Biofilm formation on dental implants with a hybrid surface microtopography: An in vitro study in a validated multispecies dynamic biofilm model.

OBJECTIVES: The objective of this study is to qualitatively and quantitatively evaluate biofilm formation on hybrid titanium implants (HS), with moderately rough and turned surface topographies. MATERIALS AND METHODS: A validated dynamic in vitro multispecies biofilm model, based on bacterial growth under flow and shear conditions resembling the oral cavity, was used to evaluate biofilm formation on the tested implant surfaces. Scanning electron microscopy (SEM) and confocal laser scanning microscopy (CLSM) were used to compare the biofilm structure and microbial biomass deposited on either the moderately rough or the turned surface of HS. Quantitative polymerase chain reaction (qPCR) was used to evaluate the total bacterial counts and counts of each specific bacterium in biofilms formed on implants with either the moderately rough or the turned surfaces, as in the hybrid titanium implants, after 24, 48 and 72 h. A general linear model was applied to compare the CLSM and qPCR results between the tested implant surfaces. RESULTS: A significantly higher bacterial biomass grew on the moderately rough implant surfaces, compared to the turned surface area of HS implants (p < .05), at all incubation times, as evidenced with both CLSM and SEM. qPCR analysis also demonstrated an important increase in the total and specific bacterial counts in moderately rough surface implants at the three incubation times. CONCLUSIONS: Implant surface topography (moderately rough versus turned) significantly influenced in vitro biofilm formation in terms of biofilm structure, bacterial biomass and quantity of the specific species selected for the model used.

Impact of the COVID pandemic on vascular access creation for haemodialysis in 16 Spanish haemodialysis centres.

Background: The coronavirus disease (COVID) pandemic has resulted in a major disruption in healthcare that has affected several medical and surgical specialties. European and American Vascular Societies have proposed deferring the creation of an elective vascular access (VA) [autologous or prosthetic arteriovenous fistula (AVF) or arteriovenous graft (AVG)] in incident patients on haemodialysis (HD) in the era of the COVID pandemic. The aim of this study is to examine the impact of the COVID pandemic on VA creation and the central venous catheter (CVC)-related hospitalizations and complications in HD patients dialyzed in 16 Spanish HD units of three different regions. Methods: We compared retrospectively two periods of time: the pre-COVID (1 January 2019-11 March 2020) and the COVID era (12 March 2020-30 June 2021) in all HD patients (prevalent and incident) dialyzed in our 16 HD centres. The variables analysed were type of VA (CVC, AVF and AVG) created, percentage of CVC in incident and prevalent HD patients, CVC-related hospitalizations and complications (infection, extrusion, disfunction, catheter removal) and percentage of CVC HD sessions that did not reach the goal of Kt (>45) as a marker of HD adequacy. Results: A total of 1791 VAs for HD were created and 905 patients started HD during the study period. Patients who underwent vascular access surgery during the COVID period compared with pre-COVID period were significantly younger, with a significant decrease in surgical activity to create AVFs and AVGs in older HD patients (>75 and >85 years of age). There was a significant increase in CVC placement (from 59.7% to 69.5%; P < 0.001) from the pre-COVID to the COVID period. During the COVID pandemic, a significantly higher number of patients started HD through a CVC (80.3% versus 69.1%; P < 0.001). The percentage of CVC in prevalent HD patients has not decreased in the 19 months since the start of the pandemic [414 CVC/1058 prevalent patients (39.4%)]. No significant changes were detected in CVC-related hospitalizations between the pre-COVID and COVID periods. In the COVID period, a significant increase in catheter replacement and the percentage of HD session that did not reach the HD dose objective (Kt > 45) was observed. Conclusions: COVID has presented a public health system crisis that has influenced VA for HD, with an increase in CVCs relative to AVFs. A decrease in HD sessions that did not reach the HD dose objective was observed in the COVID period compared with a pre-COVID period.

Evidence of Persisting Autoreactivity in Post-Acute Sequelae of SARS-CoV-2 Infection

While significant attention has been paid to the immunologic determinants of disease states associated with COVID-191,2, their contributions to post-acute sequelae of COVID-19 (PASC) remain less clear3-5. Due to the wide array of PASC presentations6, it is critical to understand if specific features of the disease are associated with discrete immune processes, and whether those processes may be therapeutically targeted. To this end, we performed wide immunologic and serological characterization of patients in the early recovery phase of COVID-19 across a breadth of symptomatic presentations. Using high-parameter proteomics screening and applied machine learning (ML), we identify clear signatures of immunologic activity between PASC patients and uncomplicated recovery, dominated by inflammatory cytokine signaling, neutrophil activity, and markers of cell death. Consistent with disease complexity, heterogeneity in plasma profiling reveals distinct PASC subsets with striking divergence in these ongoing inflammatory processes, here termed plasma quiescent (plaq) and inflammatory (infl) PASC. In addition to elevated inflammatory blood proteomics, inflPASC patients display positive clinical tests of acute inflammation including C-reactive protein and fibrinogen, increased B cell activity with extrafollicular involvement coupled with elevated targeting of viral nucleocapsid protein and clinical autoreactivity. Further, the unique plasma signatures of PASC patients allowed for the creation of refined models with high sensitivity and specificity for the positive identification of inflPASC with a streamlined assessment of 12 blood markers. Additionally, refined ML modeling highlights the unexpected significance of several markers of potential diagnostic or therapeutic use for PASC in general, including the peptide hormone, epiregulin. In all, this work identifies clear biological signatures of PASC with potential diagnostic and therapeutic potential and establishes clear disease subtypes that are both easily identifiable and highly relevant to ongoing efforts in both therapeutic targeting and epidemiological investigation of this highly complex disease.

Pathogenic neutrophilia drives acute respiratory distress syndrome in severe COVID-19 patients

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and the ensuing COVID-19 pandemic have caused [~]40 million cases and over 648,000 deaths in the United States alone. Troubling disparities in COVID-19-associated mortality emerged early, with nearly 70% of deaths confined to Black/African-American (AA) patients in some areas, yet targeted studies within this demographic are scant. Multi-omics single-cell analyses of immune profiles from airways and matching blood samples of Black/AA patients revealed low viral load, yet pronounced and persistent pulmonary neutrophilia with advanced features of cytokine release syndrome and acute respiratory distress syndrome (ARDS), including exacerbated production of IL-8, IL-1{beta}, IL-6, and CCL3/4 along with elevated levels of neutrophil elastase and myeloperoxidase. Circulating S100A12+/IFITM2+ mature neutrophils are recruited via the IL-8/CXCR2 axis, which emerges as a potential therapeutic target to reduce pathogenic neutrophilia and constrain ARDS in severe COVID-19. Graphical AbstractThe lung pathology due to severe COVID-19 is marked by a perpetual pathogenic neutrophilia, leading to acute respiratory distress syndrome (ARDS) even in the absence of viral burden. Circulating mature neutrophils are recruited to the airways via IL-8 (CXCL8)/CXCR2 chemotaxis. Recently migrated neutrophils further differentiate into a transcriptionally active and hyperinflammatory state, with an exacerbated expression of IL-8 (CXCL8), IL-1{beta} (IL1B), CCL3, CCL4, neutrophil elastase (NE), and myeloperoxidase (MPO) activity. Airway neutrophils and recruited inflammatory monocytes further increase their production of IL-8 (CXCL8), perpetuating lung neutrophilia in a feedforward loop. MdCs and T cells produce IL-1{beta} and TNF, driving neutrophils reprogramming and survival. O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=142 SRC="FIGDIR/small/446468v2_ufig1.gif" ALT="Figure 1"> View larger version (43K): org.highwire.dtl.DTLVardef@81fd3aorg.highwire.dtl.DTLVardef@181e63org.highwire.dtl.DTLVardef@172fedcorg.highwire.dtl.DTLVardef@ba55a7_HPS_FORMAT_FIGEXP M_FIG C_FIG

Sealing Socket Abutments (SSAs) in Molar Immediate Implants with a Digitalized CAD/CAM Protocol: Soft Tissue Contour Changes and Radiographic Outcomes After 2 Years.

Immediate implant placement in molar sites has the potential to improve the patient experience by reducing the number of appointments and the overall treatment time. However, primary closure remains a technical challenge. The present prospective case series evaluated the soft tissue contours and the radiographic bone levels of 17 patients who received immediate implants in molar sites and a digitally customized CAD/CAM sealing socket abutment. At the 2-year follow-up, the mean buccal tissue contours at the most coronal portion were reduced horizontally by an average of 1 mm at 1, 2, 3, and 4 mm below the gingival margin. A mean 0.53-mm apical migration of the gingival margin was seen, and the mean interproximal bone level at the 2-year follow-up was 0.89 mm. The use of CAD/CAM-generated customized healing abutments in immediate molar sites yielded minimal hard and soft tissue changes at the 2-year follow-up.

Broadly-targeted autoreactivity is common in severe SARS-CoV-2 Infection

An emerging feature of COVID-19 is the identification of autoreactivity in patients with severe disease that may contribute to disease pathology, however the origin and resolution of these responses remain unclear. Previously, we identified strong extrafollicular B cell activation as a shared immune response feature between both severe COVID-19 and patients with advanced rheumatic disease. In autoimmune settings, this pathway is associated with relaxed peripheral tolerance in the antibody secreting cell compartment and the generation of de novo autoreactive responses. Investigating these responses in COVID-19, we performed single-cell repertoire analysis on 7 patients with severe disease. In these patients, we identify the expansion of a low-mutation IgG1 fraction of the antibody secreting cell compartment that are not memory derived, display low levels of selective pressure, and are enriched for autoreactivity-prone IGHV4-34 expression. Within this compartment, we identify B cell lineages that display specificity to both SARS-CoV-2 and autoantigens, including pathogenic autoantibodies against glomerular basement membrane, and describe progressive, broad, clinically relevant autoreactivity within these patients correlated with disease severity. Importantly, we identify anti-carbamylated protein responses as a common hallmark and candidate biomarker of broken peripheral tolerance in severe COVID-19. Finally, we identify the contraction of this pathway upon recovery, and re-establishment of tolerance standards coupled with a concomitant loss of acute-derived ASCs irrespective of antigen specificity. In total, this study reveals the origins, breadth, and resolution of acute-phase autoreactivity in severe COVID-19, with significant implications in both early interventions and potential treatment of patients with post-COVID sequelae.

Critically ill SARS-CoV-2 patients display lupus-like hallmarks of extrafollicular B cell activation

Abstract/IntroductionA wide clinical spectrum has become a hallmark of the SARS-CoV-2 (COVID-19) pandemic, although its immunologic underpinnings remain to be defined. We have performed deep characterization of B cell responses through high-dimensional flow cytometry to reveal substantial heterogeneity in both effector and immature populations. More notably, critically ill patients displayed hallmarks of extrafollicular B cell activation as previously described in autoimmune settings. Extrafollicular activation correlated strongly with large antibody secreting cell expansion and early production of high levels of SARS-CoV-2-specific antibodies. Yet, these patients fared poorly with elevated inflammatory biomarkers, multi-organ failure, and death. Combined, the findings strongly indicate a major pathogenic role for immune activation in subsets of COVID-19 patients. Our study suggests that, as in autoimmunity, targeted immunomodulatory therapy may be beneficial in specific patient subpopulations that can be identified by careful immune profiling.

Rapid isolation and profiling of a diverse panel of human monoclonal antibodies targeting the SARS-CoV-2 spike protein

Antibodies are a principal determinant of immunity for most RNA viruses and have promise to reduce infection or disease during major epidemics. The novel coronavirus SARS-CoV-2 has caused a global pandemic with millions of infections and hundreds of thousands of deaths to date1,2. In response, we used a rapid antibody discovery platform to isolate hundreds of human monoclonal antibodies (mAbs) against the SARS-CoV-2 spike (S) protein. We stratify these mAbs into five major classes based on their reactivity to subdomains of S protein as well as their cross-reactivity to SARS-CoV. Many of these mAbs inhibit infection of authentic SARS-CoV-2 virus, with most neutralizing mAbs recognizing the receptor-binding domain (RBD) of S. This work defines sites of vulnerability on SARS-CoV-2 S and demonstrates the speed and robustness of new antibody discovery methodologies.

Long-term assessment of periodontal disease progression after surgical or non-surgical treatment: a systematic review

The primary aim of this systematic review was to assess the evidence on periodontal disease progression after treatment in patients receiving supportive periodontal therapy (SPT) and to identify predictors of clinical attachment level (CAL) loss. A protocol was developed to answer the following focused question: In adult patients treated for periodontitis, what is the disease progression in terms of CAL loss after surgical or non-surgical treatment? Randomized controlled clinical trials, prospective cohort studies, and longitudinal observational human studies with a minimum of 5 years of follow-up after surgical or non-surgical treatment that reported CAL and probing depth changes were selected. Seventeen publications reporting data from 14 investigations were included. Data from 964 patients with a follow-up range of 5–15 years was evaluated. When the CAL at the latest follow-up was compared to the CAL after active periodontal therapy, 10 of the included studies reported an overall mean CAL loss of ≤0.5 mm, 3 studies reported a mean CAL loss of 0.5–1 mm, and 4 studies reported a mean CAL loss of >1 mm. Based on 7 publications, the percentage of sites showing a CAL loss of ≥2 mm varied from 3% to 20%, and a high percentage of sites with CAL loss was associated with poor oral hygiene, smoking, and poor compliance with SPT. The outcomes after periodontal therapy remained stable over time. Disease progression occurred in a reduced number of sites and patients, mostly associated with poor oral hygiene, poor compliance with SPT, and smoking.

A prospective 9-month human clinical evaluation of Laser-Assisted New Attachment Procedure (LANAP) therapy.

This investigation was designed and implemented as a single-center, prospective study to evaluate the clinical response to the Laser-Assisted New Attachment Procedure (LANAP). Eight patients with advanced periodontitis were enrolled and treated with full-mouth LANAP therapy and monitored for 9 months. Fullmouth clinical measurements, including clinical attachment level (CAL), probing depth (PD), and recession, were provided at baseline and after 9 months of healing by a single calibrated examiner, including a total of 930 sites and 444 sites with initial PD equal to or greater than 5 mm. Clinical results for the 930 sites measured pre- and postoperatively revealed that mean PD was reduced from 4.62 ± 2.29 mm to 3.14 ± 1.48 mm after 9 months (P < .05). CAL decreased from 5.58 ± 2.76 mm to 4.66 ± 2.10 mm (P < .05) and recession increased from 0.86 ± 1.31 mm to 1.52 ± 1.62 after 9 months (P < .05). For the subset of 444 sites with initial PD greater than or equal to 5 mm, the PD decreased from 6.50 ± 2.07 mm to 3.92 ± 1.54 mm (P < .05) and CAL decreased from 7.42 ± 2.70 mm to 5.78 ± 2.06 mm (P < .05). As demonstrated by the clinical evaluation, the majority of treated sites demonstrated clinical improvement. LANAP therapy should be further investigated with long-term clinical trials to compare the stability of clinical results with conventional therapy.

Early bone healing around 2 different experimental, HA grit-blasted, and dual acid-etched titanium implant surfaces. A pilot study in rabbits.

PURPOSE: To compare early bone healing around different experimental titanium implant surfaces and to evaluate the role of a calcium phosphate-coated implant surface because it relates to bone-implant contact (BIC). METHODS: An experimental hydroxyapatite (HA) grit-blasted and dual acid-etched titanium surface (BAE-1) was compared to an experimental HA grit-blasted and dual acid-etched surface treated with nanometer-scale crystals of HA (BAE-2). Both experimental implant surfaces were implanted onto the tibias of 4 New Zealand white rabbits. The animals were killed at 1,6, 21, and 90 days after the implant surgery. Descriptive histology was performed at the healing responses of both implant surfaces. Quantitative morphology assessment provided measurements of BIC, number of bone multicellular units (BMUs), average penetration of BMUs, and maximum penetration of BMUs that were manually made using imaging computer software. RESULT: The overall BIC for the BAE-2 implant was higher than that for the BAE-1 implant at 21 days of healing. However, there was no significant difference at 90 days of healing. CONCLUSION: It is concluded from this animal pilot study that the bioactive BAE-2 implant surface provided a better BIC with healthy bone remodeling at 21 days of healing.

Systematic review of pre-clinical models assessing implant integration in locally compromised sites and/or systemically compromised animals.

OBJECTIVE: The aim was to systematically search the dental literature for pre-clinical models assessing implant integration in locally compromised sites (part 1) and systemically compromised animals (part 2), and to evaluate the quality of reporting of included publications. METHODS: A Medline search (1966-2011) was performed, complimented by additional hand searching. The quality of reporting of the included publications was evaluated using the 20 items of the ARRIVE (Animals in Research In Vivo Experiments) guidelines. RESULTS: One-hundred and seventy-six (part 1; mean ARRIVE score = 15.6 ± 2.4) and 104 (part 2; 16.2 ± 1.9) studies met the inclusion criteria. The overall mean score for all included studies amounted to 15.8 ± 2.2. Housing (38.3%), allocation of animals (37.9%), numbers analysed (50%) and adverse events (51.4%) of the ARRIVE guidelines were the least reported. Statistically significant differences in mean ARRIVE scores were found depending on the publication date (p < 0.05), with the highest score of 16.7 ± 1.6 for studies published within the last 2 years. CONCLUSIONS: A large number of studies met the inclusion criteria. The ARRIVE scores revealed heterogeneity and missing information for selected items in more than 50% of the publications. The quality of reporting shifted towards better-reported pre-clinical trials within recent years.