Baseline masticatory muscles' performance may predict pain relief in temporomandibular disorders.

OBJECTIVES: To compare masticatory muscles' recruitment in patients with temporomandibular disorders and asymptomatic control subjects. To evaluate if the masticatory muscles' recruitment pattern may predict symptoms' improvement after temporomandibular disorders treatment. MATERIALS AND METHODS: Standardized surface electromyography of anterior temporalis and superficial masseters muscles were recorded and compared at baseline in 26 patients with arthrogenous temporomandibular disorders (study group) and 26 asymptomatic subjects (control group). The study group was treated pharmacologically and by means of five arthrocentesis sessions. Pre-, during-, and post-treatment pain and mandibular function were assessed and compared among timepoints. Clinical improvement in terms of pain and mandibular function was correlated with pre-treatment standardized surface electromyography values. RESULTS: Temporomandibular disorders patients showed improved maximum mouth opening and pain during and after treatment with arthrocentesis compared to baseline (T-test p < 0.01). Standardized surface electromyography values were significantly different in temporomandibular disorders subjects compared to controls (T-test p < 0.05). Improvement in pain at rest after treatment was inversely correlated with pre-treatment masseters standardized surface electromyography symmetry (R-coefficient 0.3936; p < 0.05). CONCLUSIONS: Temporomandibular disorders patients showed a different muscular recruitment pattern compared to controls. The lesser the pre-treatment masseters symmetry, the greater the improvement of pain at rest after treatment.

European Cohorts of patients and schools to Advance Response to Epidemics (EuCARE): a cluster randomised interventional and observational study protocol to investigate the relationship between schools and SARS-CoV-2 infection.

BACKGROUND: Contradictory results were reported on the role of school closure/reopening on the overall SARS-CoV-2 transmission rate, as well as on which kind and level of mitigation measures implemented in schools may be effective in limiting its diffusion. Some recent studies were reassuring, showing that opening did not increase the community spread, although teachers and families are worried about the high class density. On the other hand, distance learning was associated with a negative impact on learning, sociability and psychological health, especially in vulnerable children. As it becomes clear that the SARS-CoV-2 pandemic will last for a long time, there is a high need for studies and solutions to support safe schools opening based on scientific evidence of harms and benefits. The Lolli-Methode (LM) is a strategy for epidemiological surveillance and early intervention aiming at SARS-CoV-2 outbreaks' reduction in schools, relying on polymerase chain reaction analysis of saliva samples. METHODS: In this cluster randomised trial protocol, we aim to determine whether the LM is useful to support schools opening and to reduce clusters and attack rates in schools, compared with the standard of care (SoC) surveillance by public health departments. This multicenter study will enrol 440 classes (around 8800 students, teachers and other personnel) from two countries, cluster randomised to LM or SoC. The samples from the pools will be collected and tested using PCR-based techniques. Test results will be combined with questionnaires filled in by children, parents, schoolteachers, and principals, concerning ongoing mitigation measures, their perceived psychological impact and other health and socio-economic information. An ancillary observational study will be carried out to study the prevalence of SARS-CoV-2 in schools, frequencies and size of clusters and attack rates, to compare the effectiveness of the different preventive measures adopted and to evaluate psychological issues in students and teachers in relation to the pandemic's containment measures. DISCUSSION: By the end of this study, we will have defined and characterised the applicability of the LM for SARS-CoV-2 surveillance, as well as the impact of pandemic preventive measures on children and teachers. Trial registration International Standard Randomised Controlled Trial Number: NCT05396040, 27.05.2022.

Involvement of the endocannabinoid system in current and recurrent periodontitis: A human study.

OBJECTIVE: The aim of the present study was to assess if the endocannabinoid system is involved differently in patients with recurrent and non-recurrent periodontal disease and if in sites that have a predisposition for reactivation, levels of anandamide (AEA) change after periodontal therapy. BACKGROUND: Periodontal disease (PD) may be due to a dysregulation of the endocannabinoid system. METHODS: Periodontal patients were recruited, treated for PD and monitored. Gingival samples from these patients with recurrent (n = 10) and non-recurrent (n = 10) periodontal disease were harvested before and after treatment and compared to those of periodontally healthy (n = 10) subjects. Levels of CB1 and CB2, AEA and CBs receptor activation were assessed in healthy and inflamed samples using immunohistochemistry, chromatography and autoradiography. In healed sites, AEA levels were also assessed. RESULTS: The number of CBs in inflamed sites of recurrent patients was significantly higher than in those with non-recurrent disease and also higher than those in healthy subjects. Inflamed sites of recurrent patients had significantly lower CBs receptor activation than those of healthy subjects. Levels of AEA in inflamed sites of non-recurrent patients were significantly higher than those found both in inflamed recurrent sites and in healthy sites. Otherwise, the amount of AEA in healthy subjects and recurrent inflamed sites was similar. After periodontal therapy, levels of AEA were significantly lower in both periodontal groups. In recurrent sites, they resulted significantly lower than in non-recurrent and even in healthy subjects. CONCLUSIONS: The endocannabinoid system seems involved differently in subjects with recurrent and non-recurrent periodontal disease.

Quantitative Evaluation of Inflammatory Markers in Peri-Implantitis and Periodontitis Tissues: Digital vs. Manual Analysis-A Proof of Concept Study.

Background and Objectives: In dentistry, the assessment of the histomorphometric features of periodontal (PD) and peri-implant (PI) lesions is important to evaluate their underlying pathogenic mechanism. The present study aimed to compare manual and digital methods of analysis in the evaluation of the inflammatory biomarkers in PI and PD lesions. Materials and Methods: PD and PI inflamed soft tissues were excised and processed for histological and immunohistochemical analyses for CD3+, CD4+, CD8+, CD15+, CD20+, CD68+, and CD138+. The obtained slides were acquired using a digital scanner. For each marker, 4 pictures per sample were extracted and the area fraction of the stained tissue was computed both manually using a 594-point counting grid (MC) and digitally using a dedicated image analysis software (DC). To assess the concordance between MC and DC, two blinded observers analysed a total of 200 pictures either with good quality of staining or with non-specific background noise. The inter and intraobserver concordance was evaluated using the intraclass coefficient and the agreement between MC and DC was assessed using the Bland-Altman plot. The time spent analysing each picture using the two methodologies by both observers was recorded. Further, the amount of each marker was compared between PI and PD with both methodologies. Results: The inter- and intraobserver concordance was excellent, except for images with background noise analysed using DC. MC and DC showed a satisfying concordance. DC was performed in half the time compared to MC. The morphological analysis showed a larger inflammatory infiltrate in PI than PD lesions. The comparison between PI and PD showed differences for CD68+ and CD138+ expression. Conclusions: DC could be used as a reliable and time-saving procedure for the immunohistochemical analysis of PD and PI soft tissues. When non-specific background noise is present, the experience of the pathologist may be still required.

Human Adipose-Derived Stem Cells on Rapid Prototyped Three-Dimensional Hydroxyapatite/Beta-Tricalcium Phosphate Scaffold.

In the study, we assess a rapid prototyped scaffold composed of 30/70 hydroxyapatite (HA) and beta-tricalcium-phosphate (ß-TCP) loaded with human adipose-derived stem cells (hASCs) to determine cell proliferation, differentiation toward osteogenic lineage, adhesion and penetration on/into the scaffold.In this in vitro study, hASCs isolated from fat tissue discarded after plastic surgery were expanded, characterized, and then loaded onto the scaffold. Cells were tested for: viability assay (Alamar Blue at days 3, 7 and Live/Dead at day 32), differentiation index (alkaline phosphatase activity at day 14), scaffold adhesion (standard error of the mean analysis at days 5 and 18), and penetration (ground sections at day 32).All the hASC populations displayed stemness markers and the ability to differentiate toward adipogenic and osteogenic lineages.Cellular vitality increased between 3 and 7 days, and no inhibitory effect by HA/ß-TCP was observed. Under osteogenic stimuli, scaffold increased alkaline phosphatase activity of +243% compared with undifferentiated samples. Human adipose-derived stem cells adhered on HA/ß-TCP surface through citoplasmatic extensions that occupied the macropores and built networks among them. Human adipose derived stem cells were observed in the core of HA/ß-TCP. The current combination of hASCs and HA/ß-TCP scaffold provided encouraging results. If authors' data will be confirmed in preclinical models, the present engineering approach could represent an interesting tool in treating large bone defects.

Iliac Crest Fresh-Frozen Allografts Versus Autografts in Oral Pre-Prosthetic Bone Reconstructive Surgery: Histologic and Histomorphometric Study.

PURPOSE: The aim of this work was to compare histologically bone biopsies obtained during dental implant placement in atrophic jaws reconstructed with fresh-frozen bone (FFB) or autologous bone blocks. MATERIALS AND METHODS: Twenty partially or totally edentulous patients were treated either with iliac crest FFB grafts (14 patients; group A) or with iliac crest autografts (6 patients; group B). Seven to 9 months later, dental implants were placed in the reconstructed jaws and bone biopsies were harvested. RESULTS: The reconstructive procedure was uneventful in 9 patients of group A and in all patients of group B. Samples from both groups did not show relevant differences concerning the proportion of lamellar bone, newly formed bone, and bone marrow. In group A, the decreasing proportion of blood vessels from the apical to the coronal portion, the larger amount of lacunae in the apical portion, and the fewer vessels in the coronal portion than in group B evoke an effort of the host bone to "integrate" the graft from the apical portion. CONCLUSIONS: FFB cannot be considered as successful and safe in alveolar bone reconstruction as autogenous bone grafting.

Maxillary sinus lift augmentation: A randomized clinical trial with histological data comparing deproteinized bovine bone grafting vs graftless procedure with a 5-12-year follow-up.

INTRODUCTION: Different protocols and procedures for sinus lift and implant placement are available, generally involving the use of grafts to increase the tissue volume and/or prevent the Schneiderian membrane from collapsing. Among xenografts, deproteinised bovine bone graft (DBBP) is frequently used in sinus lift procedures. Leaving an ungrafted space following membrane elevation has proven to have a bony regenerative potential as well. This study aimed to compare the clinical and histological features of sinus lift surgery performed with or without biomaterials. METHODS: Patients with severe maxillary posterior atrophy (residual bone height 2-6 mm and residual crest thickness ≥4 mm), and in need of sinus lift surgery to allow the placement of three implants were enrolled and randomly divided into two groups. They underwent sinus lifts with DBBP (control) or with a graftless technique (test) and immediate placement of two implants (a mesial and distal one). After 6 months, a bone sample was retrieved from the area between the previously inserted fixtures, and a third, central implant was placed. The collected bone samples were analyzed morphologically and histomorphometrically. The patients were provided with prosthetic restorations after 6 months and followed up for 5-12 years. RESULTS: Ten patients were enrolled in the test and nine in the control group. The 6-month follow-up showed in the control group an average augmentation of 10.31 mm (±2.12), while in the test group it was 8.5 mm (±1.41) and a success rate of 96.3% in the control and 86.7% in the test group (p > 0.05). The histological analysis evidenced the presence of new bone tissue surrounded by immature osteoid matrix in the test group, and a variable number of DBBP particles surrounded by an immature woven bone matrix in the control group. CONCLUSION: The results of the present trial indicate that, with residual bone height of 2-6 mm and residual crest thickness ≥4 mm, sinus lift surgery with or without biomaterials followed by implant restoration, produces similar clinical and histological outcomes.

COVID-19 monitoring of school personnel through molecular salivary test and dried blood spot analysis.

Background: When the coronavirus disease 2019 (COVID-19) pandemic broke out, most countries enforced school closures as a precautionary measure. Although COVID-19 is still present three years later, schools have been reopened. We aimed to test the association of molecular salivary testing (MST) and dried blood spot (DBS) analysis for community surveillance by investigating the immunological profile of a group of school staff during and following COVID-19 vaccination. Methods: We conducted the study in a school in Milan from April 2021, when school staff were administered the first dose of vaccine against SARS-CoV-2, until the school year ended in June 2022. Each participant provided samples for MST and DBS one month (T1, W1) after receiving their first dose of vaccine. Subsequently, they collected weekly MST samples for five weeks (W2-W6), plus a DBS sample in the last week (T2). Both samples were collected one (T3), four (T4), and seven months (T5) after the administration of the second vaccine dose in May 2021. A final DBS sample was collected one year (T6) after T3. Results: Sixty participants provided 327 MSTs and 251 DBSs. None of the MST samples tested positive for SARS-CoV-2 RNA during the study period. A total of 201 DBS samples tested positive for the IgG semiquantitative analysis. Negative samples were found only at T1 (20.45%) and T2 (7.32%). We observed borderline results at T1 (4.55%), T2 (7.32%), and T4 (2.70%). The anti-SARS-CoV-2 average antibody ratio increased after the second dose between T2 and T3, and the trend peaked after the third dose between T4 and T6. We performed an immunoenzymatic assay of antibodies against nucleocapsid protein on samples collected at T1 from five participants who reported having been infected before the study and from four subjects with an abnormal increase in the antibody values at T4. Two samples tested positive in the first group and two in the second one. Conclusions: Our findings show that MST and DBS could be effective tools in the active surveillance of school personnel and that schools could be considered safe settings in view of SARS-CoV-2 infection. Vaccines might have contributed to case and/or symptom reduction.

Histological findings on jaw osteonecrosis associated with bisphosphonates (BONJ) or with radiotherapy (ORN) in humans.

OBJECTIVE: To describe the histological features of bone tissue harvested from patients affected by jaw osteonecrosis associated with bisphoshponates (BONJ) or with radiotherapy (ORN), in undecalcified ground sections. MATERIALS AND METHODS: Sixteen bone tissue samples from 14 patients with BONJ and two patients with ORN were processed in order to obtain both ground, undecalcified sections and decalcified sections. The sections underwent histometric and morphometric analysis. RESULTS: Bone tissue samples obtained from patients with BONJ or ORN of the jaws shared some histological characteristics. Common histological features included the loss of bone architecture, the absence of a proper Haversian system and proper marrow spaces, the presence of necrotic spots of non-mineralized tissue, areas of empty osteocytic lacunae next to areas of hypercellularity, the presence of resorption pits with rare osteoclast-like cells and the presence of bacteria and of an inflammatory infiltrate. A violet rib of tissue characterized by large resorption pits facing was frequently observed between the mineralized bone and the inflammatory infiltrate. CONCLUSIONS: The histological features of BONJ and ORN are similar and resemble those of osteomyelitis. Even though it is not clear whether infection is the cause or consequence of bone exposure, inflammatory cells, bacteria or their products may have a massive, direct lytic effect on bone tissue challenged by bisphosphonates.

Personal Protection Equipment and Infection Control Procedures among Health Workers during the COVID-19 Pandemic.

Health workers have been particularly affected by the COVID-19 pandemic, but it is unclear which healthcare professions are more exposed. We search for information that can help identify health workers who are employed in exposure-prone professions and who, therefore, routinely wear and adopt strict infection control equipment and measures from other colleagues. Our purpose is to test the hypothesis that health professionals historically considered less exposure-prone are affected more severely by COVID-19. Taking dentists as an example of exposure-prone healthcare professionals, this study aims to analyze data on COVID-19-related deaths reported by the Italian board of doctors and dentists' database to evaluate the number of COVID-19-related deaths of doctors and dentists in Italy from the beginning of the pandemic to 31 December 2022. As of 31 December 2021, out of 364 deaths, 38 were dentists, and of the remaining 326 doctors, 140 were general practitioners (GPs). The percentage of deaths among dentists, total doctors and GPs results in 0.06%, 0.09% and 0.33%, respectively, for the whole sample. Excluding subjects over 70 years of age, the corresponding values are 0.05%, 0.06% and 0.25%. Most of the deaths occurred in Lombardia, and the geographical distribution overlaps the trend of the corresponding general Italian population. Considering the outcome of "death", dentists, despite being at high risk, are not particularly affected by COVID-19.

Dental Emergencies in an Italian Pediatric Hospital during the COVID-19 Pandemic.

Emergency rooms (ER) are largely used by patients with oral complaints, who choose the ER over private or public dental offices for oral prevention and treatment. During the COVID-19 pandemic, the activity of most dental facilities was limited, and most hospitals and ERs were dedicated to the treatment of COVID-19 patients. The present study analyzed the number of and reason for visits at the emergency room (ER) of Ospedale dei Bambini "Vittore Buzzi", the main pediatric hospital in Milano, Italy, between 2019 and 2020, with a particular focus on oral emergencies. In 2019, 25,435 children turned to the ER, compared to 16,750 in 2020. About 10% of the children were eventually admitted to the hospital in both years. The number of admissions for infectious diseases, other than COVID-19, signed an important decrease in 2020, while trauma/injuries decreased slightly in number but increased in proportion. The number and proportion of ER visits for oral complaints decreased in 2020 compared to 2019. Stomatitis was the most frequent condition, followed by traumatic injuries, which decreased in number and percentage between 2019 and 2020. Oral infections and painful caries accounted for about 15% of the cases in both 2019 and 2020. These data highlight the need to promote territorial services for the prevention and treatment of oral health complaints, including dental emergencies.

Morphological and Immunopathological Aspects of Lingual Tissues in COVID-19.

COVID-19, a recently emerged disease caused by SARS-CoV-2 infection, can present with different degrees of severity and a large variety of signs and symptoms. The oral manifestations of COVID-19 often involve the tongue, with loss of taste being one of the most common symptoms of the disease. This study aimed to detect SARS-CoV-2 RNA and assess possible morphological and immunopathological alterations in the lingual tissue of patients who died with a history of SARS-CoV-2 infection. Sixteen cadavers from 8 SARS-CoV-2 positive (COVID-19+) and 8 negative (COVID-19-) subjects provided 16 tongues, that were biopsied. Samples underwent molecular analysis through Real-Time RT-PCR for the detection of SARS-CoV-2 RNA. Lingual papillae were harvested and processed for histological analysis and for immunohistochemical evaluation for ACE2, IFN-γ and factor VIII. Real-Time RT-PCR revealed the presence of SARS-CoV-2 RNA in filiform, foliate, and circumvallate papillae in 6 out of 8 COVID-19+ subjects while all COVID-19- samples resulted negative. Histology showed a severe inflammation of COVID-19+ papillae with destruction of the taste buds. ACE2 and IFN-γ resulted downregulated in COVID-19+ and no differences were evidenced for factor VIII between the two groups. The virus was detectable in most COVID-19+ tongues. An inflammatory damage to the lingual papillae, putatively mediated by ACE2 and IFN-γ in tongues from COVID-19+ cadavers, was observed. Further investigations are needed to confirm these findings and deepen the association between taste disorders and inflammation in SARS-CoV-2 infection.

Saliva Molecular Testing for SARS-CoV-2 Surveillance in Two Italian Primary Schools.

BACKGROUND: No evidence has so far proven a significant role of schools in SARS-CoV-2 transmission, while the negative effects of their closure on children and adolescents are well documented. Surveillance, by means of frequent students and staff testing, has been advocated in order to implement school safety. Our aim was to report the results of a school surveillance program for the early detection of SARS-CoV-2 infection in pre- and asymptomatic subjects, by means of molecular salivary testing (MST). METHODS: School surveillance in two schools in Milan, Italy, was carried out for six weeks. Each participant received a saliva collection kit, to be self-performed. RESULTS: 401 students and 12 teachers were enrolled, and 5 positive children in 5 different classes were observed. All the cases were asymptomatic. Their nasopharyngeal swab was positive on the same day in four cases, while in one case it resulted negative on the same day and positive 3 days later. In one positive case, the whole family was set under surveillance. The positive child did not develop symptoms and no family member was infected. CONCLUSIONS: MST might represent an efficient way to actively survey communities in order to detect asymptomatic cases, thus limiting SARS-CoV-2 transmission.

Impact of Lifestyle Variables on Oral Diseases and Oral Health-Related Quality of Life in Children of Milan (Italy).

A large part of the Italian population doesn't receive adequate information and support on how to maintain oral health. In this observational, cross-sectional, pilot study, we investigated how some lifestyle-related variables affect oral diseases and oral health-related quality of life (OHRQoL) of children attending public-school summer services in Milan. A survey that included questions on children's oral disease, OHRQoL and lifestyle-related factors (feeding habits, oral hygiene protective behaviors, dental coaching and socio-economic and educational status), was administered to the children's caregivers. Data from 296 surveys were analyzed to assess the protective/negative effect of each variable on oral disease and OHRQoL. With respect to disease, the "never" consumption of fruit juice, the use of fluoride toothpaste, higher educational qualification and ISEE (equivalent family income) of those who filled out the form, resulted protective factors. Regarding OHRQoL, the "never" assumption/use of tea bottle, sugared pacifier and fruit juice as well as the use of fluoride toothpaste, a higher educational qualification and ISEE of those who filled out the form, resulted to have protective effects. In conclusion, protective behaviors and socio-economic status affect oral disease and OHRQoL in children of Milan.

Time sequence of bone healing around two implant systems in minipigs: preliminary histologic results.

The aim of this study was to evaluate the early phases of bone healing around two different implant surfaces. For this purpose, four minipigs were used. Implants with rough titanium surfaces (ITI sandblasted/acid-etched and Branemark TiUnite) were placed in the maxillae of the animals and sacrifice was scheduled in such a way that healing times of 3 days, 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, and 7 weeks were obtained. Histologic analysis revealed that a similar pattern was observed in the phases of bone healing around both types of implants between 3 days and 7 weeks. This pattern consisted of the replacement of blood clot and bone debris with a provisional connective tissue in the first few weeks and with mineralized tissue and marrow spaces later on. Both rough surfaces allowed for "contact osteogenesis" to take place. (Int J Periodontics Restorative Dent 2009;29:549-555.).

Tissue engineering in periodontology: Biological mediators for periodontal regeneration.

Teeth and the periodontal tissues represent a highly specialized functional system. When periodontal disease occurs, the periodontal complex, composed by alveolar bone, root cementum, periodontal ligament, and gingiva, can be lost. Periodontal regenerative medicine aims at recovering damaged periodontal tissues and their functions by different means, including the interaction of bioactive molecules, cells, and scaffolds. The application of growth factors, in particular, into periodontal defects has shown encouraging effects, driving the wound healing toward the full, multi-tissue periodontal regeneration, in a precise temporal and spatial order. The aim of the present comprehensive review is to update the state of the art concerning tissue engineering in periodontology, focusing on biological mediators and gene therapy.

Oral implants placed in bone defects treated with Bio-Oss, Ostim-Paste or PerioGlas: an experimental study in the rabbit tibiae.

OBJECTIVES: To compare the histological features of bone filled with Bio-Oss, Ostim-Paste or PerioGlas placed in defects in the rabbit tibiae by evaluating bone tissue composition and the integration of titanium implants placed in the grafted bone. MATERIAL AND METHODS: Two cylindrical bone defects, about 4 mm in diameter and 6 mm in depth, were created in the tibiae of 10 rabbits. The defects were filled with either Bio-Oss, PerioGlas, Ostim-Paste or left untreated, and covered with a collagen membrane. Six weeks later, one titanium sandblasted and acid-etched (SLA) implant was inserted at the centre of each previously created defect. The animals were sacrificed after 6 weeks of healing. RESULTS: Implants placed in bone previously grafted with Bio-Oss, PerioGlas or Ostim-Paste obtained a larger extent of osseointegration, although not statistically significant, than implants placed in non-grafted bone. The three grafting materials seemed to perform in a similar way concerning their contribution towards implant osseointegration. All grafting materials appeared to be osteoconductive, thus leading to the formation of bridges of mineralized bone extending from the cortical plate towards the implants surface through the graft scaffold. CONCLUSIONS: Grafting with the above-mentioned biomaterials did not add any advantage to the osseointegration of titanium SLA implants in a self-contained defect.

Dental treatment of a rare case of pyoderma gangrenosum with aggressive periodontal disease.

BACKGROUND AND OVERVIEW: Pyoderma gangrenosum (PG) is a rare neutrophil-mediated autoinflammatory dermatosis that can involve the oral mucosa. Dental surgery is a potential triggering factor for the onset of PG lesions. The authors describe and discuss the dental management of a rare case of aggressive periodontitis in a patient with PG, from multiple tooth extractions to prosthetic rehabilitation, including administration of systemic steroid prophylaxis before surgery to prevent the potential onset of PG-related lesions. CASE DESCRIPTION: A 22-year-old man who had a diagnosis of PG and who had aggressive periodontal disease underwent dental extractions, gingivoplastic surgery, and prosthetic rehabilitation. The patient received 8 milligrams of betamethasone intramuscularly 20 minutes before the oral surgery. The tissues healed perfectly, and no adverse effects were reported. CONCLUSIONS AND PRACTICAL IMPLICATIONS: For minor oral surgery, prophylactic corticosteroids might help reduce the risk of developing PG-related lesions. The clinician should plan the prosthetic devices to be as atraumatic as possible.