Immunohistopathology of oral mucosal chronic graft-versus-host disease severity and duration.

OBJECTIVE: Chronic graft-versus-host disease (cGVHD) is the main cause of late non-relapse mortality following hematopoietic cell transplantation. Oral mucosal (om-) cGVHD is common, but diagnosis and assessment rely on clinical interpretation and patient-reported symptoms. We investigated immunohistopathological profiles with respect to om-cGVHD severity disease duration. MATERIAL AND METHODS: Ninety-four transplant patients and 15 healthy controls (n = 212 biopsies) were investigated by quantitative immunohistochemistry for T cells (CD4, CD8, and CD5), B cells (CD19 and CD20), macrophages (CD68), and Langerhans cells (CD1a). RESULTS: We found significant increases in T (CD4, CD8) and monocytic (CD68) cells in om-cGVHD, and a notable absence of B (CD19 and CD20) cells. Histopathological activity correlated with increased CD4, CD8 and CD68. However, CD4 was associated with mild om-cGVHD, whereas CD8 and CD68 were found to be elevated in severe om-cGVHD. CD8 and CD68 levels were raised at disease onset, but during late phase, the predominant CD68 population was accompanied by CD4. CONCLUSION: Oral cGVHD is a heterogenous clinical disorder, but our knowledge of the underlying biology remains limited. We highlight the importance of CD4, CD8 and CD68 immune profiling, together with histological grading for the staging of oral cGVHD, to broaden our understanding of the biology and individual disease course.

A genomic and historical synthesis of plague in 18th century Eurasia.

Plague continued to afflict Europe for more than five centuries after the Black Death. Yet, by the 17th century, the dynamics of plague had changed, leading to its slow decline in Western Europe over the subsequent 200 y, a period for which only one genome was previously available. Using a multidisciplinary approach, combining genomic and historical data, we assembled Y. pestis genomes from nine individuals covering four Eurasian sites and placed them into an historical context within the established phylogeny. CHE1 (Chechnya, Russia, 18th century) is now the latest Second Plague Pandemic genome and the first non-European sample in the post-Black Death lineage. Its placement in the phylogeny and our synthesis point toward the existence of an extra-European reservoir feeding plague into Western Europe in multiple waves. By considering socioeconomic, ecological, and climatic factors we highlight the importance of a noneurocentric approach for the discussion on Second Plague Pandemic dynamics in Europe.

Pre-medical dental evaluation and treatment of oral infection - a survey study among hospital-affiliated dentists in Sweden.

OBJECTIVE: To examine how hospital-affiliated dentists assess risk and evaluate oral foci of infection in patients facing certain medical treatments, and whether the nature of upcoming medical treatment affects the choice of dental intervention. MATERIALS AND METHODS: A survey comprising six clinical cases (50 teeth) was sent to hospital-affiliated dentists in Sweden. A treatment option for the affected tooth/teeth in each case was selected whether the patient was facing heart valve surgery, chemotherapy, radiation therapy, intravenous bisphosphonate treatment, solid organ transplantation or was diagnosed with endocarditis. RESULTS: Consensus in choice of dental treatment was high in 62%, moderate in 32% and low in 6% of the assessments. High variability of choice of treatment was seen for eight teeth whereas the remaining 42 teeth often received the same therapy regardless of medical issue. Chemotherapy and radiotherapy were thought to entail the highest risk for oral infectious sequelae with a risk ranging from 1% to 100%. CONCLUSION: Pre-medical dental evaluations and recommended treatments are often uniform with the exception of the management of asymptomatic root canal treated teeth with persisting apical radiolucency and heavily decayed molars. In many instances, dental diagnosis has a greater impact on choice of treatment than the underlying medical issue and associated implications thereof.

Histopathological Grading of Oral Mucosal Chronic Graft-versus-Host Disease: Large Cohort Analysis.

Graft-versus-host disease (GVHD) can manifest as acute or chronic complications in patients after hematopoietic cell transplantation (HCT). Oral chronic GVHD (cGVHD) occurs in approximately 70% of HCT recipients and includes lichenoid-like mucosal reactions, restricted mouth opening, and salivary gland dysfunction. However, the underlying histopathological presentation remains to be validated in large cohorts. We characterized the histopathological features of oral mucosal cGVHD and devised a scoring model in a large patient cohort (n = 112). Oral mucosal biopsy sections (n = 303) with and without oral cGVHD were identified from archived and current HCT recipients with additional healthy controls. Histological screening was performed on hematoxylin and eosin-stained and periodic acid-Schiff-stained sections. A points-based grading tool (0 to 19, grade 0 to IV) was established based on intraepithelial lymphocytes and band-like inflammatory infiltrate, atrophic epithelium with basal cell liquefaction degeneration, including apoptosis, as well as separation of epithelium and pseudo-rete ridges. Validation involved 62 biopsy specimens, including post-HCT (n = 47) and healthy (n = 15) specimens. Remaining biopsy specimens (n = 199) were blindly graded by 3 observers. Histological severity was correlated with clinical diagnostic and distinctive features, demonstrating a spectrum of individual patient severity, including frequent signs of subclinical GVHD in healthy mucosa. However, oral cGVHD presented with significantly higher (P < .001) scores compared with HCT controls, with moderate to high positive likelihood ratios for inflammatory infiltrate, exocytosis, and basal membrane alterations. The grade II-IV biopsy specimens demonstrated a histopathological diagnosis of active mucosal lichenoid-like cGVHD, highlighting the importance of correlating clinical presentation with the dynamic histopathological processes for improved patient stratification. In addition, this tool could be used for assessing treatments, pathological processes, and immune cellular content to provide further insight into this debilitating disease.

Carvone Contact Allergy in Southern Sweden: A 21-year Retrospective Study.

Carvone (l-carvone), a mint flavour in spearmint oil, is considered a mild skin sensitizer. Carvone-sensitization may be linked to oral/perioral signs and oral lichen planus, but studies are sparse. The prevalence of patch test reactions to carvone and relevant findings from the positive group were investigated. Records for patch-tested patients at the Malmö clinic, for the period 1996 to 2016, were studied. Carvone-positive and carvone-negative patients were compared regarding patch test data from baseline series and dental series. Dental series-tested carvone-positive patients were also compared with a matched group. A total of 147 out of 4,221 referred patients had a positive patch test to carvone. Sensitized patients had higher mean age and were primarily women; 73% had oral signs and 57% had oral lichen. Concomitant patch test reactions to gold, nickel and mercury were common. In the matched group-comparison carvone-positive patients had a higher frequency of oral lichen, but no difference was found in sensitization to gold and mercury.

Quantification of l-carvone in toothpastes available on the Swedish market.

BACKGROUND: Toothpastes have widespread use in the population, and contain flavours to give a pleasant and often minty aroma. Flavours are prevalent allergens in toothpastes, and adverse reactions often present as perioral dermatitis or stomatitis. l-Carvone, a mint flavour found in spearmint oil, is one of these allergens. There are few studies on contact allergy to l-carvone, and some of them have indicated a positive relationship with oral lichenoid lesions. OBJECTIVES: To qualitatively and quantitatively investigate the presence of l-carvone in commercially available toothpastes on the Swedish market. METHODS: l-Carvone in 66 toothpastes was analysed with straight-phase high-performance liquid chromatography. The product labels were studied with regard to limonene. RESULTS: l-Carvone was found in 64 of 66 toothpastes (concentration: 0.00005-0.35%). In 10 of these, the concentration exceeded 0.1%. Higher concentrations of l-carvone were found if limonene was listed on the label. CONCLUSION: The majority of toothpastes on the Swedish market contain l-carvone, but the concentration hardly relates to the advertised flavour or labelled ingredients. It is hitherto unknown whether the found concentrations are sufficient for induction of contact allergy in individuals with healthy oral mucosa or in those with oral lichenoid lesions or other mucosal disease.

Oral ulcers, a little known adverse effect of alendronate: review of the literature.

PURPOSE: To review the published data on a hitherto not widely known adverse effect of alendronate manifesting as mucosal ulcers in the oral cavity. MATERIALS AND METHODS: The electronic database PubMed was searched for reports of this adverse effect. Publications published up to August 2010 were included. This electronic search was combined with a manual search of the reference lists of the selected publications. RESULTS: A total of 47 publications were retrieved from the electronic and manual searches. Of these, 12 were selected for the review. Mostly, the ulcers were preceded by misuse of alendronate, but they also appeared after correct administration. The appearance of the ulcers varied from a few days to several months after the start of alendronate use. Effective treatment was withdrawal of the drug or revision of the dosing and administration instructions. CONCLUSIONS: Alendronate can cause mucosal ulcerations in the oral cavity, affecting patients with intense pain and causing severe morbidity. Successful treatment of this oral pathosis is achieved by aborting the use of alendronate. This adverse effect of alendronate is a rare entity in published reports but careful monitoring of patients at risk is recommended.

Pharmacovigilance of oral bisphosphonates: adverse effects manifesting in the soft tissue of the oral cavity.

PURPOSE: It is well known that oral bisphosphonates can induce necrosis of the osseous structures of the jaws. However, there seems to be a limited awareness that oral bisphosphonates can also induce adverse effects in the soft tissues of the oral cavity, as indicated by the paucity of reported cases in the literature. Because oral bisphosphonates are widely used drugs for several skeletal conditions, it is reasonable to assume that mucosal adverse effects are more common than the small number of published cases indicates. The purpose of this study was to investigate whether this adverse effect of bisphosphonates is represented as reports from health practitioners in an adverse drug reaction database, as well as to gain knowledge about which substances are being associated with adverse drug reactions affecting the oral mucosa. MATERIALS AND METHODS: The database of the Medical Products Agency-Sweden was searched for adverse effects from oral bisphosphonates manifesting in the oral and maxillofacial region. Reports of reactions limited to the soft tissues of the oral cavity were selected and further analyzed. Only those reports showing recovery or improvement after the cessation of bisphosphonate use were included in the study. RESULTS: A total of 83 cases of adverse reactions to oral bisphosphonates were retrieved from the search. Of these, 12 were included in the study. They were associated with the use of alendronate, etidronate and risedronate, in descending order. Sixteen percent of the reports comprising the oral and maxillofacial region were limited to the oral mucosa and reported recovery or improvement after discontinuation of the drug. CONCLUSIONS: Adverse effects of oral bisphosphonates with manifestations in the soft tissue of the oral cavity seem to be more common than the small number of published cases indicates. However, considering that oral bisphosphonates are widely used drugs, the incidence is still low. These adverse drug reactions are not limited to alendronate and may also be induced by etidronate and risedronate. Still, a significant proportion of the cases are associated with alendronate. Regardless of the substance used, discontinuing the drug is an effective treatment for the mucosal lesions.

Lymphoid organisation in labial salivary gland biopsies is a possible predictor for the development of malignant lymphoma in primary Sjögren's syndrome.

OBJECTIVE: The development of non-Hodgkin's lymphoma (NHL) confers a high risk of mortality in primary Sjögren's syndrome (pSS) patients, but the sensitivity and specificity of proposed lymphoma predictors are insufficient for practical use. The performance of lymphoid organisation in the form of germinal centre (GC)-like lesions was evaluated in labial salivary gland biopsies taken at pSS diagnosis as a potential lymphoma-predicting biomarker. METHODS: Labial salivary gland tissue biopsies available from two Swedish pSS research cohorts (n=175) were re-evaluated by light microscopy in a blind study in order to identify GC-like structures as a sign of ectopic lymphoid tissue formation and organisation. A linkage study was performed with the Swedish Cancer Registry for lymphoma identification. The risk of developing NHL in GC-positive patients in comparison with GC-negative patients was evaluated using Kaplan-Meier statistics and log-rank test. Associations between GC-like structures and clinical and/or laboratory disease markers were also determined using χ(2) or Fisher's exact tests. RESULTS: At diagnosis, 25% of pSS patients had GC-like structures in their salivary glands. Seven of the 175 patients studied (14% GC+ and 0.8% GC-) developed NHL during 1855 patient-years at risk, with a median onset of 7 years following the initial diagnostic salivary gland biopsy. Six of the seven patients had GC-like structures at diagnosis; the remaining patient was GC negative at the time of diagnosis (p=0.001). CONCLUSIONS: The detection of GC-like structures by light microscopy in pSS diagnostic salivary biopsies is proposed as a highly predictive and easy-to-obtain marker for NHL development. This allows for risk stratification of patients and the possibility to initiate preventive B-cell-directed therapy.

Effects of ovariectomy and aging on tooth attachment in female mice assessed by morphometric analysis.

OBJECTIVE: Non-human primates, dogs, rats, hamsters and ferrets, have frequently been used as laboratory animals in periodontal biology and pathology. In the past, mice have been used less for this purpose, but nowadays attract a lot of interest because gene knockout and transgenic technologies utilize mice primarily. In this study, we investigate the effects of ovariectomy and aging on tooth attachment in female mice. MATERIAL AND METHODS: Eight-week-old mice (n=15) were divided into three experimental groups (control, n=5; sham-operated, n=5; ovariectomy, n=5) and ovaries removed bilaterally. Attachment level, assessed by measuring alveolar bone height and apical termination of the junctional epithelium, was determined 6 weeks post-ovariectomy by digital morphometric analysis in sagittal sections of the mandible. The plasma level of the inflammation marker serum amyloid A (SAA) was determined by ELISA. In another series of experiments, tooth attachment was determined in female mice (n=7) at 8-26 weeks of age. RESULTS: Withdrawal of female sex hormone production by ovariectomy had no effect on alveolar bone height and apical termination of the junctional epithelium. The SAA level in plasma was unaffected by removal of the ovaries, suggesting that systemic inflammation is not induced by ovariectomy. Bone height was similar in mice sacrificed at 8-26 weeks of age and apical termination of the junctional epithelium was at the cemento-enamel junction at all ages. CONCLUSIONS: Removal of ovarian production of female sex hormones by ovariectomy has no influence on tooth attachment, and further tooth attachment is preserved with age in female mice.

Expression of muscarinic receptor subtypes in salivary glands of rats, sheep and man.

In rat parotid, submandibular and sublingual glands and in ovine parotid and in human labial glands, the expression of muscarinic receptor subtypes was examined by immunoblotting and immunohistochemistry. Functional correlates were searched for in rat salivary glands. In the rat submandibular and sublingual glandular tissues clear signals of muscarinic M1 and M5 receptors could be detected in the immunoblotting and vague bands for muscarinic M3 and, in particular for, M4 receptors. The rat parotid gland differed. In this gland, the signal was less obvious for the muscarinic M1 receptor, and further, muscarinic M4 receptors appeared more strongly marked than in the submandibular glands. The results from the immunohistochemistry could be interpreted as the muscarinic M4 receptors are located on nerve fibres, since the outer layer of lobuli were densely stained. Intraglandular vessels in the rat submandibular and parotid glands showed expression of M3 receptors. In contrast to the parotid gland, the submandibular vessels also expressed M1 and M2 receptors. Occasionally M5 receptors appeared in the arteries and veins also. The functional studies in the rat confirmed muscarinic M1 receptor mediated secretion in the submandibular gland. Since the M1 receptor blockade did not affect submandibular blood flow, indirect vascular effects could not in total explain the secretory inhibition. Also in the human labial glands, muscarinic M1, M3 and M5 receptors occurred. No or low amounts of muscarinic M2 and M4 receptors could be detected. In patients with Sjögren-like symptoms an up-regulation of M3, M4 and M5 receptors was apparent in the labial glands. In ovine parotid glands all receptors could be detected, but constantly with vague bands for muscarinic M2 receptors. In conclusion, muscarinic M1 receptors seem to be expressed in seromucous/mucous glands. A secretory effect by muscarinic M5 receptors is not to be excluded, since they were expressed in all the glands examined. However, other functions, such as promotion of inflammation, cell growth and proliferation are possible as well.

Prevalence, initiating factor, and treatment outcome of medication-related osteonecrosis of the jaw-a 4-year prospective study.

OBJECTIVES: Medication-related osteonecrosis of the jaw (MRONJ) has a wide range of prevalence, and a standard therapy has not yet been established. The aim of this study was to analyze the prevalence and initiating factors of MRONJ and the outcomes of surgical therapy. STUDY DESIGN: In a prospective cohort study, all patients diagnosed with MRONJ in the Region of Skåne, in Sweden, were included. Predictor variables (comorbidity, site, stage, gender) and initiating factors (tooth extraction, periodontitis) were recorded. Surgical treatment was sequestrectomy or block resection, and the outcome variable was healing after 2 months. To estimate the prevalence, data on the use of bisphosphonate and denosumab were used. RESULTS: Fifty-five patients with MRONJ were identified. The prevalence of MRONJ was 0.043% among patients treated with oral bisphosphonates, 1.03% among those on intravenous bisphosphonates and 3.64% in those on high-dose denosumab. Periodontal disease preceded development of MRONJ in 41 patients. Fifty patients were treated surgically and followed up for at least 2 months. Remission or healing occurred in 80% of patients treated with sequestrectomy and in 92.5% of patients treated with block resection. CONCLUSIONS: The prevalence of MRONJ in Sweden is low. Periodontitis is the most common initiating factor. The outcome of treatment of MRONJ is healing in most patients treated surgically.

Autoantibodies to full body vascular cell junctions colocalize with MYZAP, ARVCF, desmoplakins I and II and p0071 in endemic pemphigus in Colombia, South America.

BACKGROUND: We previously described a new variant of endemic pemphigus foliaceus in El Bagre, Colombia (El Bagre-EPF). METHODS: Here we aimed to investigate disease autoreactivity to vessels in all body organs/systems. We compared 57 patients and 57 controls from the endemic area, matched by demographics, age, sex, and work activity. We performed immunofluorescence, immunohistochemistry, confocal microscopy, immunoblotting, indirect immune electron microscopy studies, and autometallographic studies. We performed ultrasonography on large patient arteries, investigating for vascular anomalies. In addition, we reviewed autopsies on seven patients who died affected by El Bagre-EPF. We immunoadsorbed any positive vessel immunofluorescence with desmoglein (Dsg1), investigating for new autoantigens. RESULTS: Overall, 57/57 patients affected by El Bagre-EPF displayed autoantibodies to vessels in all the organs/systems of the body via all methods (P < 0.01). The autoreactivity was polyclonal, and the patient's antibodies colocalized with commercial antibodies to desmoplakins I and II, p0071, ARVCF, and MYZAP (all from Progen Biotechnik, Germany; P < 0.01; all present at cell junctions). Immunoadsorption with Dsg1 on positive vessel immunofluorescence showed that the immune response against the vessels was directed against non-Dsg1 antigen(s). Autometallographic studies showed deposits of metals and metalloids in vessel cell junctions and in erythrocytes of 85% of patients (P < 0.01). CONCLUSIONS: Immune response to these vascular antigens is likely altering endothelial cells and vessel shapes, thus disturbing hemodynamic flow. The flow alterations likely lead to inflammation and may play a role in the atherogenesis often seen in these patients.

Clear cell variant of intraosseous calcifying epithelial odontogenic tumor: a case report and review of the literature.

CEOT is a rare benign, but locally aggressive odontogenic tumor, and some authors have claimed that the biologic behavior of the clear cell variant is even more aggressive, with greater propensity to recur. We report a rare case of clear cell calcifying epithelial odontogenic tumor (CEOT) and discuss its possibly aggressive behavior. A 40-year-old woman experienced an asymptomatic expansion of the left posterior mandible. After radiographic examination and biopsy, a block resection was performed. Histologic examination included analysis of Ki-67 reactivity as a marker of tumor growth activity. Ki-67 labeling was pronounced in the non-clear cell population in the tumor periphery but low in the central and clear cell portions. Clinical and radiographic follow-up 6 years after resection has not revealed any signs of recurrence. On the basis of a review of the literature and our own findings, there is no clear data to suggest that clear cell CEOT exhibits more aggressive behavior compared with conventional CEOT.

Osseointegration of Titanium Implants in Onlay of Cerament™, a New Ceramic Bone Substitute.

The purpose was to investigate whether a new biphasic and injectable ceramic bone substitute Cerament™ that rapidly remodels to bone, may contribute to the retention of titanium implant screws during the healing period, and to analyze the pattern of bone formation around titanium implants.Titanium screws were implanted in rat tibiae and embedded with or without Cerament™ on the cortical surface. Torsional resistance was measured after 1 day, and after 6 and 12 weeks. Implant areas without bone substitute were analyzed histologically for comparison. The torsional resistance increased over time as the screws were osseointegrated. There was no difference in resistance between screws embedded in the bone substitute and control screws. The bone apposition was more pronounced on the proximal side of the screw than on the distal side. Cerament™ is capable of conducting bone growth from a cortical bone surface. The newly formed bone in this application does not significantly add to the osseointegrative strength of the implant screw, as measured by torque resistance, during the first 12 weeks.

The influence of pH and nicotine concentration in oral moist snuff on mucosal changes and salivary pH in Swedish snuff users.

The use of Swedish oral moist snuff is a widespread habit in Sweden. In 1999, 25% of the adult male population and 3.1% of the female population were users of snuff. The aim of the present study was to evaluate how variations in pH and nicotine concentrations of snuff affect the oral mucosa, clinically and histologically, salivary pH and daily nicotine intake in 20 habitual users of loose snuff. The subjects were studied during use of their usual brand, after 12 weeks use of a snuff with lower pH and after another 12 weeks use of a snuff with both lower pH and lower nicotine concentration. Consumption data, oral soft tissue changes, salivary pH and nicotine intake were measured. Further, biopsies were taken from the central part of the clinically observed lesions and histological changes were analysed. The subjects maintained their snuff consumption during the observation periods. The average salivary pH was higher during snuff use than in the morning. Further, it was higher shortly after the snuff was removed than during snuff use. After having switched to the snuff with both lower pH and lower nicotine concentrations, they showed a significant reduction in daily nicotine intake and developed significantly less pronounced clinical and histological changes. These results indicate that nicotine is one of the substances in snuff that has a biological effect on the oral mucosa. However, there also seems to be a synergistic effect between the pH and nicotine concentration in the snuff.

Bisphosphonate-associated osteonecrosis of the auditory canal.

Only rare cases of osteonecrosis of the auditory canal associated with bisphosphonates, have been published. Our results confirm that similar reports can also be encountered in databases of adverse drug reactions.