Smooth Titanium Surfaces Research Articles

The Effect of Titanium Surface Topography on Adherent Macrophage Integrin and Cytokine Expression

Immunomodulatory biomaterials have the potential to stimulate an immune response able to promote constructive and functional tissue remodeling, as opposed to persistent inflammation and scar tissue formation. This study examined the effects of titanium surface modification on integrin expression and concurrent cytokine secretion by adherent macrophages in vitro in an attempt to delineate the molecular events involved in biomaterial-mediated immunomodulation. Non-polarised (M0) and inflammatory polarised (M1) macrophages were cultured on a relatively smooth (machined) titanium surface and two proprietary modified rough titanium surfaces (blasted and fluoride-modified) for 24 h. The physiochemical characteristics of the titanium surfaces were assessed by microscopy and profilometry, while macrophage integrin expression and cytokine secretion were determined using PCR and ELISA, respectively. After 24 h adhesion onto titanium, integrin α1 expression was downregulated in both M0 and M1 cells on all titanium surfaces. Expression of integrins α2, αM, β1 and β2 increased in M0 cells cultured on the machined surface only, whereas in M1 cells, expression of integrins α2, αM and β1 all increased with culture on both the machined and rough titanium surfaces. These results correlated with a cytokine secretory response whereby levels of IL-1β, IL-31 and TNF-α increased significantly in M1 cells cultured on the titanium surfaces. These results show that adherent inflammatory macrophages interact with titanium in a surface-dependent manner such that increased levels of inflammatory cytokines IL-1β, TNF-α and IL-31 secreted by M1 cells were associated with higher expression of integrins α2, αM and β1.

Effect of graphene-oxide-modified osteon-like concentric microgrooved surface on the osteoclastic differentiation of macrophages.

This study aimed to investigate the effect of new biomimetic micro/nano surfaces on the osteoclastic differentiation of RAW264.7 macrophages by simulating natural osteons for the design of concentric circular structures and modifying graphene oxide (GO). The groups were divided into smooth titanium surface group (SS), concentric microgrooved titanium surface group (CMS), and microgroove modified with GO group (GO-CMS). The physicochemical properties of the material surfaces were studied using scanning electron microscopy (SEM), contact-angle measurement, atomic force microscopy, X-ray photoelectron spectroscopy analysis, and Raman spectroscopy. The effect of the modified material surface on the cell biological behavior of RAW264.7 was investigated by cell-activity assay, SEM, and laser confocal microscopy. The effect on the osteoclastic differentiation of macrophages was investiga-ted by tartrate-resistant acid phosphatase (TRAP) immunofluorescence staining and quantitative real-time polymerase chain reaction (qRT-PCR) experiments. Macrophages were arranged in concentric circles along the microgrooves, and after modification with GO, the oxygen-containing groups on the surface of the material increased and hydrophilicity increased. Osteoclasts in the GO-CMS group were small in size and number and had the lowest TRAP expression. Although it promoted the proliferation of macrophages in the GO-CMS group, the expression of osteoclastic differentiation-related genes was lower than that in the SS group, and the difference was statistically significant (P<0.05). Concentric circular microgrooves restricted the fusion of osteoclasts and the formation of sealing zones. Osteomimetic concentric microgrooves modified with GO inhibited the osteoclastic differentiation of RAW 264.7 macrophages.

Antibacterial activity and cytocompatibility evaluation of the antimicrobial peptide Nal-P-113-loaded graphene oxide coating on titanium.

The initialization of the gingival crevicular barrier epithelium facing the exposed implant surface is reported to take 1-2 weeks, and peri-implant inflammation mediated by bacterial infection has been an increasing concern. In this study, we developed a novel composite coating containing graphene oxide (GO) and the antimicrobial peptide (AMP) Nal-P-113 on smooth titanium surface, and then investigated the antibacterial properties and cytocompatibility of the coating. The results showed that Nal-P-113 exhibited a slow and continuous drug release in vitro from the composite coating. The Nal-P-113-loaded GO coating demonstrated excellent antibacterial properties against both Gram-positive Streptococcus mutans (S. mutans) and Gram-negative Porphyromonas gingivalis (P. gingivalis), without no obvious cytotoxicity to human gingival fibroblast (HGF) cells. The Nal-P-113-loaded GO coating need further optimization for anti-infective and wound healing promoting purpose of peri-implant tissues.

Effects on the Titanium Implant Surface by Different Hygiene Instrumentations: A Narrative Review.

Peri-implant disease is usually caused by the accumulation of dental biofilm around the implant, and this biofilm can irradiate the gingiva tissue, which leads to inflammation and, more severely, to a deterioration of the bone structure. There is a concern regarding the removal of biofilm from the implant surface by using different hygiene instruments. Some hygiene instruments may have some effect on the dental implant surface, resulting in roughening or damage to the implant surfaces. This study reviewed the effects of titanium implant surfaces on different hygiene instruments. A literature search was conducted from PubMed, ScienceDirect, and Scopus databases for articles published from 1992 to 2021. A total of 19 full-text papers with keywords of interest that met all the eligibility criteria were selected. Surface roughness was evaluated with a scanning electron microscope and also using a profilometer, laser scanning, scanning probe, and atomic force microscopes. A metal curette produced a roughened surface on the titanium implant, but a plastic curette did not alter the surface. Instrumentation with rubber cups left the surface unchanged and appeared to smoothen the surface, whereas the air-powder abrasive instrumentation altered the surface with the presence of micro pits and pores. A conventional metal ultrasonic scaler showed significant surface topographical changes and scratches on both titanium surfaces, as a diode laser, light-emitting diode (LED), and laser treatment did not show any alteration on the rough and smooth titanium surfaces. Thus, a non-metallic instrument such as a plastic curette, rubber cups, and novel technology including diode laser, LED, and laser treatment is appropriate and can be used for debridement on smooth and machined titanium implant surfaces as well as sandblasted and acid-etched (SLA), titanium plasma-sprayed (TPS), and resorbable blasted media (RBM) surfaces. The use of metallic instruments should be avoided, and it is not recommended.

Effects of Titanium Implant Surface Topology on Bone Cell Attachment and Proliferation in vitro.

PurposeTitanium is commonly used for implants because of its corrosion resistance and osseointegration capability. It is well known that surface topology affects the response of bone tissue towards implants. In vivo studies have shown that in weeks or months, bone tissue bonds more efficiently to titanium implants with rough surfaces compared to smooth surfaces. In addition, stimulating early endosseous integration increases the long-term stability of bone-implants and hence their clinical outcome. Here, we evaluated the response of human MG-63 osteoblast-like cells to flat and solid, compared to rough and porous surface topologies in vitro 1–6 days post seeding. We compared the morphology, proliferation, and attachment of cells onto three smooth surfaces: tissue culture (TC) plastic or microscope cover glasses, machined polyether-ether-ketone (PEEK), and machined solid titanium, to cells on a highly porous (average Ra 22.94 μm) plasma-sprayed titanium surface (composite Ti-PEEK spine implants).MethodsWe used immuno-fluorescence (IF) and scanning electron microscopy (SEM), as well as Live/Dead and WST-1 cell proliferation assays.ResultsSEM analyses confirmed the rough topology of the titanium implant surface, compared to the smooth surface of PEEK, solid titanium, TC plastic and cover glasses. In addition, SEM analyses revealed that MG-63 cells seeded onto smooth surfaces (solid titanium, PEEK) adopted a flat, planar morphology, while cells on the rough titanium surface adopted an elongated morphology with numerous filopodial and lamellipodial extensions interacting with the substrate. Finally, IF analyses of focal adhesions (vinculin, focal adhesion kinase), as well as proliferation assays indicate that MG-63 cells adhere less and proliferate at a slower rate on the rough than on a smooth titanium surface.ConclusionThese observations suggest that bone-forming osteoblasts adhere less strongly and proliferate slower on rough compared to smooth titanium surfaces, likely promoting cell differentiation, which is in agreement with other porous implant materials.

Surface analysis of titanium disks with strontium coating.

Peri-implantitis is one of the most common complications after dental implant placement. Researchers have demonstrated that the peri-implantitis tends to occur around dental implants with a rough surface rather than those with a smooth surface. We aimed to investigate the ability of a smooth titanium (Ti) surface containing strontium (Sr) to enhance bone formation as a result of strontium's capacity to support osteoblast proliferation and differentiation. A thin titanium oxide film was formed on an as-mirror polished Ti surface by dipping in 5% sodium hypochlorite (NaOCl) solution for 24 h, followed by thermal treatment at 350°C. The Ti surface was then treated with 1% strontium nitrate (Sr(NO3)2) solution and turned in spin coater. The surface morphology, chemical composition, and release of strontium ions (Sr2+) were evaluated. The results demonstrate that strontium in the form of Sr2+ was successfully doped into the titanium dioxide (TiO2) film by this simple chemical treatment.

Early Biofilm Formation on Rough and Smooth Titanium Specimens: a Systematic Review of Clinical Studies.

ABSTRACTObjectivesThere is a concern whether the enhancement on implant surface roughness is responsible for higher biofilm formation, which acts as an aetiological factor for peri-implant diseases. The aim of the present systematic review was to answer the following question: “Are rough surfaces more susceptible to early biofilm formation when compared to smoother surfaces on titanium specimens?”.Material and MethodsThe research was performed on PubMed, Web of Science and Scopus, up to August 2021. Eligibility criteria included studies that analysed human biofilm formation on titanium specimens with distinct surface roughness (smooth vs minimally, moderate, or rough) over the experimental times of 1 or 3 days. Roughness average (Ra) and biofilm analysis parameters were extracted from selected articles. Risk of bias was evaluated using the Checklist for Quasi-Experimental Studies.ResultsOf 5286 papers, 5 were included and analysed. Smooth titanium surfaces included machined and anodized titanium/Ti-6Al-4V; machined and acid etched TiZr. Minimally, moderately, or rough surfaces comprised titanium and titanium alloys (TiZr, Ti-6Al-4V), that received surface treatments (anodization, acid-etching, blasting, hydroxyapatite-coating). No statistically significant difference on biofilm formation on rough and smooth titanium surfaces was reported by 3 studies, while more contamination on rough titanium surfaces was stated by 2 investigations. An isolated smooth surface has also been associated to higher contamination. Moderate to high quality methodological assessment of studies were identified.ConclusionsIt is not possible to assume that rough surfaces are more susceptible to early biofilm formation than smooth titanium surfaces. Additional studies are required to study this multifarious interaction.

Anti-inflammatory effects of the immobilization of SEMA4D on titanium surfaces in an endothelial cell/macrophage indirect coculture model

In this study, we established a procedure to prepare a Semaphorin4D (SEMA4D)-immobilized titanium surface and explored its effects on macrophage behaviors in an endothelial cell/macrophage indirect coculture model. The SEMA4D-bovine serum albumin complex was immobilized onto a preprocessed poly L-lysine titanium surface through NaOH hydrothermal treatment and self-assembly technology. All titanium specimens were examined for surface microstructure, surface element composition, and surface wettability by field emission scanning electron microscopy, x-ray photoelectron spectroscopy (XPS), and water contact angle measurement, respectively. Subsequently, we constructed an endothelial cell/macrophage indirect coculture model and evaluated the activation of NF-κB signaling pathway and the expression of proinflammatory cytokines (TNFα, IL-6, and IL-1β) in macrophages. In XPS analysis, the SEMA4D-immobilized titanium surface appeared as a loose porous structure covered with uniform film, which exhibited better hydrophilicity than the control smooth titanium surface. In the indirect coculture model, SEMA4D attenuated the activation of NF-κB signaling pathway of lipopolysaccharide-stimulated THP-1 macrophages, thereby downregulating the expression of proinflammatory cytokines in macrophages. In conclusion, SEMA4D could be immobilized on titanium surfaces through NaOH hydrothermal treatment and self-assembly technology. Meanwhile, SEMA4D immobilization altered the characteristics of the titanium surfaces, which negatively regulated macrophage behaviors in the endothelial cell/macrophage indirect coculture model.

Effectivity of homecare and professional biofilm removal procedures on initial supragingival biofilm on laser-microtextured implant surfaces in an ex vivo model

BackgroundThe aim of the current study was the evaluation of initial biofilm adhesion and development on laser-microtextured implant collar surfaces and the examination of effectivity of different biofilm management methods.MethodsInitial biofilm formation was investigated on hydrophobic machined and laser-microtextured (Laser-Lok) titanium surfaces and hydrophobic machined and laser-microtextured (Laser-Lok) titanium aluminium vanadium surfaces and compared to hydrophobic smooth pickled titanium surfaces, hydrophilic smooth and acid etched titanium surfaces, hydrophobic sandblasted large grid and acid etched titanium surfaces (titanium Promote) via erythrosine staining and subsequent histomorphometrical analysis and scanning electron microscopic investigations. After decontamination procedures, performed via tooth brushing and glycine powder blasting, clean implant surface was detected via histomorphometrical analysis.ResultsAfter 24 h mean initial plaque area was detected in the following descending order: smooth pickled titanium > titanium Promote > hydrophilic smooth and acid etched titanium > Laser-Lok titanium > Laser-Lok titanium aluminium vanadium. The same order was determined after 48 h of biofilm formation. After glycine powder blasting all samples depicted almost 100% clean implant surface. After tooth brushing, Laser-Lok titanium (67.19%) and Laser-Lok titanium aluminium vanadium (69.80%) showed significantly more clean implant surface than the other structured surfaces, hydrophilic smooth and acid etched titanium (50.34%) and titanium Promote (33.89%). Smooth pickled titanium showed almost complete clean implant surface (98.84%) after tooth brushing.ConclusionsBoth Laser-Lok surfaces showed less initial biofilm formation after 24 and 48 h than the other implant surfaces. In combination with the significant higher clean implant surfaces after domestic decontamination procedure via tooth brushing, both Laser-Lok surfaces could be a candidate for modified implant and abutment designs, especially in transmucosal areas.

The Role and Activation Mechanism of TAZ in Hierarchical Microgroove/Nanopore Topography-Mediated Regulation of Stem Cell Differentiation.

PurposeTo investigate the role and activation mechanism of TAZ in periodontal ligament stem cells (PDLSCs) perceiving hierarchical microgroove/nanopore topography.Materials and MethodsTitanium surface with hierarchical microgroove/nanopore topography fabricated by selective laser melting combined with alkali heat treatment (SLM-AHT) was used as experimental group, smooth titanium surface (Ti) and sandblasted, large-grit, acid-etched (SLA) titanium surface were employed as control groups. Alkaline phosphatase (ALP) activity assays, qRT-PCR, Western blotting, and immunofluorescence were carried out to evaluate the effect of SLM-AHT surface on PDLSC differentiation. Moreover, TAZ activation was investigated from the perspective of nuclear localization to transcriptional activity. TAZ knockdown PDLSCs were seeded on three titanium surfaces to detect osteogenesis- and adipogenesis-related gene expression levels. Immunofluorescence and Western blotting were employed to investigate the effect of the SLM-AHT surface on actin cytoskeletal polymerization and MAPK signaling pathway. Cytochalasin D and MAPK signaling pathway inhibitors were used to determine whether actin cytoskeletal polymerization and the MAPK signaling pathway were indispensable for TAZ activation.ResultsOur results showed that SLM-AHT surface had a greater potential to promote PDLSC osteogenic differentiation while inhibiting adipogenic differentiation than the other two groups. The nuclear localization and transcriptional activity of TAZ were strongly enhanced on the SLM-AHT surface. Moreover, after TAZ knockdown, the enhanced osteogenesis and decreased adipogenesis in SLM-AHT group could not be observed. In addition, SLM-AHT surface could promote actin cytoskeletal polymerization and upregulate p-ERK and p-p38 protein levels. After treatment with cytochalasin D and MAPK signaling pathway inhibitors, differences in the TAZ subcellular localization and transcriptional activity were no longer observed among the different titanium surfaces.ConclusionOur results demonstrated that actin cytoskeletal polymerization and MAPK signaling pathway activation triggered by SLM-AHT surface were essential for TAZ activation, which played a dominant role in SLM-AHT surface-induced stem cell fate decision.

MiR-181d-5p regulates implant surface roughness-induced osteogenic differentiation of bone marrow stem cells

Constructing moderate surface roughness is a widely used, non-toxic, cost-effective, and outcome-predictable approach to accelerate implant osteointegration in clinical settings. MicroRNAs (miRNAs) play vital regulatory roles in the osteogenic differentiation of bone marrow stem cells (BMSCs). However, their specific contribution to the influence of surface roughness on osteoblastic behavior remains unknown. Therefore, applying the smooth titanium surface as a control, a typical titanium surface with moderate roughness was prepared here to reveal the mechanism through which surface roughness regulates cell osteogenic behavior by altering miRNA expression. First, the morphology and roughness of two surfaces were characterized, and the enhanced osteogenic differentiation of BMSCs on rough surfaces was verified. Then, twenty-nine differentially expressed miRNAs in BMSCs cultured on different surfaces were selected via miRNA chip and corresponding functional prediction. After verifying the expression of these miRNAs using quantitative real-time polymerase chain reaction, four were considered eligible candidates. Among these, only miR-181d-5p significantly affected RUNX2 gene expression based on overexpression and knockdown experiments. From the osteogenesis-related gene and protein expression, as well as alkaline phosphatase and alizarin red experiments, we further confirmed that the downregulation of miR-181d-5p promoted osteogenic differentiation of BMSCs, and vice versa. In addition, rescue assays showed that the knockdown of miR-181d-5p improved the inferior osteogenesis observed on smooth surfaces, whereas the overexpression of miR-181d-5p suppressed the superior osteogenesis observed on rough surfaces. These results indicate that the moderate surface roughness of the implant stimulates the osteogenic differentiation of BMSCs by remarkably downregulating miR-181d-5p. These findings provide helpful information and a theoretical basis for the development of advanced implant materials for fast osteointegration.

Porous Tantalum VS. Titanium Implants: Enhanced Mineralized Matrix Formation after Stem Cells Proliferation and Differentiation.

Titanium dental implants are used routinely, with surgical procedure, to replace missing teeth. Even though they lead to satisfactory results, novel developments with implant materials can still improve implant treatment outcomes. The aim of this study was to investigate the efficiency of porous tantalum (Ta) dental implants for osseointegration, in comparison to classical titanium (Ti). Mesenchymal stem cells from the dental pulp (DPSC) were incubated on Ta, smooth titanium (STi), and rough titanium (RTi) to assess their adhesion, proliferation, osteodifferentiation, and mineralized matrix production. Cell proliferation was measured at 4 h, 24 h, 48 h with MTT test. Early osteogenic differentiation was followed after 4, 8, 12 days by alkaline phosphatase (ALP) quantification. Cells organization and matrix microstructure were studied with scanning electron microscopy (SEM) and energy dispersive X-ray spectroscopy (EDX). Collagen production and matrix mineralization were evaluated by immunostaining and histological staining. MTT test showed significantly higher proliferation of DPSC on Ta at 24 h and 48 h. However, APL quantification after 8 and 12 days was significantly lower for Ta, revealing a delayed differentiation, where cells were proliferating the more. After 3 weeks, collagen immunostaining showed an efficient production of collagen on all samples. However, Red Alizarin staining clearly revealed a higher calcification on Ta. The overall results tend to demonstrate that DPSC differentiation is delayed on Ta surface, due to a longer proliferation period until cells cover the 3D porous Ta structure. However, after 3 weeks, a more abundant mineralized matrix is produced on and inside Ta implants. Cell populations on porous Ta proliferate greater and faster, leading to the production of more calcium phosphate deposits than cells on roughened and smooth titanium surfaces, revealing a potential enhanced capacity for osseointegration.

P7. Hierarchical surface roughness produced with additive manufacturing technology significantly increases osteogenic cellular differentiation and gene expression when compared to PEEK and smooth titanium surfaces

BACKGROUND CONTEXT Spine interbody fusion devices are used to stabilize and treat diseased and degenerative spine conditions. Commonly utilized interbody fusion devices are comprised of polyetheretherketone (PEEK) or titanium (Ti). PEEK interbody devices have a hydrophobic surface that does not directly bond to bone and promotes fibrous tissue formation. In contrast, Tispinal fusion devices have favorable osseointegration properties especially for those with roughened surfaces. PURPOSE Previous studies have shown that nano-scale surface features created by subtractive grit blasting and acid etching increase osteogenic differentiation and gene expression when compared to PEEK or smooth Ti (Sm Ti) surfaces. More recently, proprietary 3D additive manufacturing processes have been used to create implants that have a hierarchical surface roughness that spans from the macro to nano-scale. The objective of this study was to evaluate whether the P3D surface increases osteogenic differentiation and gene expression when compared to PEEK or smooth Ti surfaces, as similarly seen in acid etch surfaces. STUDY DESIGN/SETTING N/A PATIENT SAMPLE N/A OUTCOME MEASURES N/A METHODS The P3D implants with hierarchical surface roughness were produced on a Q 10+ 3D printer (Arcam). Bone marrow was harvested from a BAC transgenic reporter mouse. Primary bone marrow stem cells (BMSC) were cultured under basal conditions and seeded onto P3D, Sm Ti, or PEEK constructs. Cell-constructs cultured in osteogenic condition were harvested at days 3, 7, 14 and 22. Cells were removed by accutase for fluorescence image cytometry. RNA evaluation was performed by submerging in trizol and physical scraping. cDNA synthesis and qPCR utilized sequences for 18S, osteocalcin, collagen type 1, alkaline phosphatase, osterix and PHEX. Students 2-tailed t-test was utilized to evaluate statistical differences. RESULTS Significantly more (p CONCLUSIONS Compared to PEEK and Sm Ti surfaces, the P3D rough Ti surface enabled a significantly greater proportion of BMSC to enter the osteogenic lineage and resulted in faster and significantly greater osteogenic gene expression. Similar to prior publications that demonstrated increased osteogenesis on nanotopography surfaces, P3D with macro to nano-scale surface roughness also increased osteogenesis. The increased osteogenic response from the 3DP surface features may lead to quicker bone/implant fixation which would provide a stable environment for fusion to occur and reduce the risk of complications. FDA DEVICE/DRUG STATUS This abstract does not discuss or include any applicable devices or drugs.

Biofunctionalization of Microgroove Surfaces with Antibacterial Nanocoatings.

Objectives To investigate the physical properties of the modified microgroove (MG) and antibacterial nanocoated surfaces. In addition, the biological interactions of the modified surfaces with human gingival fibroblasts (HGFs) and the antibacterial activity of the surfaces against Porphyromonas gingivalis were studied. Methods The titanium nitride (TiN) and silver (Ag) coatings were deposited onto the smooth and MG surfaces using magnetron sputtering. A smooth titanium surface (Ti-S) was used as the control. The physicochemical properties including surface morphology, roughness, and hydrophilicity were characterized using scanning electron microscopy, atomic force microscopy, and an optical contact angle analyzer. The “contact guidance” morphology was assessed using confocal laser scanning microscopy. Cell proliferation was analyzed using the Cell Counting Kit-8 assay. The expression level of the main focal adhesion-related structural protein vinculin was compared using quantitative reverse transcription PCR and Western blotting. The antibacterial activity against P. gingivalis was evaluated using the LIVE/DEAD BacLight™ Bacterial Viability Kit. Results The Ag and TiN antibacterial nanocoatings were successfully deposited onto the smooth and MG surfaces using magnetron sputtering technology. TiN coating on a grooved surface (TiN-MG) resulted in less nanoroughness and greater surface hydrophilicity than Ag coating on a smooth surface (Ag-S), which was more hydrophobic. Cell proliferation and expression of vinculin were higher on the TiN-MG surface than on the Ag-coated surfaces. Ag-coated surfaces showed the strongest antibacterial activity, followed by TiN-coated surfaces. Conclusion Nano-Ag coating resulted in good antimicrobial activity; however, the biocompatibility was questionable. TiN nanocoating on an MG surface showed antibacterial properties with an optimal biocompatibility and maintained the “contact guidance” effects for HGFs.

Epigenetic changes of osteoblasts in response to titanium surface characteristics.

We aimed to investigate the influence of titanium surface characteristics on epigenetic mechanisms and DNA damage/repair pathways. Osteoblast-like cells (MG63) were incubated on glass, smooth titanium, and minimally rough titanium discs, respectively, for 0, 1, 6, and 24 hr. The presence of double-stranded DNA damage (γH2AX), DNA repair (Chk2), and epigenetic markers (AcH3 & DNMT1) were investigated using immunofluorescence. There were no Chk2-positive cells on the minimally rough titanium surfaces at all-time points, in comparison to glass and smooth titanium. Total γH2AX-positive cells on minimally rough titanium gradually decreased as incubation time increased, on the contrary to smooth titanium. Minimally rough titanium surfaces induced cytoplasmic staining of DNMT1 up to 99% at 24 hr. For epigenetic markers related to the DNA damage/repair pathway, minimally rough titanium surfaces showed the lower percentage of AcH3-positive cells compared to glass and smooth titanium surface. The findings in the current study show that titanium surface characteristics indeed influence DNA damage and the DNA repair pathway, including epigenetic factors.

Effects of the Origin and Deacetylation Degree of Chitosan on Properties of Its Coatings on Titanium

The properties of chitosan coatings on titanium surfaces may be influenced by a variety of factors, including their chemical characteristics and the deposition method. The aim of this research was to determine the influence of a chitosan’s origin (a type of shrimp) and deacetylation degree (DD), when deposited on a very smooth titanium surface, on adhesion and biological behavior. The tests were performed using chitosan of a degree of 87% or 84% of deacetylation and that originated from armor crabs or shrimp armor. The technology of fabrication of chitosan coatings was by surface polishing to a smooth surface, oxidation in air, and immersion in a chitosan solution. The surface topographies were analyzed with an atomic force microscope and their water contact angles were measured by a falling drop method with a goniometer. The bioactivity tests were done in in vitro on osteogenic cells, type MC3T3-E1, with a biological microscope. The abrasion of the coatings was examined using a nano tribotester. The obtained results revealed that the adhesion of the coatings onto a smooth, oxidized titanium surface is appropriate as they remain sufficiently adjacent to the surface after wear tests. The source of chitin has a significant influence on biological properties, and the deacetylation degree is much less critical. The performed tests demonstrated the crucial role that the source of chitosan and the applicability of the applied surface treatment play in the preparation of chitosan coatings.

Osteoblast Responses to Titanium-Coated Subcellular Scaled Microgrooves.

Statistical data have consistently shown that implant loosening is a significant causative factor for revision surgeries. Both in vivo and in vitro studies have confirmed the positive influences of microgrooved titanium implant surfaces on improving orthopedic titanium implants compared with a smooth titanium surface. Complete cell-groove adhesion is a prerequisite for rapid and robust osseointegration. For the first time, this work has quantified the influence of the titanium groove width at the subcellular scale (5-20 μm) on osteoblast responses, using titanium-coated microgrooved silicon wafer specimens (surface roughness, Ra = ∼1.5 nm), which can avoid the latent influence of variations in surface roughness from the use of normal titanium substrates. The cell-groove adhesion increased from 53.07% to 98.55% with an increasing groove width from 5 to 20 μm. In addition, both the cell spreading area and cell width were proportional to groove width. However, no statistically significant influence (p > 0.05) of groove width was identified on cell proliferation and differentiation. An exponential model was proposed to predict the groove geometries that can facilitate complete cell-groove adhesion. The underlying mechanisms were discussed. The experimental findings of this study provide a unique basis for the design of titanium implant surfaces.

Zirconium Nitride Coating Reduced Staphylococcus epidermidis Biofilm Formation on Orthopaedic Implant Surfaces: An In Vitro Study.

One of the most commonly identified pathogens responsible for orthopaedic implant infection is Staphylococcus epidermidis, which can form biofilms on surfaces. Currently, orthopaedic implants made of various surface materials are available, each with features influencing osseointegration, biocompatibility, and adherence of bacteria to the surface, which is the first step in biofilm formation. The aim of this experimental study was to investigate the effect of a high tribologic-resistant 2.5-µm zirconium nitride top coat on an antiallergic multilayer ceramic-covered cobalt-chromium-molybdenum surface on the formation of S. epidermidis biofilm compared with other commonly used smooth and rough orthopaedic implant surface materials. (1) When evaluating the surfaces of a cobalt-chromium-molybdenum (CoCrMo) alloy with a zirconium (Zr) nitride coating, a CoCrMo alloy without a coating, titanium alloy, a titanium alloy with a corundum-blasted rough surface, and stainless steel with a corundum-blasted rough surface, does a Zr coating reduce the number of colony-forming units of S. epidermidis in an in vitro setting? (2) Is there quantitatively less biofilm surface area on Zr-coated surfaces than on the other surfaces tested in this in vitro model? To determine bacterial adhesion, five different experimental implant surface discs were incubated separately with one of 31 different S. epidermidis strains each and subsequently sonicated. Twenty test strains were obtained from orthopaedic patients undergoing emergency hip prosthesis surgeries or revision of implant infection and 10 further strains were obtained from the skin of healthy individuals. Additionally, one reference strain, S. epidermidis DSM 3269, was tested. After serial dilutions, the number of bacteria was counted and expressed as colony-forming units (CFUs)/mL. For biofilm detection, discs were stained with 0.1% Safranin-O for 15 minutes, photographed, and analyzed with computer imaging software. The lowest bacterial count was found in the CoCrMo + Zr surface disc (6.6 x 10 CFU/mL ± 4.6 x 10 SD) followed by the CoCrMo surface (1.1 x 10 CFU/mL ± 1.9 x 10 SD), the titanium surface (1.36 x 10 CFU/mL ± 1.8 x 10 SD), the rough stainless steel surface (2.65 x 10 CFU/mL ± 3.8 x 10 SD), and the rough titanium surface (2.1 x 10 CFU/mL ± 3.0 x 10 SD). The mean CFU count was lower for CoCrMo + Zr discs compared with the rough stainless steel surface (mean difference: 2.0 x 10, p = 0.021), the rough titanium alloy surface (mean difference: 1.4 x 10, p = 0.002), and the smooth titanium surface (mean difference: 7.0 x 10, p = 0.016). The results of biofilm formation quantification show that the mean covered area of the surface of the CoCrMo + Zr discs was 19% (± 16 SD), which was lower than CoCrMo surfaces (35% ± 23 SD), titanium alloy surface (46% ± 20 SD), rough titanium alloy surface (66% ± 23 SD), and rough stainless steel surface (58% ± 18 SD). These results demonstrate that a multilayer, ceramic-covered, CoCrMo surface with a 2.5-µm zirconium nitride top coat showed less S. epidermidis biofilm formation compared with other surface materials used for orthopaedic implants. CoCrMo with a 2.5-µm zirconium nitride top coat seems to be a promising surface modification technology able to reduce bacterial attachment on the surface of an implant and, hence, may further prevent implant infection with S. epidermidis biofilm formation.

Epithelial cell adhesion efficacy of a novel peptide identified by panning on a smooth titanium surface

Epithelial attachment via the basal lamina on the tooth surface provides an important structural defence mechanism against bacterial invasion in combating periodontal disease. However, when considering dental implants, strong epithelial attachment does not exist throughout the titanium-soft tissue interface, making soft tissues more susceptible to peri-implant disease. This study introduced a novel synthetic peptide (A10) to enhance epithelial attachment. A10 was identified from a bacterial peptide display library and synthesized. A10 and protease-activated receptor 4-activating peptide (PAR4-AP, positive control) were immobilized on commercially pure titanium. The peptide-treated titanium showed high epithelial cell migration ability during incubation in platelet-rich plasma. We confirmed the development of dense and expanded BL (stained by Ln5) with pericellular junctions (stained by ZO1) on the peptide-treated titanium surface. In an adhesion assay of epithelial cells on A10-treated titanium, PAR4-AP-treated titanium, bovine root and non-treated titanium, A10-treated titanium and PAR4-AP-treated titanium showed significantly stronger adhesion than non-treated titanium. PAR4-AP-treated titanium showed significantly higher inflammatory cytokine release than non-treated titanium. There was no significant difference in inflammatory cytokine release between A10-treated and non-treated titanium. These results indicated that A10 could induce the adhesion and migration of epithelial cells with low inflammatory cytokine release. This novel peptide has a potentially useful application that could improve clinical outcomes with titanium implants and abutments by reducing or preventing peri-implant disease.

In Vivo Periodontium Formation Around Titanium Implants Using Periodontal Ligament Cell Sheet.

Osseointegrated implants have been recognized as being very reliable and having long-term predictability. However, host defense mechanisms against infection have been known to be impaired around a dental implant because of the lack of a periodontal ligament (PDL). The purpose of our experimental design was to produce cementum and PDL on the implant surface adopting cell sheet technology. To this aim we used PDL-derived cells, which contain multipotential stem cells, as the cell source and we cultured them on an implant material constituted of commercially pure titanium treated with acid etching, blasting, and a calcium phosphate (CaP) coating to improve cell attachment. Implants with adhered human PDL cell sheets were transplanted into bone defects in athymic rat femurs as a xenogeneic model. Implants with adhered canine PDL-derived cell sheets were transplanted into canine mandibular bone as an autologous model. We confirmed that PDL-derived cells cultured with osteoinductive medium had the ability to induce cementum formation. The attachment of PDL cells onto the titanium surface with three surface treatments was accelerated, compared with that onto the smooth titanium surface, at 40 min after starting incubation. Results in the rat model showed that cementum-like and PDL-like tissue was partly observed on the titanium surface with three surface treatments in combination with adherent PDL-derived cell sheets. On the other hand, osseointegration was observed on almost all areas of the smooth titanium surface that had PDL-derived cell sheets, but did not have the three surface treatments. In the canine model, histological observation indicated that formation of cementum-like and PDL-like tissue was induced on the titanium surface with surface treatments and that the PDL-like tissue was perpendicularly oriented between the titanium surface with cementum-like tissue and the bone. Results demonstrate that a periodontal-like structure was formed around a titanium implant, which is similar to the environment existing around a natural tooth. The clinical application of dental implants combined with a cell sheet technique may be feasible as an alternative implant therapy. Furthermore, application of this methodology may play an innovative role in the periodontal, prosthetic, and orthodontic fields in dentistry.