Effectiveness of Photobiomodulation in Reducing Pain and Disability in Patients with Knee Osteoarthritis: A Systematic Review with Meta-Analysis.

OBJECTIVE: Photobiomodulation (PBM) is not implemented in routine clinical management for knee osteoarthritis. This study aims to systematically investigate the effects of PBM in patients with knee osteoarthritis, comparing to placebo to understand its true clinical effects. METHODS: PubMed, EMBASE, Web of Science, and Cochrane databases were searched up to October 2023. Randomized placebo-controlled trials applying PBM versus placebo were included. Study characteristics, intervention parameters, and patient-reported and physical examination outcome measures were collected. The risk of bias was judged using the Cochrane risk-of-bias tool for randomized trials (version 2) and the Grading of Recommendations Assessment, Development and Evaluation (GRADE) to interpret certainty of results. RESULTS: Ten studies were included comprising 542 participants. All studies were judged with unclear to high risk of bias. Meta-analysis for pain at rest (6 studies) showed that PBM significantly reduced pain at rest as compared to placebo (-0.7 [95% CI = -1.1 to -0.2]), moderate effect, very low certainty of evidence, whereas for the Timed "Up & Go" Test (3 studies), no significant effect was detected. Statistically significantly within-group (PBM) mean improvement was detected for pain, Lequesne Index, and gait performance outcomes, but not always clinically relevant or significant when compared to placebo. CONCLUSION: PBM reduces pain intensity in patients with knee osteoarthritis and may improve disability. However, the very low certainty of evidence does not allow to recommend its isolated use but may be used to complement other widely recommended therapies. More rigorous clinical trials and the revision of the recommended dosage guidelines are warranted to increase the strength of evidence. IMPACT: The findings indicate that photobiomodulation can reduce pain and improve disability in patients with knee osteoarthritis. However, researchers should continue to investigate isolated photobiomodulation intervention versus placebo and extend the dosage guidelines to other types of light emitters.

Devices used for photobiomodulation of the brain-a comprehensive and systematic review.

A systematic review was conducted to determine the trends in devices and parameters used for brain photobiomodulation (PBM). The revised studies included clinical and cadaveric approaches, in which light stimuli were applied to the head and/or neck. PubMed, Scopus, Web of Science and Google Scholar databases were used for the systematic search. A total of 2133 records were screened, from which 97 were included in this review. The parameters that were extracted and analysed in each article were the device design, actuation area, actuation site, wavelength, mode of operation, power density, energy density, power output, energy per session and treatment time. To organize device information, 11 categories of devices were defined, according to their characteristics. The most used category of devices was laser handpieces, which relate to 21% of all devices, while 28% of the devices were not described. Studies for cognitive function and physiological characterisation are the most well defined ones and with more tangible results. There is a lack of consistency when reporting PBM studies, with several articles under defining the stimulation protocol, and a wide variety of parameters used for the same health conditions (e.g., Alzheimer's or Parkinson's disease) resulting in positive outcomes. Standardization for the report of these studies is warranted, as well as sham-controlled comparative studies to determine which parameters have the greatest effect on PBM treatments for different neurological conditions.

Influence of Ultrasound Stimulation on the Viability, Proliferation and Protein Expression of Osteoblasts and Periodontal Ligament Fibroblasts.

Among the adjunctive procedures to accelerate orthodontic tooth movement (OTM), ultrasound (US) is a nonsurgical form of mechanical stimulus that has been explored as an alternative to the currently available treatments. This study aimed to clarify the role of US in OTM by exploring different stimulation parameters and their effects on the biological responses of cells involved in OTM. Human fetal osteoblasts and periodontal ligament fibroblasts cell lines were stimulated with US at 1.0 and 1.5 MHz central frequencies and power densities of 30 and 60 mW/cm2 in continuous mode for 5 and 10 min. Cellular proliferation, metabolic activity and protein expression were analyzed. The US parameters that significantly improved the metabolic activity were 1.0 MHz at 30 mW/cm2 for 5 min and 1.0 MHz at 60 mW/cm2 for 5 and 10 min for osteoblasts; and 1.0 MHz at 30 mW/cm2 for 5 min and 1.5 MHz at 60 mW/cm2 for 5 and 10 min for fibroblasts. By stimulating with these parameters, the expression of alkaline phosphatase was maintained, while osteoprotegerin synthesis was induced after three days of US stimulation. The US stimulation improved the biological activity of both osteoblasts and periodontal ligament fibroblasts, inducing their osteogenic differentiation.

Optimization of a Photobiomodulation Protocol to Improve the Cell Viability, Proliferation and Protein Expression in Osteoblasts and Periodontal Ligament Fibroblasts for Accelerated Orthodontic Treatment.

Numerous pieces of evidence have supported the therapeutic potential of photobiomodulation (PBM) to modulate bone remodeling on mechanically stimulated teeth, proving PBM's ability to be used as a coadjuvant treatment to accelerate orthodontic tooth movement (OTM). However, there are still uncertainty and discourse around the optimal PBM protocols, which hampers its optimal and consolidated clinical applicability. Given the differential expression and metabolic patterns exhibited in the tension and compression sides of orthodontically stressed teeth, it is plausible that different types of irradiation may be applied to each side of the teeth. In this sense, this study aimed to design and implement an optimization protocol to find the most appropriate PBM parameters to stimulate specific bone turnover processes. To this end, three levels of wavelength (655, 810 and 940 nm), two power densities (5 and 10 mW/cm2) and two regimens of single and multiple sessions within three consecutive days were tested. The biological response of osteoblasts and periodontal ligament (PDL) fibroblasts was addressed by monitoring the PBM's impact on the cellular metabolic activity, as well as on key bone remodeling mediators, including alkaline phosphatase (ALP), osteoprotegerin (OPG) and receptor activator of nuclear factor κ-B ligand (RANK-L), each day. The results suggest that daily irradiation of 655 nm delivered at 10 mW/cm2, as well as 810 and 940 nm light at 5 mW/cm2, lead to an increase in ALP and OPG, potentiating bone formation. In addition, irradiation of 810 nm at 5 mW/cm2 delivered for two consecutive days and suspended by the third day promotes a downregulation of OPG expression and a slight non-significant increase in RANK-L expression, being suitable to stimulate bone resorption. Future studies in animal models may clarify the impact of PBM on bone formation and resorption mediators for longer periods and address the possibility of testing different stimulation periodicities. The present in vitro study offers valuable insights into the effectiveness of specific PBM protocols to promote osteogenic and osteoclastogenesis responses and therefore its potential to stimulate bone formation on the tension side and bone resorption on the compression side of orthodontically stressed teeth.

Inconsistency in Shoulder Arthrometers for Measuring Glenohumeral Joint Laxity: A Systematic Review.

There is no consensus on how to measure shoulder joint laxity and results reported in the literature are not well systematized for the available shoulder arthrometer devices. This systematic review aims to summarize the results of currently available shoulder arthrometers for measuring glenohumeral laxity in individuals with healthy or injured shoulders. Searches were conducted on the PubMed, EMBASE, and Web of Science databases to identify studies that measure glenohumeral laxity with arthrometer-assisted assessment. The mean and standard deviations of the laxity measurement from each study were compared based on the type of population and arthrometer used. Data were organized according to the testing characteristics. A total of 23 studies were included and comprised 1162 shoulders. Populations were divided into 401 healthy individuals, 278 athletes with asymptomatic shoulder, and 134 individuals with symptomatic shoulder. Sensors were the most used method for measuring glenohumeral laxity and stiffness. Most arthrometers applied an external force to the humeral head or superior humerus by a manual-assisted mechanism. Glenohumeral laxity and stiffness were mostly assessed in the sagittal plane. There is substantial heterogeneity in glenohumeral laxity values that is mostly related to the arthrometer used and the testing conditions. This variability can lead to inconsistent results and influence the diagnosis and treatment decision-making.

Accuracy of Invisalign® on Upper Incisors: A Systematic Review.

The current systematic review appraises the effectiveness of the types of tooth movements performed with Invisalign® clear aligner on the maxillary incisors. An electronic literature search of published trials was performed through PubMed, LILACS, Scopus, Cochrane Library, and Web of Science databases, and selected journals, from 2009 to 2020. Out of 291 references, five relevant publications were identified for analysis: four studies were performed retrospectively and one prospectively, all non-randomized. Despite the limited set of selected articles, the sample size is significant, with 148 subjects included in the reviewed studies involving the orthodontic treatment of upper incisors. We concluded that movements with the Invisalign® clear aligner on the upper incisors present distinct accuracy, possibly related with movement complexity; intrusion of the incisors has low accuracy (in some cases, 0% of accuracy was reported when the tooth extruded), while incisor extrusion exhibit some of the highest accuracy values reported in the included studies (45%-142%, when the achieved movement was greater than the predicted). Besides, axial (i.e., torque and tip) and horizontal (i.e., translation and rotation) movements are usually effective, with accuracy values between 39%-156% and 42%-79%, respectively. Overall, we determined that the efficiency of aligner to reach the desired movements in the upper incisors was low, as often refinements were required in the included studies. The use of aligner features must be more often considered to improve movement accuracy.

Microscopic Inspection of the Adhesive Interface of Composite Onlays after Cementation on Low Loading: An In Vitro Study.

PURPOSE: This study aimed to assess the layer thickness and microstructure of traditional resin-matrix cements and flowable resin-matrix composites at dentin and enamel to composite onlay interfaces after cementation on low loading magnitude. MATERIALS AND METHODS: Twenty teeth were prepared and conditioned with an adhesive system for restoration with resin-matrix composite onlays manufactured by CAD-CAM. On cementation, tooth-to-onlay assemblies were distributed into four groups, including two traditional resin-matrix cements (groups M and B), one flowable resin-matrix composite (group G), and one thermally induced flowable composite (group V). After the cementation procedure, assemblies were cross-sectioned for inspection by optical microscopy at different magnification up to ×1000. RESULTS: The layer thickness of resin-matrix cementation showed the highest mean values at around 405 µm for a traditional resin-matrix cement (group B). The thermally induced flowable resin-matrix composites showed the lowest layer thickness values. The resin-matrix layer thickness revealed statistical differences between traditional resin cement (groups M and B) and flowable resin-matrix composites (groups V and G) (p < 0.05). However, the groups of flowable resin-matrix composites did not reveal statistical differences (p < 0.05). The thickness of the adhesive system layer at around 7 µm and 12 µm was lower at the interfaces with flowable resin-matrix composites when compared to the adhesive layer at resin-matrix cements, which ranged from 12 µm up to 40 µm. CONCLUSIONS: The flowable resin-matrix composites showed adequate flowing even though the loading on cementation was performed at low magnitude. Nevertheless, significant variation in thickness of the cementation layer was noticed for flowable resin-matrix composites and traditional resin-matrix cements that can occur in chair-side procedures due to the clinical sensitivity and differences in rheological properties of the materials.

A Scoping Review on the Polymerization of Resin-Matrix Cements Used in Restorative Dentistry.

In dentistry, clinicians mainly use dual-cured or light-cured resin-matrix cements to achieve a proper polymerization of the organic matrix leading to enhanced physical properties of the cement. However, several parameters can affect the polymerization of resin-matrix cements. The main aim of the present study was to perform a scoping review on the degree of conversion (DC) of the organic matrix, the polymerization, and the light transmittance of different resin-matrix cements used in dentistry. A search was performed on PubMed using a combination of the following key terms: degree of conversion, resin cements, light transmittance, polymerization, light curing, and thickness. Articles in the English language published up to November 2022 were selected. The selected studies' results demonstrated that restorative structures with a thickness higher than 1.5 mm decrease the light irradiance towards the resin-matrix cement. A decrease in light transmission provides a low energy absorption through the resin cement leading to a low DC percentage. On the other hand, the highest DC percentages, ranging between 55 and 75%, have been reported for dual-cured resin-matrix cements, although the polymerization mode and exposure time also influence the DC of monomers. Thus, the polymerization of resin-matrix cements can be optimized taking into account different parameters of light-curing, such as adequate light distance, irradiance, exposure time, equipment, and wavelength. Then, optimum physical properties are achieved that provide a long-term clinical performance of the cemented restorative materials.

Colorimetric Detection of SARS-CoV-2 Using Plasmonic Biosensors and Smartphones.

Low-cost, instrument-free colorimetric tests were developed to detect SARS-CoV-2 using plasmonic biosensors with Au nanoparticles functionalized with polyclonal antibodies (f-AuNPs). Intense color changes were noted with the naked eye owing to plasmon coupling when f-AuNPs form clusters on the virus, with high sensitivity and a detection limit of 0.28 PFU mL-1 (PFU stands for plaque-forming units) in human saliva. Plasmon coupling was corroborated with computer simulations using the finite-difference time-domain (FDTD) method. The strategies based on preparing plasmonic biosensors with f-AuNPs are robust to permit SARS-CoV-2 detection via dynamic light scattering and UV-vis spectroscopy without interference from other viruses, such as influenza and dengue viruses. The diagnosis was made with a smartphone app after processing the images collected from the smartphone camera, measuring the concentration of SARS-CoV-2. Both image processing and machine learning algorithms were found to provide COVID-19 diagnosis with 100% accuracy for saliva samples. In subsidiary experiments, we observed that the biosensor could be used to detect the virus in river waters without pretreatment. With fast responses and requiring small sample amounts (only 20 µL), these colorimetric tests can be deployed in any location within the point-of-care diagnosis paradigm for epidemiological control.

Photobiomodulation and visual stimulation against cognitive decline and Alzheimer's disease pathology: A systematic review.

Introduction: Given the ineffectiveness of the available drug treatment against Alzheimer disease (AD), light-based therapeutic modalities have been increasingly receiving attention with photobiomodulation (PBM) and, more recently, visual stimulation (VS) being among the most promising approaches. However, the PBM and VS light parameters tested so far, as well as their outcomes, vary a lot with conflicting results being reported. Methods: Based on Scopus, PubMed, and Web of Science databases search, this systematic review summarizes, compares, and discusses 43 cell, animal, and human studies of PBM and VS related to cognitive decline and AD pathology. Results: Preclinical work suggests that PBM with 640±30-nm light and VS at 40 Hz attenuates Aß and Tau pathology and improves neuronal and synaptic plasticity with most studies pointing towards enhancement of degradation/clearance mechanisms in the brain of AD animal models. Despite the gap of the translational evidence for both modalities, the few human studies performed so far support the use of PBM at 810-870 nm light pulsing at 40 Hz for improving brain network connectivity and memory in older subjects and AD patients, while 40 Hz VS in humans seems to improve cognition; further clinical investigation is urgently required to clarify the beneficial impact of PBM and VS in AD patients. Discussion: This review highlights PBM and VS as promising light-based therapeutic approaches against AD brain neuropathology and related cognitive decline, clarifying the most effective light parameters for further preclinical and clinical testing and use. Highlights: Light-based brain stimulation produces neural entrainment and reverts neuronal damageBrain PBM and VS attenuate AD neuropathologyPMB and VS are suggested to improve cognitive performance in AD patients and animal modelsLight stimulation represents a promising therapeutic strategy against neurodegeneration.

Light stimulation on tenocytes: A systematic review of in vitro studies.

The purpose of this systematic review is to analyze the methodologies, utilized stimulation parameters, and the main cellular outcomes obtained by in vitro studies that apply a light source on tenocyte cultures. Methods: The PubMed, Scopus, and Web of Science databases were searched up to December 9, 2019 for in vitro studies that used light sources on tenocyte cultures. A 13-item checklist was used to assess methodological quality of the studies and the risk of bias was assessed using the Risk of Bias Assessment tool for Non-randomized Studies tool. Results: Six studies were included. Tenocytes from the Achilles tendon were used by 83.3% of the studies, with 16.7% utilizing the deep digital flexor tendon, with cells in passage 2 to 5. Four studies used lasers and the other 2 used light-emitting diode or intense pulsed light, in wavelengths ranges from 530 to 1100 nm. The application of light to tenocytes resulted in positive effects reported by all studies, including an increase in cell proliferation and migration, and higher protein and gene expression of tendon biomarkers. Studies presented a lack of standardization on reporting light stimulation parameters and experimental methodologies, leading to low methodological quality. There was a high risk of selection, performance, detection, and reporting bias. Conclusions: All studies showed positive effects after light stimulation on tenocytes, regardless of the light source used. However, the lack of standardized data on light stimulation parameters, experimental setup, and the studies' main limitations hindered representative conclusions and comparisons amongst studies' main outcomes.

Engineering an optimized expression operating unit for improved recombinant protein production in Escherichia coli.

Common strategies to improve recombinant protein production in Escherichia coli often involve the test and optimization of several different variables, when using traditional expression vectors that are commercially available. Now, modern synthetic biology-based strategies allow for extensive modifications of these traditional vectors, or even construction of entirely new modular vectors, so as to permit tunable production of the recombinant proteins of interest. Herein, we describe the engineering of a new expression operating unit (EOU; 938 bp) for producing recombinant proteins in E. coli, through the combinatorial assembly of standardized and well-characterized genetic elements required for transcription and translation (promoter, operator site, RBS, junction RBS-CDS, cloning module, transcriptional terminator). We also constructed a novel T7 promoter variant with increased transcriptional activity (1.7-fold higher), when compared to the canonical wild type T7 promoter sequence. This new EOU yielded an improved production of the reporter protein superfolder GFP (sfGFP) in E. coli BL21(DE3) (relative fluorescence units/RFU = 70.62 ± 1.62 A U.) when compared to a high-producing control expression vector (plasmid BBa_I746909; RFU = 59.68 ± 1.82 A U.). The yields of purified soluble recombinant sfGFP were also higher when using the new EOU (188 mg L-1 culture vs. 108 mg L-1 in the control) and it performed similarly well when inserted into different plasmid backbones (pOPT1.0/AmpR and pOPT2.0/CmR).

Surface modification of zirconia dental implants by laser texturing.

The aim of this work was to perform an integrative literature review on the influence of laser irradiation on zirconia implants to enhance surface topographic aspects and the biological response for osseointegration. An electronic search was carried out on the PubMed database using the following search terms: "zirconia" AND "laser" AND "surface modification" OR "surface treatment" AND "dental implants" OR "bone" OR "osteoblast" OR "osseointegration." Of the identified articles, 12 studies were selected in this review. Results reported that the laser irradiation was capable of promoting changes on the zirconia surfaces regarding topographic aspects, roughness, and wettability. An increase in roughness was recorded at micro- and nano-scale and it resulted in an enhanced wettability and biological response. Also, adhesion, spreading, proliferation, and differentiation of osteogenic cells were also enhanced after laser irradiation mainly by using a femtosecond laser at 10nJ and 80 MHz. After 3 months of osseointegration, in vivo studies in dogs revealed a similar average percentage of bone-to-implant contact (BIC) on zirconia surfaces (around 47.9 ± 16%) when compared to standard titanium surfaces (61.73 ±16.27%), denoting that there is no significant difference between such different materials. The laser  approach revealed several parameters that can be used for zirconia surface modification such as irradiation intensity, time, and frequency. Laser irradiation parameters can be optimized and well-controlled to reach desirable surface morphologic aspects and biological response concerning the osseointegration process.

Rationally designed hypoallergenic mutant variants of the house dust mite allergen Der p 21.

BACKGROUND: Allergic diseases figure among the most common immune-mediated diseases worldwide, affecting more than 25% of the world's population. Allergic reactions can be triggered by house dust mite (HDM) allergens, of which the so-called group 21 of allergens is considered as clinically relevant. METHODS: Herein, we used a structural bioinformatics and immunoinformatics approach to design hypoallergenic mutant variants of the Der p 21 allergen of Dermatophagoides pteronyssinus, which were then recombinantly expressed in bacteria and tested for their IgE-reactivities. For this, we scanned the wild-type Der p 21 protein for all possible single amino acid substitutions in key IgE-binding regions that could render destabilization of the major epitope regions. RESULTS: Four main substitutions (D82P, K110G, E77G, and E87S) were selected to build mutant variants of the Der p 21 allergen, which were produced in their recombinant forms; two of these variants showed reduced reactivity with IgE. Molecular dynamic simulations and immune simulations demonstrated the overall effects of these mutations on the structural stability of the Der p 21 allergen and on the profile of immune response induced through immunotherapy. CONCLUSIONS: When produced in their recombinant forms, two of the Der p 21 mutant variants, namely proteins K110G and E87S, showed significantly reduced IgE reactivities against sera from HDM-allergic individuals (n = 20; p < 0.001). GENERAL SIGNIFICANCE: This study successfully translated a rational in silico mutagenesis design into low IgE-binding mutant variants of the allergen rDer p 21. These novel hypoallergens are promising to compose next-generation allergen-immunotherapy formulations in near future.

Biomechanical analyses of one-piece dental implants composed of titanium, zirconia, PEEK, CFR-PEEK, or GFR-PEEK: Stresses, strains, and bone remodeling prediction by the finite element method.

This work aimed to assess the biomechanics, using the finite element method (FEM), of traditional titanium Morse taper (MT) dental implants compared to one-piece implants composed of zirconia, polyetheretherketone (PEEK), carbon fiber-reinforced PEEK (CFR-PEEK), or glass fiber-reinforced PEEK (GFR-PEEK). MT and one-piece dental implants were modeled within a mandibular bone section and loaded on an oblique force using FEM. A MT implant system involving a Ti6Al4V abutment and a cp-Ti grade IV implant was compared to one-piece implants composed of cp-Ti grade IV, zirconia (3Y-TZP), PEEK, CFR-PEEK, or GFR-PEEK. Stress on bone and implants was computed and analyzed while bone remodeling prediction was evaluated considering equivalent strain. In comparison to one-piece implants, the traditional MT implant revealed higher stress peak (112 MPa). The maximum stresses on the one-piece implants reached ~80 MPa, regardless their chemical composition. MT implant induced lower bone stimulus, although excessive bone strain was recorded for PEEK implants. Balanced strain levels were noticed for reinforced PEEK implants of which CFR-PEEK one-piece implants showed proper biomechanical behavior. Balanced strain levels might induce bone remodeling at the peri-implant region while maintaining low risks of mechanical failures. However, the strength of the PEEK-based composite materials is still low for long-term clinical performance.

Multi-mechanical waves against Alzheimer's disease pathology: a systematic review.

Alzheimer's disease (AD) is the most common cause of dementia, affecting approximately 40 million people worldwide. The ineffectiveness of the available pharmacological treatments against AD has fostered researchers to focus on alternative strategies to overcome this challenge. Mechanical vibrations delivered in different stimulation modes have been associated with marked improvements in cognitive and physical performance in both demented and non-demented elderly. Some of the mechanical-based stimulation modalities in efforts are earlier whole-body vibration, transcranial ultrasound stimulation with microbubble injection, and more recently, auditory stimulation. However, there is a huge variety of treatment specifications, and in many cases, conflicting results are reported. In this review, a search on Scopus, PubMed, and Web of Science databases was performed, resulting in 37 papers . These studies suggest that mechanical vibrations delivered through different stimulation modes are effective in attenuating many parameters of AD pathology including functional connectivity and neuronal circuit integrity deficits in the brains of AD patients, as well as in subjects with cognitive decline and non-demented older adults. Despite the evolving preclinical and clinical evidence on these therapeutic modalities, their translation into clinical practice is not consolidated yet. Thus, this comprehensive and critical systematic review aims to address the most important gaps in the reviewed protocols and propose optimal regimens for future clinical application.

SARS-CoV-2 RNA Detection by a Cellphone-Based Amplification-Free System with CRISPR/CAS-Dependent Enzymatic (CASCADE) Assay.

CRISPR (Clustered regularly interspaced short palindromic repeats)-based diagnostic technologies have emerged as a promising alternative to accelerate delivery of SARS-CoV-2 molecular detection at the point of need. However, efficient translation of CRISPR-diagnostic technologies to field application is still hampered by dependence on target amplification and by reliance on fluorescence-based results readout. Herein, an amplification-free CRISPR/Cas12a-based diagnostic technology for SARS-CoV-2 RNA detection is presented using a smartphone camera for results readout. This method, termed Cellphone-based amplification-free system with CRISPR/CAS-dependent enzymatic (CASCADE) assay, relies on mobile phone imaging of a catalase-generated gas bubble signal within a microfluidic channel and does not require any external hardware optical attachments. Upon specific detection of a SARS-CoV-2 reverse-transcribed DNA/RNA heteroduplex target (orf1ab) by the ribonucleoprotein complex, the transcleavage collateral activity of the Cas12a protein on a Catalase:ssDNA probe triggers the bubble signal on the system. High analytical sensitivity in signal detection without previous target amplification (down to 50 copies µL-1) is observed in spiked samples, in ≈71 min from sample input to results readout. With the aid of a smartphone vision tool, high accuracy (AUC = 1.0; CI: 0.715 - 1.00) is achieved when the CASCADE system is tested with nasopharyngeal swab samples of PCR-positive COVID-19 patients.

In vivo cleavage of solubility tags as a tool to enhance the levels of soluble recombinant proteins in Escherichia coli.

Recombinant proteins are generally fused with solubility enhancer tags to improve the folding and solubility of the target protein of interest. However, the fusion protein strategy usually requires expensive proteases to perform in vitro proteolysis and additional chromatographic steps to obtain tag-free recombinant proteins. Expression systems based on intracellular processing of solubility tags in Escherichia coli, through co-expression of a site-specific protease, simplify the recombinant protein purification process, and promote the screening of molecules that fail to remain soluble after tag removal. High yields of soluble target proteins have already been achieved using these protease co-expression systems. Herein, we review approaches for controlled intracellular processing systems tailored to produce soluble untagged proteins in E. coli. We discuss the different genetic systems available for intracellular processing of recombinant proteins regarding system design features, advantages, and limitations of the various strategies.

The resin-matrix cement layer thickness resultant from the intracanal fitting of teeth root canal posts: an integrative review.

OBJECTIVE: The aim of this study was to perform an integrative review on the layer thickness and microstructure of resin-matrix cements around custom-made or standard teeth root intracanal posts. MATERIALS AND METHODS: An electronic search was conducted on the PubMed using a combination of the following scientific terms: intraradicular post, root intracanal post, resin cement, thickness, adaptation, endodontic post, layer thickness, fit, shape, and endodontic core. The literature selection criteria accepted articles published in the English language, up to May 2021, involving in vitro analyses, meta-analyses, randomized controlled trials, and prospective cohort studies. RESULTS: The search identified 154 studies, of which 24 were considered relevant to this study. The selected studies provided important data considering cement layer thickness, tooth preparation, endodontic post, and type of resin-matrix cement. The anatomical variability of root canal systems, such as the oval- or C-shaped, represents a challenge in dental restoration with tooth root intracanal posts. The fitting of intracanal posts to different root regions is variable resulting in thick and irregular layers of resin-matrix cement. Defects like pores, micro-cracks, and micro-gaps were detected in the resin-matrix cement microstructure and represent spots of stress concentration and fracture. Custom-made tooth root intracanal posts provide a proper fitting and decrease the layer thickness of resin-matrix cement. CONCLUSIONS: In fact, the layer thickness of resin-matrix cements depends on the fitting of endodontic posts to tooth root canals. An increase of resin cement thickness causes the appearance of defects like pores, micro-cracks, and micro-gaps that can induce stress concentration and fractures at interfaces. CLINICAL RELEVANCE: The fitting of the endodontic post into the teeth root canal determine the layer thickness of the resin-matrix cement to establish an adequate retention. However, the increase in the thickness of the resin-matrix cement layer can lead to a high number of defects like pores or cracks and therefore decrease the strength of the interface.

Biomechanical behavior of functionally graded S53P4 bioglass-zirconia dental implants: Experimental and finite element analyses.

OBJECTIVES: The aim of this work was to evaluate the biomechanical behavior of one-piece zirconia implants with a functionally graded bioglass (BG) layer as compared to monolithic zirconia and BG-coated implants, using the finite element method (FEM). METHODS: Zirconia disks were infiltrated with bioglass S53P4 and then morphologically inspected by scanning electron microscopy (SEM) followed by mechanical analyses on micro-indentation tests for further biomechanical validation using the finite element method (FEM). On modeling, zirconia dental implants anchored into mandibular bone were simulated on occlusal loading as recorded under mastication. Three types of implants were simulated: i) free of BG coating, ii) with 100 µm or 150 µm thick conventional BG coatings; and iii) with graded BG coatings involving 3 different chemical composition distributions. The stress state at both implant and bone were evaluated using the FEM. The mechanically-induced bone remodelling was analyzed through the bone strain results. RESULTS: Infiltration of BG into a zirconia structure resulted in a ∼100 µm thick layer with an exponential-like gradation of chemical composition and properties. Regarding the FEM calculations, the BG coating induced up to 30% decrease on stress in the implant body when compared to the monolithic zirconia implant. The gradient of chemical composition also improved the stresses' distribution. The stresses distribution towards the BG-coatings were significantly high and could lead to failure. Stresses on the bone were recorded down to its strength threshold, with insignificant influence of the coating layer. The bone strain values on all models indicates further bone remodelling although BG-coated and BG-graded zirconia implants showed the highest strain magnitude that may enhance the mechanical stimulation for bone maintenance. SIGNIFICANCE: Graded BG-zirconia dental implants showed enhanced overall biomechanical behaviour as compared to the BG-coated or monolithic zirconia dental implants. Also, such biomechanical improvements noticed for the BG-graded system should be considered in combination with the well-known osseointegration benefits of bioactive glasses.