Ultrastructural and immunohistochemical evaluation of hyperplastic soft tissues surrounding dental implants in fibular jaws.

In reconstructive surgery, complications post-fibula free flap (FFF) reconstruction, notably peri-implant hyperplasia, are significant yet understudied. This study analyzed peri-implant hyperplastic tissue surrounding FFF, alongside peri-implantitis and foreign body granulation (FBG) tissues from patients treated at the Department of Oral and Maxillofacial Surgery, Seoul National University Dental Hospital. Using light microscopy, pseudoepitheliomatous hyperplasia, anucleate and pyknotic prickle cells, and excessive collagen deposition were observed in FFF hyperplastic tissue. Ultrastructural analyses revealed abnormal structures, including hemidesmosome dilation, bacterial invasion, and endoplasmic reticulum (ER) swelling. In immunohistochemical analysis, unfolded protein-response markers ATF6, PERK, XBP1, inflammatory marker NFκB, necroptosis marker MLKL, apoptosis marker GADD153, autophagy marker LC3, epithelial-mesenchymal transition, and angiogenesis markers were expressed variably in hyperplastic tissue surrounding FFF implants, peri-implantitis, and FBG tissues. NFκB expression was higher in peri-implantitis and FBG tissues compared to hyperplastic tissue surrounding FFF implants. PERK expression exceeded XBP1 significantly in FFF hyperplastic tissue, while expression levels of PERK, XBP1, and ATF6 were not significantly different in peri-implantitis and FBG tissues. These findings provide valuable insights into the interconnected roles of ER stress, necroptosis, apoptosis, and angiogenesis in the pathogenesis of oral pathologies, offering a foundation for innovative strategies in dental implant rehabilitation management and prevention.

Layered structure of sialoliths compared with tonsilloliths and antroliths.

Objectives: The aim of this study was to perform a comparative analysis of the ultrastructural and chemical composition of sialoliths, tonsilloliths, and antroliths and to describe their growth pattern. Materials and Methods: We obtained 19 specimens from 18 patients and classified the specimens into three groups: sialolith (A), tonsillolith (B), and antrolith (C). The peripheral, middle, and core regions of the specimens were examined in detail by histology, micro-computed tomography (micro- CT), scanning electron microscopy (SEM), energy dispersive X-ray spectroscopy, and transmission electron microscopy (TEM). Results: In the micro-CT, group A showed alternating radiodense and radiolucent layers, while group B had a homogeneous structure. Group C specimens revealed a compact homogeneous structure. Histopathologically, group A showed a laminated, teardrop-shaped, globular structure. Group B demonstrated degrees of immature calcification of organic and inorganic materials. In group C, the lesion was not encapsulated and showed a homogeneous lamellar bone structure. SEM revealed that group A showed distinct three layers: a peripheral multilayer zone, intermediate compact zone, and the central nidus area; groups B and C did not show these layers. The main elemental components of sialoliths were O, C, Ca, N, Cu, P, Zn, Si, Zr, F, Na, and Mg. In group B, a small amount of Fe was found in the peripheral region. Group C had a shorter component list: Ca, C, O, P, F, N, Si, Na, and Mg. TEM analysis of group A showed globular structures undergoing intra-vesicular calcification. In group B, bacteria were present in the middle layer. In the outer layer of the group C antrolith, an osteoblastic rimming was observed. Conclusion: Sialoliths had distinct three layers: a peripheral multilayer zone, an intermediate compact zone and the central nidus area, while the tonsillolith and antrolith specimens lacked distinct layers and a core.

Association of heavy metal complex exposure and neurobehavioral function of children.

Background: Exposure to heavy metals is a public health concern worldwide. Previous studies on the association between heavy metal exposure and neurobehavioral functions in children have focused on single exposures and clinical manifestations. However, the present study evaluated the effects of heavy metal complex exposure on subclinical neurobehavioral function using a Korean Computerized Neurobehavior Test (KCNT). Methods: Urinary mercury, lead, cadmium analyses as well as symbol digit substitution (SDS) and choice reaction time (CRT) tests of the KCNT were conducted in children aged between 10 and 12 years. Reaction time and urinary heavy metal levels were analyzed using partial correlation, linear regression, Bayesian kernel machine regression (BKMR), the weighted quantile sum (WQS) regression and quantile G-computation analysis. Results: Participants of 203 SDS tests and 198 CRT tests were analyzed, excluding poor cooperation and inappropriate urine sample. Partial correlation analysis revealed no association between neurobehavioral function and exposure to individual heavy metals. The result of multiple linear regression shows significant positive association between urinary lead, mercury, and CRT. BMKR, WQS regression and quantile G-computation analysis showed a statistically significant positive association between complex urinary heavy metal concentrations, especially lead and mercury, and reaction time. Conclusions: Assuming complex exposures, urinary heavy metal concentrations showed a statistically significant positive association with CRT. These results suggest that heavy metal complex exposure during childhood should be evaluated and managed strictly.

Identification of biological components for sialolith formation organized in circular multi-layers.

According to the previous studies of sialolithiasis reported so far, this study is aimed to identify the biological components of sialolith, which show different ultrastructures and chemical compositions from other stones, cholelith and urolith. Twenty-two specimens obtained from 20 patients were examined histologically, and analyzed with micro-CT, scanning electron microscopy (SEM), energy dispersive X-ray spectroscopy (EDS), and transmission electron microscopy (TEM). All sialoliths (n = 22) observed in this study showed a central nidus, which was filled with organoid matrix admixed with exosome vesicles, loose calcium apatite crystals, and many bacteria. The micro-CT and SEM observation clearly defined a single or multiple central nidus(es) encircled by highly calcified compact zone. The circular compact zone showed a band-like calcification, about 1-3 mm in thickness, and usually located between the central nidus and the peripheral multilayer zone. But some sialoliths (n = 5) showed severe erosion of compact zone by expanding multilayered zone depending on the level of calcification and inflammation in sialolith. By observing TEM images, many exosome vesicles and degraded cytoplasmic organelles were found in the central nidus, and some epithelial cells were also found in the calcified matrix of peripheral multilayer zone. Particularly, EDS analysis indicated the highest Ca/P ratio in the intermediate compact zone (1.77), and followed by the central nidus area (1.39) and the peripheral multilayer zone (0.87). Taken together, these data suggest that the central nidus containing many inflammatory exosomes and degraded cytoplasmic organelles has a potential to induce a band-like calcification of compact zone, and followed by the additional multilayer deposition of exfoliated salivary epithelial cells as well as salivary materials. Thereby, the calcium apatite-based sialolith is gradually growing in its volume size, and eventually obstructs the salivary flow and provides a site for the bacterial infection.

Adverse Effects on Work and Daily Life Interference among Healthcare Workers after the First and Second ChAdOx1 and BNT162b2 COVID-19 Vaccine Doses.

In this study, we assessed the adverse effects and the work and daily life interference associated with each dose of the ChAdOx1 and BNT162b2 COVID-19 vaccines. Questionnaires were distributed to workers after they received both doses; only those who worked the day after receiving the vaccine were included in the analysis. Overall, 368 ChAdOx1-vaccinated and 27 BNT162b2-vaccinated participants were included. Among the ChAdOx1-vaccinated participants, the incidence of adverse effects was significantly lower after the second dose than after the first dose. Among the BNT162b2-vaccinated participants, however, no differences in adverse effects or work and daily life interference were found between the doses. After the first and second dose, the numeric scale score (0-10) for interference with work was 3.9 ± 2.9 and 1.6 ± 1.9 for the ChAdOx1 and 3.2 ± 2.5 and 3.6 ± 3.0 for the BNT162b2 vaccine, respectively. A similar trend was observed for interference with daily life. Factors associated with work and daily life interference in the multivariate model were age, vaccine dose (first or second), and the interaction term of vaccine type and dose. These results could be used to inform the general population of the adverse effects associated with these vaccinations.

4-Hexylresorcinol-in duced angiogenesis potential in human endothelial cells.

BACKGROUND: 4-Hexylresorcinol (4HR) is able to increase angiogenesis. However, its molecular mechanism in the human endothelial cells has not been clarified. METHODS: As endothelial cells are important in angiogenesis, we treated the human umbilical vein endothelial cells (HUVECs) with 4HR and investigated protein expressional changes by immunoprecipitation high-performance liquid chromatography (IP-HPLC) using 96 antisera. RESULTS: Here, we found that 4HR upregulated transforming growth factor-ß (TGF-ß)/SMAD/vascular endothelial growth factor (VEGF) signaling, RAF-B/ERK and p38 signaling, and M2 macrophage polarization pathways. 4HR also increased expression of caspases and subsequent cellular apoptosis. Mechanistically, 4HR increased TGF-ß1 production and subsequent activation of SMADs/VEGFs, RAF-B/ERK and p38 signaling, and M2 macrophage polarization. CONCLUSION: Collectively, 4HR activates TGF-ß/SMAD/VEGF signaling in endothelial cells and induced vascular regeneration and remodeling for wound healing.

Development of a Molecular Marker Linked to the A4 Locus and the Structure of HD Genes in Pleurotus eryngii.

Allelic differences in A and B mating-type loci are a prerequisite for the progression of mating in the genus Pleurotus eryngii; thus, the crossing is hampered by this biological barrier in inbreeding. Molecular markers linked to mating types of P. eryngii KNR2312 were investigated with randomly amplified polymorphic DNA to enhance crossing efficiency. An A4-linked sequence was identified and used to find the adjacent genomic region with the entire motif of the A locus from a contig sequenced by PacBio. The sequence-characterized amplified region marker 7-2299 distinguished A4 mating-type monokaryons from KNR2312 and other strains. A BLAST search of flanked sequences revealed that the A4 locus had a general feature consisting of the putative HD1 and HD2 genes. Both putative HD transcription factors contain a homeodomain sequence and a nuclear localization sequence; however, valid dimerization motifs were found only in the HD1 protein. The ACAAT motif, which was reported to have relevance to sex determination, was found in the intergenic region. The SCAR marker could be applicable in the classification of mating types in the P. eryngii breeding program, and the A4 locus could be the basis for a multi-allele detection marker.

Botulinum toxin A injection into the anterior belly of the digastric muscle increased the posterior width of the maxillary arch in developing rats.

BACKGROUND: The purpose of this study was to evaluate the effects of botulinum toxin A (BTX) injection into the anterior belly of the digastric muscle on a growing rat. METHODS: Ten Sprague Dawley rats were used in this study. When the rats were 13 days old, 0.5 units of BTX was injected into the anterior belly of the digastric muscle for the experimental group (n = 5). For the control, the same volume of normal saline was injected (n = 5). The rats were sacrificed at 60 days old, and the skulls were harvested for micro-computed tomography (µCT) analysis. RESULTS: In anthropometric analysis, the zygomatic arch and mandibular bi-condylar width were significantly lower in the experimental group than those in the control group (P = 0.025 and 0.027, respectively). The maxillary point width was significantly higher in the experimental group than that in the control group (P = 0.020). CONCLUSION: BTX injection into the anterior belly of the digastric muscle had effects on the maxillofacial bony width in growing rats.

Calcium pyrophosphate dihydrate deposition disease in the temporomandibular joint: diagnosis and treatment.

BACKGROUND: Calcium pyrophosphate dihydrate deposition disease (CPDD) is a rare disease in the temporomandibular joint (TMJ) space. It forms a calcified crystal mass and induces a limitation of joint movement. CASE PRESENTATION: The calcified mass in our case was occupied in the left TMJ area and extended to the infratemporal and middle cranial fossa. For a complete excision of this mass, we performed a vertical ramus osteotomy and resected the mass around the mandibular condyle. The calcified mass in the infratemporal fossa was carefully excised, and the segmented mandible was anatomically repositioned. Scanning electronic microscopy (SEM)/energy-dispersive X-ray spectroscopy (EDS) microanalysis was performed to evaluate the calcified mass. The result of SEM/EDS showed that the crystal mass was completely composed of calcium pyrophosphate dihydrate. This result strongly suggested that the calcified mass was CPDD in the TMJ area. CONCLUSIONS: CPDD in the TMJ is a rare disease and is difficult to differentially diagnose from other neoplasms. A histological examination and quantitative microanalysis are required to confirm the diagnosis. In our patient, CPDD in the TMJ was successfully removed via the extracorporeal approach. SEM/EDS microanalysis was used for the differential diagnosis.

Effect of the masseter muscle injection of botulinum toxin A on the mandibular bone growth of developmental rats.

BACKGROUND: The objective of this study was to evaluate the influence of masticatory muscle injection of botulinum toxin type A (BTX-A) on the growth of the mandibular bone in vivo. METHODS: Eleven Sprague-Dawley rats were used, and BTX-A (n = 6) or saline (n = 5) was injected at 13 days of age. All injections were given to the right masseter muscle, and the BTX-A dose was 0.5 units. All of the rats were euthanized at 60 days of age. The skulls of the rats were separated and fixed with 10% formalin for micro-computed tomography (micro-CT) analysis. RESULTS: The anthropometric analysis found that the ramus heights and bigonial widths of the BTX-A-injected group were significantly smaller than those of the saline-injected group (P < 0.05), and the mandibular plane angle of the BTX-A-injected group was significantly greater than in the saline-injected group (P < 0.001). In the BTX-A-injected group, the ramus heights II and III and the mandibular plane angles I and II showed significant differences between the injected and non-injected sides (P < 0.05). The BTX-A-injected side of the mandible in the masseter group showed significantly lower mandibular bone growth compared with the non-injected side. CONCLUSION: BTX-A injection into the masseter muscle influences mandibular bone growth.

Mandibular Reconstruction Using a Customized Three-Dimensional Titanium Implant Applied on the Lingual Surface of the Mandible.

A patient had a right mandibular defect due to resection of an ameloblastoma. Previously, the defect had been reconstructed by an iliac bone graft, and subsequently, a titanium mesh with xenograft was used. However, it was not successfully reconstructed. For the recovery of mandible continuity and rehabilitation of jaw movement, we manufactured a customized 3-dimensional titanium implant by computer-aided design and manufacturing and electron beam melting technology. This implant was designed to have a porous body structure and lingual plate. The customized implant was accurately inserted in the bony defect. As a result, the patient showed a normal range of mouth opening and jaw movement. New bone migration was observed in the porous structure of the implant. Although there was a slight plate exposure and lack of alveolar bone formation, the customized 3D titanium implant successfully reconstructed the mandibular discontinuous defect and recovered jaw movement.

Bone regeneration is associated with the concentration of tumour necrosis factor-α induced by sericin released from a silk mat.

To understand the osteogenic effect of the middle layer of the silk cocoon, sericin was examined for its cellular effects associated with tumor necrosis factor-α (TNF-α) signaling in this study. The fragmented sericin proteins in the silk mat were evaluated for the TNF-α expression level in murine macrophages. The concentration of protein released from silk mats was higher in the outermost and the innermost layers than in the middle layers, and the protein released from the silk mat was identified as sericin. The level of TNF-α in murine macrophages was dependent on the applied concentration of sericin, and the expression of genes associated with osteogenesis in osteoblast-like cells was dependent on the applied concentration of TNF-α. In animal experiments, silk mats from the middle layers led to a higher regenerated bone volume than silk mats from the innermost layer or the outermost layer. If TNF-α protein was incorporated into the silk mats from the middle layers, bone regeneration was suppressed compared with unloaded silk mats from the middle layers. Accordingly, silk mats from the silk cocoon can be considered to be a fragmented sericin-secreting carrier, and the level of sericin secretion is associated with TNF-α induction and bone regeneration.

Botulinum Toxin Conjugated With Silk Fibroin and 4-Hexylresorcinol.

PURPOSE: The objective of this study was to evaluate whether silk fibroin (SF) incorporated into 4-hexylresorcinol (4HR) could increase botulinum toxin-A (BTX-A) activity. MATERIAL AND METHODS: In total, 30 rats were used for this study. The animals were divided into 6 groups according to the injected materials (SA: saline only; SF; 4HR; B2: 2 units of BTX-A; B2 + SF + 4HR: combination of B2, SF, and 4HR; B5: 5 units of BTX-A). Serial sonography was used for the evaluation of muscle thickness after injection. Immunohistochemical staining was used for the evaluation of myosin type II (myo2) and Bcl-2 protein expression. RESULTS: The relative thickness of the masseter muscle in B2 group was 66.14% ±â€Š4.55% to the preinjection level; in B2 + SF + 4HR group was 54.59% ±â€Š4.83%, and in B5 group was 56.19% ±â€Š8.28%. Any BTX-injected group showed significantly lower value of the relative muscle thickness compared to SA, SF, or 4HR group (P < 0.001 for all). The difference of relative muscle thickness between B2 group and B2 + SF + 4HR group was statistically significant (P < 0.001). The intensity of myo2 immunostaining in B5, B2, and B2 + SF + 4HR group was significantly higher than those in the other groups (P < 0.05). CONCLUSIONS: When 2 units of BTX was incorporated to SF and 4HR, combination formula showed similar activity to those of 5 units of BTX.

The use of the buccal fat pad flap for oral reconstruction.

Many congenital and acquired defects occur in the maxillofacial area. The buccal fat pad flap (BFP) is a simple and reliable flap for the treatment of many of these defects because of its rich blood supply and location, which is close to the location of various intraoral defects. In this article, we have reviewed BFP and the associated anatomical background, surgical techniques, and clinical applications. The surgical procedure is simple and has shown a high success rate in various clinical applications (approximately 90%), including the closure of oroantral fistula, correction of congenital defect, treatment of jaw bone necrosis, and reconstruction of tumor defects. The control of etiologic factors, size of defect, anatomical location of defect, and general condition of patient could influence the prognosis after grafting. In conclusion, BFP is a reliable flap that can be applied to various clinical situations.

Accelerated biodegradation of silk sutures through matrix metalloproteinase activation by incorporating 4-hexylresorcinol.

Silk suture material is primarily composed of silk fibroin and regarded as a non-resorbable material. It is slowly degraded by proteolysis when it is implanted into the body. 4-Hexylresorcinol (4HR) is a well-known antiseptic. In this study, the biodegradability of 4HR-incorporated silk sutures were compared to that of untreated silk sutures and polyglactin 910 sutures, a commercially available resorbable suture. 4HR-incorporated silk sutures exhibited anti-microbial properties. Matrix metalloproteinase (MMP) can digest a wide spectrum of proteins. 4HR increased MMP-2, -3, and -9 expression in RAW264.7 cells. MMP-2, -3, and -9 were able to digest not only silk fibroin but also silk sutures. Consequently, 59.5% of the 4HR-incorporated silk suture material remained at 11 weeks after grafting, which was similar to that of polyglactin 910 degradation (56.4% remained). The residual amount of bare silk suture material at 11 weeks after grafting was 91.5%. The expression levels of MMP-2, -3 and -9 were high in the 4HR-incorporated silk suture-implanted site 12 weeks after implantation. In conclusion, 4HR-treated silk sutures exhibited anti-microbial properties and a similar level of bio-degradation to polyglactin 910 sutures and induced higher expression of MMP-2, -3, and -9 in macrophages.

Comparison of Bio-degradation for Ridge Preservation Using Silk Fibroin-based Grafts and a Collagen Plug.

A material for ridge preservation should have dimensional stability to resist bio-degradation. This study was designed to compare bio-degradation of ridge preservation materials. Collagen plug was used as a positive control. Untreated, ethanol-treated, and 4-hexylresorcinol (4HR)-treated silk plugs were used for the experimental group. Each material underwent a scanning electron microscopic exam and a Fourier transform infrared (FT-IR) spectroscopic exam. Bio-degradation was evaluated by analyzing cylindrical bony defects in rabbit tibias. There were no prominent differences in microstructure among the silk plug groups. FT-IR exam demonstrated that the ethanol- and 4HR-treated silk plug groups had enhanced ß-sheet structure. All silk plug groups exhibited significantly higher residual graft than the collagen plug group 4 weeks postoperative (p < 0.05). In conclusion, silk fibroin-based ridge preservation material was less bio-degradable than a collagen plug until at least 4 weeks after grafting.

Apoptotic action of botulinum toxin on masseter muscle in rats: early and late changes in the expression of molecular markers.

The purpose of this study was to compare the early or late expression levels of p65, Bcl-2, and type II myosin and the frequency of TUNEL-positive nuclei in the rat masseter muscle after injection of different concentrations of botulinum toxin-A (BTX-A). We injected either 5 U or 10 U of BTX-A into both masseter muscles of the rat. As a control group, the same volume of saline was injected. After 2 or 12 weeks, the animals were sacrificed. Subsequently, a biopsy and immunohistochemical staining of the samples were performed using a p65, Bcl-2, or type II myosin antibody. Additionally, a TUNEL assay and transmission electron microscopic analysis were performed. The expression of p65, Bcl-2, and type II myosin increased significantly with increasing concentrations of BTX-A at 2 weeks after BTX-A injection (P < 0.05). The number of TUNEL-positive nuclei was also significantly increased in the BTX-A-treated groups in comparison to the saline-treated control at 2 weeks after BTX-A injection (P < 0.05). Elevated expression of Bcl-2 was also observed in 10-unit BTX-A-treated group at 12 weeks after injection (P < 0.05). At 12 weeks after injection, the number of enlarged mitochondria was increased, and many mitochondria displayed aberrations in cristae morphology after BTX-A injection. In conclusion, BTX-A injection into the masseter muscle increased the expression level of p65, Bcl-2, and type II myosin at an early stage. The morphological changes of mitochondria were more evident at 12 weeks after injection.

Topical delivery of 4-hexylresorcinol promotes wound healing via tumor necrosis factor-α suppression.

OBJECTIVE: The objective of this study was to evaluate (1) the effect of 4-hexylresorcinol (4HR) application on TNF-α expression in RAW264.7 cells and (2) the effect of 4HR ointment on burn wound healing in a rat burn wound model. MATERIALS AND METHODS: RAW264.7 cells were cultured in the presence of different concentrations of 4HR, and the attached cells were sampled for western blot analysis at each time point. Animal studies were conducted on male Wistar rats (n=24). Three treatment groups were evaluated in this study: (1) treatment with ointment alone (negative control), (2) treatment with ointment containing 0.2wt% 4HR, and (3) treatment with ointment containing 2wt% 4HR. For the assessment of wound healing, digital photographs were taken at 1, 5, and 14 days. Animals were sacrificed at 1, 5, and 14 days for histological examination or ELISA analysis. RESULTS: Western blot analysis showed that TNF-α protein levels were reduced following treatment with 4HR at concentrations between 1 and 10µg/mL. In animal studies, the average denuded area 14 days after burn was smaller in the 2wt% 4HR treatment group compared with the control group (P=0.022). Immunohistochemical analysis revealed lower TNF-α expression in the 2wt% 4HR treatment group compared with the control group 14 days after burn (P=0.027). CONCLUSION: 4HR reduced TNF-α expression in RAW264.7 cells and in the burn wounds of rats. Burn wounds dressed with ointment containing 4HR also exhibited rapid epithelization and collagen regeneration.

Long-term follow-up of early cleft maxillary distraction.

BACKGROUND: Most of cleft lip and palate patients have the esthetic and functional problems of midfacial deficiencies due to innate developmental tendency and scar tissues from repeated operations. In these cases, maxillary protraction is required for the harmonious facial esthetics and functional occlusion. CASE PRESENTATION: A 7-year old boy had been diagnosed as severe maxillary constriction due to unilateral complete cleft lip and palate. The author tried to correct the secondary deformity by early distraction osteogenesis with the aim of avoiding marked psychological impact from peers of elementary school. From 1999 to 2006, repeated treatments, which consisted of Le Fort I osteotomy and face mask distraction, and complementary maxillary protraction using miniplates were performed including orthodontics. But, final facial profile was not satisfactory, which needs compromising surgery. CONCLUSIONS: The result of this study suggests that if early distraction treatment is performed before facial skeletal growth is completed, an orthognathic surgery or additional distraction may be needed later. Maxillofacial plastic and reconstructive surgeons should notify this point when they plan early distraction treatment for cleft maxillary deformity.