Diagnostic potential of endothelin-1 in peri-implant diseases: a cross-sectional study.

PURPOSE: This study aimed to evaluate the potential of Endothelin-1 (ET-1), a peptide derived from vascular endothelial cells, as a biomarker for diagnosing peri-implant diseases. METHODS: A cohort of 29 patients with a total of 76 implants was included in this study and subsequently divided into three groups based on peri-implant clinical parameters and radiographic examination: healthy (peri-implant health) (n = 29), mucositis (n = 22), and peri-implantitis (n = 25) groups. The levels of ET-1 (ρg/site) and interleukin (IL)-1ß (ρg/site) in peri-implant sulcus fluid (PISF) samples were determined using enzyme immunoassay. Statistical analyses were conducted using Kruskal-Wallis and Steel-Dwass tests. Logistic regression and receiver operating characteristic (ROC) curve analyses were performed to evaluate the diagnostic performance of the biomarkers. RESULTS: ET-1 levels were significantly elevated in the peri-implantitis group compared to those in the healthy group, and were highest in the peri-implant mucositis group. Additionally, IL-1ß levels were significantly higher in the peri-implantitis group than those in the healthy group. ROC curve analysis indicated that ET-1 exhibited superior area under the curve values, sensitivity, and specificity compared to those of IL-1ß. CONCLUSIONS: Our findings suggest that the presence of ET-1 in PISF plays a role in peri-implant diseases. Its significantly increased expression in peri-implant mucositis indicates its potential for enabling earlier and more accurate assessments of peri-implant inflammation when combined with conventional examination methods.

Application of anti-vascular endothelial growth factor antibody restores the function of saliva secretion in a type 2 diabetes mouse model.

OBJECTIVES: Xerostomia, a common complication of type 2 diabetes, leads to an increased risk of caries, dysphagia, and dysgeusia. Although anti-vascular endothelial growth factor (VEGF) antibodies, such as ranibizumab (RBZ), have been used to treat diabetic retinopathy, their effects on the salivary glands are unknown. This study evaluated the effects of RBZ on salivary glands to reduce inflammation and restore salivary function in a mouse model of type 2 diabetes. METHODS: Male KK-Ay mice with type 2 diabetes (10-12 weeks old) were used. The diabetes mellitus (DM) group received phosphate-buffered saline, while the DM + RBZ group received an intraperitoneal administration of RBZ (100 µg/kg) 24 h before the experiment. RESULTS: Ex vivo perfusion experiments showed a substantial increase in salivary secretion from the submandibular gland (SMG) in the DM + RBZ group. In addition, the mRNA expression levels of TNF-α and IL-1ß were considerably lower in this group. In contrast, those of aquaporin 5 were substantially higher in the DM + RBZ group, as revealed by quantitative reverse transcription PCR. Furthermore, the number of lymphocyte infiltration spots in the SMG was notably lower in the DM + RBZ group. Finally, intracellular Ca2+ signaling in acinar cells was considerably higher in the DM + RBZ group than that in the DM group. CONCLUSION: Treating a type 2 diabetic mouse model with RBZ restored salivary secretion through its anti-inflammatory effects.

Chronic kidney disease compromises structural and mechanical properties of maxillary cortical bone in a rat model.

PURPOSE: This study aimed to investigate the effects of chronic kidney disease (CKD) on the structural and mechanical properties of the maxillary and mandibular cortical bone. METHODS: The maxillary and mandibular cortical bones from CKD model rats were used in this study. CKD-induced histological, structural, and micro-mechanical alterations were assessed using histological analyses, micro-computed tomography (CT), bone mineral density (BMD) measurements, and nanoindentation tests. RESULTS: Histological analyses indicated that CKD caused an increase in the number of osteoclasts and a decrease in the number of osteocytes in the maxilla. Micro-CT analysis revealed that CKD induced a void volume/cortical volume (%) increase, which was more remarkable in the maxilla than in the mandible. CKD also significantly decreased the BMD in the maxilla. In the nanoindentation stress-strain curve, the elastic-plastic transition point and loss modulus were lower in the CKD group than that in the control group in the maxilla, suggesting that CKD increased micro fragility of the maxillary bone. CONCLUSIONS: CKD affected bone turnover in the maxillary cortical bone. Furthermore, the maxillary histological and structural properties were compromised, and micro-mechanical properties, including the elastic-plastic transition point and loss modulus, were altered by CKD.

Hepcidin expression in the trigeminal ganglion and the oral mucosa in an oral ulcerative mucositis rat model.

Severe intraoral pain induces difficulty in eating and speaking, leading to a decline in the quality of life. However, the molecular mechanisms underlying intraoral pain remain unclear. Here, we investigated gene modulation in the trigeminal ganglion and intraoral pain-related behavior in a rat model of acetic acid-induced oral ulcerative mucositis. Oral ulceration was observed on day 2 after acetic acid treatment to the oral mucosa of male Wistar rats, causing spontaneous pain and mechanical allodynia. Deoxyribonucleic acid microarray analysis of trigeminal ganglion tissue indicated that Hamp (a hepcidin gene that regulates cellular iron transport) was the most upregulated gene. In the oral ulcerative mucositis model, the upregulation of Hamp was also induced in the ulcer region but not in the liver, with no increase in hepcidin levels in the plasma and saliva, indicating that hepcidin was produced locally in the ulcer region in the model. Systemic antibiotic pretreatment did not increase the mRNA levels of Hamp in the trigeminal ganglion and ulcer regions. Hepcidin injection into the oral mucosa enhanced neuronal excitability in response to noxious mechanical stimulation of the oral mucosa in trigeminal spinal subnucleus interpolaris/caudalis neurons. These results imply that oral ulcerative mucositis induces oral mucosal pain because of infectious inflammation of the ulcerative area and potentiates Hamp, which represents anti-bacterial and anti-peptidase gene expression in the ulcer region and trigeminal ganglion. The regulation of cellular iron transport by hepcidin is likely involved in oral ulcerative mucositis-induced pain.

Functional tooth number in the posterior region associated serum ucOC levels.

PURPOSE: The purpose of this study was to investigate serum undercarboxylated osteocalcin (ucOC) levels in partially edentulous patients scheduled to receive implant treatment and determine the association between ucOC levels, vegetable intake, vitamin K, dietary fiber intake, and functional tooth number in the posterior region (p-FTN). METHODS: A total of 46 patients (20 male and 26 female, 61.9 ± 12.7 years old) were included. The association among serum ucOC levels, vegetable intake, vitamin K and dietary fiber intake was assessed using Spearman's rank correlation coefficient and binary logistic regression analysis. RESULTS: In total, 35% of patients (16/46 subjects) showed an abnormally high ucOC level (≧ 4.5 ng/mL). p-FTN showed a weak positive correlation with vegetable intake, vitamin K and dietary fiber intake (r = 0.28, 0.21, and 0.14, respectively) and a significant negative correlation with ucOC levels (r = - 0.51). Multivariate analysis demonstrated that p-FTN as well as vitamin K intake showed a significant negative association with serum ucOC levels. CONCLUSIONS: More than one-third of patients showed abnormally high ucOC levels. p-FTN showed a negative association with serum ucOC levels, which indicated the possibility that oral status affected bone quality.

A new mandibular deformation index predicts amount of bone deformation in edentulous patients treated with an implant-supported fixed prosthesis.

PURPOSE: The present study was performed to examine the mandibular deformation during mouth opening in edentulous patients, treated with an implant-supported fixed prosthesis using strain gauges, and identify factors affecting deformation. METHODS: Twenty patients with a fully edentulous mandible who received either 4 or ≥6 implants were included. The distal-most implants were placed mesial to the mental foramen (premolar region) in patients with 4 implants and distal to the mental foramen (molar region) in patients with ≥6 implants. Mandibular deformation during mouth opening was measured using strain gauges in two directions: anteroposterior direction and lateral direction between the distal-most implants on the left and right sides (arch width). The mandibular anatomy was evaluated using computed tomography. RESULTS: Arch width reduction between the left and right implants during mouth opening ranged from 47.38 to 512.80 µm; the range of deformation was 0.12 to 15.14 µm in the anteroposterior direction. Furthermore, a significant positive correlation was noted between arch width reduction in the premolar region and the ratio between the symphyseal bone height and width (P = 0.0003, r = 0.72). CONCLUSION: The reduction in arch width was higher in the molar region than in the premolar region during mouth opening. Moreover, the reduction could be high in the mandibular symphyseal bone because of its greater height and lesser width. The ratio between the symphyseal bone height and width is defined as the mandibular deformation index (MDI) and is used to predict the rate of mandibular bone deformation.

Microstructural and mechanical recovery of bone in ovariectomized rats: The effects of menaquinone-7.

The loss of bone quantity and quality in postmenopausal female patients can be a problem for dental treatment. A sufficient intake of nutrients such as calcium, magnesium, and vitamins D and K is likely correlated with the mechanical properties of bone. In particular, vitamin K2, also called menaquinone (MK), inhibits bone loss in postmenopausal women. Here we demonstrate the microstructural and mechanical properties of bone recovery in ovariectomized (OVX) rats during MK-7 administration. Bilateral ovariectomy and a sham operation were performed on 14-week-old female SPF Wistar rats. MK-4 and -7 were orally administered at 30 mg/kg daily for 12 weeks. The femur was used for the 3-point bending test and microstructural analysis of the cancellous bone by micro-CT, and the mandibular cortical bone for the evaluation of mechanical properties on a nanoscale. Micro-computed tomography revealed irregular trabecular architecture, hollow marrow cavities, and sparse trabecular bone in the femurs of the OVX group. Trabecular bone structure analysis showed that the MK-7 group had greater bone volume per tissue volume (BV/TV) and a higher trabecular number than the OVX group. The bulk-scale 3-point bending test did not allow the mechanical properties between OVX and OVX/MK7 groups to be discerned, yet at the smallest level, the elastic-plastic transition point of the nanoindentation stress-strain curve of the mandibular cortical bone was higher in the MK-7 group than in the OVX group. These findings suggest that MK-7 enables bone microstructural and mechanical recovery in the OVX model.

Local administration of HMGB-1 promotes bone regeneration on the critical-sized mandibular defects in rabbits.

Reconstruction of a critical-sized osseous defect is challenging in maxillofacial surgery. Despite novel treatments and advances in supportive therapies, severe complications including infection, nonunion, and malunion can still occur. Here, we aimed to assess the use of a beta-tricalcium phosphate (ß-TCP) scaffold loaded with high mobility group box-1 protein (HMGB-1) as a novel critical-sized bone defect treatment in rabbits. The study was performed on 15 specific pathogen-free New Zealand rabbits divided into three groups: Group A had an osseous defect filled with a ß-TCP scaffold loaded with phosphate-buffered saline (PBS) (100 µL/scaffold), the defect in group B was filled with recombinant human bone morphogenetic protein 2 (rhBMP-2) (10 µg/100 µL), and the defect in group C was loaded with HMGB-1 (10 µg/100 µL). Micro-computed tomography (CT) examination demonstrated that group C (HMGB-1) showed the highest new bone volume ratio, with a mean value of 66.5%, followed by the group B (rhBMP-2) (31.0%), and group A (Control) (7.1%). Histological examination of the HMGB-1 treated group showed a vast area covered by lamellar and woven bone surrounding the ß-TCP granule remnants. These results suggest that HMGB-1 could be an effective alternative molecule for bone regeneration in critical-sized mandibular bone defects.

Xerostomia aggravates ligation-induced peri-implantitis: A preclinical in vivo study.

OBJECTIVES: Previous studies have indicated that xerostomia is a critical factor affecting periodontitis; nonetheless, it is controversial whether xerostomia impairs peri-implant tissue. The objective of this experimental study was to evaluate the effect of xerostomia on the peri-implant hard and soft tissues in the rat model. MATERIALS AND METHODS: Implants were placed in bilateral maxillae of male Wistar rats. The animals underwent submandibular and sublingual gland resection on both sides (DRY group) or sham operation (CTR group). Silk ligatures were placed around one side of abutments, which were randomly selected in each animal. The effects of xerostomia were assessed using micro-CT, histological analysis, real-time PCR, and 16S rRNA-based metagenomic analysis. RESULTS: Ligation with silk thread caused bone resorption around implants. Although xerostomia itself did not induce bone resorption, it significantly enhanced silk ligature-mediated bone resorption around implants. Histological analysis and real-time PCR indicated that xerostomia induced inflammation and osteoclastogenesis around implants with silk ligatures. Furthermore, it altered the microbiota of the plaque on the silk thread around implants. CONCLUSION: Xerostomia accelerates mucosal inflammation and osteoclastogenesis, which aggravates bone resorption around implants.

Effects of metformin on the prevention of bisphosphonate-related osteonecrosis of the jaw-like lesions in rats.

PURPOSE: In this study, we aimed to investigate the effect of glucose metabolism on bone healing after tooth extraction in an osteoporosis rat model administered zoledronic acid (ZA) and dexamethasone (DX). METHODS: In total, 24 male Wistar rats (4 weeks old) were randomly assigned to four groups: Control (subcutaneous physiological saline), ZD (subcutaneous ZA and DX twice a week), Ins+ZD (subcutaneous insulin followed by ZD treatment), and Met+ZD (oral metformin followed by ZD treatment). Blood was collected every two weeks . Two weeks after treatment initiation, the first molar tooth on the right maxilla was extracted from all rats. Four weeks later, the rats were sacrificed, and bone healing was assessed. Maxillae samples were fixed and scanned using micro-computed tomography for quantifying areas of bone defects. Hematoxylin-eosin and tartrate-resistant acid phosphatase (TRAP) staining were performed to evaluate bone apoptosis and osteoclast number. RESULTS: In all experimental groups, body weight was statistically lower than that in the Control group, with no changes observed in uncarboxylated osteocalcin concentrations. The radiological analysis revealed that insulin or metformin administration improved healing in the tooth extraction socket (p < 0.01). Histological examination revealed that the osteonecrosis area was reduced in the Ins+ZD and Met+ZD groups (p < 0.01). TRAP staining presented increased osteoclast numbers in the ZD group when compared with that observed in the Control. CONCLUSIONS: Tooth extraction with long-term ZA and DX administration inhibited bone remodeling and induced bisphosphonate-related osteonecrosis of the jaw-like lesions. Metformin exerted protective effects ag ainst osteonecrosis of the jaw.

The effects of hyperglycaemia on peri-implant tissues after osseointegration.

PURPOSE: We assessed the effects of hyperglycaemia induced by streptozotocin (STZ) on peri-implantitis developing after implant osseointegration. METHODS: Thirty-six male Wistar rats (4 weeks old) were used. We placed titanium implants 4 weeks after extraction of the maxillary first molars. Healing abutments were attached 4 weeks later. After osseointegration was confirmed, the rats were divided into control, hyperglycaemia (STZ), and STZ with insulin (STZ+INS) groups. Hyperglycaemia was induced by a single injection of 50mg/kg STZ. Silk ligatures were placed on only the right sides (i.e. ligature sides), not on the left sides. Peri-implant tissues extracted at 4 weeks post-ligation were analysed both radiologically (via micro-computed tomography) and histologically (via toluidine blue staining). Total RNA was also extracted and analysed by quantitative PCR to detect TNF-α, IL-1ß and the receptor of advanced glycation end products (RAGE). Additionally, advanced glycation end products (AGEs) were measured by ELISA. RESULTS: Radiological and histological analyses showed that bone loss on the non-ligature sides was significantly greater in the STZ than the control group. However, on the ligature sides, bone loss was greater than on the non-ligature sides, and no significant difference was evident among the three groups. The levels of mRNAs encoding TNF-α, IL-1ß, RAGE, and AGEs on the ligature sides were significantly upregulated (all P<0.05) in the STZ compared to the control group. CONCLUSIONS: Although hyperglycaemia could be associated with bone loss around implants with increased AGE production and RAGE expression, hyperglycaemia does not become a triggering factor of ligature induced peri-implantitis.

Hyposalivation due to chemotherapy exacerbates oral ulcerative mucositis and delays its healing.

OBJECTIVES: Cancer therapy including chemotherapy causes gland atrophy, resulting in low salivary secretion in cancer patients. Since saliva plays an important role in oral health, the dysfunction may exacerbate oral ulcerative mucositis (OUM), which is another side effect. Here, we investigated the effect of hyposalivation on OUM using sialoadenectomized rats and examined the effects of anticancer drugs on the salivary glands. DESIGN: As models for hyposalivation, the bilateral submandibular and sublingual glands except (2EXT) or together with (3EXT) the parotid glands were extracted. At 16 days after the procedure, OUM was experimentally developed by topical acetic acid treatment on the labial fornix region of the inferior incisors, and the severity and bacterial loading level were evaluated. The salivary gland weights and histology were analyzed after administration of the representative anticancer drugs 5-fluorouracil or cisplatin. RESULTS: The severity of OUM was greater in both the 3EXT and 2EXT rats and delayed the healing process compared with that in sham rats without salivary gland extraction. The healing process in the 3EXT rats was longer than that in the 2EXT rats. The number of colony-forming units in the ulcerative region from the 3EXT rats was 10-fold greater than that in the sham rats. Both 5-fluorouracil and cisplatin reduced glands weights and damaged the salivary glands. CONCLUSIONS: These results suggest that chemotherapy-induced hyposalivation exacerbates OUM and delays healing, most likely due to loss of salivary clearance and antimicrobial functions. This study illustrates the significance of oral health care for cancer patients undergoing chemotherapy.

Implant-supported fixed prosthesis improves nutrient intake in patients with partial edentulous posterior regions.

PURPOSE: This study investigated changes in food and nutrient intake after implant-supported fixed prosthesis treatment in patients with partial edentulous posterior regions. METHODS: This study included 30 patients who received implant treatment with fixed prostheses in the posterior region. Food and nutrient intake was evaluated using a brief self-administered diet history questionnaire at baseline and post-implant treatment, and the results were statistically analyzed. RESULTS: Treatment with implant-supported fixed prostheses in patients with posterior edentulous conditions tended to increase the amounts of soy products and vegetables consumed: in particular, intake of carrot and squash was significantly increased. The total energy, protein, lipid, and carbohydrate intakes were comparable between baseline and post-implant treatment. On the other hand, the vegetable protein, α-carotene, daidzein, and genistein intakes were significantly increased, and dietary fiber and ß-carotene intakes tended to be increased in patients with implant-supported fixed prostheses. CONCLUSIONS: Implant-supported fixed prostheses in patients with posterior edentulous conditions affected food intake, resulting in improved nutrient intake.

Prostanoid-dependent spontaneous pain and PAR2-dependent mechanical allodynia following oral mucosal trauma: involvement of TRPV1, TRPA1 and TRPV4.

Abstract: During dental treatments, intraoral appliances frequently induce traumatic ulcers in the oral mucosa. Such mucosal injury-induced mucositis leads to severe pain, resulting in poor quality of life and decreased cooperation in the therapy. To elucidate mucosal pain mechanisms, we developed a new rat model of intraoral wire-induced mucositis and investigated pain mechanisms using our proprietary assay system for conscious rats. A thick metal wire was installed in the rats between the inferior incisors for one day. In the mucosa of the mandibular labial fornix region, which was touched with a free end of the wire, traumatic ulcer and submucosal abscess were induced on day 1. The ulcer was quickly cured until next day and abscess formation was gradually disappeared until five days. Spontaneous nociceptive behavior was induced on day 1 only, and mechanical allodynia persisted over day 3. Antibiotic pretreatment did not affect pain induction. Spontaneous nociceptive behavior was sensitive to indomethacin (cyclooxygenase inhibitor), ONO-8711 (prostanoid receptor EP1 antagonist), SB-366791, and HC-030031 (TRPV1 and TRPA1 antagonists, respectively). Prostaglandin E2 and 15-deoxyΔ12,14-prostaglandin J2 were upregulated only on day 1. In contrast, mechanical allodynia was sensitive to FSLLRY-NH2 (protease-activated receptor PAR2 antagonist) and RN-1734 (TRPV4 antagonist). Neutrophil elastase, which is known as a biased agonist for PAR2, was upregulated on days 1 to 2. These results suggest that prostanoids and PAR2 activation elicit TRPV1- and TRPA1-mediated spontaneous pain and TRPV4-mediated mechanical allodynia, respectively, independently of bacterial infection, following oral mucosal trauma. The pathophysiological pain mechanism suggests effective analgesic approaches for dental patients suffering from mucosal trauma-induced pain.

Distinct TRPV1- and TRPA1-based mechanisms underlying enhancement of oral ulcerative mucositis-induced pain by 5-fluorouracil.

In many patients with cancer, chemotherapy-induced severe oral ulcerative mucositis causes intractable pain, leading to delays and interruptions in therapy. However, the pain mechanism in oral ulcerative mucositis after chemotherapy has not been extensively studied. In this study, we investigated spontaneous pain and mechanical allodynia in a preclinical model of oral ulcerative mucositis after systemic administration of the chemotherapy drug 5-fluorouracil, using our proprietary pain assay system for conscious rats. 5-Fluorouracil caused leukopenia but did not induce pain-related behaviors. After 5-fluorouracil administration, oral ulcers were developed with topical acetic acid treatment. Compared with saline-treated rats, 5-fluorouracil-exposed rats showed more severe mucositis with excessive bacterial loading due to a lack of leukocyte infiltration, as well as enhancements of spontaneous pain and mechanical allodynia. Antibacterial drugs, the lipid A inhibitor polymyxin B and the TRPV1/TRPA1 channel pore-passing anesthetic QX-314, suppressed both the spontaneous pain and the mechanical allodynia. The cyclooxygenase inhibitor indomethacin and the TRPV1 antagonist SB-366791 inhibited the spontaneous pain, but not the mechanical allodynia. In contrast, the TRPA1 antagonist HC-030031 and the N-formylmethionine receptor FPR1 antagonist Boc MLF primarily suppressed the mechanical allodynia. These results suggest that 5-fluorouracil-associated leukopenia allows excessive oral bacterial infection in the oral ulcerative region, resulting in the enhancement of spontaneous pain through continuous TRPV1 activation and cyclooxygenase pathway, and mechanical allodynia through mechanical sensitization of TRPA1 caused by neuronal effects of bacterial toxins. These distinct pain mechanisms explain the difficulties encountered with general treatments for oral ulcerative mucositis-induced pain in patients with cancer and suggest more effective approaches.