Antiresorptive-Type and Discontinuation-Timing Affect ONJ Burden.

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.

Zoledronate Impairs Socket Healing after Extraction of Teeth with Experimental Periodontitis.

Osteonecrosis of the jaws (ONJ) is a rare but severe complication of antiresorptive medications, such as bisphosphonates, used in the treatment of bone malignancy or osteoporosis. Tooth extraction and dental disease have been strongly associated with ONJ development. Here, we investigated molecular and cellular markers of socket healing after extraction of healthy or teeth with experimental periodontitis (EP) in Wistar-Han rats treated with zoledronic acid (ZA). We included 4 experimental groups: vehicle-treated animals with extraction of healthy teeth or teeth with ligature-induced EP and ZA-treated animals with extraction of healthy teeth or teeth with EP. Animals were pretreated with vehicle or ZA for a week, and EP was induced. Four weeks later, the second maxillary molars were extracted; sockets were allowed to heal for 4 wk; animals were euthanized; and maxillae were isolated. Radiographically, extraction sockets in groups 1, 2, and 3 demonstrated normal healing. Contrary incomplete socket healing was noted after extraction of teeth with EP in ZA-treated rats of group 4. Histologically, persistent inflammation and extensive osteonecrosis were seen in group 4. Disorganization of the collagen network, collagen type III predominance, and lack of collagen fiber insertion in the necrotic bone were associated with impaired socket healing. Cells positive for MMP-9, MMP-13, and α-SMA expression were present at the areas of epithelial invagination and adjacent to osteonecrotic bone. Importantly, human biopsies from patients with ONJ showed similar findings. Our data emphasize the importance of dental disease and tooth extraction in ONJ pathogenesis and help delineate an altered profile in wound-healing markers during ONJ development.

Susceptibility of different mouse strains to peri-implantitis.

BACKGROUND AND OBJECTIVE: Peri-implantitis (PI) is an inflammatory condition that affects the tissues surrounding dental implants. Although the pathogenesis of PI is not fully understood, evidence suggests that the etiology is multifactorial and may include a genetic component. The aim of this study was to investigate the role of genetics in the development of peri-implantitis. MATERIAL AND METHODS: Four-week-old C57BL/6J, C3H/HeJ and A/J male mice had their left maxillary molars extracted. Implants were placed in the healed extraction sockets. Upon osseointegration, ligatures were placed around the implant head for 1 or 4 weeks to induce PI. Micro-computed tomography scanning was used to measure volumetric bone loss. Histological analyses were also performed to evaluate collagen organization and the presence of neutrophils and osteoclasts. RESULTS: Radiographically, comparing the ligature-treated mice, C57BL/6J displayed the greatest amount of bone loss, followed by C3H/HeJ and A/J mice at 1 and 4 weeks. Histologically, at 1 week, C57BL/6J mice presented with the highest numbers of neutrophils and osteoclasts. At 4 weeks, C57BL/6J mice presented with the most active bone remodeling compared with the other two strains. CONCLUSION: There were significant differences in the severity of peri-implantitis among the different mouse strains, suggesting that the genetic framework can affect implant survival and success. Future work is needed to dissect the genetic contribution to the development of peri-implantitis.

Heritability of periodontal bone loss in mice.

BACKGROUND: Periodontitis is an inflammatory disease of the periodontal tissues that compromises tooth support and can lead to tooth loss. Although bacterial biofilm is central in disease pathogenesis, the host response plays an important role in the progression and severity of periodontitis. Indeed, clinical genetic studies indicate that periodontitis is 50% heritable. In this study, we hypothesized that lipopolysaccharide (LPS) injections lead to a strain-dependent periodontal bone loss pattern. MATERIAL AND METHODS: We utilized five inbred mouse strains that derive the recombinant strains of the hybrid mouse diversity panel. Mice received Porphyromonas gingivalis-LPS injections for 6 wk. RESULTS AND CONCLUSION: Micro-computed tomography analysis demonstrated a statistically significant strain-dependent bone loss. The most susceptible strain, C57BL/6J, had a fivefold higher LPS-induced bone loss compared to the most resistant strain, A/J. More importantly, periodontal bone loss revealed 49% heritability, which closely mimics periodontitis heritability for patients. To evaluate further the functional differences that underlie periodontal bone loss, osteoclast numbers of C57BL/6J and A/J mice were measured in vivo and in vitro. In vitro analysis of osteoclastogenic potential showed a higher number of osteoclasts in C57BL/6J compared to A/J mice. In vivo LPS injections statistically significantly increased osteoclast numbers in both groups. Importantly, the number of osteoclasts was higher in C57BL/6J vs. A/J mice. These data support a significant role of the genetic framework in LPS-induced periodontal bone loss and the feasibility of utilizing the hybrid mouse diversity panel to determine the genetic factors that affect periodontal bone loss. Expanding these studies will contribute in predicting patients genetically predisposed to periodontitis and in identifying the biological basis of disease susceptibility.

Ligature-induced peri-implantitis in mice.

BACKGROUND AND OBJECTIVE: Peri-implantitis has a prevalence of 11-47%, involves destruction of peri-implant bone and may lead to implant loss. A detailed understanding of the pathogenesis of peri-implantitis is lacking. The objective of this study was to develop a murine model of experimental peri-implantitis. MATERIAL AND METHODS: Machined, smooth-surface, screw-shaped titanium implants were placed in the healed alveolar bone of the left maxillary molars of C57BL/6J male mice, 8 wk after tooth extraction. Peri-implantitis was induced by securing silk ligatures around the head of the implant fixtures. Implant survival and peri-implant bone levels were analyzed by micro-computed tomography (micro-CT) scans and histology, 12 wk after ligature placement. RESULTS: Implant survival was 60% (six of 10) for implants with ligatures and 100% (eight of eight) for controls. Micro-CT revealed significantly greater bone loss around the implants that received ligatures and that survived, compared with controls. The radiographic findings were confirmed via histology and toluidine blue staining. CONCLUSION: This study describes a murine model of experimental peri-implantitis around screw-shaped titanium implants placed in the edentulous alveolar bone. This model should be a useful tool to dissect pathogenic mechanisms of peri-implantitis and evaluate potential treatment interventions.

Prostaglandins E(2) and F(2alpha) enhance differentiation of cementoblastic cells.

BACKGROUND: The prostaglandins (PG) E(2) and PGF(2alpha) are important cytokines in periodontal physiology and pathology. PGE(2) and PGF(2alpha) alter cell function by binding and activating the plasmamembrane G-protein-coupled PG receptors. In this study, we examined the PGE(2) and PGF(2alpha) effects on the immortalized cementoblastic OCCM cells. METHODS: Confluent OCCM cells were treated with PGE(2), PGF(2alpha), specific activators/inhibitors of the EP prostanoid receptors, a specific activator of the FP prostanoid receptor, and direct activators/inhibitors of the protein kinase C (PKC) signaling pathway. Mineral nodule formation was assessed by the von Kossa stain. RESULTS: PGE(2) and PGF(2alpha) significantly increased mineralization of OCCM cells. The EP1 and EP3 PG receptor activators 16,16-dimethyl-prostaglandin E(2) and sulprostone, also increased mineralization. In contrast, specific activators of the EP2 or the EP2/EP3/EP4 receptors did not have any effect. Fluprostenol, a specific activator of the FP receptor, significantly increased mineralization of OCCM cells. FP and EP (1 or 3) receptors signal through activation of the protein kinase C (PKC) pathway. Indeed, phorbol 12-myristate 13-acetate (PMA), a direct activator of the PKC pathway, significantly increase OCCM mineralization, while pre-treatment of OCCM cells with the PKC inhibitor GF109203x (bisindolylmaleimide) significantly decreased mineralization. CONCLUSION: We conclude that PGE(2) and PGF(2alpha) exert an anabolic effect on OCCM mineralization through activation of PKC signaling.

Prostaglandins E2 and F2α Enhance Differentiation of Cementoblastic Cells.

BACKGROUND: The prostaglandins (PG) E2 and PGF2α are important cytokines in periodontal physiology and pathology. PGE2 and PGF2α alter cell function by binding and activating the plasmamembrane G-protein-coupled PG receptors. In this study, we examined the PGE2 and PGF2α effects on the immortalized cementoblastic OCCM cells. METHODS: Confluent OCCM cells were treated with PGE2 , PGF2α , specific activators/inhibitors of the EP prostanoid receptors, a specific activator of the FP prostanoid receptor, and direct activators/inhibitors of the protein kinase C (PKC) signaling pathway. Mineral nodule formation was assessed by the von Kossa stain. RESULTS: PGE2 and PGF2α significantly increased mineralization of OCCM cells. The EP1 and EP3 PG receptor activators 16,16-dimethyl-prostaglandin E2 and sulprostone, also increased mineralization. In contrast, specific activators of the EP2 or the EP2/EP3/EP4 receptors did not have any effect. Fluprostenol, a specific activator of the FP receptor, significantly increased mineralization of OCCM cells. FP and EP (1 or 3) receptors signal through activation of the protein kinase C (PKC) pathway. Indeed, phorbol 12-myristate 13-acetate (PMA), a direct activator of the PKC pathway, significantly increase OCCM mineralization, while pre-treatment of OCCM cells with the PKC inhibitor GF109203× (bisindolylmaleimide) significantly decreased mineralization. CONCLUSION: We conclude that PGE2 and PGF2α exert an anabolic effect on OCCM mineralization through activation of PKC signaling. J Periodontol 2005;76:303-309.