In vitroanalysis of insoluble salt formation mechanism associated with Mg corrosion-variations depending on the diffusion environment in model tissue.

Magnesium (Mg) alloys have attracted attention as biodegradable metals, but the details of their corrosion behavior under biological environment have not been elucidated. Previous studies have suggested that diffusion through blood flow may influence Mg corrosion. Therefore, to understand the degradation behaviors of Mg, we analyzed insoluble salt precipitation associated with Mg corrosion in model tissue with different diffusion rates. A pure Mg specimen was immersed into a model tissue prepared with cell culture medium supplemented by a thickener at a different concentration (0.2%-0.5%) to form the gel. Micro-focus x-ray computed tomography of the gel was performed to observe gas cavity formation around the specimen. The insoluble salt layer formed on the specimen surface were analyzed by scanning electron microscopy with energy-dispersive x-ray spectroscopy, and Raman spectroscopy. As results, gas cavity formation was observed for all specimens. At day 7, the gas cavity volume was the highest at 0.5% thickener gel followed by 0.3% thickener gel. The insoluble salts were classified into three types based on their morphology; plate-like, granular-like, and crater-like salts. The crater-like salts were observed to cover 16.8 ± 3.9% of the specimen surface immersed in the 0.5% thickener gel, at the specimen area contacted to the gas cavity. The crater-like salts were composed by Mg hydroxide and carbonate from the deepest to the top layer. In plate-like or granular-like salts, Mg carbonate was formed in the deepest layer, but phosphates and carbonates, mainly containing calcium not Mg, were formed on the surface layer. In conclusion, the increase in the thickener concentration increased the gas cavity volume contacting to the specimen surface, resulting in the increase in precipitation of Mg hydroxide and carbonate, composing crater-like salts. Mg hydroxide and carbonate precipitation suggests the local increase in OH-concentration, which may be attributed to the decrease in diffusion rate.

Advantages of a High-Powered Ultrasonic Vibration Cutting Device for Orthognathic Surgeries.

This study aimed to evaluate the advantages of using a high-powered ultrasonic vibration cutting device (HP-UVCD) in orthognathic surgeries. We analyzed 22 consecutive patients who underwent Le FortⅠosteotomy and bilateral sagittal split osteotomies for cleft lip and palate. We used conventional power instruments, such as reciprocating saws and rotating burrs in the first 10 patients, and an HP-UVCD in the remaining 12 patients. Surgical time, intraoperative blood loss, and postoperative inferior alveolar nerve affection were retrospectively investigated. The surgical time in the ultrasonic group (223±12.217 min) was significantly shorter than that in the conventional group (278±11.153 min, P =0.008). The amount of intraoperative blood loss in the ultrasonic group was an average of 230±45.154 g, and that in the conventional group was an average of 343±49.463 g. Although the blood loss in the ultrasonic group was reduced compared with that in the conventional group, this difference was not statistically significant ( P =0.118). All patients in both groups recovered normal sensation by 6 months postoperatively. Our findings suggest that the HP-UVCD can be used in place of a reciprocating saw or rotating burrs in orthognathic surgeries, resulting in reduced surgical time and minimal blood loss. The use of an ultrasonic bone-cutting device may allow for a safer and less invasive orthognathic surgery.

Impact of palatoplasty techniques on tympanic membrane findings and hearing prognosis in children with cleft palate.

OBJECTIVE: Children with cleft palate (CP) are at high risk of developing otitis media with effusion (OME) due to Eustachian tube (ET) dysfunction. Palatoplasty has been reported to decrease the frequency of middle ear disease and improve ET function, and although various techniques have been developed, there is no consensus on the differences in the impact of different techniques on the middle ear. The purpose of this study was to determine the differential effects of palatoplasty on middle ear function and hearing. METHODS: We performed a retrospective observational survey of pediatric patients who underwent palatoplasty for CP between June 2010 and October 2018 at Tohoku University Hospital. Cases were divided into three groups depending on the palatoplasty procedures performed: the push-back palatoplasty group, the two-flap palatoplasty group, and the Furlow double-opposing Z-plasty group. We examined the differences in clinical characteristics between patients who underwent each procedure. The primary outcome variable was tympanic membrane (TM) findings, and the secondary outcome was hearing test results. RESULTS: Children who underwent the two-flap palatoplasty had a higher tympanostomy tube (TT) insertion rate and a higher total number of TT insertions than those who underwent the Furlow double-opposing Z-plasty or the push-back palatoplasty. The TM retraction rate tended to be lower in the Furlow double-opposing Z-plasty group than in the push-back palatoplasty group or the two-flap palatoplasty group. The hearing test results at the last visit were not significantly different among the three groups. CONCLUSIONS: Children who underwent the two-flap palatoplasty had a higher rate of TT insertions, potentially increasing the risk of TM perforation. In contrast, the Furlow double-opposing Z-plasty group had a lower tendency for TM regression, possibly due to improved ET function and reduced incidence of OME. It is important to understand the advantages and disadvantages of each technique before selecting one suitable for the child's cleft and arch width. Additionally, it is important to conduct regular follow-up of TM findings and hearing test results even after palatoplasty.

CARD9-mediated macrophage responses and collagen fiber capsule formation caused by textured-type breast implants.

BACKGROUND: An increasing number of women are undergoing breast implantation for cosmetic purposes and for reconstructive purposes after breast excision. The surface morphology of the breast implant is one of the key factors associated with the induction of capsule contraction. The effect of surface morphology on the inflammatory response following implant insertion remains unclear, however. This study conducted comparative analyses to determine the effect of the textured and smooth surface morphology of silicone sheets. METHODS: Each type of silicone sheet was inserted into the subcutaneous pocket below the panniculus carnosus in C57BL/6 mice and mice with genetic disruption of CARD9, Dectin-1, Dectin-2, or Mincle. We also analyzed the collagen fiber capsule thickness, histological findings, and macrophage inflammatory response, including TGF-ß synthesis. RESULTS: We found that textured surface morphology contributed to the formation of collagen fiber capsules and the accumulation of fibroblasts and myofibroblasts, and was accompanied by the accumulation of TGF-ß-expressing macrophages and foreign-body giant cells. CARD9 deficiency attenuated collagen fiber capsule formation, macrophage responses, and TGF-ß synthesis, although the responsible C-type lectin receptors (CLRs) remain to be clarified. CONCLUSIONS: These results suggest that CARD9 may have a strong impact on silicone sheet insertion through the regulation of macrophage responses.

Cutaneous wound healing promoted by topical administration of heat-killed Lactobacillus plantarum KB131 and possible contribution of CARD9-mediated signaling.

Optimal conditions for wound healing require a smooth transition from the early stage of inflammation to proliferation, and during this time alternatively activated (M2) macrophages play a central role. Recently, heat-killed lactic acid bacteria (LAB), such as Lactobacillus plantarum (L. plantarum) have been reported as possible modulators affecting the immune responses in wound healing. However, how signaling molecules regulate this process after the administration of heat-killed LAB remains unclear. In this study, we examined the effect of heat-killed L. plantarum KB131 (KB131) administration on wound healing and the contribution of CARD9, which is an essential signaling adaptor molecule for NF-kB activation upon triggering through C-type lectin receptors, in the effects of this bacterium. We analyzed wound closure, histological findings, and inflammatory responses. We found that administration of KB131 accelerated wound closure, re-epithelialization, granulation area, CD31-positive vessels, and α-SMA-positive myofibroblast accumulated area, as well as the local infiltration of leukocytes. In particular, M2 macrophages were increased, in parallel with CCL5 synthesis. The acceleration of wound healing responses by KB131 was canceled in CARD9-knockout mice. These results indicate that the topical administration of KB131 accelerates wound healing, accompanying increased M2 macrophages, which suggests that CARD9 may be involved in these responses.

Surgical treatment of nonuremic calciphylaxis: a case report and review of literature.

Calciphylaxis is characterized by extremely painful skin ulcers and develops in patients with or without severe kidney diseases. Herein, a female patient without renal dysfunction developed calciphylaxis in the right lower extremity and underwent successful surgical debridement and split-thickness skin grafting. Nevertheless, prompt wound evaluation and proper surgical approaches are essential.

Magnetoencephalographic evaluation of repaired lip sensation in patients with cleft lip.

BACKGROUND: Cleft lip is the most common congenital anomaly worldwide. Nevertheless, lip somatosensory characteristics of patients with cleft lip after cheiloplasty have not yet been determined. The present study used magnetoencephalography to objectively evaluate the lip sensation in patients with unilateral cleft lip to establish a new objective evaluation method. METHODS: Participants were 15 patients with unilateral cleft lip after cheiloplasty (UCL group), and 30 healthy young subjects (control group). Five points of the upper and lower lips were stimulated electrically to measure somatosensory evoked magnetic fields (SEFs). The sources of the magnetic fields were modeled as single equivalent current dipoles (ECDs). ECDs located on the central sulcus by superimposition on magnetic resonance images were analyzed. Latency and intensity at 50-75 ms (cP60m) observed in the UCL group were compared with those in the control group. Thresholds of tactile stimuli in both groups were obtained using Semmes-Weinstein monofilaments for subjective sensory evaluation. RESULTS: No significant difference was found in the intensity of the cP60m or subjective evaluation between the groups. However, the latency of the cP60m was significantly longer in the upper lip of the UCL group than in the control group. CONCLUSIONS: SEFs showed a difference in lip sensation between the UCL group and the control group, suggesting that longer latency might be caused by the effects of surgical scarring on the neurotransmission pathway. These results suggest SEFs as useful for the objective evaluation of lip sensations. This study might improve future surgical procedures and lip functions of patients with cleft lip.

Somatosensory evoked magnetic fields induced by electrical palate stimulation in patients with unilateral cleft lip and palate after palatoplasty.

Palatal sensation is important for articulation, feeding, and swallowing. However, palatal sensation in patients with cleft palate (CP) after palatoplasty has been investigated only inadequately because of the complexity and high costs of objective evaluation. This study compared the somatosensory evoked magnetic fields (SEFs) induced by electrical stimulation of the palates of patients with CP after palatoplasty and the palatal sensory thresholds (PSTs) of the stimulation with those of healthy (control) subjects. The CP group comprised 12 patients with unilateral cleft lip and palate (UCLP). The control group comprised 31 control subjects. No significant difference in intensity was found between them. Nevertheless, the PSTs in the UCLP group were significantly higher than those in the control group at all sites (p < 0.05). We infer that the electrical signals transmitted from palatal sensory receptors of patients with CP might be amplified by a compensation mechanism in the central nervous system. SEFs provide an effective method for objective evaluation of palatal sensation in patients with CP after palatoplasty. Evaluation of SEFs during palatal sensory stimulation in patients with CP after palatoplasty might lead to better corrective surgical methods that also preserve palatal sensation.

Pilot study of factors contributing to canine impaction after secondary alveolar bone grafting in unilateral cleft lip and palate patients.

Alveolar bone grafting (ABG) is broadly performed for cleft lip and palate patients. The rate of canine impaction post-ABG is much higher than the prevalence of canine impaction in non-cleft patients. This pilot study was designed to investigate factors involved in canine eruption failure after ABG and to predict the possibility of canine impaction in unilateral cleft lip and palate (UCLP) patients. This retrospective observational study examined 45 patients with UCLP (mean age 7.9 years) classified into an impacted group (n = 9) and a spontaneously erupted group (n = 36). From 3D images, we measured lateral incisor presence or absence, lateral incisor position on the cleft side, canine position, movement change, cleft volume, and canine and first premolar overlap-area. Multivariate logistic regression analysis using independent variables indicated significant differences in results, selecting highly relevant items. Multivariate analysis indicated a significant association between the overlap-area between the canine and the first premolar at pre-ABG (p = 0.038) and the distance between the cleft side cusp tips of canine and the lateral cleft margin of pre-ABG (p = 0.005). Results suggest that canine impaction is predictable at an early stage in pre-ABG and show the possibility of comprehensive diagnosis of canine impaction using computed tomography.

Topical Administration of Heat-Killed Enterococcus faecalis Strain KH2 Promotes Re-Epithelialization and Granulation Tissue Formation during Skin Wound-Healing.

Lactic acid bacteria (LAB) are known to have beneficial effects on immune responses when they are orally administered as bacterial products. Although the beneficial effects of LAB have been reported for the genera Lactobacillus and Lactococcus, little has been uncovered on the effects of the genus Enterococcus on skin wound-healing. In this study, we aimed to clarify the effect of heat-killed Enterococcus faecalis KH2 (heat-killed KH2) strain on the wound-healing process and to evaluate the therapeutic potential in chronic skin wounds. We analyzed percent wound closure, re-epithelialization, and granulation area, and cytokine and growth factor production. We found that heat-killed KH2 contributed to the acceleration of re-epithelialization and the formation of granulation tissue by inducing tumor necrosis factor-α, interleukin-6, basic fibroblast growth factor, transforming growth factor (TGF)-ß1, and vascular endothelial growth factor production. In addition, heat-killed KH2 also improved wound closure, which was accompanied by the increased production of TGF-ß1 in diabetic mice. Topical administration of heat-killed KH2 might have therapeutic potential for the treatment of chronic skin wounds in diabetes mellitus. In the present study, we concluded that heat-killed KH2 promoted skin wound-healing through the formation of granulation tissues and the production of inflammatory cytokines and growth factors.

Contribution of Invariant Natural Killer T Cells to the Clearance of Pseudomonas aeruginosa from Skin Wounds.

Chronic infections are considered one of the most severe problems in skin wounds, and bacteria are present in over 90% of chronic wounds. Pseudomonas aeruginosa is frequently isolated from chronic wounds and is thought to be a cause of delayed wound healing. Invariant natural killer T (iNKT) cells, unique lymphocytes with a potent regulatory ability in various inflammatory responses, accelerate the wound healing process. In the present study, we investigated the contribution of iNKT cells in the host defense against P. aeruginosa inoculation at the wound sites. We analyzed the re-epithelialization, bacterial load, accumulation of leukocytes, and production of cytokines and antimicrobial peptides. In iNKT cell-deficient (Jα18KO) mice, re-epithelialization was significantly decreased, and the number of live colonies was significantly increased, when compared with those in wild-type (WT) mice on day 7. IL-17A, and IL-22 production was significantly lower in Jα18KO mice than in WT mice on day 5. Furthermore, the administration of α-galactosylceramide (α-GalCer), a specific activator of iNKT cells, led to enhanced host protection, as shown by reduced bacterial load, and to increased production of IL-22, IL-23, and S100A9 compared that of with WT mice. These results suggest that iNKT cells promote P. aeruginosa clearance during skin wound healing.

Osteogenic response under the periosteum by magnesium implantation in rat tibia.

This study was designed to examine osteoconductive effects of Mg in rats tibia. The animals were sacrificed after 1, 2, and 8 weeks. The elemental analysis was performed using SEM/EDX at week 1. Following X-ray micrography at weeks 2 and 8, samples were embedded in paraffin. The expression of osteocalcin was observed by immunohistochemical staining. The element concentrations of fibrous capsules around the specimens were also measured by ICP-MS. The concentrations of Ca and P on the surface of the Mg specimen increased in SEM/EDX. The tissue specimen showed new bone formation on the bone surface near the implanted area. The concentrations of Mg, Ca, and P were high in the fibrous capsules surrounding Mg. Implantation induced differentiation of osteoblasts, and this process was considered to be associated with new bone formation. Induction of cell differentiation may be influenced by corrosion products in addition to corroding magnesium.

Distinct Roles for Dectin-1 and Dectin-2 in Skin Wound Healing and Neutrophilic Inflammatory Responses.

C-type lectin receptors recognize microbial polysaccharides. The C-type lectin receptors such as dendritic cell-associated C-type lectin (Dectin)-1 and Dectin-2, which are triggered by ß-glucan and α-mannan, respectively, contribute to upregulation of the inflammatory response. Recently, we demonstrated that activation of the Dectin-2 signal delayed wound healing; in previous studies, triggering the Dectin-1 signal promoted this response. However, the precise roles of these C-type lectin receptors in skin wound healing remain unclear. This study was conducted to determine the roles of Dectin-1 and Dectin-2 in skin wound healing, with a particular focus on the kinetics of neutrophilic inflammatory response. Full-thickness wounds were created on the backs of C57BL/6 mice, and the effects of Dectin-1 or Dectin-2 deficiency and those of ß-glucan or α-mannan administration were examined. We also analyzed wound closure, histological findings, and neutrophilic inflammatory response, including neutrophil extracellular trap formation at the wound sites. We found that Dectin-1 contributed to the acceleration of wound healing by inducing early-phase neutrophil accumulation, whereas Dectin-2 was involved in prolonged neutrophilic responses and neutrophil extracellular trap formation, leading to delayed wound healing. Dectin-2 deficiency also improved collagen deposition and TGF-ß1 expression. These results suggest that Dectin-1 and Dectin-2 have different roles in wound healing through their different effects on the neutrophilic response.

Congenital Lateral Palatal Synechia Associated With Cleft Palate: A Case Report With Long-Term Follow-Up and Review of the Literature.

Lateral palatal synechiae are rare congenital adhesions running from the free borders of the cleft palate to the lateral parts of the tongue or the oral cavity floor, typically found in cleft palate lateral synechiae syndrome. We present a case of congenital lateral palatal synechia associated with a cleft palate that we treated and followed up for 10 years. We present the long-term prognosis. We also discuss variations in intraoral synechiae associated with cleft palate and the etiology of lateral palatal synechiae through a literature review.

Defect of Interferon γ Leads to Impaired Wound Healing through Prolonged Neutrophilic Inflammatory Response and Enhanced MMP-2 Activation.

Interferon (IFN)-γ is mainly secreted by CD4+ T helper 1 (Th1), natural killer (NK) and NKT cells after skin injury. Although IFN-γ is well known regarding its inhibitory effects on collagen synthesis by fibroblasts in vitro, information is limited regarding its role in wound healing in vivo. In the present study, we analyzed how the defect of IFN-γ affects wound healing. Full-thickness wounds were created on the backs of wild type (WT) C57BL/6 and IFN-γ-deficient (KO) mice. We analyzed the percent wound closure, wound breaking strength, accumulation of leukocytes, and expression levels of COL1A1, COL3A1, and matrix metalloproteinases (MMPs). IFN-γKO mice exhibited significant attenuation in wound closure on Day 10 and wound breaking strength on Day 14 after wound creation, characteristics that are associated with prolonged neutrophil accumulation. Expression levels of COL1A1 and COL3A1 mRNA were lower in IFN-γKO than in WT mice, whereas expression levels of MMP-2 (gelatinase) mRNA were significantly greater in IFN-γKO than in WT mice. Moreover, under neutropenic conditions created with anti-Gr-1 monoclonal antibodies, wound closure in IFN-γKO mice was recovered through low MMP-2 expression levels. These results suggest that IFN-γ may be involved in the proliferation and maturation stages of wound healing through the regulation of neutrophilic inflammatory responses.

Long-term Prognosis of Mandibular Distraction in 3 Cases of Hypoglossia-Hypodactyly Syndrome without Extremity Anomalies.

Hypoglossia-hypodactyly, or aglossia-adactylia syndrome with or without limb anomalies, is an extremely rare congenital condition. It is characterized by a narrow, V-shaped mandibular dental arch with micrognathia, and is typically challenging to treat. We have previously reported 3 patients with hypoglossia-hypodactyly syndrome without limb anomalies who were treated with transverse distraction osteogenesis at the mandibular symphysis during childhood. In this report, we present the long-term prognoses of these 3 cases, until 18 years of age. Of the 3 total cases, sufficient and stable results in occlusion, speech, mastication, and facial appearance were obtained in 2 cases with subsequent orthodontic treatments. Similar results were not observed in the remaining case when continuous orthodontic treatments were not performed. Based on these results, we propose that the transverse distraction osteogenesis procedure at the mandibular symphysis during childhood could be a reliable method to correct the V-shaped mandibular arch of hypoglossia-hypodactyly syndrome, when subsequent orthodontic treatments are provided during the patient's growing period.

The Birth Prevalence of Cleft Lip and/or Cleft Palate After the 2011 Tohoku Earthquake and Tsunami.

OBJECTIVE: This study examined whether the 2011 Tohoku earthquake and tsunami affected the birth prevalence of cleft lip and/or cleft palate (CL/P) in Miyagi Prefecture, where the earthquake and tsunami caused severe damage. DESIGN: This was a retrospective cohort study. SETTING: The study was conducted at university and children's hospitals in Miyagi Prefecture. PARTICIPANTS: The annual and monthly numbers of infants born with CL/P were obtained from medical records. The affected period of birth was defined as 1 year from December 1, 2011, to November 30, 2012. The control period was 5 years from January 1, 2006, to December 31, 2010. The annual and monthly numbers of births in Miyagi Prefecture were obtained using e-Stat, which is a portal for Japanese government statistics. MAIN OUTCOME MEASURES: Main outcome measures were birth prevalence of CL/P during the control and affected periods. RESULTS: There were no significant differences between the control and affected periods in the prevalence (per 10 000 live births) of cleft lip with or without palate (13.8 vs 16.7; P = .342), isolated cleft palate (5.2 vs 3.2; P = .267), or overall CL/P (19.0 vs 19.9; P = .799). CONCLUSIONS: We did not observe that the 2011 Tohoku earthquake and tsunami affected the birth prevalence of CL/P in Miyagi Prefecture, even though it severely impacted human health in the area.

Dectin-2-Mediated Signaling Leads to Delayed Skin Wound Healing through Enhanced Neutrophilic Inflammatory Response and Neutrophil Extracellular Trap Formation.

Dendritic cell-associated C-type lectin-2 (i.e., dectin-2) recognizes fungal polysaccharides, including α-mannan. Dectin-2-mediated recognition of fungi, such as Candida albicans, leads to NF-κB activation, which induces production of inflammatory cytokines. However, the role of dectin-2 in skin wound healing remains unclear. In this study, we sought to determine how dectin-2 deficiency and the administration of α-mannan affected the wound healing process. Full-thickness wounds were created on the backs of wild type C57BL/6 and dectin-2-deficient mice. We analyzed wound closure, histological findings, and re-epithelialization. We also examined the neutrophilic inflammatory responses and neutrophil extracellular trap (NET)-osis at the wound sites after administration of α-mannan. The percent wound closure and re-epithelialization was significantly accelerated in dectin-2-knockout mice compared with wild-type mice on days 3 and 5 after wounding. In contrast, administration of α-mannan delayed wound closure in wild-type mice, and these responses were canceled in dectin-2-knockout mice. Furthermore, mice administered α-mannan, neutrophil infiltration was prolonged, and the expression of citrullinated histone, an indicator of NETosis, at the wound sites was accelerated. Administration of a neutrophil elastase inhibitor significantly improved the delayed wound healing caused by α-mannan. These results suggest that dectin-2 may have a deep impact on the skin wound healing process through regulation of neutrophilic responses.