Sustained Exosome-Guided Macrophage Polarization Using Hydrolytically Degradable PEG Hydrogels for Cutaneous Wound Healing: Identification of Key Proteins and MiRNAs, and Sustained Release Formulation.

Macrophages (Mφs) are characterized by remarkable plasticity, an essential component of chronic inflammation. Thus, an appropriate and timely transition from proinflammatory (M1) to anti-inflammatory (M2) Mφs during wound healing is vital to promoting resolution of acute inflammation and enhancing tissue repair. Herein, exosomes derived from M2-Mφs (M2-Exos), which contain putative key regulators driving Mφ polarization, are used as local microenvironmental cues to induce reprogramming of M1-Mφs toward M2-Mφs for effective wound management. As an injectable controlled release depot for exosomes, hydrolytically degradable poly(ethylene glycol) (PEG) hydrogels (Exogels) are designed and employed for encapsulating M2-Exos to maximize their therapeutic effects in cutaneous wound healing. The degradation time of the hydrogels is adjustable from 6 days or up to 27 days by controlling the crosslinking density and tightness. The localization of M2-Exos leads to a successful local transition from M1-Mφs to M2-Mφs within the lesion for more than 6 days, followed by enhanced therapeutic effects including rapid wound closure and increased healing quality in an animal model for cutaneous wound healing. Collectively, the hydrolytically degradable PEG hydrogel-based exosome delivery system may serve as a potential tool in regulating local polarization state of Mφs, which is crucial for tissue homeostasis and wound repair.

Ultrathin Nanostructured Films of Hyaluronic Acid and Functionalized ß-Cyclodextrin Polymer Suppress Bacterial Infection and Capsular Formation of Medical Silicone Implants.

A type of ultrathin films has been developed for suppressing capsule formation induced by medical silicone implants and hence reducing the inflammation response to such formation and the differentiation to myofibroblasts. The films were each fabricated from hyaluronic acid (HA) and modified ß-cyclodextrin (Mod-ß-CyD) polymer which was synthesized with a cyclodextrin with partially substituted quaternary amine. Ultrathin films comprising HA and Mod-ß-CyD or poly(allylamine hydrochloride) (PAH) were fabricated by using a layer-by-layer dipping method. The electrostatic interactions produced from the functional groups of Mod-ß-CyD and HA influenced the surface morphology, wettability, and bio-functional activity of the film. Notably, medical silicone implants coated with PAH/HA and Mod-ß-CyD multilayers under a low pH condition exhibited excellent biocompatibility and antibiofilm and anti-inflammation properties. Implantation of these nanoscale film-coated silicones showed a reduced capsular thickness as well as reduced TGFß-SMAD signaling, myofibroblast differentiation, biofilm formation, and inflammatory response levels. We expect our novel coating system to be considered a strong candidate for use in various medical implant applications in order to decrease implant-induced capsule formation.

Novel Characterization of Constipation Phenotypes in ICR Mice Orally Administrated with Polystyrene Microplastics.

Indirect evidence has determined the possibility that microplastics (MP) induce constipation, although direct scientific proof for constipation induction in animals remains unclear. To investigate whether oral administration of polystyrene (PS)-MP causes constipation, an alteration in the constipation parameters and mechanisms was analyzed in ICR mice, treated with 0.5 µm PS-MP for 2 weeks. Significant alterations in water consumption, stool weight, stool water contents, and stool morphology were detected in MP treated ICR mice, as compared to Vehicle treated group. Also, the gastrointestinal (GI) motility and intestinal length were decreased, while the histopathological structure and cytological structure of the mid colon were remarkably altered in treated mice. Mice exposed to MP also showed a significant decrease in the GI hormone concentration, muscarinic acetylcholine receptors (mAChRs) expression, and their downstream signaling pathway. Subsequent to MP treatment, concentrations of chloride ion and expressions of its channel (CFTR and CIC-2) were decreased, whereas expressions of aquaporin (AQP)3 and 8 for water transportation were downregulated by activation of the mitogen-activated protein kinase (MAPK)/nuclear factor (NF)-κB signaling pathway. These results are the first to suggest that oral administration of PS-MP induces chronic constipation through the dysregulation of GI motility, mucin secretion, and chloride ion and water transportation in the mid colon.

Immunogenicity of low-dose MF59-adjuvanted 2009 influenza A/H1N1 vaccine in dialysis patients.

BACKGROUND: In response to the pandemic 2009 A/H1N1 virus, monovalent MF59-adjuvanted vaccines were prepared. Recently, single 3.75-µg doses of MF59-adjuvanted vaccines have shown good immunogenicity in young adults. However, the immunogenicity of these vaccines has not been evaluated in dialysis patients. METHODS: Dialysis patients received a single 3.75-µg dose of MF59-adjuvanted vaccine by intramuscular injection. For immunogenicity assays, serum samples were obtained before vaccination and 28 days after vaccination. All sera were tested by hemagglutination inhibition assays. RESULTS: Overall, 48 hemodialysis (HD) patients and 34 peritoneal dialysis (PD) patients were included in immunogenicity analysis. In HD patients, geometric mean titers (GMTs) were significantly increased compared with baseline GMTs in both young (aged 18-60 years) and elderly (aged ≥ 60 years) patients (51.2 ± 51.4 vs. 14.1 ± 20.7 in young patients, P = 0.012; 37.9 ± 73.9 vs. 6.8 ± 8.0 in elderly patients, P = 0.018, respectively). The rates of seroprotection and seroconversion were 27.6 and 17.2 % in young patients and 31.6 and 26.3 % in elderly patients, respectively. Among PD patients, GMTs were increased only in young patients (39.8 ± 51.4 vs. 6.8 ± 5.0, P = 0.001). The rates of seroprotection and seroconversion were 36.0 and 36.0 % in young patients and 11.1 and 0.0 % in elderly patients, respectively. CONCLUSION: A single 3.75-µg dose of MF59-adjuvanted vaccine was suboptimal to elicit protective antibody response in dialysis patients. Antibody responses against vaccine were compromised especially in elderly PD patients. Trials of different vaccination protocols such as a two-dose schedule or a higher hemagglutinin antigen dose of MF59-adjuvanted vaccine are necessary for improving antibody response in dialysis patients.

Vessel-Derived Decellularized Extracellular Matrices (VdECM): Novel Bio-Engineered Materials for the Wound Healing.

BACKGROUND: Decellularized extracellular matrix (dECM) is a non-cellular scaffold with various functions in tissue engineering and regenerative medicine. Elastin is related to tissue elasticity and scarless wound healing, abundantly found in lung and blood vessel tissues. We studied the characteristics of blood vessel-derived dECM (VdECM) and its effect in wound healing. METHODS: VdECM was prepared from porcine blood vessel tissue. Weight percentages of elastin of VdECM and atelocollagen were analyzed. Migratory potential of VdECM was tested by scratch assay. VdECM in hydrogel form was microscopically examined, tested for fibroblast proliferation, and examined for L/D staining. Cytokine array of various growth factors in adipocyte-derived mesenchymal stem cell (ASC) media with VdECM was done. Animal wound model showed the wound healing effect of VdECM hydrogel in comparison to other topical agents. RESULTS: VdECM contained 6.7 times more elastin than atelocollagen per unit weight. Microscopic view of 0.35% VdECM hydrogel showed consistent distribution. Compared to 3% atelocollagen, 0.35% VdECM showed superior results in fibroblast migration. Fluorescent microscopic findings of L/D assay had highest percentage of cell survival in 1% VdECM compared to atelocollagen. Growth factor expression was drastically amplified when VdECM was added to ASC media. In the animal study model, epithelialization rate in the VdECM group was higher than that of control, oxytetracycline, and epidermal growth factor ointments. CONCLUSION: VdECM contains a high ratio of elastin to collagen and amplifies expressions of many growth factors. It promotes fibroblast migration, proliferation, and survival, and epithelialization comparable to other topical agents.

Homodimeric SV40 NLS peptide formed by disulfide bond as enhancer for gene delivery.

Recently, cysteine residue incorporation increased liposome-mediated transfection compared to unmodified peptide. Therefore, we designed novel modified SV40 NLS peptides, homodimeric (NLS-CTHD, NLS-NTHD) and closed structure (cyclic NLS), simply using disulfide bond between cysteines to develop more efficient and safe non-viral gene delivery system. The simple mix of NLS-CTHD among these novel transfection enhancing peptides with DNA increased the gene transfer potency of cationic liposomes more efficiently with no additional cytotoxicity.

Bone Regeneration with 3D-Printed Hybrid Bone Scaffolds in a Canine Radial Bone Defect Model.

BACKGROUND: The repair of large bone defects remains a significant challenge in clinical practice and requires bone grafts or substitute materials. In this study, we developed a unique hybrid bone scaffold comprising a three dimensional (3D)-printed metal plate for weight bearing and a biodegradable polymer tube serving as bone conduit. We assessed the long-term effect of the hybrid bone scaffold in repairing radial bone defects in a beagle model. METHODS: Bone defects were created surgically on the radial bone of three beagle dogs and individually-tailored scaffolds were used for reconstruction with or without injection of autologous bone and decellularized extracellular matrix (dECM). The repaired tissue was evaluated by X-ray, micro-computed tomography, and histological observation 6 months after surgery. The functional integrity of hybrid bone scaffold-mediated reconstructions was assessed by gait analysis. RESULTS: In vivo analysis showed that the hybrid bone scaffolds maintained the physical space and bone conductivity around the defect. New bone was formed adjacent to the scaffolds. Addition of autologous bone and dECM in the polymer tube improved healing by enhancing bone induction and osteoconduction. Furthermore, the beagles' gait appeared normal by 4 months. CONCLUSION: The future of bone healing and regeneration is closely related to advances in tissue engineering. Bone production using autologous bone and dECM loaded on 3D-printed hybrid bone scaffolds can successfully induce osteogenesis and provide mechanical force for functional bone regeneration, even in large bone defects.

Promoting Effects of Titanium Implants Coated with Dipterocarpus tuberculatus Extract on Osseointegration.

Titanium (Ti) is the most commonly used biomaterial for dental implants. When inserting Ti implants into jawbones, the main issue is the lack of strong bonding between the Ti implant and the host bone (osseointegration). Inspired by the outstanding adhesion performance of natural phenolic compounds on metal substrates and promoting effect for cell adhesion, we coated a natural plant extract, Dipterocarpus tuberculatus (MED), on Ti implants. We tested three groups of Ti plates and screw-shaped fixtures: nontreated Ti as commercially produced, ozone-treated Ti as commonly used surface modification for dental implants, and MED-coated Ti. Interestingly, the MED coating on the Ti plate preserved the surface wetting property for 20 days, whereas the hydrophilic wetting of ozone-treated Ti was readily transformed to hydrophobic within a day. Computerized tomography and histopathological analysis revealed that MED coating enhanced new bone tissue formation and regeneration. The gene expression level of integrin as a bone cell adhesion receptor and its downstream key regulators was significantly increased than that of ozone-treated Ti. Therefore, we suggest considering MED-mediated cell signaling pathways in screening natural products for cell adhesion and osseointegration, and MED as a suitable coating agent for improving Ti implantation.

Age estimation based on 3D post-mortem computed tomography images of mandible and femur using convolutional neural networks.

Age assessment has attracted increasing attention in the field of forensics. However, most existing works are laborious and requires domain-specific knowledge. Modern computing power makes it is possible to leverage massive amounts of data to produce more reliable results. Therefore, it is logical to use automated age estimation approaches to handle large datasets. In this study, a fully automated age prediction approach was proposed by assessing 3D mandible and femur scans using deep learning. A total of 814 post-mortem computed tomography scans from 619 men and 195 women, within the age range of 20-70, were collected from the National Forensic Service in South Korea. Multiple preprocessing steps were applied for each scan to normalize the image and perform intensity correction to create 3D voxels that represent these parts accurately. The accuracy of the proposed method was evaluated by 10-fold cross-validation. The initial cross-validation results illustrated the potential of the proposed method as it achieved a mean absolute error of 5.15 years with a concordance correlation coefficient of 0.80. The proposed approach is likely to be faster and potentially more reliable, which could be used for age assessment in the future.

A transport system based on a quantum dot-modified nanotracer is genetically and developmentally stable in pregnant mice.

The use of nanoscale materials (NMs) could cause problems such as cytotoxicity, genomic aberration, and effects on human health, but the impacts of NM exposure during pregnancy remain uncharacterized in the context of clinical applications. It was sought to determine whether nanomaterials pass through the maternal-fetal junction at any stage of pregnancy. Quantum dots (QDs) coated with heparinized Pluronic 127 nanogels and polyethyleneimine (PEI) were administered to pregnant mice. The biodistribution of QDs, as well as their biological impacts on maternal and fetal health, was evaluated. Encapsulation of QDs with a nanogel coating produces a petal-like nanotracer (PNt), which could serve as a nano-carrier of genes or drugs. PNts were injected through the tail vein and accumulated in the liver, kidneys, and lungs. QD accumulation in reproductive organs (uterus, placenta, and fetus) differed among phases of pregnancy. In phase I (7 days of pregnancy), the QDs did not accumulate in the placenta or fetus, but by phase III (19 days) they had accumulated at high levels in both tissues. Karyotype analysis revealed that the PNt-treated pups did not have genetic abnormalities when dams were treated at any phase of pregnancy. PNts have the potential to serve as carriers of therapeutic agents for the treatment of the mother or fetus and these results have a significant impact on the development and application of QD-based NPs in pregnancy.

Clinical characteristics and complications associated with mesiodentes.

PURPOSE: The purpose of the present study was to analyze the clinical characteristics and complications of patients with mesiodens. PATIENTS AND METHODS: Using the radiographic reviews of patients who visited Seoul National University Dental Hospital from January 2005 through January 2008, 919 patients were identified as having mesiodens (total 1,200 mesiodentes). The number, location, position, eruption status, shape, direction, and complications of the mesiodens and gender and age were collected and analyzed. RESULTS: The mean patient age was 11.05 years (median 7.58) with a male/female ratio of 2.74:1. The results showed that 71.38% of the patients had 1 mesiodens, 27.75% had 2, 0.65% had 3, and 0.22% had 4 mesiodentes. Complications occurred in 46.92% of the mesiodentes. Of the complications, midline diastema was observed in 35.34%, delayed eruption in 20.60%, displacement in 16.60%, rotation in 11.02%, root resorption of adjacent teeth in 7.58%, cystic changes or cyst formation in 5.29%, and nasal eruption in 3.58%. CONCLUSIONS: Complications were found in approximately one half of the patients with a mesiodens. Early detection and timely surgical intervention should be emphasized to prevent unwanted complications.

Nonsyndromic multiple mandibular supernumerary premolars.

PURPOSE: The purpose of this study was to estimate the frequency of and describe the clinical characteristics and complications of patients with nonsyndromic multiple mandibular supernumerary premolars. PATIENTS AND METHODS: Among 110,500 patients seen between July 2003 and January 2007, 32 cases of multiple (2 or more) mandibular supernumerary premolars were found. Data including age, gender, number, distribution, and location of supernumerary premolars, extraction procedure, and related complications (such as cystic changes, root resorption, or adjacent teeth eruption disturbance) were collected. RESULTS: Six of 32 patients were female; the male to female ratio was 4.3:1. Prevalence of non-syndromic multiple mandibular supernumerary premolars was 0.029%. Mean age was 17.8 years. Root resorption of an adjacent tooth was found in 1 patient. Eruption anomalies were found in 5 patients and included impaction of a permanent tooth, prolonged retention of a deciduous tooth, and displacement of an adjacent tooth. Supernumerary teeth locations were predominantly lingual except for 4 teeth. CONCLUSION: Prevalence found in this study was lower compared with previous studies. However, the rate of related complications was higher, even though the mean age was relatively low. Surgical intervention should be considered to prevent unwanted sequelae for any signs of complication.

Physiologic facial muscle uptake on 18F-FDG PET/CT by chewing-like habitual movement in patient with Sjögren syndrome.

An 84-year-old female patient with known Sjögren syndrome underwent 18F-FDG PET/CT to detect recurrence of uterine cervix cancer. Sjögren syndrome is autoimmune disease that typically produces symptoms of dry mouth and eyes. We report a case of physiologic 18F-FDG uptake on facial muscles by chewing-like habitual movement, which was confused with salivary retention at first. The physiologic FDG uptake in oral cavity and facial muscles has to be reviewed carefully not to be confused with abnormal uptake.

Electrospun polystyrene-poly(styrene-co-maleic anhydride) nanofiber as a new aptasensor platform.

Here, we report the use of an aptamer-immobilized electrospun polystyrene-poly(styrene-co-maleic anhydride) (PS-PSMA) nanofiber as a new aptasensor platform for protein detection. Two thrombin-binding aptamers (TBA29 and TBA15) were used as a model platform to facilitate efficient detection of thrombin in a sandwich manner. Thrombin concentration was measured by fluorescence microscopy and spectroscopy, in which aptamers were labeled with either fluorescein dye or quantum dots. The results indicated that thrombin was captured uniformly on the surface of the nanofiber. Using this sandwich-type biosensor, the minimum detectable concentration of thrombin was 10 pM, with a dynamic range of 0.1-50 nM, when quantum dots were used for labeling. In contrast, the limit of detection was 1 nM, with a dynamic range of 10-200 nM, when using fluorescein dye labeling. This aptamers-on-nanofiber-based biosensor showed 2500-fold higher sensitivity than a 96-microwell plate format, attributed mainly to the large surface area of the nanofibers. In addition, this novel platform also exhibited similar high sensitivity in the detection of exogenously added thrombin in diluted human serum. This aptamers-on-nanofiber system, which is competitive with other sensing platforms and clinically meaningful in terms of its detection limit, is expected to be useful for the detection of various other targets because of its ease of application and manipulation.