Medication-related osteonecrosis of the jaws after tooth extraction in senescent female mice treated with zoledronic acid: microtomographic, histological and immunohistochemical characterization

AbstractTreatment with cumulative dosages of zoledronic acid (ZA) in elderly patients is a risk factor for the development of medication-related osteonecrosis of the jaws (MRONJ), mainly related to surgical triggers such as tooth extraction. However, animal models for the investigation and understanding of MRONJ pathophysiology in senescent and postmenopausal stages remains to be developed and characterized. The aim of this study was to analyze MRONJ development in senescent female mice treated with cumulative dosages of ZA. For this purpose, twenty 129/Sv female mice, 64 weeks old, were treated with 0.9% saline solution as Control group (n=10), and with ZA at 250µg/Kg (n=10), once a week, starting 4 weeks before the upper right incisor extraction and until the end of the experimental time points (7 days and 21 days). At 7 and 21 days, specimens were harvested for microCT, histological, birefringence and immunohistochemical analysis. Clinically, an incomplete epithelialization was observed in ZA group at 7 days and a delayed bone matrix mineralization and collagen maturation at 7 and 21 days compared to the controls. Controls revealed sockets filled with mature bone at 21 days as observed by microCT and birefringence, while ZA group presented delayed bone deposition at 7 and 21 days, as well increased leukocyte infiltration and blood clot at 7 days, and increased bone sequestrum and empty osteocyte lacunae at 21 days (p<0.05). Also, ZA group presented decreased quantity TGFb+ and Runx-2+ cells at 7 days, and decreased quantity of TRAP+ osteoclasts compared to the control at 21 days (p<0.05). Togheter, these data demonstrate the usefulness of this model to understanding the pathophysiology of MRONJ.

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Medication-related osteonecrosis of the jaws after tooth extraction in senescent female mice treated with zoledronic acid: Microtomographic, histological and immunohistochemical characterization

PLoS ONE ◽

10.1371/journal.pone.0214173 ◽

2019 ◽

Vol 14 (6) ◽

pp. e0214173 ◽

Cited By ~ 2

Author(s):

Claudia Cristina Biguetti ◽

André Hergesel De Oliva ◽

Kent Healy ◽

Ramez Hassan Mahmoud ◽

Isabela Do Carmo Custódio ◽

...

Keyword(s):

Zoledronic Acid ◽

Tooth Extraction ◽

Female Mice ◽

Osteonecrosis Of The Jaws

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BMP-2/β-TCP Local Delivery for Bone Regeneration in MRONJ-Like Mouse Model

International Journal of Molecular Sciences ◽

10.3390/ijms21197028 ◽

2020 ◽

Vol 21 (19) ◽

pp. 7028

Author(s):

Akihiro Mikai ◽

Mitsuaki Ono ◽

Ikue Tosa ◽

Ha Thi Thu Nguyen ◽

Emilio Satoshi Hara ◽

...

Keyword(s):

Bone Formation ◽

Bone Remodeling ◽

Tooth Extraction ◽

Pathological Condition ◽

Immunohistochemical Analysis ◽

Osteonecrosis Of The Jaw ◽

Control Group ◽

Extraction Socket ◽

Osteoclast Activity ◽

Prevention Studies

Medication-related osteonecrosis of the jaw (MRONJ) is a severe pathological condition associated mainly with the long-term administration of bone resorption inhibitors, which are known to induce suppression of osteoclast activity and bone remodeling. Bone Morphogenetic Protein (BMP)-2 is known to be a strong inducer of bone remodeling, by directly regulating osteoblast differentiation and osteoclast activity. This study aimed to evaluate the effects of BMP-2 adsorbed onto beta-tricalcium phosphate (β-TCP), which is an osteoinductive bioceramic material and allows space retention, on the prevention and treatment of MRONJ in mice. Tooth extraction was performed after 3 weeks of zoledronate (ZA) and cyclophosphamide (CY) administration. For prevention studies, BMP-2/β-TCP was transplanted immediately after tooth extraction, and the mice were administered ZA and CY for an additional 4 weeks. The results showed that while the tooth extraction socket was mainly filled with a sparse tissue in the control group, bone formation was observed at the apex of the tooth extraction socket and was filled with a dense connective tissue rich in cellular components in the BMP-2/β-TCP transplanted group. For treatment studies, BMP-2/β-TCP was transplanted 2 weeks after tooth extraction, and bone formation was followed up for the subsequent 4 weeks under ZA and CY suspension. The results showed that although the tooth extraction socket was mainly filled with soft tissue in the control group, transplantation of BMP-2/β-TCP could significantly accelerate bone formation, as shown by immunohistochemical analysis for osteopontin, and reduce the bone necrosis in tooth extraction sockets. These data suggest that the combination of BMP-2/β-TCP could become a suitable therapy for the management of MRONJ.

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Osteopromotion Capacity of Bovine Cortical Membranes in Critical Defects of Rat Calvaria: Histological and Immunohistochemical Analysis

International Journal of Biomaterials ◽

10.1155/2020/6426702 ◽

2020 ◽

Vol 2020 ◽

pp. 1-9 ◽

Cited By ~ 1

Author(s):

Carolina Ferrairo Danieletto-Zanna ◽

Vinícius Ferreira Bizelli ◽

Guilherme André Del Arco Ramires ◽

Tamires Melo Francatti ◽

Paulo Sérgio Perri de Carvalho ◽

...

Keyword(s):

Blood Clot ◽

Immunohistochemical Analysis ◽

Negative Control Group ◽

Repair Process ◽

Positive Control ◽

Negative Control ◽

Control Group ◽

Collagen Membrane ◽

Rat Calvaria ◽

Experimental Group

Membranes that aid the guided bone regeneration (GBR) process have been the subject of studies of compatible biomaterials that contribute to this repair process. The present study compared different membranes used in critical-size defects of rat calvaria by assessing GBR as well as histological, histomorphometric, and immunohistochemical reactions. Forty-eight male albino Wistar rats were randomly allocated into four groups (n = 12 each), namely, C: membrane-free control group (only blood clot, negative control group); BG: porcine collagen membrane group (Bio-Gide®, positive control group); GD: bovine cortical membrane group (first experimental group); and GDF: thicker bovine cortical membrane group (second experimental group). Rats were euthanized at 30 and 60 days postoperatively. Quantitative data from the histometric analysis were submitted to two-way ANOVA and Tukey’s posttest when p<0.05. Histomorphometric results of the thicker bovine cortical membrane at 30 and 60 days were promising, showing improved new bone formation values (p<0.05), and the CD group presented similar results in both analysis periods, being surpassed only by the GDF group (p<0.05). The immunohistochemical results were associated with the histomorphometric data. A less-thick membrane also assisted in GBR. All membranes promoted GBR, especially the positive control and experimental groups.

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Antiresorptive-Type and Discontinuation-Timing Affect ONJ Burden

Journal of Dental Research ◽

10.1177/0022034520986804 ◽

2021 ◽

pp. 002203452098680

Author(s):

D. Hadaya ◽

A. Soundia ◽

I. Gkouveris ◽

O. Bezouglaia ◽

S.M. Dry ◽

...

Keyword(s):

Zoledronic Acid ◽

Experimental Design ◽

Oral Cavity ◽

Tooth Extraction ◽

Treatment Options ◽

Severe Side Effect ◽

Oral Surgical Procedures ◽

Affected Area ◽

Osteonecrosis Of The Jaws ◽

Clinical Scenarios

Osteonecrosis of the jaws (ONJ), a severe side effect of antiresorptive medications, is characterized by exposed, nonhealing bone in the oral cavity. Treatment options for ONJ range from management of symptomology to surgical resection of the affected area. Antiresorptive discontinuation, often termed a “drug holiday,” has been used for managing ONJ patients. Antiresorptives can be discontinued prior to oral surgical procedures, such as tooth extraction, to prevent ONJ development or in patients with established ONJ to accelerate healing. Here, our objective was to test these clinical scenarios using the potent bisphosphonate, zoledronic acid (ZA), and the denosumab surrogate for rodents, OPG-Fc, in a rat model of ONJ. Animals were pretreated with antiresorptives or saline, after which we induced ONJ using periapical disease and tooth extraction. In our first experimental design, antiresorptives were discontinued 1 wk prior to tooth extraction, and animals were evaluated 4 wk later for clinical, radiographic, and histologic features of ONJ. In the second experiment, ONJ was established and antiresorptives were discontinued for 4 wk. Discontinuation of OPG-Fc, but not ZA, prior to tooth extraction ameliorated subsequent ONJ development. In contrast, discontinuation of either ZA or OPG-Fc in rats with established ONJ did not lead to ONJ resolution. In conclusion, our findings suggest that antiresorptive discontinuation is dependent on both the type of antiresorptive and the timing of discontinuation.

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Annual intravenous zoledronic acid for three years increased cancellous bone matrix mineralization beyond normal values in the HORIZON biopsy cohort

Journal of Bone and Mineral Research ◽

10.1002/jbmr.1780 ◽

2013 ◽

Vol 28 (3) ◽

pp. 442-448 ◽

Cited By ~ 24

Author(s):

Barbara M Misof ◽

Paul Roschger ◽

Daniela Gabriel ◽

Eleftherios P Paschalis ◽

Erik F Eriksen ◽

...

Keyword(s):

Zoledronic Acid ◽

Cancellous Bone ◽

Normal Values ◽

Bone Matrix ◽

Matrix Mineralization ◽

Bone Matrix Mineralization

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Evaluation of the Use of an Inorganic Bone Matrix in the Repair of Bone Defects in Rats Submitted to Experimental Alcoholism

Materials ◽

10.3390/ma13030695 ◽

2020 ◽

Vol 13 (3) ◽

pp. 695

Author(s):

Iris Jasmin Santos German ◽

Karina Torres Pomini ◽

Ana Carolina Cestari Bighetti ◽

Jesus Carlos Andreo ◽

Carlos Henrique Bertoni Reis ◽

...

Keyword(s):

Blood Clot ◽

Bone Matrix ◽

Experimental Period ◽

Chronic Alcoholism ◽

Bone Defects ◽

Ethanol Solution ◽

Liquid Diet ◽

Male Rats ◽

Control Group ◽

Significant Difference

To assess the effects of chronic alcoholism on the repair of bone defects associated with xenograft. Forty male rats were distributed in: control group (CG, n = 20) and experimental group (EG, n = 20), which received 25% ethanol ad libitum after a period of adaptation. After 90 days of liquid diet, the rats were submitted to 5.0-mm bilateral craniotomy on the parietal bones, subdividing into groups: CCG (control group that received only water with liquid diet and the defect was filled with blood clot), BCG (control group that received only water with liquid diet and the defect was filled with biomaterial), CEG (alcoholic group that received only ethanol solution 25% v/v with liquid diet and the defect was filled with blood clot), and BEG (alcoholic group that received only ethanol solution 25% v/v with liquid diet and the defect was filled with biomaterial). In the analysis of body mass, the drunk animals presented the lowest averages in relation to non-drunk animals during the experimental period. Histomorphologically all groups presented bone formation restricted to the defect margins at 60 days, with bone islets adjacent to the BCG biomaterial particles. CEG showed significant difference compared to BEG only at 40 days (17.42 ± 2.78 vs. 9.59 ± 4.59, respectively). In the birefringence analysis, in early periods all groups showed red-orange birefringence turning greenish-yellow at the end of the experiment. The results provided that, regardless of clinical condition, i.e., alcoholic or non-alcoholic, in the final period of the experiment, the process of bone defect recomposition was similar with the use of xenograft or only clot.

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MKP1-dependent PTH modulation of bone matrix mineralization in female mice is osteoblast maturation stage specific and involves P-ERK and P-p38 MAPKs

Journal of Endocrinology ◽

10.1530/joe-12-0372 ◽

2012 ◽

Vol 216 (3) ◽

pp. 315-329 ◽

Cited By ~ 13

Author(s):

Chandrika D Mahalingam ◽

Bharat Reddy Sampathi ◽

Sonali Sharma ◽

Tanuka Datta ◽

Varsha Das ◽

...

Keyword(s):

Bone Matrix ◽

Matrix Gla Protein ◽

Maturation Stage ◽

Limited Information ◽

Matrix Mineralization ◽

P38 Inhibitor ◽

Female Mice ◽

Osteoblast Maturation ◽

P38 Mapks

Limited information is available on the role of MAPK phosphatase 1 (MKP1) signaling in osteoblasts. We have recently reported distinct roles for MKP1 during osteoblast proliferation, differentiation, and skeletal responsiveness to parathyroid hormone (PTH). As MKP1 regulates the phosphorylation status of MAPKs, we investigated the involvement of P-ERK and P-p38 MAPKs in MKP1 knockout (KO) early and mature osteoblasts with respect to mineralization and PTH response. Calvarial osteoblasts from 9–14-week-old WT and MKP1 KO male and female mice were examined. Western blot analysis revealed downregulation and sustained expressions of P-ERK and P-p38 with PTH treatment in differentiated osteoblasts derived from KO males and females respectively. Exposure of early osteoblasts to p38 inhibitor, SB203580 (S), markedly inhibited mineralization in WT and KO osteoblasts from both genders as determined by von Kossa assay. In osteoblasts from males, ERK inhibitor U0126 (U), not p38 inhibitor (S), prevented the inhibitory effects of PTH on mineralization in early or mature osteoblasts. In osteoblasts from KO females, PTH sustained mineralization in early osteoblasts and decreased mineralization in mature cells. This effect of PTH was attenuated by S in early osteoblasts and by U in mature KO cells. Changes in matrix Gla protein expression with PTH in KO osteoblasts did not correlate with mineralization, indicative of MKP1-dependent additional mechanisms essential for PTH action on osteoblast mineralization. We conclude that PTH regulation of osteoblast mineralization in female mice is maturation stage specific and involves MKP1 modulation of P-ERK and P-p38 MAPKs.

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Augmentation of Demineralized Bone Matrix Post-Tooth Extraction Increases The Density of Gingival Collagen Fiber of Rabbit Mandible

The Indonesian Journal of Dental Research ◽

10.22146/theindjdentres.10174 ◽

2010 ◽

Vol 1 (1) ◽

pp. 9 ◽

Cited By ~ 1

Author(s):

Regina TC Tandelilin

Keyword(s):

Wound Healing ◽

Tooth Extraction ◽

Collagen Fiber ◽

Demineralized Bone Matrix ◽

Healing Process ◽

Bone Matrix ◽

Control Group ◽

Wound Healing Process ◽

Absorbable Sutures ◽

Demineralized Bone

Most cases report that the abnormality of bone defect is engendered by tooth extraction. The powder of demineralized bone matrix (DBM) is required to fill alveolus bone for reconstructing material or preventingtissues defect after tooth extraction. The aim of this study was to determine the density of gingival collagen fiber on wound healing after the augmentation of DBM following the extraction of incisivus tooth. In this study, 36 male rabbits aged 2.5-3 months weighing 900-1,100 grams were randomly divided into two groups: control and treated rabbits. The incisivus teeth of mandibles of treated rabbits were extracted and augmented with the allograft DBM powder. The gingival was sutured with non-absorbable sutures. The same procedures were employed to the control group, except that these rabbits were augmented with DBM powder. Subsequently, the rabbits were sacrificed on days 1, 3, 5, 7, 10, and 14 after surgery, and each observation was represented by three rabbits in the sample. The gingival (ca. 0.5-1cm) was cut and fixed immediately in 10% paraformaldehide. The staining was done using van Geison. In the treated rabbits, the density of gingival collagen fiber significantly increases in all observation days except on first day, indicating that the allograft powder of DBM successfully accelerates the wound healing process.

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Increased Osteocyte Lacunae Density in the Hypermineralized Bone Matrix of Children with Osteogenesis Imperfecta Type I

International Journal of Molecular Sciences ◽

10.3390/ijms22094508 ◽

2021 ◽

Vol 22 (9) ◽

pp. 4508

Author(s):

Matthias Mähr ◽

Stéphane Blouin ◽

Martina Behanova ◽

Barbara M. Misof ◽

Francis H. Glorieux ◽

...

Keyword(s):

Trabecular Bone ◽

Bone Metabolism ◽

Bone Mineralization ◽

Bone Biopsy ◽

Bone Matrix ◽

Type I ◽

Osteogenesis Imperfecta Type ◽

Bone Homeostasis ◽

Matrix Mineralization ◽

Osteocyte Lacunae

Osteocytes are terminally differentiated osteoblasts embedded within the bone matrix and key orchestrators of bone metabolism. However, they are generally not characterized by conventional bone histomorphometry because of their location and the limited resolution of light microscopy. OI is characterized by disturbed bone homeostasis, matrix abnormalities and elevated bone matrix mineralization density. To gain further insights into osteocyte characteristics and bone metabolism in OI, we evaluated 2D osteocyte lacunae sections (OLS) based on quantitative backscattered electron imaging in transiliac bone biopsy samples from children with OI type I (n = 19) and age-matched controls (n = 24). The OLS characteristics were related to previously obtained, re-visited histomorphometric parameters. Moreover, we present pediatric bone mineralization density distribution reference data in OI type I (n = 19) and controls (n = 50) obtained with a field emission scanning electron microscope. Compared to controls, OI has highly increased OLS density in cortical and trabecular bone (+50.66%, +61.73%; both p < 0.001), whereas OLS area is slightly decreased in trabecular bone (−10.28%; p = 0.015). Correlation analyses show a low to moderate, positive association of OLS density with surface-based bone formation parameters and negative association with indices of osteoblast function. In conclusion, hyperosteocytosis of the hypermineralized OI bone matrix associates with abnormal bone cell metabolism and might further impact the mechanical competence of the bone tissue.

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