Non-touch, Quick Removal of an Occluding Intratracheal Balloon Using High Intensity Focused Ultrasound and Limonene Emulsion

In recent years, fetal endoscopic tracheal occlusion （FETO） using a balloon has been clinically employed for promoting prenatal lung growth to ameliorate postnatal respiratory failure caused by severe in utero lung hypoplasia. After a successful FETO, in some limited fetal centers, planned or emergency balloon removal using another fetoscopy is performed to release the tracheal occlusion immediately before delivery. To overcome this additional fetoscopy, we previously reported an innovative simple procedure to remove the occluding balloon by bursting it with a pre-planned sequence of high intensity focused ultrasound （HIFU） irradiation. In that previous study, which used rabbits euthanized and submerged in degassed water, we inflated the balloon by injecting a mixture of perfluorocarbon and ultrasound contrast medium through a fetoscopically-guided catheter. The rate of successful balloon burst and deflation using HIFU irradiation was high enough （100％）, but the mode and timing of tracheal reopening （i.e., sudden burst or slow shrinkage of the balloon） was rather varied and collateral damage to the dermal/tracheal tissue was identified in 72.7％ of the experimental animals. Accordingly, to standardize the HIFU irradiation sequence and to achieve a reliable and secure balloon burst, we conducted another series of animal experiments in which the mixture of perfluorocarbon was replaced with “limonene emulsion” （D-limonene micelle emulsified in physiological saline） as the balloon injection fluid. In all 6 animals, we succeeded in reopening the airway by achieving an instantaneous and timely balloon burst without definite skin/tracheal damage. We conclude that HIFU irradiation together with injecting the balloon with “limonene emulsion” is an improved method for safe tracheal release from a balloon occlusion

Piezo1-pannexin-1-P2X3 axis in odontoblasts and neurons mediates sensory transduction in dentinal sensitivity

According to the “hydrodynamic theory,” dentinal pain or sensitivity is caused by dentinal fluid movement following the application of various stimuli to the dentin surface. Recent convergent evidence in Vitro has shown that plasma membrane deformation, mimicking dentinal fluid movement, activates mechanosensitive transient receptor potential (TRP)/Piezo channels in odontoblasts, with the Ca2+ signal eliciting the release of ATP from pannexin-1 (PANX-1). The released ATP activates the P2X3 receptor, which generates and propagates action potentials in the intradental Aδ afferent neurons. Thus, odontoblasts act as sensory receptor cells, and odontoblast-neuron signal communication established by the TRP/Piezo channel-PANX-1-P2X3 receptor complex may describe the mechanism of the sensory transduction sequence for dentinal sensitivity. To determine whether odontoblast-neuron communication and odontoblasts acting as sensory receptors are essential for generating dentinal pain, we evaluated nociceptive scores by analyzing behaviors evoked by dentinal sensitivity in conscious Wistar rats and Cre-mediated transgenic mouse models. In the dentin-exposed group, treatment with a bonding agent on the dentin surface, as well as systemic administration of A-317491 (P2X3 receptor antagonist), mefloquine and 10PANX (non-selective and selective PANX-1 antagonists), GsMTx-4 (selective Piezo1 channel antagonist), and HC-030031 (selective TRPA1 channel antagonist), but not HC-070 (selective TRPC5 channel antagonist), significantly reduced nociceptive scores following cold water (0.1 ml) stimulation of the exposed dentin surface of the incisors compared to the scores of rats without local or systemic treatment. When we applied cold water stimulation to the exposed dentin surface of the lower first molar, nociceptive scores in the rats with systemic administration of A-317491, 10PANX, and GsMTx-4 were significantly reduced compared to those in the rats without systemic treatment. Dentin-exposed mice, with somatic odontoblast-specific depletion, also showed significant reduction in the nociceptive scores compared to those of Cre-mediated transgenic mice, which did not show any type of cell deletion, including odontoblasts. In the odontoblast-eliminated mice, P2X3 receptor-positive A-neurons were morphologically intact. These results indicate that neurotransmission between odontoblasts and neurons mediated by the Piezo1/TRPA1-pannexin-1-P2X3 receptor axis is necessary for the development of dentinal pain. In addition, odontoblasts are necessary for sensory transduction to generate dentinal sensitivity as mechanosensory receptor cells

Alveolar bone grafting in association with polyostotic fibrous dysplasia and bisphosphonate-induced abnormal bone turnover in a bilateral cleft lip and palate patient: a case report

A case is presented of extensive alveolar bone grafting in a patient with bilateral cleft lip and palate and polyostotic fibrous dysplasia. The patient previously underwent bisphosphonate therapy. Because of an abnormal and often decreased bone turnover caused by the fibrous dysplasia and the bisphosphonate therapy, bone grafting in such a patient poses several potential difficulties. In addition, the histomorphometric analysis of the bone grafts showed markedly decreased bone turnover. However, alveolar bone grafting using the iliac crest was performed successfully. Sufficient occlusion was achieved by postoperative low-loading orthodontic treatment

A New Regenerative Approach to Fetal Myelomeningocele by Cell Sheet Transplantation

Myelomeningocele (MMC) is the most common form of congenital neural tube defect. Current fetal surgical repair performed to prevent the exposed spinal cord from being injured until delivery cannot reverse those injuries already inflicted in utero. “Cell sheet” technology has been adopted successfully for the regeneration of diverse organs and tissues, although this promising modality has not yet been used for fetal therapy. This study thus tested our hypothesis that fetal MMC tissue histologically injured in utero could be regenerated using cell sheet technology. We used the L6 myoblast cell line derived from rat skeletal muscle for the cell sheet engineering. A fetal MMC model was also obtained from pregnant Sprague-Dawley (SD) rats fed orally with retinoic acid (60mg/kg, embryonic day 10). Cell sheets were then transplanted onto the fetal MMC lesion (embryonic day 19) following maternal anesthesia, laparotomy, and hysterotomy. The incisional wound of the uterus was kept open for 4 hours under anesthesia with the MMC lesion maintained outside the body with the transplanted cell sheet. Subsequently, the experimental fetuses were sacrificed for histological (HE stain) and immunohistochemical studies to evaluate viability and differentiation potential based on cell markers of the transplanted L6 myoblasts. Immunohistochemical studies clearly demonstrated cell-sheet markers specific to neurons, skeletal muscle, and myoblasts within the treated MMC lesions, confirming that the cell sheet was biologically implanted within 4 hours after the procedure. Cell sheet technology could be useful for intrauterine regeneration of the fetal rat spinal cord injured by an associated MMC

Functional expression of TRPM8 and TRPA1 channels in rat odontoblasts.

Odontoblasts produce dentin during development, throughout life, and in response to pathological conditions by sensing stimulation of exposed dentin. The functional properties and localization patterns of transient receptor potential (TRP) melastatin subfamily member 8 (TRPM8) and ankyrin subfamily member 1 (TRPA1) channels in odontoblasts remain to be clarified. We investigated the localization and the pharmacological, biophysical, and mechano-sensitive properties of TRPM8 and TRPA1 channels in rat odontoblasts. Menthol and icilin increased the intracellular free Ca(2+) concentration ([Ca(2+)]i). Icilin-, WS3-, or WS12-induced [Ca(2+)]i increases were inhibited by capsazepine or 5-benzyloxytriptamine. The increase in [Ca(2+)]i elicited by allyl isothiocyanate (AITC) was inhibited by HC030031. WS12 and AITC exerted a desensitizing effect on [Ca(2+)]i increase. Low-temperature stimuli elicited [Ca(2+)]i increases that are sensitive to both 5-benzyloxytriptamine and HC030031. Hypotonic stimulation-induced membrane stretch increased [Ca(2+)]i; HC030031 but not 5-benzyloxytriptamine inhibited the effect. The results suggest that TRPM8 channels in rat odontoblasts play a role in detecting low-temperature stimulation of the dentin surface and that TRPA1 channels are involved in sensing membrane stretching and low-temperature stimulation. The results also indicate that odontoblasts act as mechanical and thermal receptor cells, detecting the stimulation of exposed dentin to drive multiple cellular functions, such as sensory transduction

FGFR3 down-regulates PTH/PTHrP receptor gene expression by mediating JAK/STAT signaling in chondrocytic cell line

The signaling axis comprised by the parathyroid hormone (PTH)-related peptide (PTHrP), the PTH/PTHrP receptor and the fibroblast growth factor receptor 3 (FGFR3) plays a central role in chondrocyte proliferation. The Indian hedgehog (IHH) gene is normally expressed in early hypertrophic chondrocytes, and its negative feedback loop was shown to regulate PTH/PTHrP receptor signaling. In this study, we examined the regulation of PTH/PTHrP receptor gene expression in a FGFR3-transfected chondrocytic cell line, CFK2. Expression of IHH could not be verified on these cells, with consequent absence of hypertrophic differentiation. Also, expression of the PTH/PTHrP receptor (75% reduction of total mRNA) and the PTHrP (50% reduction) genes was reduced in CFK2 cells transfected with FGFR3 cDNA. Interestingly, we verified significant reduction in cell growth and increased apoptosis in the transfected cells. STAT1 was detected in the nuclei of the CFK2 cells transfected with FGFR3 cDNA, indicating predominance of the JAK/STAT signaling pathway. The reduction in PTH/PTHrP receptor gene in CFK2 cells over-expressing FGFR3 was partially blocked by treatment with an inhibitor of JAK3 (WHI-P131), but not with an inhibitor of MAPK (SB203580) or JAK2 (AG490). Altogether, these findings suggest that FGFR3 down-regulates PTH/PTHrP receptor gene expression via the JAK/STAT signaling in chondrocytic cells