Targeting phosphatidylinositol-3-kinase for inhibiting maxillary bone resorption.

Previous studies have suggested a role of phosphatidylinositol-3-kinase gamma (PI3Kγ) in bone remodeling, but the mechanism remains undefined. Here, we explored the contribution of PI3Kγ in the resorption of maxillary bone and dental roots using models of orthodontic tooth movement (OTM), orthodontic-induced inflammatory root resorption, and rapid maxillary expansion (RME). PI3Kγ-deficient mice (PI3Kγ-/- ), mice with loss of PI3Kγ kinase activity (PI3KγKD/KD ) and C57BL/6 mice treated with a PI3Kγ inhibitor (AS605240) and respective controls were used. The maxillary bones of PI3Kγ-/- , PI3KγKD/KD , and C57BL/6 mice treated with AS605240 showed an improvement of bone quality compared to their controls, resulting in reduction of the OTM and RME in all experimental groups. PI3Kγ-/- mice exhibited increased root volume and decreased odontoclasts counts. Consistently, the pharmacological blockade or genetic deletion of PI3K resulted in increased numbers of osteoblasts and reduction in osteoclasts during OTM. There was an augmented expression of Runt-related transcription factor 2 (Runx2) and alkaline phosphatase (Alp), a reduction of interleukin-6 (Il-6), as well as a lack of responsiveness of receptor activator of nuclear factor kappa-Β (Rank) in PI3Kγ-/- and PI3KγKD/KD mice compared to control mice. The maxillary bones of PI3Kγ-/- animals showed reduced p-Akt expression. In vitro, bone marrow cells treated with AS605240 and cells from PI3Kγ-/- mice exhibited significant augment of osteoblast mineralization and less osteoclast differentiation. The PI3Kγ/Akt axis is pivotal for bone remodeling by providing negative and positive signals for the differentiation of osteoclasts and osteoblasts, respectively.

Klotho in Osx+-mesenchymal progenitors exerts pro-osteogenic and anti-inflammatory effects during mandibular alveolar bone formation and repair.

Maxillofacial bone defects are commonly seen in clinical practice. A clearer understanding of the regulatory network directing maxillofacial bone formation will promote the development of novel therapeutic approaches for bone regeneration. The fibroblast growth factor (FGF) signalling pathway is critical for the development of maxillofacial bone. Klotho, a type I transmembrane protein, is an important components of FGF receptor complexes. Recent studies have reported the presence of Klotho expression in bone. However, the role of Klotho in cranioskeletal development and repair remains unknown. Here, we use a genetic strategy to report that deletion of Klotho in Osx-positive mesenchymal progenitors leads to a significant reduction in osteogenesis under physiological and pathological conditions. Klotho-deficient mensenchymal progenitors also suppress osteoclastogenesis in vitro and in vivo. Under conditions of inflammation and trauma-induced bone loss, we find that Klotho exerts an inhibitory function on inflammation-induced TNFR signaling by attenuating Rankl expression. More importantly, we show for the first time that Klotho is present in human alveolar bone, with a distinct expression pattern under both normal and pathological conditions. In summary, our results identify the mechanism whereby Klotho expressed in Osx+-mensenchymal progenitors controls osteoblast differentiation and osteoclastogenesis during mandibular alveolar bone formation and repair. Klotho-mediated signaling is an important component of alveolar bone remodeling and regeneration. It may also be a target for future therapeutics.

Sutural fibroblasts exhibit the function of vascular endothelial cells upon mechanical strain.

Midfacial hypoplasia is a type of facial dysplasia. The technique of trans-sutural distraction osteogenesis promotes midface growth so as to ameliorate this symptom. In the process of distraction osteogenesis, the fiber matrix in the suture acts as a mechanical sensor. Compared with osteogenesis, the formation of collagen fibers by fibroblasts is significant in the early stage of sutural distraction. However the transformation of fibroblasts during sutural bone formation induced by tensile force is poorly characterized. Here, we used single-cell RNA sequencing to define the cell classification of the zygomatic maxillary suture and the changes of cell clusters in the suture before and after seven-day distraction. We identified twenty-nine cell subsets spanning monocyte/macrophages, neutrophils, red blood cells, B cells and fibroblasts. Compared with the control group, Monocle analysis revealed the emergence of a unique fibroblast subset (Cdh5+, Col4a1+, Fat1-, and Acta2-) (cluster 27) that expressed vascular endothelial cell genes within the distracted zygomatic maxillary suture. We constructed the differentiation trajectories of the fibroblast population (cluster 23, 27) in the suture before and after distraction. In addition, we clarified that a subset of fibroblasts (cluster 27) lost expression of Fat1, an upregulator of the Hippo pathway, and upregulated Cyr61, a downstream gene of the Hippo pathway, during the distraction process. Further enrichment analysis suggests that cells of the new subset (cluster 27) are undergoing conversion of their identity into a vascular endothelial cell-like state in response to mechanical stimulation, associated with upregulation of angiogenesis genes along the single-cell trajectory. Further immunofluorescence staining confirmed this phenomenon. A combined general transcriptome RNA sequencing data analysis demonstrated that the fibroblasts expressed a number of extracellular matrix-related genes under mechanical strain. These data together provide a new view of the role of fibroblasts in tension-induced sutural angiogenesis via interaction with the Hippo pathway.

In-bone protein digestion followed by LC-MS/MS peptide analysis as a new way towards the routine proteomic characterization of human maxillary and mandibular bone tissue in oral surgery.

Proteomic characterization of alveolar bones in oral surgery represents an analytical challenge due to their insoluble character. The implementation of a straightforward technique could lead to the routine use of proteomics in this field. This work thus developed a simple technique for the characterization of bone tissue for human maxillary and mandibular bones. It is based on the direct in-bone tryptic digestion of proteins in both healthy and pathological human maxillary and mandibular bone samples. The released peptides were then identified by the LC-MS/MS. Using this approach, a total of 1120 proteins were identified in the maxillary bone and 1151 proteins in the mandibular bone. The subsequent partial least squares-discrimination analysis (PLS-DA) of protein data made it possible to reach 100% discrimination between the samples of healthy alveolar bones and those of the bone tissue surrounding the inflammatory focus. These results indicate that the in-bone protein digestion followed by the LC-MS/MS and subsequent statistical analysis can provide a deeper insight into the field of oral surgery at the molecular level. Furthermore, it could also have a diagnostic potential in the differentiation between the proteomic patterns of healthy and pathological alveolar bone tissue. Data are available via ProteomeXchange with the identifier PXD026775.

Effect of VEGFC on lymph flow and inflammation-induced alveolar bone loss.

The lymphatic system plays a crucial role in the maintenance of tissue fluid homeostasis and the immunological response to inflammation. The effects of lymphatic drainage dysfunction on periodontitis have not been well studied. Here we show that lymphatic vessel endothelial receptor 1 (LYVE1)+ /podoplanin (PDPN)+ lymphatic vessels (LVs) are increased in the periodontal tissues, with accumulation close to the alveolar bone surface, in two murine periodontitis models: rheumatoid arthritis (RA)-associated periodontitis and ligature-induced periodontitis. Further, PDPN+ /alpha-smooth muscle actin (αSMA)- lymphatic capillaries are increased, whereas PDPN+ /αSMA+ collecting LVs are decreased significantly in the inflamed periodontal tissues. Both mouse models of periodontitis have delayed lymph flow in periodontal tissues, increased TRAP-positive osteoclasts, and significant alveolar bone loss. Importantly, the local administration of adeno-associated virus for vascular endothelial growth factor C, the major growth factor that promotes lymphangiogenesis, increases the area and number of PDPN+ /αSMA+ collecting LVs, promotes local lymphatic drainage, and reduces alveolar bone loss in both models of periodontitis. Lastly, LYVE1+ /αSMA- lymphatic capillaries are increased, whereas LYVE1+ /αSMA+ collecting LVs are decreased significantly in gingival tissues of patients with chronic periodontitis compared with those of clinically healthy controls. Thus, our findings reveal an important role of local lymphatic drainage in periodontal inflammation-mediated alveolar bone loss. © 2020 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.

Localized juvenile spongiotic gingival hyperplasia: A report of 27 cases.

BACKGROUND: Localized juvenile spongiotic gingival hyperplasia (LJSGH) is a poorly understood but distinctive inflammatory hyperplasia occurring in children and young adults. Fewer than 100 cases have been reported since its initial description. METHODS: During the period of 2015 to 2018, cases of LJSGH were identified, retrieved and their clinical and histopathological data reviewed. RESULTS: There were 27 cases, with a median age of 13 years (range 7-72 years). Twenty-four of 27 patients were less than 20 years old, and in three cases the patients were over 60 years of age. The most commonly affected site was the anterior maxillary gingiva presenting as a solitary, red, and papillated lesion. Typical microscopic findings included elevated areas of variably acanthotic, spongiotic nonkeratinized epithelium with elongated rete ridges, accompanied by a neutrophilic-rich infiltrate. An abrupt transition between epithelium affected by LJSGH and normal mucosa was characteristic. LJSGH typically exhibited full-thickness epithelial expression of CK19 without expression of estrogen and progesterone receptors. CONCLUSIONS: The clinical and histopathologic characteristics of LJSGH are unique and consistent. Despite the name, the condition is not limited to juveniles and can occur in adults. LJSGH in adults and juveniles shares the same spectrum of histopathologic and immunohistochemical findings.

Study of bone remodeling in corticotomy-assisted orthodontic tooth movement in rats.

The goal of this study was to determine the structure change of the alveolar bone and the expression of a group of bone remodeling-related factors. Sixty healthy male Wistar rats were randomly divided into three groups. Selective alveolar decortication (SAD), tooth movement (TM), and "combined therapy" (SAD+TM) was performed in group I, II, and III, respectively. On days 0, 7, 14, 21, and 42, a Micro-CT scan was performed on the maxillary alveolar bone and tooth. In addition, on days 0, 7, 14, 21, 28, and 42, some of the rats were killed by cervical dislocation and tissues were harvested. Analysis of scan data revealed a significant decrease in bone density of the alveolar bone at 14 days post-surgery, and increased at 42 days post-surgery to a level higher than that before the surgery. Microarray and bioinformatics analysis were performed to explore gene expression profile in three groups (SAD, TM, and SAD+TM), and a large number of differentially expressed genes were identified. In addition, real-time polymerase chain reaction was performed to determine the expression of bone remodeling-related factors. The expression of osteoblast-related cytokines, including osteopontin, bone sialoprotein, and osteocalcin, and osteoclast regulators macrophage-colony stimulating factor (M-CSF) and RANKL (activator of nuclear factor KB receptor ligand) were increased in group III, suggesting that there was increased bone synthesis and activation of bone absorption. Moreover, group III had a unique alveolar bone remodeling pattern: RANKL and osteoprotegerin-promoted alveolar remodeling. In conclusion, during the early stage of orthodontic tooth movement, corticotomy can accelerate the movement of teeth, modulate the state of bone metabolism, and activate osteogenesis and osteoclast, which support the theory of regional acceleratory phenomenon.

Therapeutic effect of localized vibration on alveolar bone of osteoporotic rats.

OBJECTIVES: Vibration, in the form of high frequency acceleration (HFA), stimulates alveolar bone formation under physiologic conditions and during healing after dental extractions. It is not known if HFA has an anabolic effect on osteoporotic alveolar bone. Our objective is to determine if HFA has a regenerative effect on osteoporotic alveolar bone. METHODS AND MATERIALS: Adult female Sprague-Dawley rats were divided into five groups: 1) Ovariectomized Group (OVX), 2) Sham-OVX Group that received surgery without ovariectomy, 3) OVX-HFA Group that was ovariectomized and treated daily with HFA, 4) OVX+Static Force Group that was ovariectomized and received the same force as HFA, but without vibration, and 5) Control Group that did not receive any treatment. All animals were fed a low mineral diet for 3 months. Osteoporosis was confirmed by micro-CT of the fifth lumbar vertebra and femoral head. HFA was applied to the maxillary first molar for 5 minutes/day for 28 and 56 days. Maxillae were collected for micro-CT, histology, fluorescent microscopy, protein and RNA analysis, and three-point bending mechanical testing. RESULTS: Micro-CT analysis revealed significant alveolar bone osteoporosis in the OVX group. Vibration restored the quality and quantity of alveolar bone to levels similar to the Sham-OVX group. Animals exposed to HFA demonstrated higher osteoblast activity and lower osteoclast activity. Osteogenic transcription factors (RUNX2, Foxo1, Osterix and Wnt signaling factors) were upregulated following vibration, while RANKL/RANK and Sclerostin were downregulated. HFA did not affect serum TRAcP-5b or CTx-1 levels. The osteogenic effect was highest at the point of HFA application and extended along the hemimaxillae this effect did not cross to the contra-lateral side. CONCLUSIONS: Local application of vibration generated gradients of increased anabolic metabolism and decreased catabolic metabolism in alveolar bone of osteoporotic rats. Our findings suggest that HFA could be a predictable treatment for diminished alveolar bone levels in osteoporosis patients.

Sclerostin injection enhances orthodontic tooth movement in rats.

OBJECTIVE: It was aimed to investigate the in vivo effects of local injection of sclerostin protein on orthodontic tooth movement. DESIGN: A total of 48 rats underwent orthodontic mesialization of the maxillary first molars on both sides. Local injection was given at the compression side in the alveolar bone on both maxillary sides, with sclerostin protein carried by hydrogel on one side, and the same volume of normal saline carried by hydrogel on the other side serving as the control. After two weeks, the tooth movement amount and effects on the periodontium were assessed through micro-computed tomography (µCT) analysis, tartrate-resistant acid phosphatase (TRAP) staining and immunohistochemistry (IHC) analysis. RESULTS: After two weeks of intervention, tooth movement was significantly greater in the 4 µg/kg and 20 µg/kg sclerostin injection groups, compared to the control. Analysis of the furcation area of the maxillary first molar showed that the 20 µg/kg group had significantly decreased BV/TV. At the compression side, the number of TRAP-positive osteoclasts was significantly increased in 20 µg/kg group compared to the control. The expression of RANKL was statistically higher in all the sclerostin groups, while the expression of OPG was statistically lower in the 4 µg/kg and 20 µg/kg groups, compared to the control. At the tension side, the expression of RUNX2 and COL-1 was statistically higher in the 20 µg/kg group compared to the control. CONCLUSIONS: Local injection of sclerostin protein in the alveolar bone at the compression side accelerates OTM in rats by promoting osteoclastogenesis.

Induction of maxillary and mandibular squamous epithelial cell proliferation in mink (Neovison vison) by ß-naphthoflavone.

A jaw lesion reported in mink exposed to 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) and TCDD-like chemicals is considered a potential indicator of exposure to these chemicals. Many of the effects of TCDD-like chemicals are induced through interaction with the aryl hydrocarbon receptor. The present study indicates that mink dosed with ß-naphthoflavone, which is an aryl hydrocarbon receptor ligand but not a TCDD-like chemical, also develop the lesion. Environ Toxicol Chem 2019;38:460-463. © 2018 SETAC.

Do Implant Surgical Guides Allow an Adequate Zone of Keratinized Tissue for Flapless Surgery?

PURPOSE: A major advantage of guided implant surgery using 3-dimensionally printed guides is the ability to perform accurate flapless surgery. A drawback of a flapless technique is the inability to manipulate soft tissue to ensure sufficient gingiva around the implant. The purpose of this study was to determine how often flapless surgery using surgical guides results in less than 2 mm of keratinized tissue surrounding the implant. MATERIALS AND METHODS: This retrospective analysis included 27 maxillary and 27 mandibular implant sites that underwent treatment planning for implant-guided surgery using 3Shape Implant Studio (3Shape, Copenhagen, Denmark). Intraoral scan images were used to measure the width of the keratinized tissue on the buccal aspect of each implant site in both arches and the lingual aspect in the mandibular arch. Three examiners measured the amount of buccal and lingual keratinized tissue in millimeters at each implant site. Analysis of variance (P < .05) and correlation coefficients were used to determine statistically significant differences in keratinized tissue among sites. RESULTS: No statistically significant difference was found either between the widths of buccal keratinized tissue in the maxillary anterior (4.06 ± 1.42 mm) and posterior (4.93 ± 2.54 mm) areas (P = .293) or between the amounts of buccal and lingual keratinized tissue in the mandible (P = .995). The keratinized tissue width in the maxillary buccal area was significantly different (4.48 ± 2.04 mm) from that in the mandibular posterior buccal (1.98 ± 1.41 mm) and lingual (1.98 ± 1.23 mm) areas (P < .001). Over 77% of maxillary implant sites had greater than 3 mm of gingiva, and just over 20% had sufficient gingiva in the mandible. CONCLUSIONS: Adequate keratinized tissue was found in most of the planned maxillary implant sites, whereas most of the mandibular posterior implant sites had inadequate keratinized tissue. Therefore, elevation of a flap to preserve and reposition existing keratinized tissue around implants should be considered when planning to use tooth-borne surgical guides in the posterior mandible.

CCR2 Contributes to F4/80+ Cells Migration Along Intramembranous Bone Healing in Maxilla, but Its Deficiency Does Not Critically Affect the Healing Outcome.

Bone healing depends of a transient inflammatory response, involving selective migration of leukocytes under the control of chemokine system. CCR2 has been regarded as an essential receptor for macrophage recruitment to inflammation and healing sites, but its role in the intramembranous bone healing on craniofacial region remains unknown. Therefore, we investigated the role of CCR2 on F4/80+ cells migration and its consequences to the intramembranous healing outcome. C57BL/6 wild-type (WT) and CCR2KO mice were subjected to upper right incisor extraction, followed by micro-computed tomography, histological, immunological, and molecular analysis along experimental periods. CCR2 was associated with F4/80+ cells influx to the intramembranous bone healing in WT mice, and CCR2+ cells presented a kinetics similar to F4/80+ and CCR5+ cells. By contrast, F4/80+ and CCR5+ cells were significantly reduced in CCR2KO mice. The absence of CCR2 did not cause major microscopic changes in healing parameters, while molecular analysis demonstrated differential genes expression of several molecules between CCR2KO and WT mice. The mRNA expression of TGFB1, RUNX2, and mesenchymal stem cells markers (CXCL12, CD106, OCT4, NANOG, and CD146) was decreased in CCR2KO mice, while IL6, CXCR1, RANKL, and ECM markers (MMP1, 2, 9, and Col1a2) were significantly increased in different periods. Finally, immunofluorescence and FACS revealed that F4/80+ cells are positive for both CCR2 and CCR5, suggesting that CCR5 may account for the remaining migration of the F4/80+ cells in CCR2KO mice. In summary, these results indicate that CCR2+ cells play a primary role in F4/80+ cells migration along healing in intramembranous bones, but its deficiency does not critically impact healing outcome.

970 nm low-level laser affects bone metabolism in orthodontic tooth movement.

OBJECTIVE: During orthodontic tooth movement (OTM), the speed of movement depends on the rate of bone turnover. In this study, we used a rat model to investigate the effect of 970 nm low-level laser therapy (LLLT) on OTM under different dose and frequency protocols. METHODS: We first compared the OTM rates between the OTM only control and the OTM + LLLT group (1250 J/cm2) in Experiment 1 and showed that LLLT significantly increased OTM. In Experiment 2, we employed 3 different LLLT protocols: the low-dose group and the high-dose group receiving 5 doses of 750 J/cm2 and 15,000 J/cm2 of LLLT every 3 days, respectively, and the early high-dose group which received 5 daily doses at 15,000 J/cm2 at the beginning of the experiments. The OTM-only control group received no LLLT. Tooth movement rate was measured through sequential silicone impressions. MicroCT was also performed to evaluate bone de-mineralization rate. Bone histmorphometry was used to compare the bone turnover rate between LLLT group and control group. Finally, TRAP, Osteocalcin, and VEGF expression is evaluated by immunohistochemistry (IHC) in tissue sections. RESULTS: When LLLT treatment was given every three days, both the 1250 J/cm2 and 15,000 J/cm2 groups showed significantly increased OTM compared to the control group. No significant difference was observed in the 750 J/cm2 group, or in the early irradiation group, when compared with controls, although 750 J/cm2 showed the same trend of accelerating OTM. The MicroCT result of rat maxilla demonstrated that LLLT increased bone remodeling and showed decreased bone mineral density and bone volume/total volume in the furcation areas of the maxillary first molars at the end of experiment. LLLT without OTM increased bone turnover as evidenced by fluorochrome incorporation. Immunohistochemistry analyses revealed high osteocalcin expression at later stages of OTM in the LLLT group, while VEGF expression was highly induced in the LLLT + OTM group at an early stage. CONCLUSION: Our results suggest that the 970 nm LLLT increases the rate of OTM in a dose-sensitive and frequency-dependent manner. Further animal and human studies are needed to determine the optimal timing and dosage of LLLT for OTM acceleration.

Sclerostin-Neutralizing Antibody Enhances Bone Regeneration Around Oral Implants.

BACKGROUND: Dental implants are an important option for replacement of missing teeth. A major clinical challenge is how best to accelerate bone regeneration and reduce the healing time for functional restoration after implant placement. A sclerostin-neutralizing antibody (Scl-Ab) has been shown to enhance alveolar bone formation and fracture repair. The aim of this study was to investigate the effects of systemic administration of Scl-Ab on dental implant osseointegration and bone regeneration in an experimental alveolar ridge tooth extraction model. MATERIALS AND METHODS: To investigate the effects of Scl-Ab on bone regeneration and dental implant osseointegration, an experimental alveolar bone osteotomy rat model was adopted. One month after extraction of maxillary right first molars, osteotomy defects were created at the coronal aspect of each of the extraction sites, and 1 × 2-mm custom titanium implants were installed into the osteotomies. Coincident with implant placement, Scl-Ab was administered subcutaneously at a dose of 25 mg/kg twice weekly for 10-28 days and compared with a vehicle control. Animals were sacrificed 10, 14, and 28 days after surgery, and maxillae were harvested and analyzed by microcomputed tomography (microCT), histology, and histomorphometry. RESULTS: microCT analysis demonstrated that the maxillary bone volume fraction was approximately 2- to 2.5-fold greater in Scl-Ab-treated animals compared with vehicle alone at days 14 and 28. Consistent with those findings, two-dimensional bone fill percentages within the coronal osteotomy sites were highest in Scl-Ab treatment groups at 28 days. In addition, bone-implant contact at 28 days was approximately twofold greater in the Scl-Ab group compared with the vehicle control. CONCLUSIONS: These results indicate that systemic Scl-Ab administration enhances osseointegration and bone regeneration around dental implants. This approach offers potential as a treatment modality for patients with low bone mass or bone defects to achieve more predictable bone regeneration at alveolar bone defects and to enhance dental implant osseointegration.

Maxillary Bone Regeneration Based on Nanoreservoirs Functionalized ε-Polycaprolactone Biomembranes in a Mouse Model of Jaw Bone Lesion.

Current approaches of regenerative therapies constitute strategies for bone tissue reparation and engineering, especially in the context of genetical diseases with skeletal defects. Bone regeneration using electrospun nanofibers' implant has the following objectives: bone neoformation induction with rapid healing, reduced postoperative complications, and improvement of bone tissue quality. In vivo implantation of polycaprolactone (PCL) biomembrane functionalized with BMP-2/Ibuprofen in mouse maxillary defects was followed by bone neoformation kinetics evaluation using microcomputed tomography. Wild-Type (WT) and Tabby (Ta) mice were used to compare effects on a normal phenotype and on a mutant model of ectodermal dysplasia (ED). After 21 days, no effect on bone neoformation was observed in Ta treated lesion (4% neoformation compared to 13% in the control lesion). Between the 21st and the 30th days, the use of biomembrane functionalized with BMP-2/Ibuprofen in maxillary bone lesions allowed a significant increase in bone neoformation peaks (resp., +8% in mutant Ta and +13% in WT). Histological analyses revealed a neoformed bone with regular trabecular structure, areas of mineralized bone inside the membrane, and an improved neovascularization in the treated lesion with bifunctionalized membrane. In conclusion, PCL functionalized biomembrane promoted bone neoformation, this effect being modulated by the Ta bone phenotype responsible for an alteration of bone response.

Clastic cells are absent around the root surface in pulp-exposed periapical periodontitis lesions in mice.

INTRODUCTION: Clastic cells, originating from the monocyte-macrophage lineage, resorb mineralized tissues. In periapical periodontitis, alveolar bone around the tooth apex becomes resorbed; however, the roots of the teeth are often left intact by yet unknown mechanisms. Here, we examined the status of clastic cells in a periapical periodontitis model in mice. METHODS: Periapical periodontitis was induced by performing pulp exposure on the maxillary first molar. The contralateral maxillary first molar was used as a control. The maxillae were harvested, fixed, and subjected to µCT scanning and three-dimensional volumetric analysis. TRAP staining was performed, and osteoclasts were quantified. Immunohistochemical staining was performed for RANKL, OPG, and F4/80, a marker for macrophages. RESULTS: At the apex of the tooth, pulp exposure resulted in periapical radiolucency with mineralized tissues at the surrounding bone surfaces but not on the root surfaces. Histologically, clastic cells were present on the bone surfaces but absent around the root surfaces. Expression of F4/80 and RANKL was not found at close proximity to the root surfaces, but OPG was globally expressed. CONCLUSION: The absence of clastic cells around the root surface of pulp-exposed teeth, in part, is associated with the lack of macrophages and RANKL expression.

Lactation induces increases in the RANK/RANKL/OPG system in maxillary bone.

The underlying causes of maxillary bone loss during lactation remain poorly understood. We evaluated the impact of lactation on physiological and mechanically-induced alveolar bone remodeling. Nulliparous non-lactating (N-LAC) and 21-day lactating (LAC) mice underwent mechanically-induced bone remodeling by orthodontic tooth movement (OTM). Micro-computed tomography (microCT) was performed in the maxilla, femur and vertebra. Tartrate-resistant-acid phosphatase (TRAP) and Masson's trichrome labelling was performed in the maxillary bone and gene expression was determined in the periodontal ligament. The effect of prolactin on osteoclast (OCL) and osteoblast (OBL) differentiation was also investigated in N-LAC and LAC mice. Lactation increased alveolar bone loss in the maxilla, femur and vertebra, while OTM was enhanced. The number of OCL and OBL was higher in the maxilla of LAC mice. OTM increased OCL in both groups; while OBL was increased only in N-LAC but not in LAC mice, in which cell numbers were already elevated. The alveolar bone loss during lactation was associated with increased expression of receptor activator of nuclear factor-KappaB (RANK), RANK ligand (RANKL), and osteoprotegerin (OPG) in the maxilla. OTM induced the same responses in N-LAC mice, whereas it had no further effect in LAC mice. Lactation enhanced differentiation of OCL and OBL from bone marrow cells, and prolactin recapitulated OCL differentiation in N-LAC mice. Thus, lactation increases physiological maxillary bone remodeling and OTM, and both require activation of RANK/RANKL/OPG system. These findings expand our knowledge of lactation-induced osteopenia and have possible impact on clinical practice regarding orthodontic treatments and dental implants in lactating women.

Regeneration of the cementum and periodontal ligament using local BDNF delivery in class II furcation defects.

Periodontal furcation defects are usually addressed by the placement of a physical barrier which may limit the regenerative potential of periodontal wounds. This study morphometrically quantified the regenerative effect of brain-derived neurotrophic factor (BDNF) in furcation defects in a non-human primate model. Grade II furcation defects (with and without induced inflammation prior to surgery) were created on the first and second molars of eight non-human primates. Defects were treated with open flap debridement and subsequently filled with either: Group A; BDNF (500 µg mL-1 ) in high-molecular weight-hyaluronic acid (HMW-HA), Group B; BDNF (50 µg mL-1 ) in HMW-HA, Group C; HMW-HA acid only, Group D; unfilled defect, or Group E; BDNF (500 µg mL-1 ) in saline. Periodontal wound healing was observed every 2 weeks by computed-tomography. At 11 weeks all animals were sacrificed and maxillary and mandibular block biopsies were referred for nondecalcified histology. Linear measurements of new cementum (cellular and acellular) and periodontal ligament (PDL) formation were performed. Computerized-tomography reconstruction and software quantification demonstrated successful bone fill for all groups. However, histometric assessment demonstrated significantly higher level of total periodontal regeneration for the 500 µg mL-1 BDNF HMW-HA relative to all other groups. No significant differences in cementogenesis were observed among groups. Significantly higher acellular cementum formation was observed for sites where inflammation was not induced prior to surgical procedures. While all groups experienced similar bone fill and cementogenesis, the 500 µg mL-1 BDNF HMW-HA appeared to most effectively repair PDL (minimum increase of ∼22% relative to all groups; over 200% relative to unfilled defects). © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 106B: 1611-1617, 2018.

Oxygen Saturation in the Dental Pulp of Maxillary and Mandibular Molars - Part 2

Abstract This study determined the oxygen saturation (SaO2) in dental pulp of healthy maxillary and mandibular molars. Mean of SaO2 was evaluated in 112 maxillary and mandibular molars using pulse oximetry. Quantitative variables were described by mean and standard deviation. Variables with symmetric distribution were compared by Student t test and Mann-Whitney test. Pearson's correlation coefficient was used to correlate quantitative variables. Analysis of variance was used to assess differences in SaO2 levels between the molar groups, followed by post-hoc Tukey. The significance level established at p<0.05. Mean of oxygen saturation for the 112 molar dental pulps was 85.09%. There was no significant correlation (r=-0.007; p=0.977) between the mean of SaO2 of molar pulps with patient´s indicator finger (92.89%). There was a significant difference (p=0.037) between the mean of SaO2 of the first (85.76%) and second maxillary molars (81.87%), and it was not significant (p=0.1775) between the first and second mandibular molars. Maxillary molars had lower pulpal SaO2 (83.59%) than mandibular molars (86.89%) (p=0.018). The mean of the patient's response time to the cold stimulus was 1.12 s (maxillary molars 1.25 s and mandibular molars 0.99 s)(p=0.052). There was no significant correlation between the time response of the patient to the cold stimulus and the SaO2 for molars. The mean oxygen saturation level was 85.09%. The mandibular molars presented higher SaO2 level than maxillary molars.

Resumo Este estudo determinou o nível de saturação de oxigênio (SaO2) em polpas dentais hígidas de molares. O nível de SaO2 foi avaliado em 112 molares superiores e inferiores usando oxímetro de pulso. As variáveis quantitativas foram descritas pela média e desvio padrão. As variáveis com distribuição simétrica foram comparadas pelo teste t de Student e teste de Mann-Whitney. O coeficiente de correlação de Pearson foi utilizado para correlacionar variáveis quantitativas. A análise de variância foi utilizada para avaliar as diferenças nos níveis de SaO2 entre os grupos de molares, seguido de Tukey pós-hoc. A significância foi estabelecida em 0,05. O nível médio de SaO2 para as polpas de 112 molares foi de 85,09%, não havendo correlação com a média de SaO2 do dedo indicador do paciente (92,89%). Houve diferença significativa entre o nível médio de SaO2 dos primeiros molares superiores (85,76%) e os segundos molares superiores (81,87%) e não foi significativo entre os primeiros e os segundos molares inferiores. Os molares superiores apresentaram menor nível de SaO2 (83,59%) do que os molares inferiores (86,89%). A média do tempo de resposta do paciente ao estímulo com frio foi de 1,12 s (molares superiores 1,25 segundos e molares inferiores 0,99 segundos). Não houve correlação significativa entre o tempo de resposta do paciente ao estímulo com frio e o nível de saturação de oxigênio para os molares. Em resumo, o nível médio de saturação de oxigênio foi de 85,09%. Os molares inferiores apresentaram maior nível de SaO2 do que os molares superiors

Oxygen Saturation in the Dental Pulp of Maxillary Premolars in Different Age Groups - Part 1

Abstract The aim of this study was to determine oxygen saturation levels in the dental pulp of maxillary premolars in different age groups. A total of 120 human maxillary premolars with normal dental pulps were selected covering the following age groups: 20-24, 25-29, 30-34, 35-39 and 40-44 years (n=24 each group). Oxygen saturation was assessed using pulse oximetry. Analysis of variance was used to assess differences in oxygen saturation levels and Tukey's test was used to identify the age groups that differed from each other. Significance was set at 0.05. Mean oxygen saturation of 120 premolars was 86.20% considering all age groups. Significantly reduced levels were found in the oldest group compared to the other groups: 40 to 44 years - 80.00% vs. 89.71, 87.67, 88.71, and 84.80% for age groups 20-24, 25-29, 30-34, 35-39 years, respectively. The mean oxygen saturation levels were similar between 20 and 39 years of age (86.20%) in the whole sample, but reduced significantly in the 40-44-year age group, suggesting that older patients present lower oxygen saturation results even in the absence of pulp tissue injury.

Resumo Este estudo determinou os níveis de saturação de oxigênio (SaO2) em polpas dentárias de pré-molares superiores em diferentes faixas etárias. Foram selecionados 120 pré-molares superiores humanos com polpas dentárias normais, abrangendo os seguintes grupos etários: 20-24, 25-29, 30-34, 35-39 e 40-44 anos (n=24 para cada grupo). A saturação de oxigênio foi avaliada utilizando oximetria de pulso. A análise de variância foi utilizada para avaliar diferenças nos níveis de saturação de oxigênio, e o teste de Tukey foi utilizado para identificar os grupos etários que diferiam uns dos outros. A significância foi estabelecida em 0,05. A saturação média de oxigênio foi de 86,20% considerando todos os grupos etários. Níveis significativamente reduzidos foram encontrados no grupo de indivíduos de maior idade em comparação aos outros grupos: 40 a 44 anos - 80,00% vs. 89,71, 87,67, 88,71 e 84,80% para os grupos etários 20-24, 25-29, 30-34, 35-39 anos. Os níveis médios de saturação de oxigênio foram semelhantes entre os 20 e os 39 anos de idade (86,20%), mas reduziram-se significativamente na faixa etária de 40-44 anos, sugerindo que os pacientes mais idosos apresentam menor saturação de oxigênio mesmo na ausência de lesão do tecido pulpar.