Bacillus subtilis spore vaccines displaying protective antigen induce functional antibodies and protective potency.

BACKGROUND: Bacillus anthracis is the causative agent of anthrax, a disease of both humans and various animal species, and can be used as a bioterror agent. Effective vaccines are available, but those could benefit from improvements, including increasing the immunity duration, reducing the shot frequency and adverse reactions. In addition, more sophisticated antigen delivery and potentiation systems are urgently required. The protective antigen (PA), one of three major virulence factors associated with anthrax was displayed on the surface of Bacillus subtilis spores, which is a vaccine production host and delivery vector with several advantages such as a low production cost, straightforward administration as it is safe for human consumption and the particulate adjuvanticity. Mice were immunized orally (PO), intranasally (IN), sublingually (SL) or intraperitoneally (IP) with the PA displaying probiotic spore vaccine. Clinical observation, serological analysis and challenge experiment were conducted to investigate the safety and efficacy of the vaccine. RESULTS: A/J mice immunized with the PA spore vaccine via PO, IN, SL, and IP were observed to have increased levels of active antibody titer, isotype profiles and toxin neutralizing antibody in sera, and IgA in saliva. The immunized mice were demonstrated to raise protective immunity against the challenge with lethal B. anthracis spores. CONCLUSIONS: In this study, we developed a B. subtilis spore vaccine that displays the PA on its surface and showed that the PA-displaying spore vaccine was able to confer active immunity to a murine model based on the results of antibody isotype titration, mucosal antibody identification, and a lethal challenge experiment.

Effects of 3D-Printed Polycaprolactone/ß-Tricalcium Phosphate Membranes on Guided Bone Regeneration.

This study was conducted to compare 3D-printed polycaprolactone (PCL) and polycaprolactone/ß-tricalcium phosphate (PCL/ß-TCP) membranes with a conventional commercial collagen membrane in terms of their abilities to facilitate guided bone regeneration (GBR). Fabricated membranes were tested for dry and wet mechanical properties. Fibroblasts and preosteoblasts were seeded into the membranes and rates and patterns of proliferation were analyzed using a kit-8 assay and by scanning electron microscopy. Osteogenic differentiation was verified by alizarin red S and alkaline phosphatase (ALP) staining. An in vivo experiment was performed using an alveolar bone defect beagle model, in which defects in three dogs were covered with different membranes. CT and histological analyses at eight weeks after surgery revealed that 3D-printed PCL/ß-TCP membranes were more effective than 3D-printed PCL, and substantially better than conventional collagen membranes in terms of biocompatibility and bone regeneration and, thus, at facilitating GBR.

Vital signal sensing and manipulation of a microscale organ with a multifunctional soft gripper.

Soft grippers that incorporate functional materials are important in the development of mechanically compliant and multifunctional interfaces for both sensing and stimulating soft objects and organisms. In particular, the capability for firm and delicate grasping of soft cells and organs without mechanical damage is essential to identify the condition of and monitor meaningful biosignals from objects. Here, we report a millimeter-scale soft gripper based on a shape memory polymer that enables manipulating a heavy object (payload-to-weight ratio up to 6400) and grasping organisms at the micro/milliscale. The silver nanowires and crack-based strain sensor embedded in this soft gripper enable simultaneous measurement of the temperature and pressure on grasped objects and offer temperature and mechanical stimuli for the grasped object. We validate our miniaturized soft gripper by demonstrating that it can grasp a snail egg while simultaneously applying a moderate temperature stimulation to induce hatching process and monitor the heart rate of a newborn snail. The results present the potential for widespread utility of soft grippers in the area of biomedical engineering, especially in the development of conditional or closed-loop interfacing with microscale biotissues and organisms.

Synergistic Effects of Beta Tri-Calcium Phosphate and Porcine-Derived Decellularized Bone Extracellular Matrix in 3D-Printed Polycaprolactone Scaffold on Bone Regeneration.

Bone-derived extracellular matrix (ECM) is widely used in studies on bone regeneration because of its ability to provide a microenvironment of native bone tissue. However, a hydrogel, which is a main type of ECM application, is limited to use for bone graft substitutes due to relative lack of mechanical properties. The present study aims to fabricate a scaffold for guiding effective bone regeneration. A polycaprolactone (PCL)/beta-tricalcium phosphate (ß-TCP)/bone decellularized extracellular matrix (dECM) scaffold capable of providing physical and physiological environment are fabricated using 3D printing technology and decoration method. PCL/ß-TCP/bone dECM scaffolds exhibit excellent cell seeding efficiency, proliferation, and early and late osteogenic differentiation capacity in vitro. In addition, outstanding results of bone regeneration are observed in PCL/ß-TCP/bone dECM scaffold group in the rabbit calvarial defect model in vivo. These results indicate that PCL/ß-TCP/bone dECM scaffolds have an outstanding potential as bone graft substitutes for effective bone regeneration.

Efficacy of rhBMP-2 Loaded PCL/ß-TCP/bdECM Scaffold Fabricated by 3D Printing Technology on Bone Regeneration.

This study was undertaken to evaluate the effect of 3D printed polycaprolactone (PCL)/ß-tricalcium phosphate (ß-TCP) scaffold containing bone demineralized and decellularized extracellular matrix (bdECM) and human recombinant bone morphogenetic protein-2 (rhBMP-2) on bone regeneration. Scaffolds were divided into PCL/ß-TCP, PCL/ß-TCP/bdECM, and PCL/ß-TCP/bdECM/BMP groups. In vitro release kinetics of rhBMP-2 were determined with respect to cell proliferation and osteogenic differentiation. These three reconstructive materials were implanted into 8 mm diameter calvarial bone defect in male Sprague-Dawley rats. Animals were sacrificed four weeks after implantation for micro-CT, histologic, and histomorphometric analyses. The findings obtained were used to calculate new bone volumes (mm3) and new bone areas (%). Excellent cell bioactivity was observed in the PCL/ß-TCP/bdECM and PCL/ß-TCP/bdECM/BMP groups, and new bone volume and area were significantly higher in the PCL/ß-TCP/bdECM/BMP group than in the other groups (p < .05). Within the limitations of this study, bdECM printed PCL/ß-TCP scaffolds can reproduce microenvironment for cells and promote adhering and proliferating the cells onto scaffolds. Furthermore, in the rat calvarial defect model, the scaffold which printed rhBMP-2 loaded bdECM stably carries rhBMP-2 and enhances bone regeneration confirming the possibility of bdECM as rhBMP-2 carrier.

Efficient digitalization method for dental restorations using micro-CT data.

The objective of this study was to demonstrate the feasibility of using micro-CT scan of dental impressions for fabricating dental restorations and to compare the dimensional accuracy of dental models generated from various methods. The key idea of the proposed protocol is that dental impression of patients can be accurately digitized by micro-CT scan and that one can make digital cast model from micro-CT data directly. As air regions of the micro-CT scan data of dental impression are equivalent to the real teeth and surrounding structures, one can segment the air regions and fabricate digital cast model in the STL format out of them. The proposed method was validated by a phantom study using a typodont with prepared teeth. Actual measurement and deviation map analysis were performed after acquiring digital cast models for each restoration methods. Comparisons of the milled restorations were also performed by placing them on the prepared teeth of typodont. The results demonstrated that an efficient fabrication of precise dental restoration is achievable by use of the proposed method.

Endodontic Treatment of an Anomalous Anterior Tooth with the Aid of a 3-dimensional Printed Physical Tooth Model.

Endodontic treatment of tooth formation anomalies is a challenge to clinicians and as such requires a complete understanding of the aberrant root canal anatomy followed by careful root canal disinfection and obturation. Here, we report the use of a 3-dimensional (3D) printed physical tooth model including internal root canal structures for the endodontic treatment of a challenging tooth anomaly. A 12-year-old boy was referred for endodontic treatment of tooth #8. The tooth showed class II mobility with swelling and a sinus tract in the buccal mucosa and periapical radiolucency. The tooth presented a very narrow structure between the crown and root by distal concavity and a severely dilacerated root. Moreover, a perforation site with bleeding and another ditching site were identified around the cervical area in the access cavity. A translucent physical tooth model carrying the information on internal root canal structures was built through a 3-step process: data acquisition by cone-beam computed tomographic scanning, virtual modeling by image processing, and manufacturing by 3D printing. A custom-made guide jig was then fabricated to achieve a safe and precise working path to the root canal. Endodontic procedures including access cavity preparation were performed using the physical tooth model and the guide jig. At the 7-month follow-up, the endodontically treated tooth showed complete periapical healing with no clinical signs and symptoms. This case report describes a novel method of endodontic treatment of an anomalous maxillary central incisor with the aid of a physical tooth model and a custom-made guide jig via 3D printing technique.

Forgotten denture in a hemiplegic patient.

A 74-yr-old man developed left hemiplegia because of a right middle cerebral artery territory infarction and also had clinical features of dysphagia and speech difficulty. At that time, he complained of neck pain, but the symptom was ignored because he had a nasogastric tube and had been diagnosed with a huge epiglottic cyst that had already shown several symptoms such as severe hoarseness and throat discomfort. A videofluoroscopic swallowing study was planned to find out the cause and the type of dysphagia. On the videofluoroscopic swallowing study, a foreign body was found at the hypopharynx. Surprisingly, it was confirmed as a denture. After removing the denture, the patient's swallowing and speech difficulty were significantly improved. This case emphasizes the need for elderly stoke patients presenting with dysphagia or communication problems to receive more careful history taking and a more complete physical examination, with the cooperation of several clinical departments.

A case of hypersensitivity pneumonitis with giant cells in a female dental technician.

OBJECTIVES: Dental technicians are exposed to methyl methacrylate(MMA) and hard metal dusts while working, and several cases of hypersensitivity pneumonitis caused by the exposure have been reported. The authors experienced a case of hypersensitivity pneumonitis in a female dental technician who had 10 years' work experience and report the case with clinical evidence. METHOD: The patient's work, personal, social, and past and present medical histories were investigated based on patient questioning and medical records. Furthermore, the workplace conditions and tools and materials the patient worked with were also evaluated. Next, the pathophysiology and risk factors of pneumonitis were studied, and studies on the relationship between hypersensitivity pneumonitis and a dental technician's exposure to dust were reviewed. Any changes in the clinical course of her disease were noted for evaluation of the work-relatedness of the disease. RESULTS: The patient complained of cough and sputum for 1 year. In addition, while walking up the stairs, the patient was not able to ascend without resting due to dyspnea. She visited our emergency department due to epistaxis, and secondary hypertension was incidentally suspected. Laboratory tests including serologic, electrolyte, and endocrinologic tests and a simple chest radiograph showed no specific findings, but chest computed tomography revealed a centrilobular ground-glass pattern in both lung fields. A transbronchial biopsy was performed, and bronchoalveolar washing fluid was obtained. Among the findings of the laboratory tests, microcalcification, noncaseating granuloma containing foreign body-type giant cells, and metal particles within macrophages were identified histologically. Based on these results, hypersensitivity pneumonitis was diagnosed. The patient stopped working due to admission, and she completely quit her job within 2 months of restarting work due to reappearance of the symptoms. CONCLUSION: In this study, the patient did not have typical radiologic findings, but pathological evaluation of the lung biopsy from the bronchoscope led to the suspicion of pneumonitis. Under the microscope, the sample contained fibrotic changes in the lung, multinucleated giant cells, and particles in macrophages and was diagnosed as dental technician pneumoconiosis by the pathology. Working as a dental technician had directly exposed her to light metal dust and MMA, and her clinical symptoms and radiologic findings subsided after withdrawal from exposure to the workplace. These outcomes led to the diagnosis of hypersensitity pneumonitis due to MMA exposure and strong work-relatedness.