Titanium Discs Research Articles

Tissue-Safe Low-Temperature Plasma Treatment for Effective Management of Mature Peri-Implantitis Biofilms on Titanium Surfaces.

The unique screw-shape design and microstructure of implants pose a challenge for mechanical debridement in removing biofilms. Biofilms exhibit increased resistance to antimicrobials relative to single planktonic cells, emphasizing the need for effective biofilm removal during periodontal therapy for peri-implantitis treatment. To tackle this issue, our team evaluated the effectiveness of low-temperature plasma (LTP) for disinfecting titanium discs contaminated with multispecies biofilms associated with peri-implantitis, specifically focusing on biofilms matured for 14 and 21 days as well as biofilms that had formed on StraumannⓇ Ti-SLA implants for 21 days. The biofilms included Actinomyces naeslundii, Porphyromonas gingivalis, Streptococcus oralis, and Veillonella dispar, which were grown in anaerobic conditions. These biofilms were subjected to LTP treatment for 1, 3, and 5 min, using distances of 3 or 10 mm from the LTP nozzle to the samples. Control groups included biofilms formed on Ti discs or implants that received no treatment, exposure to argon flow at 3 or 10 mm of distance for 1, 3, or 5 min, application for 1 min of 14 μg/mL amoxicillin, 140 μg/mL metronidazole, or a blend of both, and treatment with 0.12% chlorhexidine (CHX) for 1 min. For the implants, 21-day-old biofilms were treated with 0.12% CHX 0.12% for 1 min and LTP for 1 min at a distance of 3 mm for each quadrant. Biofilm viability was assessed through bacterial counting and confocal laser scanning microscopy. The impact of LTP was investigated on reconstituted oral epithelia (ROE) contaminated with P. gingivalis, evaluating cytotoxicity, cell viability, and histology. The results showed that a 1 min exposure to LTP at distances of 3 or 10 mm significantly lowered bacterial counts on implants and discs compared to the untreated controls (p < 0.017). LTP exposure yielded lower levels of cytotoxicity relative to the untreated contaminated control after 12 h of contamination (p = 0.038), and cell viability was not affected by LTP (p ≥ 0.05); thus, LTP-treated samples were shown to be safe for tissue applications, with low cytotoxicity and elevated cell viability post-treatment, and these results were validated by qualitative histological analysis. In conclusion, the study's results support the effectiveness of 1 min LTP exposure in successfully disinfecting mature peri-implantitis multispecies biofilms on titanium discs and implants. Moreover, it validated the safety of LTP on ROE, suggesting its potential as an adjunctive treatment for peri-implantitis.

Effects of i-PRF, A-PRF+, and EMD on Osteogenic Potential of Osteoblasts on Titanium.

The study evaluates three biologically active substances with known bone-inductive potential on previously decontaminated titanium (Ti) discs. Rough and smooth Ti surfaces were contaminated with a multispecies biofilm and cleaned with a chitosan brush. Discs were treated either with injectable-platelet-rich fibrin (i-PRF), advanced platelet-rich fibrin (A-PRF+), or enamel matrix derivatives (EMDs) before osteoblast seeding. Biocompatibility, adhesion, migration, and gene expression of runt-related transcription factor 2 (RUNX2), collagen Type I Alpha 2 (COL1a2), alkaline phosphatase (ALP), osteocalcin (OC), and osteonectin (ON) were performed. All the tested biologic agents similarly increased cell viability. Specifically, osteoblasts seeded over i-PRF and EMD-treated surfaces showed improvement in adhesion and migration and significantly increased ALP, OC, ON, RUNX-2, and COL1a2 mRNA levels up to 2.8 fold (p < 0.05) with no differences between Ti surfaces. i-PRF and EMD possess beneficial bioactive properties that enhance tissue healing and promote regeneration on thoroughly sterilized surfaces. Biologically active materials may hold the potential to influence the process of implant re-osseointegration, which warrants more research since sterilization of the affected surfaces under clinical conditions is still not reliably possible and remains one of the greatest challenges.

The effect of titanium surface treatment by application of constant potential or current on the viability of pre-osteoblast cells: an in-vitro study.

The aim of this study was to investigate the impact of electrochemical treatment of a titanium surface employing constant current and potential on the viability of the tissue cells attached to the surface and determining the safety limits for this type of treatment. Pre-osteoblast cells (pOB) were cultured and seeded onto titanium discs. The cell-seeded discs were then exposed to a range of contant direct electrical potentials (-6V-6V) or contant direct electrical currents (-12.5mA, -25mA, or -50mA) using a three-electrode system connected to a potentiostat. Cell viability was assessed using live/dead assay and fluorescence microscopy. Exposure of cells to high negative potentials caused cell detachment, while exposure to positive ones led to cell death on the cpTi surfaces. However, cellular viability was preserved when the electrical potentials were kept between -3 and +3V. Cells retained 80% viability when subjected to -12.5mA currents with an initial pOB cell count of 5 × 104. However, when the initial cell count was elevated to 1 × 105, the cells demonstrated the ability to withstand an even greater current (-25mA) while preserving their vitality at the same level. Treatment of a titanium dental implant surface employing constant potential or current can harm cells surrounding dental implants. However, this damage can be minimized by keeping the potential within a safety limit.

Low toxicity of dissolved silver from silver-coated titanium dental implants to human primary osteoblast cells

Objectivesthe controlled release of silver as a biocide from Ag-coated medical implants is desirable. However, the biocompatibility of Ag leachates is poorly understood. This study investigated the toxicity of silver released from the silver plated titanium implants to human primary osteoblast cells; and the effect of cell culture medium on the silver speciation and bioavailability. Methods: Ti6Al4V discs were coated with Ag nanoparticles (NPs), silver plus hydroxyapatite (HA) nanoparticles (Ag+nHA), or Ag NPs plus microparticles (Ag+mHA). Primary human osteoblast cells were exposed to the leachates from the various discs for up to 7 days. Results: the total Ag concentrations released as leachate from the silver-plated titanium discs were 0.7–1.6 mg L−1, regardless of treatment. Cumulative silver release over 7 days was approximately 3 mg L−1 in all treatments. The concentration of total Ag in the cell homogenates from all the Ag-containing treatments was modest, ∼ 0.1 µg mg protein−1 or less at day 7. Cells showed normal healthy morphology with no appreciable leak of LDH or ALP activity into the external media compared to the reference control. Similarly, there was no significant differences (Kruskal Wallis, p > 0.05) in the LDH or ALP activity in the cell homogenate between treatments. Conclusions: overall, there was a controlled release of Ag into the external media, but this remained biocompatible with no deleterious effects on the osteoblast cells, which means that the released silver to the peri-implant environment is not toxic making the coating potential for clinical use

Probabilistic risk assessment method considering machining-induced random residual stress

The aero-engine disks inevitably have manufacturing-induced anomalies and machining-induced random residual stress (RS) in localized and critical areas, which cause a severe threat to the safety of the aircraft. Traditional structural design of the disks fails to establish a quantitative correlation between the machining process and the failure risk. Therefore, this paper proposes a probabilistic model considering random RS to quantify the influence of machining RS subjected to low-cycle fatigue. The RS dispersion is quantified using a scaling parameter, obtained by X-ray diffraction measurements and orthogonal cutting simulations. The crack life database under varying RSs is established for efficient probability calculations. Results show that the coefficient of variation (COV) of the RS on the same machined surface with the same processing parameters is 7.62 % in the local area and 13.74 % in the whole machined surface. The risk results show that the probability of failure (POF) considering the deterministic RS is 2–4 % lower than the POF without RS, owing to the extension of fatigue life by compressive RS. Furthermore, the POF considering the random RS is almost the same (difference <0.6 %) as the POF considering the deterministic RS because the depth of the machining RS is around 0.2 mm. The proposed method predicts the POF more accurately and is thus valuable for the safety assessment of an aero-engine titanium disk.

Effect of Er,Cr:YSGG Laser Irradiation on the Surface Modification and Cell Adhesion on Titanium Discs: An In Vitro Study

This study evaluates the potential of erbium, chromium-doped:yttrium, scandium, gallium, and garnet (Er,Cr:YSGG) laser irradiation to modify the titanium surface for optimal seeding of fibroblasts and osteoblasts in the treatment of peri-implantitis. Titanium discs were treated using the Er,Cr:YSGG laser, an ultrasonic device with a stainless tip, or titanium scalers. Changes in surface properties were analyzed by profilometer and scanning electron microscopy (SEM). Murine fibroblast and osteoblast adhesion and proliferation were evaluated qualitatively and quantitatively at 24 and 72 h. Profilometric surface topography and SEM showed that titanium scalers and ultrasonic debridement techniques significantly changed the structure of the machined and rough titanium surfaces. The Er,Cr:YSGG laser irradiation, on the other hand, did not alter titanium microstructures. The Er,Cr:YSGG laser irradiation with the 40 Hz group showed a significantly higher attached fibroblast cell numbers than the titanium scaler group at 72 h after treatment (p = 0.023). Additionally, the number of the attached osteoblasts in the Er,Cr:YSGG laser irradiation with the 40 Hz group was significantly higher than that of the no-treatment groups 24 h after treatment (p = 0.045). The Er,Cr:YSGG laser effectively promoted adherence of fibroblasts and osteoblasts to the titanium surface without significantly altering the titanium surface, suggesting its superiority for treating peri-implantitis.

CO2-Laser Irradiation Affects Positively Surface Energy of Rough Titanium Surfaces.

The purpose of this study was to investigate the free surface energy before and after CO2 irradiation on rough titanium surfaces. Sandblasted, large grit, acidetched (SLA) implant surfaces were used for this study. Contact angle evaluations were performed for saline, albumin, and artificial blood using a contact angle goniometer in 25°C and 37°C (20 droplets in each group, 120 in total). The titanium disks were then irradiated with a CO2 laser (10,600 nm) using a non-contact, defocused, 2W continuous wave for 30 seconds. Contact angle measurements for saline, albumin and artificial blood were evaluated immediately after irradiation (new 120 in total contact angle measurements) and compared with the measurements before laser irradiation. Paired t-test using SPSS was performed for 5% statistical significance level. Lower numbers of contact angle measurements were found and changes after CO2-laser irradiation were statistically significant (p < 0.05) for all groups except for albumin at 25°C, as follows: 25℃: (saline) (Pre) 111.97° ±13.67°, (Post) 79.29° ±10.83° / p<0.0001; (albumin) (Pre) 89.04° ±12.02°, (Post) 83.29° ±20.71° / p&#61;0.297; (blood) (Pre) 97.79° ±14.44°, (Post) 86.29° ±10.82° / p&#61;0.019; 37℃: (saline) (Pre) 104.12° ±13.138°, (Post) 92.66° ±4.777° / p&#61;0.001; (albumin) (Pre) 89.04° ±12.02°, (Post) 83.29° ±20.71° / p&#61;0.297 (blood) (Pre) 80.42° ±19.00°, (Post) 58.62° ±24.34° / p&#61;0.012. The CO2 laser irradiation in general positively affected the wettability and hydrophilicity of rough titanium (SLA) surfaces.

In Vitro Comparison of Titanium Disc Surface Roughness and Bacterial Colonization After Ultrasonic Instrumentation With Three Different Tips.

During implant maintenance, preserving a smooth surface on the machined transmucosal abutment is critical to reduce biofilm attachment and colonization. The present study compared the surface roughness and bacterial colonization of machined titanium surfaces after instrumentation with various materials. Forty-four machined grade 23 titanium discs were instrumented with a round polyether ether ketone (PEEK) tip, a plastic curette tip, or a pure titanium curette tip with piezoelectric devices. Before and after instrumentation, the surface roughness (Ra and Rz) values were analyzed with a profilometer and scanning electron microscopy (SEM). Streptococcus sanguinis was cultured and incubated for 24 hours on the instrumented discs, and colony-forming units per milliliter were obtained for each group. Samples instrumented with the metal ultrasonic tip significantly increased surface roughness compared with the other groups. This resulted in greater colonization by S. sanguinis than surfaces instrumented with PEEK tips or the negative control. Samples instrumented with PEEK and plastic tips did not exhibit any statistically significant increase in surface roughness, and SEM analysis revealed a significantly rougher surface of discs instrumented with metal compared with discs instrumented with plastic or PEEK tips despite the possibility of debris from tip dissolution. Our results suggest that instrumentation with metal ultrasonic tips with piezoelectric devices significantly increased machined titanium's surface roughness and elicited higher biofilm formation in vitro. Meanwhile, instrumentation of machined titanium with PEEK or plastic ultrasonic tips did not affect the surface roughness or bacterial adhesion.

Comparison In Vitro Study on the Interface between Skin and Bone Cell Cultures and Microporous Titanium Samples Manufactured with 3D Printing Technology Versus Sintered Samples.

Percutaneous implants osseointegrated into the residuum of a person with limb amputation need to provide mechanical stability and protection against infections. Although significant progress has been made in the biointegration of percutaneous implants, the problem of forming a reliable natural barrier at the level of the surface of the implant and the skin and bone tissues remains unresolved. The use of a microporous implant structure incorporated into the Skin and Bone Integrated Pylon (SBIP) should address the issue by allowing soft and bone tissues to grow directly into the implant structure itself, which, in turn, should form a reliable barrier to infections and support strong osseointegration. To evaluate biological interactions between dermal fibroblasts and MC3T3-E1 osteoblasts in vitro, small titanium discs (with varying pore sizes and volume fractions to achieve deep porosity) were fabricated via 3D printing and sintering. The cell viability MTT assay demonstrated low cytotoxicity for cells co-cultured in the pores of the 3D-printed and sintered Ti samples during the 14-day follow-up period. A subsequent Quantitative Real-Time Polymerase Chain Reaction (RT-PCR) analysis of the relative gene expression of biomarkers that are associated with cell adhesion (α2, α5, αV, and β1 integrins) and extracellular matrix components (fibronectin, vitronectin, type I collagen) demonstrated that micropore sizes ranging from 200 to 500 µm of the 3D printed and sintered Ti discs were favorable for dermal fibroblast adhesion. For example, for representative 3D-printed Ti sample S6 at 72 h the values were 4.71 ± 0.08 (α2 integrin), 4.96 ± 0.08 (α5 integrin), 4.71 ± 0.08 (αV integrin), and 1.87 ± 0.12 (β1 integrin). In contrast, Ti discs with pore sizes ranging from 400 to 800 µm demonstrated the best results (in terms of marker expression related to osteogenic differentiation, including osteopontin, osteonectin, osteocalcin, TGF-β1, and SMAD4) for MC3T3-E1 cells. For example, for the representative 3D sample S4 on day 14, the marker levels were 11.19 ± 0.77 (osteopontin), 7.15 ± 0.29 (osteonectin), and 6.08 ± 0.12 (osteocalcin), while for sintered samples the levels of markers constituted 5.85 ± 0.4 (osteopontin), 4.45 ± 0.36 (osteonectin), and 4.46 ± 0.3 (osteocalcin). In conclusion, the data obtained show the high biointegrative properties of porous titanium structures, while the ability to implement several pore options in one structure using 3D printing makes it possible to create personalized implants for the best one-time integration with both skin and bone tissues.

Diamond-like carbon (DLC) surface treatment decreases biofilm burden by S. aureus on titanium alloy in vitro- a pilot study.

Periprosthetic joint infection is a complication of total joint arthroplasty with treatment costs over $1.6billion dollars per year in the US with high failure rates. Therefore, generation of coatings that can prevent infection is paramount. Diamond-like carbon (DLC) is an ideal coating for implants as they are wear-resistant, corrosion-resistant, inert, and have a low friction coefficient. The purpose of this study was to test the efficacy of DLC surface treatment in prevention of biofilm on titanium discs infected with Staphylococcus aureus in vitro. Titanium alloy discs (n = 4 non-coated and n = 4 DLC-coated) were infected with 5 × 105 colony-forming units (CFU) of S. aureus for 2 weeks then analysed via crystal violet and scanning electron microscopy (SEM). Crystal violet analysis yielded differences in the appearance of biofilm on implant surface where DLC-coated had a clumpier appearance but no difference in biofilm quantification. Interestingly, this clumpy appearance did lead to differences in SEM biofilm coverage where significantly less biofilm coverage was found on DLC-coated discs (81.78% vs. 54.17%, p < 0.003). DLC-coated titanium alloy implants may have preventative properties in S. aureus infection. Observing differences in biofilm coverage does warrant additional testing including CFU titration and biofilm kinetics with eventual use in an animal model of periprosthetic joint infection.

S. aureus develops biofilms on titanium within 2 weeks in vitro.

The formation of biofilms, characterized by cell aggregation and extracellular polymeric substance (EPS) production, is a common feature of periprosthetic joint infections (PJI). The current study aimed to investigate the development of biofilm features in vitro within less than 3 weeks by Staphylococcus aureus isolated from PJIs. Biofilms were grown on sandblasted titanium discs, and fluorescence spectroscopy and microscopy were used to observe biofilm maturation for 21 days. DNA mass decreased initially, then increased from day 5 onwards, and decreased again after day 7. The proportion of living to dead bacteria oscillated until day 7 and increased at day 10 for strain A and day 14 for strain B. EPS mass decreased initially and then continuously increased. Multilayer bacterial organization was observed at day 7. Cell aggregation occurred during the first week, followed by EPS production in the second week, and characteristic biofilm features were observed within 1 to 2 weeks.

Surface Decontamination of Titanium Dental Implants Subjected to Implantoplasty by Treatment with Citric Acid Solutions

Implantoplasty is one of the most common techniques to remove peri-implantitis from the surface of dental implants. It is a process of mechanization of the titanium surface, causing the loss of the roughness of the dental implant, which leads to difficulty in tissue regeneration. The aim of this research is to apply a decontaminant based on citric acid and add collagen and magnesium cations to promote tissue formation and have a bactericidal character. Eighty commercially pure grade 3 titanium discs were used to perform the implantoplasty protocol, like the one used in dental clinics. They were treated with four different solutions: 25% citric acid, 25% citric acid with the addition of collagen 0.25 g/L, 25% citric acid with the addition of 0.50 g/L and the latter with the addition of 1% Mg (NO3)2. The roughness was determined by confocal microscopy, the contact angle, adhesion and proliferation of HFFs fibroblasts, proliferation of SaOS-2 osteoblasts and bactericidal behavior by culturing very common bacteria in the oral cavity, Gram-positive Streptococcus sanguinis and gordonii and as Gram-negative Pseudomonas aeruginosa. The results showed that the treatment with citric acid slightly increases the roughness and decreases the contact angle from 78 to 13°, making the surface superhydrophilic. Fibroblast proliferation studies show a very significant increase at 24 h, the most favorable solution being the one containing 0.50 g/L of collagen with the presence of magnesium in a 25% citric acid solution. This same solution shows the highest cytocompatibility and osteoblastic proliferation with statistically significant differences with respect to the control and the rest of the solutions. Microbiological studies show a bactericidal effect due to the presence of citric acid, which is especially effective on Gram-positive bacteria. The results allow us to have mouthwashes that can be applied in the patient’s mouth, which will help the regeneration of tissues and avoid new bacterial colonization.

Macrophage-to-osteocyte communication: Impact in a 3D in vitro implant-associated infection model

Macrophages and osteocytes are important regulators of inflammation, osteogenesis and osteoclastogenesis. However, their interactions under adverse conditions, such as biomaterial-associated infection (BAI) are not fully understood. We aimed to elucidate how factors released from macrophages modulate osteocyte responses in an in vitro indirect 3D co-culture model. Human monocyte-derived macrophages were cultured on etched titanium disks and activated with either IL-4 cytokine (anti-inflammatory M2 phenotype) or Staphylococcus aureus secreted virulence factors to simulate BAI (pro-inflammatory M1 phenotype). Primary osteocytes in collagen gels were then stimulated with conditioned media (CM) from these macrophages. The osteocyte response was analyzed by gene expression, protein secretion, and immunostaining. M1 phenotype macrophages were confirmed by IL-1β and TNF-α secretion, and M2 macrophages by ARG-1 and MRC-1.Osteocytes receiving M1 CM revealed bone inhibitory effects, denoted by reduced secretion of bone formation osteocalcin (BGLAP) and increased secretion of the bone inhibitory sclerostin (SOST). These osteocytes also downregulated the pro-mineralization gene PHEX and upregulated the anti-mineralization gene MEPE. Additionally, exhibited pro-osteoclastic potential by upregulating pro-osteoclastic gene RANKL expression. Nonetheless, M1-stimulated osteocytes expressed a higher level of the potent pro-osteogenic factor BMP-2 in parallel with the downregulation of the bone inhibitor genes DKK1 and SOST, suggesting a compensatory feedback mechanisms. Conversely, M2-stimulated osteocytes mainly upregulated anti-osteoclastic gene OPG expression, suggesting an anti-catabolic effect. Altogether, our findings demonstrate a strong communication between M1 macrophages and osteocytes under M1 (BAI)-simulated conditions, suggesting that the BAI adverse effects on osteoblastic and osteoclastic processes in vitro are partly mediated via this communication. Statement of significanceBiomaterial-associated infections are major challenges and the underlying mechanisms in the cellular interactions are missing, especially among the major cells from the inflammatory side (macrophages as the key cell in bacterial clearance) and the regenerative side (osteocyte as main regulator of bone). We evaluated the effect of macrophage polarization driven by the stimulation with bacterial virulence factors on the osteocyte function using an indirect co-culture model, hence mimicking the scenario of a biomaterial-associated infection. The results suggest that at least part of the adverse effects of biomaterial associated infection on osteoblastic and osteoclastic processes in vitro are mediated via macrophage-to-osteocyte communication.

Kinetics of Oxygen Catalyst Based on Pyrochlore Oxide Nano-Particles for Low Temperature Water Electrolysis

Alkaline water electrolysis is carried out at 60 to 80 oC using nickel-based electrode materials and highly concentrated KOH solutions, which make highly corrosive environment against the electrodes, the separator, the current collector, and the others of the cell. Reduction in operating temperature would be preferable to suppress corrosion, although it would also make the catalytic activity for OER and HER down, making the cell voltage up. Since oxygen evolution is known as a high overpotential reaction which is more than 400 mV with nickel-based oxides at 60 oC or more, and alternative materials with lower overpotential and higher stability for OER in AWE have been needed. This paper presents a novel oxygen catalyst nano-particles based on bismuth ruthenium oxide, in which ruthenium at the B site in the pyrochlore structure is partly replaced with manganese1).Mn-doped bismuth ruthenium oxide (MBRO) was prepared by precipitation of metal hydroxide precursor, which had been obtained by addition of NaOH solution into the metal salt solution containing Bi(III) and Ru(III) with Mn(II), followed by calcination at 600 oC. XRD, EDX, EXAFS, ICP-AES, and RBS were used for structural and compositional analyses of MBRO. The obtained oxide was added into distilled water and the solution was treated by ultrasonic vibration to make a suspension, which was then dropped on a titanium disk (4 mm diam.) and dried at RT for 24 h, resulting in oxide particles-coated titanium disk, in which no binders or ionomers were used in the process. The disk was mounted in a commercial rotating disk electrode and was used as the working electrode with a platinum counter electrode, an Hg/HgO reference electrode, and 0.1 mol/L KOH solution. Polarization curves were measured by LSV for OER, of which the results were analyzed to obtain the onset potential, Tafel slope, exchange current density. All measurements were performed at RT (25 oC).EDX and EXAFS studies of MBRO showed that Ru(IV) was replaced with Mn(IV) at the B-site of pyrochlore structure, and this doping made a single phase of pyrochlore structure oxide without sub-product up to 30 at% replacement . Linear sweep voltammetry revealed the overpotential of MBRO for OER was less than 200 mV and the Tafel slope analysis of the LSV curve gave 40 mV/dec or less. The onset potential and the Tafel slope were almost the same when Ru:Mn atomic ratio of MBRO was changed from 100:0 to 70:30 at%. Further analysis of the polarization curve indicated the exchange current density slightly decreased with increasing Mn ratio. These results suggest that Ru is the active site for OER but Mn is as well. The Tafel slope of OER catalysts reported in literatures is mostly 60 mV/dec or more, including more than 200 mV/dec of nickel-based oxides2,3). Therefore, MBRO is a hyper catalytic oxide for OER in alkaline media and can significantly reduce the overpotential of the anode in AWE even at ambient temperature.Ref.1) M. Morimitsu, Patent US.2022085387.A1.2) T. Shinagawa, A. T. Garcia-Esparza, K. Takanabe, Scientific Reports, Vol. 5, Article No.13801 (2015).3) S. Yagi, et al., Nature Communications, Vol. 6, Article No. 8249 (2015).

Anti-biofilm activity of 445nm and 970nm diode laser on mixed species colonies of- aggregatibacter actinomycetemcomitans and porphyromonas gingivalis cultured on titanium discs -an in vitro study.

To assess and compare the anti-microbial efficacy of 445nm and 970nm diode laser on mixed species biofilm of Aggregatibacter actinomycetemcomitans [A.a] and Porphyromonas gingivalis [P.g] cultured on machined pure titanium discs. A total of 65 machined pure titanium discs with no surface modifications with a 10-mm diameter and a 2-mm height were sterilized by autoclaving at 121°C for 15min and incubated with the commercially available bacterial strains ATCC(American Type Culture Collection- P.g 33277 and A.a 29522)mixture of Aggregatibacter actinomycetemcomitans(A.a) and Porphyromonas gingivalis(P.g).After a 2-week incubation period with the mixture of bacteria to develop a mixed species biofilm, the discs were divided into three groups: (1) no treatment (control), (2) 445nm laser (test), (3) 970nm laser (test). For each laser wavelength (445 and 970nm), the discs were exposed to 1.0W and 2.0W in continuous wave mode for the times points of 15, 30, and 60s. The antimicrobial efficacy was assessed by qPCR. A significant reduction in the levels of both species of bacteria was observed between control and the laser intervention groups. A higher efficacy for the 445nm diode laser against Porphyromonas gingivalis and a similar efficacy against Aggregatibacter actinomycetemcomitans was observed as compared to the 970nm group. 445nm wavelength represents a potential and effective laser wavelength which can be used for the management of peri-implant infection. The present study findings also need to be further validated through clinical interventional trials.

Roughness affects the response of human fibroblasts and macrophages to sandblasted abutments

BackgroundA strong seal of soft-tissue around dental implants is essential to block pathogens from entering the peri-implant interface and prevent infections. Therefore, the integration of soft-tissue poses a challenge in implant-prosthetic procedures, prompting a focus on the interface between peri-implant soft-tissues and the transmucosal component. The aim of this study was to analyse the effects of sandblasted roughness levels on in vitro soft-tissue healing around dental implant abutments. In parallel, proteomic techniques were applied to study the interaction of these surfaces with human serum proteins to evaluate their potential to promote soft-tissue regeneration.ResultsGrade-5 machined titanium discs (MC) underwent sandblasting with alumina particles of two sizes (4 and 8 μm), resulting in two different surface types: MC04 and MC08. Surface morphology and roughness were characterised employing scanning electron microscopy and optical profilometry. Cell adhesion and collagen synthesis, as well as immune responses, were assessed using human gingival fibroblasts (hGF) and macrophages (THP-1), respectively. The profiles of protein adsorption to the surfaces were characterised using proteomics; samples were incubated with human serum, and the adsorbed proteins analysed employing nLC–MS/MS. hGFs exposed to MC04 showed decreased cell area compared to MC, while no differences were found for MC08. hGF collagen synthesis increased after 7 days for MC08. THP-1 macrophages cultured on MC04 and MC08 showed a reduced TNF-α and increased IL-4 secretion. Thus, the sandblasted topography led a reduction in the immune/inflammatory response. One hundred seventy-six distinct proteins adsorbed on the surfaces were identified. Differentially adsorbed proteins were associated with immune response, blood coagulation, angiogenesis, fibrinolysis and tissue regeneration.ConclusionsIncreased roughness through MC08 treatment resulted in increased collagen synthesis in hGF and resulted in a reduction in the surface immune response in human macrophages. These results correlate with the changes in protein adsorption on the surfaces observed through proteomics.

Healthy and diabetic primary human osteoblasts exhibit varying phenotypic profiles in high and low glucose environments on 3D-printed titanium surfaces.

The revolution of orthopedic implant manufacturing is being driven by 3D printing of titanium implants for large bony defects such as those caused by diabetic Charcot arthropathy. Unlike traditional subtractive manufacturing of orthopedic implants, 3D printing fuses titanium powder layer-by-layer, creating a unique surface roughness that could potentially enhance osseointegration. However, the metabolic impairments caused by diabetes, including negative alterations of bone metabolism, can lead to nonunion and decreased osseointegration with traditionally manufactured orthopedic implants. This study aimed to characterize the response of both healthy and diabetic primary human osteoblasts cultured on a medical-grade 3D-printed titanium surface under high and low glucose conditions. Bone samples were obtained from six patients, three with Type 2 Diabetes Mellitus and three without. Primary osteoblasts were isolated and cultured on 3D-printed titanium discs in high (4.5 g/L D-glucose) and low glucose (1 g/L D-Glucose) media. Cellular morphology, matrix deposition, and mineralization were assessed using scanning electron microscopy and alizarin red staining. Alkaline phosphatase activity and L-lactate concentration was measured in vitro to assess functional osteoblastic activity and cellular metabolism. Osteogenic gene expression of BGLAP, COL1A1, and BMP7 was analyzed using reverse-transcription quantitative polymerase chain reaction. Diabetic osteoblasts were nonresponsive to variations in glucose levels compared to their healthy counterparts. Alkaline phosphatase activity, L-lactate production, mineral deposition, and osteogenic gene expression remained unchanged in diabetic osteoblasts under both glucose conditions. In contrast, healthy osteoblasts exhibited enhanced functional responsiveness in a high glucose environment and showed a significant increase in osteogenic gene expression of BGLAP, COL1A1, and BMP7 (p<.05). Our findings suggest that diabetic osteoblasts exhibit impaired responsiveness to variations in glucose concentrations, emphasizing potential osteoblast dysfunction in diabetes. This could have implications for post-surgery glucose management strategies in patients with diabetes. Despite the potential benefits of 3D printing for orthopedic implants, particularly for diabetic Charcot collapse, our results call for further research to optimize these interventions for improved patient outcomes.

The Addition of Hydroxyapatite Nanoparticles on Implant Surfaces Modified by Zirconia Blasting and Acid Etching to Enhance Peri-Implant Bone Healing.

This study investigated the impact of adding hydroxyapatite nanoparticles to implant surfaces treated with zirconia blasting and acid etching (ZiHa), focusing on structural changes and bone healing parameters in low-density bone sites. The topographical characterization of titanium discs with a ZiHa surface and a commercially modified zirconia-blasted and acid-etched surface (Zi) was performed using scanning electron microscopy, profilometry, and surface-free energy. For the in vivo assessment, 22 female rats were ovariectomized and kept for 90 days, after which one implant from each group was randomly placed in each tibial metaphysis of the animals. Histological and immunohistochemical analyses were performed at 14 and 28 days postoperatively (decalcified lab processing), reverse torque testing was performed at 28 days, and histometry from calcified lab processing was performed at 60 days The group ZiHa promoted changes in surface morphology, forming evenly distributed pores. For bone healing, ZiHa showed a greater reverse torque, newly formed bone area, and bone/implant contact values compared to group Zi (p < 0.05; t-test). Qualitative histological and immunohistochemical analyses showed higher features of bone maturation for ZiHa on days 14 and 28. This preclinical study demonstrated that adding hydroxyapatite to zirconia-blasted and acid-etched surfaces enhanced peri-implant bone healing in ovariectomized rats. These findings support the potential for improving osseointegration of dental implants, especially in patients with compromised bone metabolism.

Development of Microstructure on Titanium Implant Surface Using CO2 Laser Processing

Background: Dental biomaterials made of titanium are commonly used. It depends on the implant surface texture to improve fixation and prevent the unwanted adhesion of bone cells. This study aimed to investigate whether continuous laser beam carbon dioxide (CNC - CO2) lasers produce specific textures on titanium surfaces with micrometer-sized indentations that influence cell behavior. Materials and method: (CNC - CO2) red laser device; with a fundamental wavelength of λ=10600 nm and power pulses of 34 W were applied, and textures on the surface of titanium discs were achieved. Results: Excellent degrees of uniformity and repeatability were achieved for the desired portions of the surface by creating different surface textures. The surface topography and chemical composition of the specimens were investigated by scanning electron microscopy, electron dispersive spectroscopy, X-ray diffraction, and surface roughness measurements. Also, a laser power of 34 watts raised the surface roughness, Ra (1.71 nm), and Rz (1.99 nm). Conclusion: Titanium surface textures with unique qualities can be formed in response to an increased heat input. When excessive laser power was used, the measured roughness increased because of instantaneous re-melting. The use of a right continuous-wave (CNC - CO2) laser on titanium used in dental implants can form specific surface textures.

Increased biofilm formation in dual-strain compared to single-strain communities of Cutibacterium acnes

Cutibacterium acnes is a known opportunistic pathogen in orthopedic implant-associated infections (OIAIs). The species of C. acnes comprises distinct phylotypes. Previous studies suggested that C. acnes can cause single- as well as multi-typic infections, i.e. infections caused by multiple strains of different phylotypes. However, it is not known if different C. acnes phylotypes are organized in a complex biofilm community, which could constitute a multicellular strategy to increase biofilm strength and persistency. Here, the interactions of two C. acnes strains belonging to phylotypes IB and II were determined in co-culture experiments. No adverse interactions between the strains were observed in liquid culture or on agar plates; instead, biofilm formation in both microtiter plates and on titanium discs was significantly increased when combining both strains. Fluorescence in situ hybridization showed that both strains co-occurred throughout the biofilm. Transcriptome analyses revealed strain-specific alterations of gene expression in biofilm-embedded cells compared to planktonic growth, in particular affecting genes involved in carbon and amino acid metabolism. Overall, our results provide first insights into the nature of dual-type biofilms of C. acnes, suggesting that strains belonging to different phylotypes can form biofilms together with additive effects. The findings might influence the perception of C. acnes OIAIs in terms of diagnosis and treatment.