Age as a decisive factor in general anaesthesia use in paediatric proton beam therapy.

Proton therapy for paediatric cancer patients is an effective treatment; however, young children have may have difficulties staying still during irradiation. This study investigated the indication of general anaesthesia in paediatric proton therapy. Background information and anaesthesia/treatment protocols were retrospectively extracted from the medical records of cancer patients under 15 years who underwent proton therapy at Southern TOHOKU General Hospital, Fukushima, Japan between April 2016 and December 2018. The anaesthesia and non-anaesthesia groups were compared to evaluate factors determining the need for general anaesthesia. Thirty-two patients who received 285 irradiations were analysed. The median age was 5 years old (range: 1-15), and 13 patients (40.6%) were female. Twelve (37.5%) patients received general anaesthesia. In the general anaesthesia group, airway management using a laryngeal mask was performed in 11 patients (91.6%). Patient age was significantly lower in the general anaesthesia group than in the non-anaesthetised group (p < 0.001). Considering all background factors, only age was strongly associated with anaesthesia in the univariate logistic regression model (odds ratio 0.55 [95% confidence interval 0.35-0.86]; P < 0.01). Thus, age is one of the most important factors determining the need for general anaesthesia during proton therapy in children.

Multifaceted Therapeutic Benefits of Factors Derived From Dental Pulp Stem Cells for Mouse Liver Fibrosis.

: Chronic liver injury from various causes often results in liver fibrosis (LF). Although the liver possesses endogenous tissue-repairing activities, these can be overcome by sustained inflammation and excessive fibrotic scar formation. Advanced LF leads to irreversible cirrhosis and subsequent liver failure and/or hepatic cancer. Here, using the mouse carbon tetrachloride (CCl4)-induced LF model, we showed that a single intravenous administration of stem cells derived from human exfoliated deciduous teeth (SHEDs) or of SHED-derived serum-free conditioned medium (SHED-CM) resulted in fibrotic scar resolution. SHED-CM suppressed the gene expression of proinflammatory mediators, such as TNF-α, IL-1ß, and iNOS, and eliminated activated hepatic stellate cells by inducing their apoptosis, but protected parenchymal hepatocytes from undergoing apoptosis. In addition, SHED-CM induced tissue-repairing macrophages that expressed high levels of the profibrinolytic factor, matrix metalloproteinase 13. Furthermore, SHED-CM suppressed the CCl4-induced apoptosis of primary cultured hepatocytes. SHED-CM contained a high level of hepatocyte growth factor (HGF). Notably, HGF-depleted SHED-CM (dHGF-CM) did not suppress the proinflammatory response or resolve fibrotic scarring. Furthermore, SHED-CM, but not dHGF-CM, inhibited CCl4-induced hepatocyte apoptosis. These results suggest that HGF plays a central role in the SHED-CM-mediated resolution of LF. Taken together, our findings suggest that SHED-CM provides multifaceted therapeutic benefits for the treatment of LF. SIGNIFICANCE: This study demonstrated that a single intravenous administration of stem cells from human exfoliated deciduous teeth (SHEDs) or of the serum-free conditioned medium (CM) derived from SHEDs markedly improved mouse liver fibrosis (LF). SHED-CM suppressed chronic inflammation, eliminated activated hepatic stellate cells by inducing their apoptosis, protected hepatocytes from undergoing apoptosis, and induced differentiation of tissue-repairing macrophages expressing high levels of the profibrinolytic factor matrix metalloproteinase 13. Furthermore, hepatocyte growth factor played a central role in the SHED-CM-mediated resolution of LF. This is the first report demonstrating the multifaceted therapeutic benefits of secreted factors derived from SHEDs for LF.

Conditioned Medium from the Stem Cells of Human Exfoliated Deciduous Teeth Ameliorates Experimental Autoimmune Encephalomyelitis.

Multiple sclerosis (MS) is a major neuroinflammatory demyelinating disease of the CNS. Current MS treatments, including immunomodulators and immunosuppressants, do not result in complete remission. Stem cells from human exfoliated deciduous teeth (SHEDs) are mesenchymal stem cells derived from dental pulp. Both SHED and SHED-conditioned medium (SHED-CM) exhibit immunomodulatory and regenerative activities and have the potential to treat various diseases. In this study, we investigated the efficacy of SHED-CM in treating experimental autoimmune encephalomyelitis (EAE), a mouse model of MS. EAE mice treated with a single injection of SHED-CM exhibited significantly improved disease scores, reduced demyelination and axonal injury, and reduced inflammatory cell infiltration and proinflammatory cytokine expression in the spinal cord, which was associated with a shift in the microglia/macrophage phenotype from M1 to M2. SHED-CM also inhibited the proliferation of myelin oligodendrocyte glycoprotein-specific CD4(+) T cells, as well as their production of proinflammatory cytokines in vitro. Treatment of EAE mice with the secreted ectodomain of sialic acid-binding Ig-like lectin-9, a major component of SHED-CM, recapitulated the effects of SHED-CM treatment. Our data suggest that SHED-CM and secreted ectodomain of sialic acid-binding Ig-like lectin-9 may be novel therapeutic treatments for autoimmune diseases, such as MS.

[A Case of Anaphylaxis during Anesthesia Caused Twice by Rocuronium Bromide.]

A 49-year-old woman was scheduled for endoscopic cholecystectomy under general anesthesia in another hospital. The anesthesia was performed by a surgeon, but the operation was cancelled because of anaphylac- tic shock, and the surgeon performed emergency treatment Afterward, the surgeon declined operation, and she could not receive medical treatment. She visited our hospital for complete examination and 'operation. Drug-induced lymphocyte stimulation test (DLST) and standard perioperative test results were almost normal, and we could not find the cause of anaphylaxis preoperatively. After induction of anesthesia, the erythema ap- peared with hypotension and tachycardia. She was responsive to symptomatic treatment such as transfu- sion, antihistamine agent and streroid administration. After the recovery from the shock state, the operation proceeded without complications and she recovered from anesthesia uneventfully. She had no major post- operative complication.

Conditioned medium from the stem cells of human dental pulp improves cognitive function in a mouse model of Alzheimer's disease.

Alzheimer's disease (AD) is a progressive, neurodegenerative disease characterized by a decline in cognitive abilities and the appearance of ß-amyloid plaques in the brain. Although the pathogenic mechanisms associated with AD are not fully understood, activated microglia releasing various neurotoxic factors, including pro-inflammatory cytokines and oxidative stress mediators, appear to play major roles. Here, we investigated the therapeutic benefits of a serum-free conditioned medium (CM) derived from the stem cells of human exfoliated deciduous teeth (SHEDs) in a mouse model of AD. The intranasal administration of SHEDs in these mice resulted in substantially improved cognitive function. SHED-CM contained factors involved in multiple neuroregenerative mechanisms, such as neuroprotection, axonal elongation, neurotransmission, the suppression of inflammation, and microglial regulation. Notably, SHED-CM attenuated the pro-inflammatory responses induced by ß-amyloid plaques, and generated an anti-inflammatory/tissue-regenerating environment, which was accompanied by the induction of anti-inflammatory M2-like microglia. Our data suggest that SHED-CM may provide significant therapeutic benefits for AD.

Involvement of glial activation in trigeminal ganglion in a rat model of lower gingival cancer pain.

Glial cells were investigated to elucidate their involvement in mechanisms underlying oral cancer pain. Squamous cell carcinoma (SCC-158) was inoculated into the lower gingiva of male Fisher rats. Pharmacological and immunohistochemical studies were performed to examine the roles played by TRPV1 and TRPV2 expressed in neurons and satellite glia in trigeminal ganglia (TG), and microglia and astrocytes in trigeminal spinal nucleus caudalis. Inoculation of SCC-158 into the lower gingiva induced marked mechanical allodynia in the whisker-pad skin area on days 16 through 28, and in the submandibular skin area on days 10 through 20. Cutaneous allodynia was diminished by systemic morphine administration. The number of TRPV1 and TRPV2-positive neurons in trigeminal ganglia increased in the medium and large cell groups on day 14 after tumor inoculation. The number of satellite glial cells encircling the medium and large trigeminal ganglion neurons increased on day 28 after tumor inoculation. In this gingival cancer pain model, microglia and astrocytes in trigeminal spinal nucleus caudalis were not activated, although they were reported to be activated in neuropathic and inflammatory pain models. These results suggest that TRPV1 and TRPV2 upregulation in trigeminal ganglion neurons may play an important role in inducing the mechanical allodynia observed in experimental models of oral squamous cell carcinoma. In addition, activation of satellite cells seems to be involved in the maintenance of mechanical allodynia, which could be the potential therapeutic target for oral cancer pain.

Effects of the permeability of shields with autologous bone grafts on bone augmentation.

PURPOSE: The objective of this study was to histologically evaluate and compare the effects of the permeability of shields on bone augmentation in a rabbit calvarial model. MATERIALS AND METHODS: Twelve adult male Japanese white rabbits were used for the study. Each received four titanium cylinders, which were placed into perforated slits made in the outer cortical bone of the calvaria and filled with autologous iliac bone. The tops of the cylinders were randomly covered with the following test materials: (1) uncovered (control), (2) a titanium mesh, (3) an expanded polytetrafluoroethylene (e-PTFE) membrane, or (4) a titanium plate. After 8 weeks, the animals were sacrificed, and ground sections were obtained for histomorphometric analysis. RESULTS: There was no significant difference in augmented bone volume among all groups. However, the distribution of augmented bone in the cylinders differed among the groups. In the uncovered control, there was significantly less augmented bone in the upper third of the cylinder than in the middle or lower thirds. Findings were similar for the titanium mesh group and the e-PTFE membrane group, with significantly less augmented bone in the upper third than in the middle or lower thirds. In the titanium plate group, there was no significant difference in augmented bone among the upper, middle, and lower thirds. The differences among the upper, middle, and lower thirds of the cylinder were smaller in the order of titanium plate, e-PTFE membrane, titanium mesh, and uncovered control. CONCLUSION: The use of low-permeability shields resulted in small differences in the distribution of bone structure in the present bone augmentation model.

Effects of self-assembling peptide hydrogel scaffold on bone regeneration with recombinant human bone morphogenetic protein-2.

PURPOSE: The objective of this pilot study was to histologically evaluate bone regeneration using a self-assembling peptide hydrogel scaffold with recombinant human bone morphogenetic protein-2 (rhBMP-2) in a rabbit calvaria model. MATERIALS AND METHODS: Five adult New Zealand White rabbits were used for the study. Each received four titanium cylinders, which were placed into perforated slits made in the outer cortical bone of the calvaria. The cylinders were filled with the following test materials: (1) unfilled control; (2) rhBMP-2; (3) PuraMatrix (PM), a synthetic self-assembling peptide (RADA16-I) consisting of a 16-amino acid sequence and with a three-dimensional structure; and (4) PM/rhBMP-2. Each cylinder was covered with a titanium lid. After 8 weeks, the animals were sacrificed, and ground sections were obtained for histomorphometric analysis. RESULTS: Histomorphometric analysis showed that regenerated tissue in the cylinder with PM/rhBMP-2 was significantly increased compared to the empty control. The mean area values of regenerated tissue in the cylinders were 35.80% ± 10.35% (control), 47.94% ± 5.65% (rhBMP-2), 48.94% ± 11.33% (PM), and 58.06% ± 14.84% (PM/rhBMP-2). The mean area values of newly formed bone in the cylinders were 9.39% ± 4.34% (control), 14.03% ± 2.25% (rhBMP-2), 13.99% ± 2.15% (PM), and 16.61% ± 3.79% (PM/rhBMP-2). Neither rhBMP-2 nor PM alone significantly enhanced bone regeneration compared to the empty control cylinder. CONCLUSIONS: PM with rhBMP-2 significantly enhanced bone regeneration on the bone augmentation model in a rabbit. PM promises to be a useful alternative synthetic material as a carrier for rhBMP-2 for bone regeneration.

Stem cells from human exfoliated deciduous tooth-derived conditioned medium enhance recovery of focal cerebral ischemia in rats.

Regenerative therapy using stem cells is a promising approach for the treatment of stroke. Recently, we reported that dental pulp stem cells (DPSC) ameliorated ischemic tissue injury in the rat brain and accelerated functional recovery after middle cerebral artery occlusion (MCAO). In this study, we investigated the effects of stem cells from human exfoliated deciduous tooth (SHED)-derived conditioned medium (SHED-CM) on permanent MCAO (pMCAO). Adult male Sprague-Dawley rats were subjected to pMCAO. SHED-CM were then administered intranasally, and the motor function and infarct volume were evaluated. Neurogenesis and vasculogenesis were determined using immunochemical markers. The SHED-CM group had more positive signals than the Dulbecco's modified Eagle's medium group, with doublecortin (DCX), neurofilament H, neuronal nuclei, and rat endothelial cell antigen observed in the peri-infarct area. Migration of neuronal progenitor cells (NPC) with DCX from the subventricular zone to the peri-infarct area was observed on days 6 and 16, with migration on day 6 being the most prominent. In conclusion, SHED-CM promoted the migration and differentiation of endogenous NPC, induced vasculogenesis, and ameliorated ischemic brain injury after pMCAO as well as transplantation of DPSC.

Lewis y antigen is expressed in oral squamous cell carcinoma cell lines and tissues, but disappears in the invasive regions leading to the enhanced malignant properties irrespective of sialyl-Lewis x.

Expression and implication of carbohydrate antigens in squamous cell carcinomas (SCCs) in oral cavity was examined. In the cell lines, type 2H and Lewis y antigens were markedly expressed. In the tissues from SCC patients and benign disorders, type 2H was highly expressed in hyperplasia (96.4 %), displasia (92.9 %) and SCC (100 %). Lewis y was, in turn, expressed mainly in cancer tissues (91.3 %), suggesting that Lewis y is a cancer-associated antigen. Normal oral mucosa showed no expression of these blood group antigens. Surprisingly, Lewis y antigen disappeared in the invasion sites where Ki-67 was definitely stained. Over-expression of Lewis y with manipulation of a fucosyltransferase cDNA resulted in suppression of cell growth and invasion, and knockdown of Lewis y also brought about increased cell growth and invasion. In either situations, no changes in the expression of sialyl-Lewis x could be found. Lowered tumor growth and invasion into surrounding tissues were also shown in Lewis y-positive SCC grafts in nu/nu mice. All these results together with alternative staining between Lewis y and Ki-67 in cancer tissues and FUT1 transfectants suggested that loss of Lewis y is a crucial event for the late stage of SCCs.

Enhancement of malignant properties of human osteosarcoma cells with disialyl gangliosides GD2/GD3.

The expression and implications of gangliosides in human osteosarcomas have not been systematically analyzed. In this study, we showed that gangliosides GD3 and GD2 are highly expressed in the majority of human osteosarcoma cell lines derived from oral cavity regions. Introduction of GD3 synthase cDNA into a GD3/GD2-negative (GD3/GD2-) human osteosarcoma subline resulted in the establishment of GD3/GD2+ transfectant cells. They showed increased cell migration and invasion activities in wound healing and Boyden chamber invasion assays, respectively, compared to the control cells. When treated with serum, GD3/GD2+ cells showed stronger tyrosine phosphorylation of p130Cas, focal adhesion kinase, and paxillin than GD3/GD2- cells. In particular, paxillin underwent much stronger phosphorylation, suggesting its role in cell motility. Furthermore, we tried to dissect the roles of GD3 and GD2 in the malignant properties of the transfectant cells by establishing single ganglioside-expressing cells, that is, either GD3 or GD2. Although GD3/GD2+ cells showed the most malignant properties, GD2+ cells showed almost equivalent levels to GD3/GD2+ cells in invasion and migration activities, and in the intensities of tyrosine phosphorylation of paxillin. Among Src family kinases, Lyn was expressed predominantly, and was involved in the invasion and motility of GD3- and/or GD2-expressing transfectants. Furthermore, it was elucidated by gene silencing that Lyn was located in a different pathway from that of FAK to eventually lead paxillin activation. These results suggested that GD2/GD3 are responsible for the enhancement of the malignant features of osteosarcomas, and might be candidate targets in molecular-targeted therapy.

Functional activation of Src family kinase yes protein is essential for the enhanced malignant properties of human melanoma cells expressing ganglioside GD3.

The possible roles of Src family kinases in the enhanced malignant properties of melanomas related to GD3 expression were analyzed. Among Src family kinases only Yes, not Fyn or Src, was functionally involved in the increased cell proliferation and invasion of GD3-expressing transfectant cells (GD3+). Yes was located upstream of p130Cas and paxillin and at an equivalent level to focal adhesion kinase. Yes underwent autophosphorylation even before serum treatment and showed stronger kinase activity in GD3+ cells than in GD3- cells following serum treatment. Coimmunoprecipitation experiments revealed that Yes bound to focal adhesion kinase or p130Cas more strongly in GD3+ cells than in GD3- cells. As a possible mechanism for the enhancing effects of GD3 on cellular phenotypes, it was shown that majority of Yes was localized in glycolipid-enriched microdomain/rafts in GD3+ cells even before serum treatment, whereas it was scarcely detected in glycolipid-enriched microdomain/rafts in GD3- cells. An in vitro kinase assay of Yes revealed that coexistence of GD3 with Yes in membranous environments enhances the kinase activity of GD3- cell-derived Yes toward enolase, p125, and Yes itself. Knockdown of GD3 synthase resulted in the alleviation of tumor phenotypes and reduced activation levels of Yes. Taken together, these results suggest a role of GD3 in the regulation of Src family kinases.

Dental pulp-derived CD31⁻/CD146⁻ side population stem/progenitor cells enhance recovery of focal cerebral ischemia in rats.

Regenerative therapy using stem cells is a promising approach for the treatment of stroke. Recently, we reported that CD31⁻/CD146⁻ side population (SP) cells from porcine dental pulp exhibit highly vasculogenic potential in hindlimb ischemia. In this study, we investigated the influence of CD31⁻/CD146⁻ SP cells after transient middle cerebral artery occlusion (TMCAO). Adult male Sprague-Dawley rats were subjected to 2 h of TMCAO. Twenty-four hours after TMCAO, CD31⁻/CD146⁻ SP cells were transplanted into the brain. Motor function and infarct volume were evaluated. Neurogenesis and vasculogenesis were determined with immunochemical markers, and the levels of neurotrophic factors were assayed with real-time reverse transcription-polymerase chain reaction. In the cell transplantation group, the number of doublecortin-positive cells increased twofold, and the number of NeuN-positive cells increased eightfold, as compared with the control phosphate-buffered saline group. The vascular endothelial growth factor level in the ischemic brain with transplanted cells was 28 times higher than that in the normal brain. In conclusion, CD31⁻/CD146⁻ SP cells promoted migration and differentiation of the endogenous neuronal progenitor cells and induced vasculogenesis, and ameliorated ischemic brain injury after TMCAO.

Stability of a locking plate and self-drilling screws as orthodontic skeletal anchorage in the maxilla: a retrospective study.

PURPOSE: The aim of this retrospective study was to determine the success rate of an orthodontic skeletal anchorage system consisting of a locking plate and 2 self-drilling screws to intrude the upper molars and detect factors that contribute to its stability. PATIENTS AND METHODS: The subjects were 32 orthodontic and generally healthy patients who had skeletal anchorage plates placed supraperiosteally and unilaterally or bilaterally. The anchorage plate was considered successful if the plate remained stable throughout the period of intrusion of the upper molar without any movement, persistent pain, or infection and was then retrieved without difficulty. The success rates of the anchorage plate were statistically analyzed on the basis of clinically categorized variables. RESULTS: The 32 patients comprised 6 male and 26 female individuals with ages ranging from 11.4 to 35.1 years. The overall success rate of the total 61 plates was 93.4%. No significant differences were observed among the respective success rates analyzed in accordance with gender, age, side of placement, and length of the screws. The thickness of the bony walls that supported the screws was significantly greater in the success group (mean 1.6 +/- SD, 0.2 vs 1.0 +/- 0.1 mm, P < .001). CONCLUSION: Bone thickness is a critical factor in supporting the self-drilling screws and locking plate. Skeletal anchorage combining the plate and 2 screws promises a higher success rate with a thicker bone than with the threshold value of thickness that exists within the 1.1 to 1.4 mm range in the maxillary walls.