Effect of inorganic fillers on the light transmission through traditional or flowable resin-matrix composites for restorative dentistry.

OBJECTIVES: The aim of this in vitro study was to evaluate the light transmission through five different resin-matrix composites regarding the inorganic filler content. METHODS: Resin-matrix composite disc-shaped specimens were prepared on glass molds. Three traditional resin-matrix composites contained inorganic fillers at 74, 80, and 89 wt. % while two flowable composites revealed 60 and 62.5 wt. % inorganic fillers. Light transmission through the resin-matrix composites was assessed using a spectrophotometer with an integrated monochromator before and after light curing for 10, 20, or 40s. Elastic modulus and nanohardness were evaluated through nanoindentation's tests, while Vicker's hardness was measured by micro-hardness assessment. Chemical analyses were performed by FTIR and EDS, while microstructural analysis was conducted by optical microscopy and scanning electron microscopy. Data were evaluated using two-way ANOVA and Tukey's test (p < 0.05). RESULTS: After polymerization, optical transmittance increased for all specimens above 650-nm wavelength irradiation since higher light exposure time leads to increased light transmittance. At 20- or 40-s irradiation, similar light transmittance was recorded for resin composites with 60, 62, 74, or 78-80 wt. % inorganic fillers. The lowest light transmittance was recorded for a resin-matrix composite reinforced with 89 wt. % inorganic fillers. Thus, the size of inorganic fillers ranged from nano- up to micro-scale dimensions and the high content of micro-scale inorganic particles can change the light pathway and decrease the light transmittance through the materials. At 850-nm wavelength, the average ratio between polymerized and non-polymerized specimens increased by 1.6 times for the resin composite with 89 wt. % fillers, while the composites with 60 wt. % fillers revealed an increased ratio by 3.5 times higher than that recorded at 600-nm wavelength. High mean values of elastic modulus, nano-hardness, and micro-hardness were recorded for the resin-matrix composites with the highest inorganic content. CONCLUSIONS: A high content of inorganic fillers at 89 wt.% decreased the light transmission through resin-matrix composites. However, certain types of fillers do not interfere on the light transmission, maintaining an optimal polymerization and the physical properties of the resin-matrix composites. CLINICAL SIGNIFICANCE: The type and content of inorganic fillers in the chemical composition of resin-matrix composites do affect their polymerization mode. As a consequence, the clinical performance of resin-matrix composites can be compromised, leading to variable physical properties and degradation.

Mechanical properties and cellular content of leukocyte- and platelet-rich fibrin membranes of patients on antithrombotic drugs.

INTRODUCTION: The aim of this study was to examine the potential influence of antithrombotics on leukocyte- and platelet-rich fibrin (L-PRF) membranes. METHODS: Tensile tests and cell counts were performed with L-PRF membranes originating from patients on anticoagulants and antiplatelets versus patients not taking antithrombotics. RESULTS: For the tensile tests, 13 control patients, 12 on anticoagulants, and 10 on antiplatelets donated blood. Compared to controls, membranes from anticoagulated donors were weaker (strength 0.57 ± 0.24 MPa vs. 0.80 ± 0.27 MPa, p = .03) and could not be stretched as far (1.8 ± 0.3 vs. 2.1 ± 0.3 times the initial length, p = .01). For the cell counting, 23 control patients, 16 on anticoagulants, and 16 on antiplatelets donated blood. The percentage of platelets was ±50% in the three groups. The percentage of leukocytes was lower in the anticoagulant group compared with controls (69 ± 10% vs. 78 ± 8%, p = .04). However, because of the unknown error of method, it is questionable whether the statistical significance is meaningful. There was no difference between membranes from the control group and the group on antiplatelets. CONCLUSION: Our results indicate that L-PRF membranes originating from patients on anticoagulants are weaker, stretch less far, and contain less leukocytes than L-PRF membranes of patients not taking these drugs.

qDNase assay: A quantitative method for real-time assessment of DNase activity on coated surfaces.

DNase coatings show great potential to prevent biofilm formation in various applications of the medical implant, food and marine industry. However, straightforward and quantitative methods to characterize the enzymatic activity of these coatings are currently not available. We here introduce the qDNase assay, a quantitative, real-time method to characterize the activity of DNase coatings. The assay combines (1) the use of an oligonucleotide probe, which fluoresces upon cleavage by coated DNases, and (2) the continuous read-out of the fluorescent signal within a microplate fluorometer format. The combination of these two properties results in a real-time fluorescent signal that is used to directly quantify the activity of DNase coatings. As a proof of concept, bovine DNase I coatings were immobilized on titanium by means of chemical grafting and their activity was estimated at 3.87 × 10-4 U. To our knowledge, the qDNase assay provides the first approach to report the activity of a DNase coating in absolute DNase activity units. This assay will not only serve to compare existing DNase coating methods more accurately, but will also enable the rational design of new DNase coating methods in the future.

Ultrafast laser selective phase removal for surface modification of nanocomposite materials.

Ultrafast laser processing of zirconia/alumina nanocomposite ceramics, the current gold standard material for ceramic bearing components in orthopedics, was investigated. Instead of considering the substrate as a homogeneous material, as commonly assumed in laser micromachining, the damage behavior of different phases around the laser ablation threshold upon ultrafast laser irradiation was investigated. Under appropriate experimental conditions, the zirconia phase was selectively ablated while the alumina phase remained intact. The origin of this selective ablation behavior and its relationship with the material band gaps were discussed. Due to the nonlinear absorption mechanisms under ultrafast laser irradiation, the zirconia phase, with its band gap of 5.8 eV, can absorb more laser energy than the alumina phase which has a larger band gap of 8.8 eV. The negligible heat diffusion length ensures that the absorbed laser energy remains confined in the individual phases, leading to the selective ablation of zirconia phase under the given laser fluence. Based on this observation, an ultrafast laser selective phase removal method which can be used to modify the surface composition of nanocomposite materials consisting of phases with different band gaps was proposed.

In vitro polymicrobial inter-kingdom three-species biofilm model: influence of hyphae on biofilm formation and bacterial physiology.

Biofilms are an important medical burden, notably for patients with orthopaedic device-related infections. When polymicrobial, these infections are more lethal and recalcitrant. Inter-kingdom biofilm infections are poorly understood and challenging to treat. Here, an in vitro three-species model including Staphylococcus aureus, Escherichia coli and Candida albicans was developed, to represent part of the diversity observed in orthopaedic infections or other clinical contexts. The importance of fungal hyphae for biofilm formation and virulence factor expression was explored. Two protocols were set up, allowing, or not, for hyphal formation. Culturable cells and biomass were characterised in both models, and biofilms were imaged in bright-field, confocal and electron microscopes. The expression of genes related to virulence, adhesion, exopolysaccharide synthesis and stress response was analysed in early-stage and mature biofilms. It was found that biofilms enriched in hyphae had larger biomass and showed higher expression levels of genes related to bacterial virulence or exopolysaccharides synthesis.

Mechanical and structural properties of leukocyte- and platelet-rich fibrin membranes: An in vitro study on the impact of anticoagulant therapy.

OBJECTIVE AND BACKGROUND: Little is known about structural and mechanical properties of leukocyte- and platelet-rich fibrin (L-PRF) membranes and even less about the influence of antithrombotic drugs on L-PRF. The aim of this in vitro study is therefore to investigate mechanical properties, fibrin structure and cell content of L-PRF membranes and the impact of anticoagulant therapy on L-PRF. METHODS: Blood samples were obtained from 12 volunteers and supplemented with either no, 1.25 IU, 2.5 IU, 5 IU, or 10 IU enoxaparin. L-PRF membranes were characterized with tensile testing, scanning electron microscopy, and measurement of platelets and leukocytes. Control and enoxaparin-supplemented L-PRF membranes were compared. RESULTS: At 10 IU enoxaparin, no L-PRF membranes could be generated, whereas the low doses of 1.25 and 2.5 IU had no influence on L-PRF properties. The mechanical properties, fibrin networks, and number of platelets and leukocytes of 5 IU supplemented membranes were unlike the control membranes, but were not found to be significantly different because of limited sampling and inter- and intra-variability. CONCLUSION: Low doses of the anticoagulant enoxaparin do not affect mechanical properties, fibrin network, nor cellular content of L-PRF, whereas high doses impair L-PRF generation.

Covalent immobilization of antimicrobial agents on titanium prevents Staphylococcus aureus and Candida albicans colonization and biofilm formation.

OBJECTIVES: Biofilm-associated implant infections represent a serious public health problem. Covalent immobilization of antimicrobial agents on titanium (Ti), thereby inhibiting biofilm formation of microbial pathogens, is a solution to this problem. METHODS: Vancomycin (VAN) and caspofungin (CAS) were covalently bound on Ti substrates using an improved processing technique adapted to large-scale coating of implants. Resistance of the VAN-coated Ti (VAN-Ti) and CAS-coated Ti (CAS-Ti) substrates against in vitro biofilm formation of the bacterium Staphylococcus aureus and the fungal pathogen Candida albicans was determined by plate counting and visualized by confocal laser scanning microscopy. The efficacy of the coated Ti substrates was also tested in vivo using an adapted biomaterial-associated murine infection model in which control-Ti, VAN-Ti or CAS-Ti substrates were implanted subcutaneously and subsequently challenged with the respective pathogens. The osseointegration potential of VAN-Ti and CAS-Ti was examined in vitro using human bone marrow-derived stromal cells, and for VAN-Ti also in a rat osseointegration model. RESULTS: In vitro biofilm formation of S. aureus and C. albicans on VAN-Ti and CAS-Ti substrates, respectively, was significantly reduced compared with biofilm formation on control-Ti. In vivo, we observed over 99.9% reduction in biofilm formation of S. aureus on VAN-Ti substrates and 89% reduction in biofilm formation of C. albicans on CAS-Ti substrates, compared with control-Ti substrates. The coated substrates supported osseointegration in vitro and in vivo. CONCLUSIONS: These data demonstrate the clinical potential of covalently bound VAN and CAS on Ti to reduce microbial biofilm formation without jeopardizing osseointegration.

Fungal ß-1,3-glucan increases ofloxacin tolerance of Escherichia coli in a polymicrobial E. coli/Candida albicans biofilm.

In the past, biofilm-related research has focused mainly on axenic biofilms. However, in nature, biofilms are often composed of multiple species, and the resulting polymicrobial interactions influence industrially and clinically relevant outcomes such as performance and drug resistance. In this study, we show that Escherichia coli does not affect Candida albicans tolerance to amphotericin or caspofungin in an E. coli/C. albicans biofilm. In contrast, ofloxacin tolerance of E. coli is significantly increased in a polymicrobial E. coli/C. albicans biofilm compared to its tolerance in an axenic E. coli biofilm. The increased ofloxacin tolerance of E. coli is mainly biofilm specific, as ofloxacin tolerance of E. coli is less pronounced in polymicrobial E. coli/C. albicans planktonic cultures. Moreover, we found that ofloxacin tolerance of E. coli decreased significantly when E. coli/C. albicans biofilms were treated with matrix-degrading enzymes such as the ß-1,3-glucan-degrading enzyme lyticase. In line with a role for ß-1,3-glucan in mediating ofloxacin tolerance of E. coli in a biofilm, we found that ofloxacin tolerance of E. coli increased even more in E. coli/C. albicans biofilms consisting of a high-ß-1,3-glucan-producing C. albicans mutant. In addition, exogenous addition of laminarin, a polysaccharide composed mainly of poly-ß-1,3-glucan, to an E. coli biofilm also resulted in increased ofloxacin tolerance. All these data indicate that ß-1,3-glucan from C. albicans increases ofloxacin tolerance of E. coli in an E. coli/C. albicans biofilm.

How well do antimicrobial mouth rinses prevent dysbiosis in an in vitro periodontitis biofilm model?

BACKGROUND: Periodontal diseases are associated with dysbiosis in the oral microbial communities. Managing oral biofilms is therefore key for preventing these diseases. Management protocols often include over-the-counter antimicrobial mouth rinses, which lack data on their effects on the oral microbiome's ecology, bacterial composition, metabolic activity, and dysbiosis resilience. This study examined the efficacy of antimicrobial mouth rinses to halt dysbiosis in in vitro oral biofilms under periodontitis-simulating conditions. METHODS: Multispecies oral biofilms were grown on hydroxyapatite discs (HADs) and rinsed daily with one of six mouth rinses. Positive and negative controls were included. After three rinses, biofilms were analyzed with viability quantitative polymerase chain reaction and visualized using scanning electron microscopy. Supernatants of rinsed biofilms were used for metabolic activity analysis. In addition, human oral keratinocytes were exposed to rinsed biofilms to assess their inflammatory response. All outputs were analyzed for correlation using Spearman coefficient. RESULTS: Product-related changes were observed in the rinsed biofilms. Three of the six tested mouth rinses could significantly prevent dysbiosis with ≥30% reduction in pathobiont abundance relative to the control. These biofilms had lower metabolic activity, and the exposed human oral keratinocyte produced less interleukin-8. Interleukin-8 production correlated to both pathobiont quantity and the metabolic activity of the biofilms. CONCLUSION: Some mouth rinses could support biofilm resilience and stop dysbiosis evolution in the biofilm model, with a clear product-related effect. Such mouth rinses can be considered for patients under maintenance/supportive periodontal therapy to prevent/delay disease recurrence. Others are more useful for different periodontal therapy stages.

Development and characterization of colloidal pNIPAM-methylcellulose microgels with potential application for drug delivery in dentoalveolar tissue engineering strategies.

Incorporation of growth factors, signaling molecules and drugs can be vital for the success of tissue engineering in complex structures such as the dentoalveolar region. This has led to the development of a variety of drug release systems. This study aimed to develop pNIPAM-methylcellulose microgels with different synthesis parameters based on a 23 full factorial design of experiments for this application. Microgel properties, including volume phase transition temperature (VPTT), hydrodynamic size, drug loading and release, and cytocompatibility were systematically evaluated. The results demonstrated successful copolymerization and development of the microgels, a hydrodynamic size ranging from ∼200 to ∼500 nm, and VPTT in the range of 34-39 °C. Furthermore, loading of genipin, capable of inducing odontoblastic differentiation, and its sustained release over a week was shown in all formulations. Together, this can serve as a solid basis for the development of tunable drug-delivering pNIPAM-methylcellulose microgels for specific tissue engineering applications.

pH-Triggered Controlled Release of Chlorhexidine Using Chitosan-Coated Titanium Silica Composite for Dental Infection Prevention.

Peri-implantitis is a growing pathological concern for dental implants which aggravates the occurrence of revision surgeries. This increases the burden on both hospitals and the patients themselves. Research is now focused on the development of materials and accompanying implants designed to resist biofilm formation. To enhance this endeavor, a smart method of biofilm inhibition coupled with limiting toxicity to the host cells is crucial. Therefore, this research aims to establish a proof-of-concept for the pH-triggered release of chlorhexidine (CHX), an antiseptic commonly used in mouth rinses, from a titanium (Ti) substrate to inhibit biofilm formation on its surface. To this end, a macroporous Ti matrix is filled with mesoporous silica (together referred to as Ti/SiO2), which acts as a diffusion barrier for CHX from the CHX feed side to the release side. To limit release to acidic conditions, the release side of Ti/SiO2 is coated with crosslinked chitosan (CS), a pH-responsive and antimicrobial natural polymer. Scanning electron microscopy coupled with energy dispersive X-ray spectroscopy (SEM/EDX) and Fourier transform infrared (FTIR) spectroscopy confirmed successful CS film formation and crosslinking on the Ti/SiO2 disks. The presence of the CS coating reduced CHX release by 33% as compared to non-coated Ti/SiO2 disks, thus reducing the antiseptic exposure to the environment in normal conditions. Simultaneous differential scanning calorimetry and thermogravimetric analyzer (SDT) results highlighted the thermal stability of the crosslinked CS films. Quartz crystal microbalance with dissipation monitoring (QCM-D) indicated a clear pH response for crosslinked CS coatings in an acidic medium. This pH response also influenced CHX release through a Ti/SiO2/CS disk where the CHX release was higher than the average trend in the neutral medium. Finally, the antimicrobial study revealed a significant reduction in biofilm formation for the CS-coated samples compared to the control sample using viability quantitative polymerase chain reaction (v-qPCR) measurements, which were also corroborated using SEM imaging. Overall, this study investigates the smart triggered release of pharmaceutical agents aimed at inhibiting biofilm formation, with potential applicability to implant-like structures.

Powdered Cross-Linked Gelatin Methacryloyl as an Injectable Hydrogel for Adipose Tissue Engineering.

The tissue engineering field is currently advancing towards minimally invasive procedures to reconstruct soft tissue defects. In this regard, injectable hydrogels are viewed as excellent scaffold candidates to support and promote the growth of encapsulated cells. Cross-linked gelatin methacryloyl (GelMA) gels have received substantial attention due to their extracellular matrix-mimicking properties. In particular, GelMA microgels were recently identified as interesting scaffold materials since the pores in between the microgel particles allow good cell movement and nutrient diffusion. The current work reports on a novel microgel preparation procedure in which a bulk GelMA hydrogel is ground into powder particles. These particles can be easily transformed into a microgel by swelling them in a suitable solvent. The rheological properties of the microgel are independent of the particle size and remain stable at body temperature, with only a minor reversible reduction in elastic modulus correlated to the unfolding of physical cross-links at elevated temperatures. Salts reduce the elastic modulus of the microgel network due to a deswelling of the particles, in addition to triple helix denaturation. The microgels are suited for clinical use, as proven by their excellent cytocompatibility. The latter is confirmed by the superior proliferation of encapsulated adipose tissue-derived stem cells in the microgel compared to the bulk hydrogel. Moreover, microgels made from the smallest particles are easily injected through a 20G needle, allowing a minimally invasive delivery. Hence, the current work reveals that powdered cross-linked GelMA is an excellent candidate to serve as an injectable hydrogel for adipose tissue engineering.

Light transmittance through resin-matrix composite onlays adhered to resin-matrix cements or flowable composites.

OBJECTIVE: The aim of this study was to evaluate the influence of the thickness of resin-matrix composite blocks manufactured by CAD-CAM on the light transmittance towards different resin-matrix cements or flowable composites. METHODS: Sixty specimens of resin-matrix composite CAD-CAM blocks reinforced with 89 wt% inorganic fillers were cross-sectioned with 2 or 3 mm thicknesses. The specimens were conditioned with adhesive system and divided in groups according to the luting material, namely: two dual-cured resin-matrix cements, two traditional flowable resin-matrix composites, and one thermal-induced flowable resin-matrix composite. Specimens were light-cured at 900 mW/cm2 for 40s. Light transmittance assays were preformed using a spectrophotometer with an integrated monochromator before and after light-curing. Microstructural analysis was performed by optical and scanning electron microscopy (SEM). Nanoindentation tests were performed to evaluate mechanical properties for indirect evaluation of degree of monomers conversion. RESULTS: Optical and SEM images revealed low thickness values for the cementation interfaces for the traditional flowable resin-matrix composite. The cement thickness increased with the size and content of inorganic fillers. The highest light transmittance was recorded for the onlay blocks cemented with the traditional flowable resin-matrix composites while a group cemented with the dual-cured resin-matrix cement revealed the lowest light transmittance. The elastic modulus and hardness increased for specimens with high content of inorganic fillers as well as it increased in function of the light transmittance. CONCLUSIONS: The light transmittance of flowable resin-matrix composites was higher than that for resin-matrix cement after cementation to resin-matrix composites blocks. The type, size, and content of inorganic fillers of the luting material affected the thickness of the cement layer and light transmittance through the materials. CLINICAL RELEVANCE: On chair-side light curing, the transmission of visible light can be interfered by the chemical composition and viscosity of the luting materials. The increase in size and content of inorganic fillers of resin-matrix composites and luting materials can decrease the light transmittance leading to inefficient polymerization.

Exploiting phage-antibiotic synergies to disrupt Pseudomonas aeruginosa PAO1 biofilms in the context of orthopedic infections.

IMPORTANCE: Biofilm-related infections are among the most difficult-to-treat infections in all fields of medicine due to their antibiotic tolerance and persistent character. In the field of orthopedics, these biofilms often lead to therapeutic failure of medical implantable devices and urgently need novel treatment strategies. This forthcoming article aims to explore the dynamic interplay between newly isolated bacteriophages and routinely used antibiotics and clearly indicates synergetic patterns when used as a dual treatment modality. Biofilms were drastically more reduced when both active agents were combined, thereby providing additional evidence that phage-antibiotic combinations lead to synergism and could potentially improve clinical outcome for affected patients.

Oral Biofilm Composition, Dissemination to Keratinocytes, and Inflammatory Attenuation Depend on Probiotic and Synbiotic Strain Specificity.

Several inflammatory diseases are characterized by a disruption in the equilibrium between the host and its microbiome. Due to the increase in resistance, the use of antibiotics for the widespread, nonspecific killing of microorganisms is at risk. Pro-microbial approaches focused on stimulating or introducing beneficial species antagonistic toward pathobionts may be a viable alternative for restoring the host-microbiome equilibrium. Unfortunately, not all potential probiotic or synbiotic species and even subspecies (to strain level) are equally effective for the designated pathology, leading to conflicting accounts of their efficacy. To assess the extent of these species- and strain-specific effects, 13 probiotic candidates were evaluated for their probiotic and synbiotic potential with glycerol on in vitro oral biofilms, dissemination from biofilms to keratinocytes, and anti-inflammatory activity. Species- and strain-specific effects and efficacies were observed in how they functioned as probiotics or synbiotics by influencing oral pathobionts and commensals within biofilms and affected the dissemination of pathobionts to keratinocytes, ranging from ineffective strains to strains that reduced pathobionts by 3 + log. In addition, a minority of the candidates exhibited the ability to mitigate the inflammatory response of LPS-stimulated monocytes. For a comprehensive assessment of probiotic therapy for oral health, a judicious selection of fully characterized probiotic strains that are specifically tailored to the designated pathology is required. This approach aims to challenge the prevailing perception of probiotics, shifting the focus away from "form over function." Rather than using unproven, hypothetical probiotic strains from known genera or species, one should choose strains that are actually functional in resolving the desired pathology before labelling them probiotics.

Biomimetic Periodontal Ligament Transplantation Activated by Gold Nanoparticles Protects Alveolar Bone.

Stem cell therapy might be a promising method to stimulate alveolar bone regeneration, which is currently a major clinical challenge. However, its therapeutic features largely depend on pretreatment and transplantation preparation. Herein, a novel biomimetic periodontal ligament transplantation composed of human periodontal ligament stem cells (hPDLSCs) pretreated with gold nanocomplexes (AuNCs) and embedded in a type-I collagen hydrogel scaffold is developed to protect alveolar bone from resorption. AuNCs are readily absorbed by primary hPDLSCs, with limited cytotoxicity, and promote osteogenic differentiation of hPDLSCs effectively in vitro. In addition, the AuNCs-induced hPDLSCs are encapsulated with type-I collagen hydrogel scaffold to mimic their native physiological niche, and then are transplanted into a rat model of alveolar bone resorption. Both micro-computed tomography (micro-CT) and immunohistochemical assays demonstrate that alveolar bone loss is significantly prevented. Furthermore, the underlying therapeutic mechanism is elucidated, in which transplantation-activated osteogenesis is associated with autophagy, which enables bone remodeling and regeneration. This study provides critical insight into the role of PDLSCs in bone homeostasis and proposes an innovative AuNCs-based strategy for stem cell therapy in bone regeneration.

3D-printed porous Ti6Al4V scaffolds for long bone repair in animal models: a systematic review.

BACKGROUND: Titanium and its alloys have been widely employed for bone tissue repair and implant manufacturing. The rapid development of three-dimensional (3D) printing technology has allowed fabrication of porous titanium scaffolds with controllable microstructures, which is considered to be an effective method for promoting rapid bone formation and decreasing bone absorption. The purpose of this systematic review was to evaluate the osteogenic potential of 3D-printed porous Ti6Al4V (Ti64) scaffold for repairing long bone defects in animal models and to investigate the influential factors that might affect its osteogenic capacity. METHODS: Electronic literature search was conducted in the following databases: PubMed, Web of Science, and Embase up to September 2021. The SYRCLE's tool and the modified CAMARADES list were used to assess the risk of bias and methodological quality, respectively. Due to heterogeneity of the selected studies in relation to protocol and outcomes evaluated, a meta-analysis could not be performed. RESULTS: The initial search revealed 5858 studies. Only 46 animal studies were found to be eligible based on the inclusion criteria. Rabbit was the most commonly utilized animal model. A pore size of around 500-600 µm and porosity of 60-70% were found to be the most ideal parameters for designing the Ti64 scaffold, where both dodecahedron and diamond pores optimally promoted osteogenesis. Histological analysis of the scaffold in a rabbit model revealed that the maximum bone area fraction reached 59.3 ± 8.1% at weeks 8-10. Based on micro-CT assessment, the maximum bone volume fraction was found to be 34.0 ± 6.0% at weeks 12. CONCLUSIONS: Ti64 scaffold might act as a promising medium for providing sufficient mechanical support and a stable environment for new bone formation in long bone defects. Trail registration The study protocol was registered in the PROSPERO database under the number CRD42020194100.

Hydrolytic Enzymes as Potentiators of Antimicrobials against an Inter-Kingdom Biofilm Model.

Biofilms are recalcitrant to antimicrobials, partly due to the barrier effect of their matrix. The use of hydrolytic enzymes capable to degrade matrix constituents has been proposed as an alternative strategy against biofilm-related infections. This study aimed to determine whether hydrolytic enzymes could potentiate the activity of antimicrobials against hard-to-treat interkingdom biofilms comprising two bacteria and one fungus. We studied the activity of a series of enzymes alone or in combination, followed or not by antimicrobial treatment, against single-, dual- or three-species biofilms of Staphylococcus aureus, Escherichia coli, and Candida albicans, by measuring their residual biomass or culturable cells. Two hydrolytic enzymes, subtilisin A and lyticase, were identified as the most effective to reduce the biomass of C. albicans biofilm. When targeting interkingdom biofilms, subtilisin A alone was the most effective enzyme to reduce biomass of all biofilms, followed by lyticase combined with an enzymatic cocktail composed of cellulase, denarase, and dispersin B that proved previously active against bacterial biofilms. The subsequent incubation with antimicrobials further reduced the biomass. Enzymes alone did not reduce culturable cells in most cases and did not interfere with the cidal effects of antimicrobials. Therefore, this work highlights the potential interest of pre-exposing interkingdom biofilms to hydrolytic enzymes to reduce their biomass besides the number of culturable cells, which was not achieved when using antimicrobials alone. IMPORTANCE Biofilms are recalcitrant to antimicrobial treatments. This problem is even more critical when dealing with polymicrobial, interkingdom biofilms, including both bacteria and fungi, as these microorganisms cooperate to strengthen the biofilm and produce a complex matrix. Here, we demonstrate that the protease subtilisin A used alone, or a cocktail containing lyticase, cellulase, denarase, and dispersin B markedly reduce the biomass of interkingdom biofilms and cooperate with antimicrobials to act upon these recalcitrant forms of infection. This work may open perspectives for the development of novel adjuvant therapies against biofilm-related infections.

High rates of implant fracture of a generic polished tapered femoral stem.

OBJECTIVES: Cemented polished tapered stems have demonstrated excellent long-term outcomes. Based on this concept, many generic tapered stems have been released into the market. The aim of this study was to evaluate implant-related complications of 1 specific stem design. METHODS: Between 2010 and 2017, 315 total hip replacements were performed using a Fortress stem (Biotechni, La Ciotat, France). Patient records and radiology were retrospectively reviewed for implant-related complications. A failure analysis was performed on the failed Fortress stems in order to determine the cause of premature failure. RESULTS: 7 (2.2%) patients sustained a fracture of the neck of the implant after a mean of 5 years (range 50-81 months). All fractures were atraumatic, originating at the introducer inlet of the stem. All fractured occurred in obese patients (BMI >33 kg/m2) with a small sized prosthesis. Of these, there were 5 135° and 2 125° stems. Fracture risk was 23% (7/30) for patients with a small sized stem and a BMI >30 kg/m2. All cases were revised using a cement-in-cement technique or a cementless modular revision stem. Failure analysis on the retrieved stems revealed a stress riser at the bottom of the introducer inlet. CONCLUSIONS: An alarmingly high rate of early implant fractures was seen using this specific type of cemented stem, particularly when using smaller implant sizes in obese patients. Although based on a proven design, a specific modification led to a stress riser in the neck area, which resulted in a high incidence of implant failure. This series underlines the importance of a stepwise introduction into the market of new orthopaedic devices even when based on established concepts. Generic stems may not behave as the original stem upon which it was designed.

Mechanical behavior of Ti6Al4V produced by laser powder bed fusion with engineered open porosity for dental applications.

Implant failure due to biofilm formation is a substantial problem in the field of dental prosthetics. A solution has been proposed in the form of implants with a built-in drug reservoir, but combining sufficient strength and longevity with controlled release capability has proven difficult. This work investigates the feasibility of using laser powder bed fusion to create Ti6Al4V structures with open pore channels while maintaining their mechanical stability. These interconnected pore channels are generated by increasing the distance between consecutive melt pools, denominated as oversized hatch spacing. The impact of varying hatch spacing, laser power and scan speed on the degree of porosity was examined, with both an increase in hatch spacing and a decrease in energy density leading to higher porosity. The pore channels were found to be fully interconnected at total porosity values of 14% or more. The compressive modulus, yield strength and ultimate compressive strength are shown to be strongly related to the density of the structure. Based on the minimal strength and full interconnectivity requirements, the optimal additive manufacturing building conditions were determined. The fatigue properties of the resulting samples were investigated under uniaxial and under inclined compression-compression testing according to ISO 14801, which indicated an endurance limit of 217 MPa in the heat treated state. The results indicate that the use of an oversized hatch spacing is suitable for engineering open porous networks.