Multimodal and Multiscale Characterization of the Bone-Bacteria Interface in a Case of Medication-Related Osteonecrosis of the Jaw.

Medication-related osteonecrosis of the jaw (MRONJ) is a known side effect of bisphosphonates (BPs). Although bacterial infection is usually present, the etiology of MRONJ remains unknown. Here we apply a multimodal and multiscale (micro-to-nano) characterization approach to investigate the interface between necrotic bone and bacteria in MRONJ. A non-necrotic bone sample was used as control. Both necrotic and non-necrotic bone samples were collected from the jaw of a female individual affected by MRONJ after using BPs for 23 years. For the first time, resin cast etching was used to expose bacteria at the necrotic site. The bone-bacteria interface was also resolved at the nanoscale by scanning transmission electron microscopy (STEM). Nanosized particulates, likely corresponding to degraded bone mineral, were often noted in close proximity to or enclosed by the bacteria. STEM also revealed that the bone-bacteria interface is composed of a hypermineralized front fading into a highly disordered region, with decreasing content of calcium and phosphorus, as assessed by electron energy loss spectroscopy (EELS). This, combined with the variation in calcium, phosphorus, and carbon across the necrotic bone-bacteria interface evaluated by scanning electron microscopy (SEM)-energy dispersive X-ray spectroscopy (EDX) and the lower mineral-to-matrix ratio measured by micro-Raman spectroscopy in necrotic bone, indicates the absence of a mineralization front in MRONJ. It appears that the bone-bacteria interface originates not only from uncontrolled mineralization but also from the direct action of bacteria degrading the bone matrix. © 2022 The Authors. JBMR Plus published by Wiley Periodicals LLC on behalf of American Society for Bone and Mineral Research.

Osteonecrosis of the jaw among patients with cancer treated with denosumab or zoledronic acid: Results of a regulator-mandated cohort postauthorization safety study in Denmark, Norway, and Sweden.

BACKGROUND: Osteonecrosis of the jaw (ONJ) is an adverse effect of antiresorptive treatment. This study estimated incidence proportions and incidence rates of ONJ in cancer patients with bone metastases from solid tumors treated for the prevention of skeletal-related events in routine clinical practice. METHODS: This cohort study in Denmark, Norway, and Sweden in 2011-2018 included 3 treatment cohorts: a denosumab inception cohort (DEIC), a zoledronic acid inception cohort (ZAIC), and a denosumab-switch cohort (DESC). The authors estimated 1- to 5-year incidence proportions and incidence rates of ONJ overall, by cancer site (breast, prostate, or other solid tumor), and by country. ONJ diagnoses were confirmed by adjudication. RESULTS: There were 1340 patients in the DEIC, 1352 in the ZAIC, and 408 in the DESC. The median ages of the 3 cohorts were 70, 69, and 70 years, respectively; the proportions of men were 72.6%, 53.8%, and 48.3%, respectively; and the median follow-up was 19.8, 12.9, and 13.3 months, respectively. The 5-year incidence proportions of ONJ were 5.7% (95% confidence interval [CI], 4.4%-7.3%) in the DEIC, 1.4% (95% CI, 0.8%-2.3%) in the ZAIC, and 6.6% (95% CI, 4.2%-10.0%) in the DESC. The corresponding ONJ incidence rates per 100 person-years were 3.0 (95% CI, 2.3-3.7), 1.0 (95% CI, 0.6-1.5), and 4.3 (95% CI, 2.8-6.3). Incidence proportions and incidence rates were highest in patients with prostate cancer and in Denmark. CONCLUSIONS: This study provides estimates of the risk of medically confirmed ONJ among patients initiating denosumab or zoledronic acid in routine clinical practice in 3 Scandinavian countries. The results varied by cancer site and by country. LAY SUMMARY: Denosumab and zoledronic acid reduce the risk of bone fractures, pain, and surgery in patients with advanced cancers involving bone. Osteonecrosis of the jaw (ONJ)-death of a jawbone-is a known side effect of treatment with denosumab or zoledronic acid. The authors examined almost 2900 denosumab- or zoledronic acid-treated patients with cancer in Denmark, Norway, and Sweden. Over the course of 5 years, ONJ developed in 5.7% of the patients whose initial treatment was denosumab, in 1.4% of the patients whose initial treatment was zoledronic acid, and in 6.6% of the patients who switched from zoledronic acid to denosumab.

Uncertainty managing patients treated with antiresorptive drugs: a cross-sectional study of attitudes and self-reported behavior among dentists in Sweden.

Objective: To evaluate the level of knowledge and attitudes among Swedish general dentists regarding the behavior and management of patients treated with bisphosphonates and denosumabMaterials and Methods: A questionnaire was sent to 656 dentists. The web-based survey included questions on demographics, risk perception of osteonecrosis of the jaws (ONJ), the experience of managing patients treated with bisphosphonates and denosumab and requests to acquire new knowledge. Independence and associations were tested using Fisher's exact test and the chi-square test function.Results: The overall response rate was 57.6%. Most of the dentists, 69.7%, had been in the profession for more than 5 years. The criteria for ONJ were not known by 43.2% and 86.9% did not feel comfortable with their current level of knowledge when managing the patients in question. The rest of the respondents felt uncertain and more than 70% were unaware of the different stages of ONJ, when to refer a patient to a specialist and when to prescribe antibiotics.Conclusion: Dentists practicing in Sweden express a strong need for an improved level of knowledge when managing patients treated with bisphosphonates and denosumab. The results demonstrate a need for the necessary advancement of education and clinical training at dental schools in order to prepare dentists to provide safe, modern care within the healthcare sector.

A Case Report on Gardner Syndrome With Dental Implant Treatment and a Long-Term Follow-Up.

A case of Gardner syndrome (GS) in a 37-year-old woman is presented in which rehabilitation with dental implant treatment was followed for 7 years. The course of diagnostics and treatment is followed through an overview of 45 tissue biopsy and cytology samples during a period of 30 years. The patient was diagnosed with GS after a routine review of an oral panoramic radiograph presenting with multiple osteomas and multiple unerupted supernumerary teeth. Biopsy results and surgical procedures with histopathologic diagnoses from 1986 to 2016 are presented. Histologic analysis of the sampled jawbone showed a picture similar to an osteoid osteoma. The installed implant was functionally stable, with no clinical or radiographic events observed at yearly visits, for 7 years. In 1991, benign tumors appeared; in 2014, multiple adenomas were found in the small intestine and rectum. This case report shows the potential for dental implant treatment in a patient with GS.

[Osteonecrosis of the jaws (ONJ) associated to antiresorptive treatment]. / Kunskap om läkemedelsrelaterad käkbensnekros behöver spridas.

Osteonecrosis of the jaws (ONJ) has been associated to antiresorptive treatment and is an increasing problem all over the world. ONJ is a severe adverse effect of antiresorptive treatment with bisphosphonate and denosumab used for treatment of osteoporosis, metastases from certain malignant conditions and as an adjuvant treatment in postmenopausal women with breast cancer, and of treatment with certain chemotherapeutic drugs. In this paper the epidemiology, symptoms, diagnostic features, clinical findings, and treatment are reviewed. Guidelines are presented for dental treatment, surgery and referral of patients on antiresorptive treatment.

Micrometer-Sized Magnesium Whitlockite Crystals in Micropetrosis of Bisphosphonate-Exposed Human Alveolar Bone.

Osteocytes are contained within spaces called lacunae and play a central role in bone remodelling. Administered frequently to prevent osteoporotic fractures, antiresorptive agents such as bisphosphonates suppress osteocyte apoptosis and may be localized within osteocyte lacunae. Bisphosphonates also reduce osteoclast viability and thereby hinder the repair of damaged tissue. Osteocyte lacunae contribute to toughening mechanisms. Following osteocyte apoptosis, the lacunar space undergoes mineralization, termed "micropetrosis". Hypermineralized lacunae are believed to increase bone fragility. Using nanoanalytical electron microscopy with complementary spectroscopic and crystallographic experiments, postapoptotic mineralization of osteocyte lacunae in bisphosphonate-exposed human bone was investigated. We report an unprecedented presence of ∼80 nm to ∼3 µm wide, distinctly faceted, magnesium whitlockite [Ca18Mg2(HPO4)2(PO4)12] crystals and consequently altered local nanomechanical properties. These findings have broad implications on the role of therapeutic agents in driving biomineralization and shed new insights into a possible relationship between bisphosphonate exposure, availability of intracellular magnesium, and pathological calcification inside lacunae.

Design and methods for a Scandinavian pharmacovigilance study of osteonecrosis of the jaw and serious infections among cancer patients treated with antiresorptive agents for the prevention of skeletal-related events.

OBJECTIVE: Osteonecrosis of the jaw (ONJ) is a recognized complication of potent antiresorptive therapies, especially at the doses indicated to prevent skeletal complications for cancer patients with bone metastases. This paper describes the rationale and methods for a prospective, post-authorization safety study of cancer patients treated with antiresorptive therapies. METHODS: As part of a comprehensive pharmacovigilance plan, developed with regulators' input, the study will estimate incidence of ONJ and of serious infections among adult cancer patients with bone metastases treated with denosumab (120 mg subcutaneously) or zoledronic acid (4 mg intravenously, adjusted for renal function). Patients will be identified using routinely collected data combined with medical chart review in Denmark, Sweden, and Norway. Followup will extend from the first administration of antiresorptive treatment to the earliest of death, loss-to-follow-up, or 5 years after therapy initiation. Results will be reported for three treatment cohorts: denosumab-naïve patients, zoledronic acid-naïve patients, and patients who switch from bisphosphonate treatment to denosumab. ONJ cases will be identified in three newly established national ONJ databases and adjudicated by the committee that functioned during the XGEVA(®) clinical trials program. CONCLUSION: This study will provide a real world counterpart to the clinical trial-estimated risks for ONJ and serious infections for cancer patients initiating denosumab or zoledronic acid. The establishment of ONJ databases in the three Scandinavian countries will have potential benefits outside this study for the elucidation of ONJ risk factors and the evaluation of ONJ treatment strategies.

Antimicrobial Effect of a Single Dose of Amoxicillin on the Oral Microbiota.

PURPOSE: Amoxicillin is commonly used in oral surgery for antimicrobial prophylaxis against surgical-site infection and bacteremia because of its effect on oral streptococci. The aim of this study was to determine whether amoxicillin reaches the break-point concentrations in saliva and has any effect on the salivary microbiota, colonizing bacteria on mucosal membranes and on the gingival crevice after a single dose of amoxicillin. MATERIAL AND METHODS: Twenty subjects received 2 g of amoxicillin, per os. The facultative and strictly anaerobic microflora, as well as the streptococcal microflora specifically, were followed from baseline and after 1, 4, and 24 hours. Samples were taken for microbial analysis from saliva, the dorsum of the tongue, and the gingival crevice, and were inoculated and cultured. Plasma samples and saliva samples were analyzed for amoxicillin concentrations (free and protein bound) using liquid chromatography and mass-spectrometry. RESULTS: Amoxicillin was detected in concentrations over the break-point (>2 µg/mL) of amoxicillin in plasma after 1 and 4 hours but not after 24 hours. The dose had a significant effect on the streptococci in the gingival crevice. CONCLUSION: A single dose given as prophylaxis to prevent a surgical-site infection results in a significant reducing effect on the oral streptococcal microflora in the gingival crevice and may have an impact on bacteria spreading into tissues and the bacteremia of streptococci.

Jaw Bone Samples From Bisphosphonate-Treated Patients: A Pilot Cohort Study.

UNLABELLED: Osteonecrosis of the jaw (ONJ) is a severe complication of bisphosphonate treatment. PURPOSE: A detailed characterization of sampled peri-necrotic jawbone from bisphosphonate-treated patients was performed at tissue and cellular level (histological analyses and gene expression). MATERIALS AND METHODS: Alveolar bone samples were collected from patients with (n = 5) and without ONJ (n = 5). Healthy patients served as controls (n = 10). RESULTS: The histological analysis demonstrated low to moderate inflammation, displaying areas of inflammatory infiltrate in the bone marrow. Multinuclear giant cells and osteoclasts were found in both groups. Markers of bone formation (alkaline phosphatase, Col1a1, and osteocalcin), bone resorption (receptor activator of NF-kappaB ligand [RANKL], osteoprotegerin [OPG], tartrate-resistant acid phosphatase, and cathepsin K), inflammation (tumor necrosis factor-alpha, interleukin [IL]-1ß, and IL-6), angiogenesis (vascular endothelial growth factor A), and apoptosis (Casp3, Casp8, p53, and Smac) were evaluated. Nonparametric statistical tests were used to identify differences between the groups. In patients with ONJ, the expression level of the proinflammatory marker IL-1ß was strongly up-regulated compared with controls (p = .040). CONCLUSIONS: A down-regulated expression of Casp8 compared with controls was observed (p = .014) in patients treated with bisphosphonates. The RANKL/OPG ratios were similar in the three groups. The results indicate a need to further investigate the molecular mechanisms involved in the course of ONJ related to antiresorptive treatment.

Existing data sources for clinical epidemiology: Scandinavian Cohort for osteonecrosis of the jaw - work in progress and challenges.

Osteonecrosis of the jaw (ONJ) is a severe side effect associated with antiresorptive treatment. Monitoring of ONJ using routine databases in Scandinavian countries is a challenge owing to lack of valid algorithms and to heterogeneous referral practices. The aim of this paper is to describe the process of establishing a Scandinavian ONJ Cohort enrolling all ONJ cases related to antiresorptive treatment arising in Denmark, Norway, and Sweden between 2011 and 2019. The initial purpose of the cohort is to support an ongoing pharmacovigilance study of denosumab and zoledronic acid in Denmark, Norway, and Sweden. The three countries, with their 199 clinics, departments, and units of oral and maxillofacial surgery, both hospital-based and freestanding, differ somewhat in referral practices of the ONJ patients. By directly contacting all providers of care to ONJ patients in the three countries, we established a network for reporting incident cases to each country's research database directly or through a member of the Scandinavian ONJ task force as a liaison. The task force includes a Scandinavian coordinator and three national coordinators collaborating directly with the clinics. A uniform ONJ registration form has been developed, and the relevant medical community has been informed either directly or through presentations at professional meetings. A website with study information is published in each country, and data entry is ongoing. This large-scale systematic uniform registration of ONJ cases in Denmark, Norway, and Sweden, with an underlying total population of more than 20 million people, merged into the Scandinavian ONJ Cohort, will contribute to better knowledge and understanding of this challenging group of patients, and ultimately, help improve patient care. The Scandinavian ONJ Cohort as a whole and its component national ONJ research databases may offer the potential for large-scale multinational intervention and safety studies in the future.

A 5-year retrospective case series evaluating Brånemark Integration BioHelix (™) dental implants placed in a private practice by a specialist.

OBJECTIVE: To evaluate, in a case series, survival rate and complications of Brånemark Integration BioHelix(™) dental implants, placed according to conventional procedures in patients treated consecutively in a Swedish specialist private practice after 5 years. MATERIALS AND METHODS: Eighty-three consecutively-treated patients received 89 final fixed prostheses supported by 310 implants placed according to 'conventional' procedure, i.e. no implants shorter than 10 mm, no immediate post-extraction implants and no bone-grafting procedures. In 70 patients, implants were left to heal submerged, whereas 13 patients were treated according to the one-stage protocol. All implants were incorporated in a prosthetic restoration. Probing was only performed when clinical signs of inflammation were present and this was then evaluated further with intra-oral radiographic examination. Outcome measures were implant survival and prosthetic complications. RESULTS: Five years after implant placement, two fixtures were removed because of loosening. One fixture was lost after 12 months in the lower jaw in one patient and one fixture was lost in the upper jaw in another patient after 3 years, both inserted using the two-stage technique. No other prosthetic complications occurred, giving a 99.3% cumulative survival rate. CONCLUSIONS: Brånemark Integration BioHelix(™) dental implants placed according to one- or two-stage procedures in patients in a private practice produced excellent 5-year results. Randomized clinical trials with suitable controls are suggested to confirm these results.