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Analysis of tooth extraction site in hematoxylin and eosin (H&E). Complete wound repair with epithelial and connective tissue (a) (×100), bone exposure (b) (×100) where non-vital bone is being circumscribed by soft tissue (c) (×200) with epithelial cell proliferation and inflammatory infiltrate (d) (×400). Vital bone showing neoformation (e) (×400) and non-vital bone with inflammatory infiltrate and adjacent epithelial cell proliferation (f) (×400)
Source publication
Objective
This study aimed to investigate the effect of hyperbaric oxygen therapy (HBOT) on tooth extraction sites in rats treated with bisphosphonate.
Materials and methods
Rats were treated with zoledronic acid, subjected to tooth extractions and allocated into groups: (1) 7 days of HBOT, (2) 14 days of HBOT, (3) 7-day control, and (4) 14-day con...
Citations
... The inflammatory pathway of receptor activator of nuclear factor NF-kB ligand and macrophage colony-stimulating factor are involved in the pathogenesis of this condition. These factors are necessary for the survival and proliferation of osteoclast precursors; in fact, they signal the cell via their receptor c-Fms, which in turn activates extracellular signal-regulated kinase and phosphoinositide 3-kinase/Akt [72][73][74][75][76][77][78][79]. Non-coding RNAs have been implicated in the pathophysiology of mandibular osteonecrosis, and this information is helpful for diagnosis because these small RANs may be biomarkers of apoptotic activity in bone. ...
Osteonecrosis of the jaw is the progressive loss and destruction of bone affecting the maxilla or mandible in patients treated with antiresorptive and antiangiogenic agents without receiving prior radiation therapy. The pathogenesis involves the inflammatory pathway of receptor activator of nuclear factor NF-kB ligand and the macrophage colony-stimulating factor, essential for osteoclast precursors survival and proliferation and acting through its receptor c-Fms. Evidence has shown the role of non-coding RNAs in the pathogenesis of osteonecrosis of the jaw and this finding might be useful in diagnosis since these small RNAs could be considered as biomarkers of apoptotic activity in bone. Interestingly, it has been proved that miR-29 and miR-31-5p, acting on specific targets such as CALCR and RhoA, promote programmed-cell death and consequently the necrosis of bone tissue. Specific long non-coding RNAs, instead, have been detected both at reduced levels in patients with multiple myeloma and osteonecrosis, and associated with suppression of osteoblast differentiation, with consequences in the progression of mandible lesions. Among non-coding genic material, circular RNAs have the capability to modify the expression of specific mRNAs responsible for the inhibition of bisphosphonates activity on osteoclastogenesis.
... However, tooth extraction is reported as the most common trigger for the condition occurring with a prevalence of 41% in patients using bisphosphonate (URADE et al., 2011) Although several treatment protocols have been proposed, there is still no consensus on the therapeutic approach to BRONJ, due to the limited effectiveness of these treatments. Several new therapeutic proposals have been tested for the prevention and treatment of this condition (AZARPAZHOOH, and LIMEBACK, 2008;CEPONIS et al., 2017;INCHINGOLO et al., 2017;NØRHOLT and HARTLEV, 2016;SILVA et al., 2017;SOYDAN and UCKAN, 2014). In this context, Acid Electrolyzed Water (AEA) that has already been investigated for wound healing and as a microbial agent (BUI et al., 2017;OKUBO, URAKAMI, TAMURA, 1999;TAMAKI et al., 2014;YAHAGI et al., 2000) may show promise as an adjunct to the prevention of BRONJ. ...
Bisphosphonate-related osteonecrosis of the jaws (BRONJ) is a complication associated with dental intervention and prolonged use of bisphosphonates (BF). Acidified Electrolyzed Water (AEA) is a substance that seems to improve tissue regeneration. To evaluate the effectiveness of (AEA) as an adjuvant in the treatment of BRONJ after lower molar extractions in Wistar rats. A split-mouth study was carried out in twelve Wistar rats (6 females and 6 males) received weekly intravenous injection of 0.3 mL of zoledronate (80 μg/kg in PBS) for 9 weeks and underwent tooth extraction. mandibular first and second molars in the eighth and ninth week, respectively. The Acidified Electrolyzed Water (AAG) group (right side of the mandible) was irrigated with AEA every three days while the control group (CG) (right side of the mandible) received no treatment after extractions. The animals were euthanized 6 weeks after the second extraction, the mandibles were dissected for clinical analysis with the data obtained submitted to statistical analysis with a significance level of 0.05. (Fischer's test). Two males were lost during AEA therapy, leaving 10 animals, 6 females and 4 males. Inflammation was absent in all alveoli; bone sequestration was noted in all alveoli; suppuration was present in 3 sites of the GC group, epithelialization present in 5 GAEA alveoli and 2 GC. Comparison of bone exposure did not show statistically significant results. The results obtained were slightly favorable to the use of AEA, however, without statistical significance.
... The rats were anesthetized with a mixture of 70 mg/kg ketamine hydrochloride (Syntec, Cotiá, SP, Brazil) and 7 mg/kg xylazine hydrochloride (Syntec) administered intraperitoneally (IP) (Luvizuto et al., 2010). Extraction of the first and second upper right molars was performed with an adapted 3s carver (SSWhite, Duflex, Rio de Janeiro, RJ, Brazil) and pediatric forceps (Edlo, Canoas, RS, Brazil) adapted to the size of the teeth [24,25]. Immediately after tooth extraction, a groove osteotomy was made at the extraction site with a 1.8-mm diameter ball bur under water irrigation, obtaining a standardized bone defect (1.8 mm x 1.8 mm x 6 mm). ...
... The blood samples collected were centrifuged after clot formation, and the serum was stored at −20°C. Maxillae were dissected, and the site of the tooth extractions was cut in a coronal direction by using an extra fine diamond disk (KG Sorensen, Cotia, SP, Brazil) and divided into two fragments, both showing the area of interest on the cut surface [24,25]. The specimens were immersed in 10% buffered formalin for 24 h. ...
Objective
Considering the chemical and structural properties of dentin, this study was aimed at evaluating the effect of dentin matrix alone or combined with mesenchymal stromal cells (MSC) on postextraction alveolar bone regeneration.Material and methodsWistar rats were subjected to tooth extraction with osteotomy and allocated into groups according to the graft inserted: (1) Gelita-Spon®, (2) Bio-Oss®, (3) Dentin, (4) MSC, (5) Dentin/MSC, and (6) Control. Maxillae were analyzed by means of hematoxylin and eosin (H&E) staining, immunohistochemical (IHC) analysis, microcomputed tomography (micro-CT), and scanning electron microscopy (SEM). Serum levels of calcium and phosphorus were quantified.ResultsThe Bio-Oss group showed less bone than Gelita-Spon and Dentin/MSC; no other significant differences were seen in H&E analysis. The Bio-Oss group showed higher expression of collagen type I compared to the Dentin and Dentin/MSC groups and also higher osteocalcin expression than the Dentin/MSC group. There was a tendency of higher expression of osteopontin in the MSC, Dentin, and Dentin/MSC groups and higher VEGF in the MSC group. On micro-CT analysis, the Bio-Oss and the Dentin/MSC groups exhibited greater bone volume than the Control. Serum calcium and phosphorus levels did not significantly differ between the groups. SEM analysis depicted particles of Bio-Oss and dentin in the respective groups, as well as significant cellularity in the MSC group.Conclusion
Autogenous nondemineralized dentin is an alternative for alveolar bone grafting, which can be improved by combination with MSC.Clinical relevanceThis work provides support for the clinical applicability of dentin graft alone or combined with MSC.
... Studies have shown that hyperbaric oxygen therapy can increase the effective oxygen content of tissue, promote the proliferation of capillaries, accelerate the formation of collateral circulation, and then promote osteogenesis, so as to accelerate the repair and healing of lesions in BRONJ patients. 41,42 Platelet-rich plasma (PRP) has been clinically used for the prevention of BRONJ. e Immunohistochemistry of CD31. ...
The significant clinical feature of bisphosphonate-related osteonecrosis of the jaw (BRONJ) is the exposure of the necrotic jaw. Other clinical manifestations include jaw pain, swelling, abscess, and skin fistula, which seriously affect the patients’ life, and there is no radical cure. Thus, new methods need to be found to prevent the occurrence of BRONJ. Here, a novel nanoparticle, tFNA-KLT, was successfully synthesized by us, in which the nanoparticle tetrahedral framework nucleic acid (tFNA) was used for carrying angiogenic peptide, KLT, and then further enhanced angiogenesis. TFNA-KLT possessed the same characteristics as tFNA, such as simple synthesis, stable structure, and good biocompatibility. Meanwhile, tFNA enhanced the stability of KLT and carried more KLT to interact with endothelial cells. First, it was confirmed that tFNA-KLT had the superior angiogenic ability to tFNA and KLT both in vitro and in vivo. Then we apply tFNA-KLT to the prevention of BRONJ. The results showed that tFNA-KLT can effectively prevent the occurrence of BRONJ by accelerating angiogenesis. In summary, the prepared novel nanoparticle, tFNA-KLT, was firstly synthesized by us. It was also firstly confirmed by us that tFNA-KLT significantly enhanced angiogenesis and can effectively prevent the occurrence of BRONJ by accelerating angiogenesis, thus providing a new avenue for the prevention of BRONJ and a new choice for therapeutic angiogenesis.
... OPG, the soluble decoy receptor for RANKL, inhibits RANKL binding to RANK and prevents bone resorption [26]. Hypoxia and hyperoxia could modulate the RANKL/OPG ratio [27,28]. However, we did not find any HBOTinduced changes in serum OPG or RANKL levels. ...
Introduction:
Oxygen is emerging as an important factor in the local regulation of bone remodeling. Some preclinical data suggest that hyperoxia may have deleterious effects on bone cells. However, its clinical relevance is unclear. Hence, we studied the effect of hyperbaric oxygen therapy (HBOT) on serum biomarkers reflecting the status of the Wnt and receptor activator of NF-κB ligand (RANKL) pathways, two core pathways for bone homeostasis.
Materials and methods:
This was a prospective study of 20 patients undergoing HBOT (mean age 58 yrs., range 35-82 yrs.) because of complications of radiotherapy or chronic anal fissure. Patients were subjected to HBOT (100% oxygen; 2.4 atmospheres absolute for 90 min). The average number of HBOT sessions was 20 ± 5 (range 8-31). Serum hypoxia-inducible factor 1-α (HIF1-α), osteoprotegerin (OPG), RANKL, and the Wnt inhibitors sclerostin and dickkopf-1 (DKK1) were measured at baseline and after HBOT by using specific immunoassays.
Results:
HIF-1α in eight patients with measurable serum levels increased from 0.084 (0.098) ng/mL at baseline to 0.146 (0.130) ng/mL after HBOT (p = 0.028). However, HBOT did not induce any significant changes in the serum levels of OPG, RANKL, sclerostin or DKK1. This was independent of the patients' diagnosis, either neoplasia or benign.
Conclusion:
Despite the potential concerns about hyperoxia, we found no evidence that HBOT has any detrimental effect on bone homeostasis.
... Of the 54 studies investigating preventive and curative treatments, 41 involved preventive therapies (Supplemental Table 1). Some of the investigated preventive treatments included PTH [105,[122][123][124][125][126][127], laser therapy [128][129][130]; photodynamic therapy [131,132], hyperbaric oxygen therapy [133,134], resveratrol [84,135], local chelation of the alveolar bone matrix using cadmium or EDTA [114,136], and local transplantation of mesenchymal stem cells (MSCs) or the application of derived products from MSCs [137][138][139]. Interestingly, other preventive modalities investigated the effects of discontinuation of pARS (OPG-Fc or ZOL) before or after tooth extraction [91,140]. ...
Medication-related osteonecrosis of the jaw (MRONJ) is a potentially severe adverse event affecting patients with cancer and patients with osteoporosis who have been treated with powerful antiresorptives (pARs) or angiogenesis inhibitors (AgIs). pARs, including nitrogen-containing bisphosphonates (N-BPs; e.g., zoledronic acid, alendronate) and anti-RANKL antibodies (e.g., denosumab), are used to manage bone metastases in patients with cancer or to prevent fragility fractures in patients with osteoporosis.
Though significant advances have been made in understanding MRONJ, its pathophysiology is still not fully elucidated. Multiple species have been used in preclinical MRONJ research, including the rat, mouse, rice rat, rabbit, dog, sheep, and pig. Animal research has contributed immensely to advancing the MRONJ field, particularly, but not limited to, in developing models and investigating risk factors that were first observed in humans. MRONJ models have been developed using clinically relevant doses of systemic risk factors, like N-BPs, anti-RANKL antibodies, or AgIs. Specific local oral risk factors first noted in humans, including tooth extraction and inflammatory dental disease (e.g., periodontitis, periapical infection, etc.), were then added. Research in rodents, particularly the rat, and, to some extent, the mouse, across multiple laboratories, has contributed to establishing multiple relevant and complementary preclinical models. Models in larger species produced accurate clinical and histopathologic outcomes suggesting a potential role for confirming specific crucial findings from rodent research. We view the current state of animal models for MRONJ as good. The rodent models are now reliable enough to produce large numbers of MRONJ cases that could be applied in experiments testing treatment modalities. The course of MRONJ, including stage 0 MRONJ, is characterized well enough that basic studies of the molecular or enzyme-level findings in different MRONJ stages are possible.
This review provides a current overview of the existing models of MRONJ, their more significant features and findings, and important instances of their application in preclinical research.
... Animals were anesthetized with a single intraperitoneal injection (IP) of a mixture of ketamine hydrochloride at 70 mg/kg (Syntec, Cotiá, SP, Brazil) and xylazine hydrochloride at 7 mg/kg (Syntec) [27]. The three upper right molars were extracted using an adapted 3s spatula (SSWhite, Duflex, Rio de Janeiro, RJ, Brazil) for luxation and pediatric forceps (Edlo, Canoas, RS, Brazil) adapted to the size of the teeth [26,29,30]. ...
... Euthanasia, macroscopic analysis, and specimen processing Afterwards, the maxilla was cut into two fragments in the coronal direction, using an extra fine diamond disk (KG Sorensen, Cotia, SP, Brazil) [29,30]. The maxilla fragments designated for histological analysis were fixed for 24 h in 10% buffered formalin, and those designated for electron microscopy analysis were fixed in 2.5% glutaraldehyde. ...
... In H&E staining, five fields were captured in each slide aiming to cover the entire tooth extraction area; in immunohistochemical analysis, four fields were captured per slide, including bone and the adjacent connective tissue at the tooth extraction site. The captured images were stored in TIFF format (uncompressed) [29]. ...
Objective:
The aim of this study was to evaluate morphological and immunohistochemical features of tooth extraction sites in rats subjected to different antiresorptive drugs.
Materials and methods:
Wistar rats were allocated into 4 groups according to the treatment: (1) alendronate, (2) raloxifene, (3) strontium ranelate, and (4) control. The animals underwent tooth extraction (60th day of treatment) and afterwards were euthanized (90th day of treatment). Tooth extraction sites were analyzed by means of scanning electron microscopy (SEM), hematoxylin-eosin staining (H&E), and immunohistochemical staining (RANKL and OPG).
Results:
On H&E analysis, the alendronate group showed greater amounts of non-vital bone, biofilm, inflammatory infiltrate and root fragment, and smaller amount of vital bone. The strontium ranelate group showed great amount of non-vital bone. This group also had lower levels of OPG, while the alendronate group showed lower OPG and RANKL than the other groups. On SEM analysis, the alendronate group showed a considerable number of microcracks on the alveolar bone surface and few Howship lacunae and lack of bone cells as well. The raloxifene, strontium ranelate, and control groups showed a large number of bone cells and Howship lacunae on the bone surface and few microcracks.
Conclusion:
Alendronate therapy is associated with macro- and microscopic features of medication-related osteonecrosis of the jaw at tooth extraction sites, whereas raloxifene therapy is not, and strontium ranelate therapy is associated with non-vital bone.
Clinical relevance:
Osteonecrosis of the jaws is a serious side effect of alendronate therapy, where tooth extraction is a major risk factor. Considering the significant number of patients undergoing antiresorptive therapies worldwide, the present study investigated whether raloxifene and strontium ranelate interfere with bone repair after tooth extraction in a similar way to bisphosphonates.
... In other studies, this phase was affected. For example, in one study proliferative genes of bone cells in rat teeth were deceased after extraction [22]. Irradiated salivary glands of mice showed increased EGF levels [23]. ...
Background: Hyperbaric oxygen (HBO2) therapy can
have a positive effect on wound healing, angiogenesis and
blood flow. No prior study has described the effects of HBO2
therapy and gene expression of this process. The goal of our
research was to show the effects of HBO2 and its impact at
the molecular level on angiogenesis, proliferation, differentiation,
oxidative stress, inflammation, and extracellular
matrix formation. Live animal subjects were used for
simulating the process of wound healing under standard
conditions and under the influence of HBO2.
Methods: Two experimental groups were created using
injured rabbits (N=24), one group (N=12) treated with
hyperbaric therapy twice a day and one (N=12) with
standard wound care management. Wounds were surgical,
uninfected, and in healthy animal test subjects. We compared
the whole genomic analysis of the transcriptome with the use
of microarray technology at three intervals during treatment.
Results: The induction of the wounds in rabbit skin
increased expression of hundreds of genes in both
treatment groups. The numbers of elevated and decreased
genes gradually reduced as the wound healed. Gene
expression analysis showed elevated expression of
several genes associated with inflammation in both groups
of injured animals. Genes connected to the process of
angiogenesis, proliferation, differentiation, oxidative stress
and extracellular matrix formation were without statistically
significant changes.
Conclusion: The evidence did not support that HBO2
had any significant effect on gene expression during wound
healing. Additionally, there was no evidence to support that
there were changes in gene expression in either treatment
group.
... recently, hyperbaric oxygen therapy (HBo) was used to optimize bone repair. the increase in oxygen availability to the tissues induced by HBo allows better wound repair, [10][11][12][13][14][15] since this therapy promotes reduction in hypoxia, promotes angiogenesis, and the differentiation of fibroblasts. 10,11,16,17 More recently, studies have determined that HBo also generates reactive oxygen species (roS) which, in turn, interfere with the synthesis of inflammation mediators, antioxidants and growth factors resulting in improved repair. ...
... 10,11,16,17 More recently, studies have determined that HBo also generates reactive oxygen species (roS) which, in turn, interfere with the synthesis of inflammation mediators, antioxidants and growth factors resulting in improved repair. [12][13][14] in oral and maxillofacial surgery, HBo can be used as adjunctive therapy in reconstructive procedures. 3, 18 after maxillofacial trauma, ruptured blood vessels lead to the formation of a hypoxic zone. ...
Introduction:
The aim of this literature review was to determine the benefits of hyperbaric oxygen therapy after bone reconstruction procedures in humans and identify information that may be useful for the development of optimal protocols for hyperbaric oxygen therapy to stimulate bone healing.
Evidence acquisition:
We searched the electronic database PubMed/Medline for studies published between January 1999 and December 2018, using the key words: 'bone' or 'bone graft' and 'mandible reconstruction' or 'jaw reconstruction' and 'hyperbaric oxygen' or 'HBO'. First, the titles and abstracts of the studies found were evaluated and those that corresponded to the aims of this review were pre-selected for analysis of the full text. Subsequently, the full texts were analyzed, and those that met the eligibility criteria were pre- selected for the review. The full texts of studies whose abstracts did not provide enough data for decision were also evaluated. Two examiners independently assessed eligibility, risk of bias and extracted data.
Evidence synthesis:
A total of 2237 studies were found according to pre- established criteria for data collection, of which only 5 studies were included in this systematic review. Although we observed positive results in the included studies, there are still few standardized clinical studies in the literature, assessing hyperbaric oxygen therapy after extensive bone reconstructive procedures.
Conclusions:
It is difficult to compare results found in different studies due to the variety of methodological and clinical conditions assessed.
... 11,13 Hence, many in vivo studies have focused on the healing of extraction sockets in studies related to MRONJ. 9,10,11,12,14,15,16 The treatment of MRONJ is challenging; 14 hence, prevention is crucial for its management. 2 Furthermore, the AAOMS promoted animal studies to validate improved strategies for the prevention of MRONJ as one of its future research areas. 1 Moreover, the Japanese Allied Committee reported that an understanding of the effects of antiresorptive agents at a cellular level was needed, and that these agents play a critical role in the closure of tooth extraction sockets to protect the alveolar bone from exposure to the oral cavity. 6 Various animal studies have been reported recently, 9,14,17,18 but few studies on the prevention methods for MRONJ have been conducted in animal models. ...
... 9,10,11,12,14,15,16 The treatment of MRONJ is challenging; 14 hence, prevention is crucial for its management. 2 Furthermore, the AAOMS promoted animal studies to validate improved strategies for the prevention of MRONJ as one of its future research areas. 1 Moreover, the Japanese Allied Committee reported that an understanding of the effects of antiresorptive agents at a cellular level was needed, and that these agents play a critical role in the closure of tooth extraction sockets to protect the alveolar bone from exposure to the oral cavity. 6 Various animal studies have been reported recently, 9,14,17,18 but few studies on the prevention methods for MRONJ have been conducted in animal models. The authors attempted to address two questions, based on the collagen sponge, which has been commonly used for socket preservation, and bone morphogenetic protein (BMP), which has been very effective for bone regeneration in the clinic: Does closure of the extraction socket with a collagen sponge have a positive effect on the prevention of alveolar bone exposure? ...
The aim of the present study was to evaluate the possible use of a commercial absorbed collagen sponge and bone morphogenetic protein (BMP) for the prevention of bisphosphonate-related osteonecrosis of the jaw (BRONJ) in rats. Twenty rats received intraperitoneal injections of 0.1-mg/kg of zoledronic acid three times a week for eight weeks before the extraction of both maxillary first molars after eight weeks. A collagen sponge (experimental group 1) and a collagen sponge with recombinant human BMP-2 (experimental group 2) were applied to the right extraction sockets of ten rats each. The 20 left extraction sockets (control groups 1 and 2) were left unprotected. After eight weeks, all rats were euthanized. Macroscopic analysis, micro-computed tomography (CT) analysis, and histological analysis were performed. There was a significant difference in the bone density between the control and experimental groups on micro-CT analysis. Impaired healing of the extraction sockets, indicating BRONJ, was observed in 80% of control group 1, 90% of control group 2, 30% of experimental group 1, and 20% of experimental group 2. The collagen sponge with/without BMP used for protecting the extraction socket had the potential for a positive effect in reducing the incidence of bisphosphonate-related osteonecrosis of the jaw in rats.
