amounts of BCP was delayed, suggesting that incorporated BCP may facilitate the switch from proliferation to differentiation of the osteoblasts. The incorporation of BCP enhanced the expression of osteogenic genes, as well as induced calcium deposition by the osteoblasts in the extracellular matrix(ECM) after 21 days of culture on the PLGA/BCP composite nanofibers. Overall, these results can provide evidence of the potential of BCP incorporation into the biomaterials for effective bone regeneration.

Effects of nano-surface properties on initial osteoblast adhesion and Ca/P adsorptionability for titanium alloys

Titanium alloys (Ti6Al4V), while subjected to high temperature surface treatment,experience altered nano-surface characteristics. The effects of such surface treatments areexamined, including the initial adhesion force experienced by osteoblasts, the Ca/Padsorption capability, and the nano-surface properties, including the amounts ofamphoteric Ti–OH groups, surface topography, and surface roughness. The initial adhesionforce is considered a quantitative indicator of cyto-compatibility in vitro. Previously, acyto-detacher was applied in a pioneer attempt measuring the initial adhesionforce of fibroblasts on a metal surface. Presently, the cyto-detacher is furtherapplied to evaluate the initial adhesion force of osteoblasts. Results reveal that(1) titanium alloys subjected to heat treatment could promote the adsorption capabilityof Ca and P; (2) titanium alloys subjected to heat treatment could have higherinitial osteoblast adhesion forces; (3) the adhesion strength of osteoblasts, rangingfrom 38.5 to 58.9 nN (nanonewtons), appears stronger for rougher surfaces. It isconcluded that the heat treatment could have impacted the biocompatibility interms of the initial osteoblast adhesion force and Ca/P adsorption capability.

Increased osteoblast adhesion on nanograined Ti modified with KRSR

Peptide sequences such as lysine-arginine-serine-arginine (KRSR) selectively bind transmembrane proteoglycans (e.g. heparin sulfate) of osteoblasts (bone-forming cells) and are, therefore, actively being investigated for orthopedic applications. Further, nanophase materials (or materials with grain or particle sizes less than 100 nm) are promising new materials that promote new bone growth more than compared to conventional (that is, micron grain or particle size) materials. To combine the above two promising approaches for improving orthopedic implants, the objective of this in vitro study was to functionalize titanium (Ti) surfaces (both nanophase and conventional) with KRSR peptides and study their osteoblast cell adhesive properties. Materials were characterized by X-ray photoelectron spectroscopy, scanning electron microscopy, and atomic force microscopy. Results of this in vitro study provided evidence of increased osteoblast adhesion on nanophase compared to conventional Ti whether functionalized with KRSR or not. Results further showed that the immobilization of KRSR onto Ti (both nanophase and conventional) increased osteoblast adhesion compared to respective nonfunctionalized Ti and those functionalized with the negative control peptide KSRR. Most importantly, osteoblast adhesion on nonfunctionalized nanophase Ti increased compared to conventional Ti functionalized with KRSR. Further, select initial osteoblast adhesion was observed to occur at particle boundaries for any type of nanophase and conventional Ti formulated in this study. In summary, results provided evidence that not only should nonfunctionalized nanophase Ti be further studied for improved orthopedic applications but so should nanophase Ti functionalized with KRSR.

Influence of surface pretreatment of titanium- and cobalt-based biomaterials on covalent immobilization of fibrillar collagen

Collagen type-I is a major component of the extracellular matrix of most tissues and it is increasingly utilized for surface engineering of biomaterials to accelerate receptor-mediated cell adhesion. In the present study, coatings with layers of fibrillar type-I collagen were prepared on titanium, titanium alloy, and cobalt alloy to improve initial osteoblast adhesion and implant–tissue integration. To suppress the quick in vivo degradation rate of collagen the deposited layers were covalently immobilized at the metal surfaces as well as chemically cross-linked. The application of different oxidation techniques to the metallic substrates resulted in surfaces with varying hydroxyl group contents, which directly influenced the amount of immobilized silane coupling agents. It was found that a high density of surface-bound coupling agents increased the stability of the covalently linked collagen layers. After coating of metallic biomaterials with a cross-linked collagen layer, an improved cellular response of human osteoblast-like cells (MG-63) in vitro could be recognized.