Spatial mapping of mobile genetic elements and their bacterial hosts in complex microbiomes.

The exchange of mobile genetic elements (MGEs) facilitates the spread of functional traits including antimicrobial resistance within bacterial communities. Tools to spatially map MGEs and identify their bacterial hosts in complex microbial communities are currently lacking, limiting our understanding of this process. Here we combined single-molecule DNA fluorescence in situ hybridization (FISH) with multiplexed ribosomal RNA-FISH to enable simultaneous visualization of both MGEs and bacterial taxa. We spatially mapped bacteriophage and antimicrobial resistance (AMR) plasmids and identified their host taxa in human oral biofilms. This revealed distinct clusters of AMR plasmids and prophage, coinciding with densely packed regions of host bacteria. Our data suggest spatial heterogeneity in bacterial taxa results in heterogeneous MGE distribution within the community, with MGE clusters resulting from horizontal gene transfer hotspots or expansion of MGE-carrying strains. Our approach can help advance the study of AMR and phage ecology in biofilms.

Utilizing Individualized Titanium Frames for Protected Alveolar Bone Augmentation: A Feasibility Case Series.

Despite the various treatments proposed with barrier membranes, one of the main challenges for guided bone regeneration (GBR) is maintaining space for large defects and ensuring an adequate blood supply. The presented feasibility case series aims to introduce an original titanium frame (TF) design, customized for each defect, as a modification of well-known principles and materials for GBR to achieve an enhanced and more predictable horizontal and vertical bone augmentation. Three patients with significant horizontal defects were treated with pre-trimmed TFs to create needed space, and then a 50/50 mixture of autograft and bovine xenograft was placed and covered with a collagen membrane. After 8 months of healing, the sites were reopened, and the titanium screws were removed with the frame. An average of 8.0 ± 1.0 mm of horizontal and 3.0 ± 0.0 mm of vertical bone gain were achieved at the time of reentry and implant placement surgery. Bone core biopsy sample was obtained during the implant placement. Histomorphometric analysis revealed that 42.8% of the sample was new vital bone, 18.8% was residual bone graft particles, and 38.4% was bone marrow-like structures. After 3 to 4 months from implant placement, the implants were restored with provisional crowns and then finalized with zirconia screw-retained crowns. This case series suggests that GBR utilizing TFs with or without collagen membranes can be considered a suitable approach for horizontal and vertical bone augmentation. However, based on only three reported cases, the results should be carefully interpreted.

Utilizing Individualized Titanium Frames (ITFs) for Protected Alveolar Bone Augmentation: A feasibility case series.

Despite the various barrier membranes proposed, one of the main challenges for guided bone regeneration (GBR) is space maintenance for large defects as well as ensure adequate blood supply. The presented feasibility case series aims to introduce an original titanium frame (TF) design, customized for each defect, as a modification of well-known principles and materials for GBR, for an enhanced and more predictable horizontal and vertical bone augmentation. Three patients with significant horizontal defects were treated with pre-trimmed TFs to create needed space, a 50%-50% mixture of autograft and bovine xenograft was placed, and then covered with collagen membrane. After 8 months of healing, the sites were reopened, the titanium screws were removed with the frame. An average of 8.0 ± 1.0mm horizontal and 3.0 ± 0.0mm vertical bone gain was achieved at the time of re-entry and implant placement surgery. Bone core biopsy was obtained during the implant placement. Histomorphometric analysis revealed that 42.8% of the sample was new vital bone, 18.8% was residual bone graft particles, and 38.4% was bone marrow like structures. After 3-4 months from implant placement, the implants were restored with provisional crowns and then finalized with zirconia screw-retained crowns. This case series suggests that GBR utilizing TFs with or without collagen membranes can be considered a suitable approach for horizontal and vertical bone augmentation. However, based on only three reported cases, the result should be carefully interpreted.

Partial Extraction Therapy (Part 1): Applications in Full-Arch Dental Implant Therapy.

Partial extraction therapy (PET) is a set of surgical techniques that preserve a portion of the patient's own root structure to maintain blood supply derived from the periodontal ligament complex in order to maintain the periodontium and peri-implant tissues during restorative and implant therapy. PET includes the socket shield technique (SST), proximal shield technique (PrST), pontic shield technique (PtST), and root submergence technique (RST). In a traditional hybrid technique, total extraction and full-arch dental implant therapy often require significant bone reduction and palatal/lingual implant placement. In addition, postextraction preservation of the ridge architecture is a major challenge. This case series demonstrates the use of a combination of PET techniques with digital implant planning and guided implant surgery to achieve highly esthetic outcomes in full-arch implant therapy.

Partial Extraction Therapy (Part 2): Complication Management in Full-Arch Dental Implant Therapy.

Partial extraction therapy (PET) is a group of surgical techniques that preserve the periodontium and peri-implant tissues during restorative and implant therapy by conserving a portion of the patient's own root structure to maintain the blood supply, derived from the periodontal ligament complex. PET includes the socket shield technique (SST), proximal shield technique (PrST), pontic shield technique (PtST), and root submergence technique (RST). Although their clinical success and benefits have been demonstrated, several studies report possible complications. The focus of this article is to highlight management strategies for the most common complications associated with PET, including internal root fragment exposure, external root fragment exposure, and root fragment mobility.

Hydrogel viscoelasticity modulates migration and fusion of mesenchymal stem cell spheroids.

Multicellular spheroids made of stem cells can act as building blocks that fuse to capture complex aspects of native in vivo environments, but the effect of hydrogel viscoelasticity on cell migration from spheroids and their fusion remains largely unknown. Here, we investigated the effect of viscoelasticity on migration and fusion behavior of mesenchymal stem cell (MSC) spheroids using hydrogels with a similar elasticity but different stress relaxation profiles. Fast relaxing (FR) matrices were found to be significantly more permissive to cell migration and consequent fusion of MSC spheroids. Mechanistically, inhibition of ROCK and Rac1 pathways prevented cell migration. Moreover, the combination of biophysical and biochemical cues provided by fast relaxing hydrogels and platelet-derived growth factor (PDGF) supplementation, respectively, resulted in a synergistic enhancement of migration and fusion. Overall, these findings emphasize the important role of matrix viscoelasticity in tissue engineering and regenerative medicine strategies based on spheroids.

A Digital Approach to Immediate-Load, Full-Arch Implant Dentistry: A Case Report.

Conventional approaches to full-arch implant dentistry require a verified master model created by luting together impression jigs. This process involves numerous steps and is sometimes prone to errors that require subsequent correction. A novel approach involving an extraoral scanning technique using an Imetric 4D Imaging system demonstrates an alternative for same-day delivery of printed full-arch prosthetics. Advantages include the ability to offer a same-day provisional restoration without needing to verify an analog master cast.

Treatment of Multiple Gingival Recessions Using Modified Tunnel Technique with V-reverse Sutures: A Report of Three Cases.

AIM: The clinical case series presents a minimally invasive modified tunnel procedure with autogenous connective tissue graft (CTG) using a V-reverse sutures to treat multiple gingival recessions. BACKGROUND: In periodontal and peri-implant plastic procedures, proper graft and flap stabilization are crucial in the outcomes. The coronally advanced flap allows for better access with the possibility of suturing the graft to the de-epithelialized papillae of the periosteum; there is little evidence with using the V-reverse sutures technique in stabilizing the graft and the flap when performing tunnel techniques (TUN). The following case series presents a minimally invasive modified tunnel procedure with autogenous CTG using V-reverse sutures to treat gingival recessions. CASE DESCRIPTION: Three patients with Miller Class I maxillary buccal gingival recessions defects were selected for this study. All subjects were treated with the minimally invasive modified tunnel technique with autogenous subepithelial CTG. V-reverse sutures technique was performed to further improve the stability of the graft at the recipient site. Clinical parameters, including mean recession depth and root coverage esthetic score (RES), were recorded at baseline, 1 week, 2 weeks, 1 month, 3 months, 6 months, and 1-year postoperative follow-up visits. CONCLUSION: At the 1-year follow-up, complete root coverage was achieved in multiple gingival recessions defect sites. In conclusion, this technique represents an alternative treatment for Miller Class I gingival recessions defects with clinical and esthetically satisfactory outcomes. CLINICAL SIGNIFICANCE: Combining the advantages of V-reverse sutures and CTG in the treatment of gingival recessions is feasible and noninvasive.

Viscoelastic Biomaterials for Tissue Regeneration.

The extracellular matrix (ECM) mechanical properties regulate key cellular processes in tissue development and regeneration. The majority of scientific investigation has focused on ECM elasticity as the primary mechanical regulator of cell and tissue behavior. However, all living tissues are viscoelastic, exhibiting both solid- and liquid-like mechanical behavior. Despite increasing evidence regarding the role of ECM viscoelasticity in directing cellular behavior, this aspect is still largely overlooked in the design of biomaterials for tissue regeneration. Recently, with the emergence of various bottom-up material design strategies, new approaches can deliver unprecedented control over biomaterial properties at multiple length scales, thus enabling the design of viscoelastic biomaterials that mimic various aspects of the native tissue ECM microenvironment. This review describes key considerations for the design of viscoelastic biomaterials for tissue regeneration. We provide an overview of the role of matrix viscoelasticity in directing cell behavior toward regenerative outcomes, highlight recent strategies utilizing viscoelastic hydrogels for regenerative therapies, and outline remaining challenges, potential solutions, and emerging applications for viscoelastic biomaterials in tissue engineering and regenerative medicine. Impact statement All living tissues are viscoelastic. As we design viscoelastic biomaterials for tissue engineering and regenerative medicine, we must understand the effect of matrix viscoelasticity on in vitro cell behavior and in vivo regenerative outcomes. Engineering the next generation of biomaterials with tunable viscoelasticity to direct cell and tissue behavior will contribute to the development of in vitro tissue models and in vivo regenerative therapies to address unmet clinical needs.

Partial Extraction Therapy: A Review of Human Clinical Studies.

Partial extraction therapy (PET) is a collective concept encompassing a group of surgical techniques including socket shield, root membrane, proximal shield, pontic shield, and root submergence. PET uses the patient's own root structure to maintain blood supply derived from the periodontal ligament complex to preserve the periodontium and peri-implant tissues during restorative and implant therapy. This review aims to summarize the current knowledge regarding PET techniques and present a comprehensive evaluation of human clinical studies in the literature. Two independent reviewers conducted electronic and manual searches until January 1, 2021, in the following electronic bibliographic databases: PubMed, EMBASE, and Dentistry & Oral Sciences Source. Gray literature was searched to identify additional candidates for potential inclusion. Articles were screened by a group of 4 reviewers using the Covidence software and synthesized. A systematic search of the literature yielded 5714 results. Sixty-four articles were selected for full-text assessment, of which 42 eligible studies were included in the review. Twelve studies were added to the synthesis after a manual search of the reference lists. A total of 54 studies were examined in this review. In sum, PET techniques offer several clinical advantages: (1) preservation of buccal bone postextraction and limitation of alveolar ridge resorption, (2) mitigation of the need for invasive ridge augmentation procedures, and (3) soft-tissue dimensional stability and high esthetic outcomes. Further randomized clinical studies with larger sample sizes are needed to improve the understanding of the long-term clinical outcomes of PET.

Polymeric Scaffolds for Dental, Oral, and Craniofacial Regenerative Medicine.

Dental, oral, and craniofacial (DOC) regenerative medicine aims to repair or regenerate DOC tissues including teeth, dental pulp, periodontal tissues, salivary gland, temporomandibular joint (TMJ), hard (bone, cartilage), and soft (muscle, nerve, skin) tissues of the craniofacial complex. Polymeric materials have a broad range of applications in biomedical engineering and regenerative medicine functioning as tissue engineering scaffolds, carriers for cell-based therapies, and biomedical devices for delivery of drugs and biologics. The focus of this review is to discuss the properties and clinical indications of polymeric scaffold materials and extracellular matrix technologies for DOC regenerative medicine. More specifically, this review outlines the key properties, advantages and drawbacks of natural polymers including alginate, cellulose, chitosan, silk, collagen, gelatin, fibrin, laminin, decellularized extracellular matrix, and hyaluronic acid, as well as synthetic polymers including polylactic acid (PLA), polyglycolic acid (PGA), polycaprolactone (PCL), poly (ethylene glycol) (PEG), and Zwitterionic polymers. This review highlights key clinical applications of polymeric scaffolding materials to repair and/or regenerate various DOC tissues. Particularly, polymeric materials used in clinical procedures are discussed including alveolar ridge preservation, vertical and horizontal ridge augmentation, maxillary sinus augmentation, TMJ reconstruction, periodontal regeneration, periodontal/peri-implant plastic surgery, regenerative endodontics. In addition, polymeric scaffolds application in whole tooth and salivary gland regeneration are discussed.

Regenerative Medicine Technologies to Treat Dental, Oral, and Craniofacial Defects.

Additive manufacturing (AM) is the automated production of three-dimensional (3D) structures through successive layer-by-layer deposition of materials directed by computer-aided-design (CAD) software. While current clinical procedures that aim to reconstruct hard and soft tissue defects resulting from periodontal disease, congenital or acquired pathology, and maxillofacial trauma often utilize mass-produced biomaterials created for a variety of surgical indications, AM represents a paradigm shift in manufacturing at the individual patient level. Computer-aided systems employ algorithms to design customized, image-based scaffolds with high external shape complexity and spatial patterning of internal architecture guided by topology optimization. 3D bioprinting and surface modification techniques further enhance scaffold functionalization and osteogenic potential through the incorporation of viable cells, bioactive molecules, biomimetic materials and vectors for transgene expression within the layered architecture. These computational design features enable fabrication of tissue engineering constructs with highly tailored mechanical, structural, and biochemical properties for bone. This review examines key properties of scaffold design, bioresorbable bone scaffolds produced by AM processes, and clinical applications of these regenerative technologies. AM is transforming the field of personalized dental medicine and has great potential to improve regenerative outcomes in patient care.

Promotion mechanism for the growth of CO2 hydrate with urea using molecular dynamics simulations.

The role of urea as a kinetic promoter for the growth of CO2 hydrates is revealed for the first time using molecular dynamics simulations. Analysis of simulation trajectories shows that urea plays two important roles in the growth process: increasing mass transport of CO2 and catalyzing cage formation at the solid-liquid interface.

Electric-Field-Driven Assembly of Dipolar Spheres Asymmetrically Confined between Two Electrodes.

Externally applied electric fields have previously been utilized to direct the assembly of colloidal particles confined at a surface into a large variety of colloidal oligomers and nonclose-packed honeycomb lattices (J. Am. Chem. Soc. 2013, 135, 7839-7842). The colloids under such confinement and fields are observed to spontaneously organize into bilayers near the electrode. To extend and better understand how particles can come together to form quasi-two-dimensional materials, we have performed Monte Carlo simulations and complementary experiments of colloids that are strongly confined between two electrodes under an applied alternating current electric field, controlling field strength and particle area fraction. Of particular importance, we control the fraction of particles in the upper vs lower plane, which we describe as asymmetric confinement, and which effectively modulates the coordination number of particles in each plane. We model the particle-particle interactions using a Stockmayer potential to capture the dipolar interactions induced by the electric field. Phase diagrams are then delineated as a function of the control parameters, and a theoretical model is developed in which the energies of several idealized lattices are calculated and compared. We find that the resulting theoretical phase diagrams agree well with simulation. We have not only reproduced the structures observed in experiments using parameters that are close to experimental conditions but also found several previously unobserved phases in the simulations, including a network of rectangular bands, zig zags, and a sigma lattice, which we were then able to confirm in experiment. We further propose a simple way to precisely control the number ratio of particles between different planes, that is, superimposing a direct current electric field with the alternating current electric field, which can be implemented conveniently in experiments. Our work demonstrates that a diverse collection of materials can be assembled from relatively simple ingredients, which can be analyzed effectively through comparison of simulation, theory, and experiment. Our model further explains possible pathways between different phases and provides a platform for examining phases that have yet to be observed in experiments.

COVID-19's impact on private practice and academic dentistry in North America.

The COVID-19 pandemic is a major public health crisis for countries around the world. In response to this global outbreak, the World Health Organization declared a public health emergency of international concern. Dental professionals are especially at high risk of contracting the COVID-19 virus due to the unique nature of dentistry, more specifically, exposure to aerosols and droplets. When it comes to dental emergencies, it was crucial to maintain urgent dental care services operational to help reduce the burden on our healthcare system and hospitals already under pressure. The COVID-19 pandemic has significantly impacted how dentistry is practiced in North America in both the private practice and academic settings. This article shares the perspectives of dentists practicing in private practice and clinician-researchers in academic dental institutions. More specifically, we discuss about measures implemented to minimize risks of disease transmission, challenges in emergency dental care, impact on patients, as well as impact on the professional and personal lives of the dental team during the COVID-19 crisis.

The impact of COVID-19 on dental education in North America-Where do we go next?

During the COVID-19 pandemic, the dental education community faced unprecedented challenges. In this commentary, we share the perspectives of faculty clinicians, residents and students in academic dental institutions in the United States and Canada. We discuss COVID-19's impact on various aspects of academic dentistry including patient care, education, research and raise key concerns regarding the future of dental education post-pandemic.

Hydrate Growth on Methane Gas Bubbles in the Presence of Salt.

Methane bubble dispersions in a water column can be observed in both vertical subsea piping as well as subsea gas seepages. Hydrate growth has been shown to occur at the gas-water interface under flowing conditions, yet the majority of the current literature is limited to quiescent systems. Gas hydrate risks in subsea piping have been shown to increase in late life production wells with increased water content and with gas-in-water bubble dispersions. The dissolution of subsea methane seepages into seawater, or methane release into the atmosphere, can be affected by hydrate film growth on rising bubbles. A high-pressure water tunnel (HPWT), was used to generate a turbulent, continuous water flow system representative of a vertical jumper line to study the relationship between bulk methane hydrate growth and bubble size during a production-well restart. The HPWT comprises a flow loop of 19.1 mm inner diameter and 4.9 m length, with a vertical section containing an optical window to enable visualization of the bubble and hydrate flow dynamics via a high-speed, high-resolution video camera. Additional online monitoring includes the differential pressure drop, viscosity, temperature, flow rates, and gas consumption. Experimental conditions were maintained at 275 K and 6.2 MPa during hydrate formation and 298 K and 1.4 MPa during hydrate dissociation. Hydrate growth using freshwater and saltwater (3.5 wt % NaCl) was measured at four flow velocities (0.8, 1.2, 1.6, and 1.9 m s-1). The addition of salt is shown in this work to alter the surface properties of bubbles, which introduces changes to bubble dynamics of dispersion and coalescence. Hydrate volume fractions and growth rates in the presence of salt were on average ∼32% lower compared to that in freshwater. This was observed and validated to be due to bubble size and dynamic factors and not due to the 1.5 K thermodynamic inhibition effect of salt. Throughout hydrate growth, methane bubbles in pure freshwater maintained larger diameters (2.4-4.2 mm), whereas the presence of salt promoted fine gas bubble dispersions (0.1-0.7 mm), increasing gas-water interfacial area. While gas bubble coalescence was observed in all freshwater experiments, the addition of salt limited coalescence between gas bubbles and reduced bubble size. Consequently, earlier formation of solid hydrate shells in saltwater produced early mass-transfer barriers reducing hydrate growth rates. While primarily directed toward flow assurance, the observed relationship between hydrates, bubble size, and saltwater also applies to broader research fields in subsea gas seepages and naturally occurring hydrates.

Cage occupancies, lattice constants, and guest chemical potentials for structure II hydrogen clathrate hydrate from Gibbs ensemble Monte Carlo simulations.

In this paper, equilibrium properties of structure II hydrates of hydrogen were determined from Monte Carlo simulations in the isothermal-isobaric Gibbs ensemble. Water and hydrogen molecules are described by the TIP4P/Ice and Silvera-Goldman models, respectively. The use of the Gibbs ensemble has many key advantages for the simulation of hydrates. By the separation of hydrogen vapor and hydrate phases into their own domains, coupled with transfer moves of hydrogen molecules between domains, cage occupancies were determined. Furthermore, the choice of this ensemble also allows equilibrium lattice constants and guest molecule chemical potentials to be straightforwardly estimated. Results for hydrogen mass fractions indicate reasonable agreement with prior simulation data and theoretical models, while detailed analysis of cage occupancy distributions and neighboring cage pair occupancy combinations gives valuable insight into the behavior of this hydrate at the inter-cage scale. These results will aid in the construction of theoretical models, for which knowledge of the occupancy of neighboring cages is of great importance. In support of previous experimental and theoretical works, we also find evidence of double occupancy of a few small cages inside of the hydrate stability zone, albeit at very high pressures; approximately 0.1% of small cages are doubly occupied at 300 MPa, for temperatures of 225 K and 250 K.