MicroRNAs as biomarkers for dental diseases.

MicroRNAs (miRNAs) are short, noncoding RNAs that act as key regulators of diverse biological processes by mediating translational repression or mRNA degradation of target genes. Recent studies discovered miRNAs in saliva, and these miRNAs are promising candidates for use as biomarkers of dental diseases. In this review, the results of miRNA studies in the dental field are presented, and a brief overview of the current progress, limitations, and perspectives regarding miRNA biomarkers for dental diseases is given.

Vertical Ridge Augmentation with Customized Titanium Mesh Using a 3D-Printing Model: A Prospective Study in Humans.

PURPOSE: To evaluate the usefulness of ridge augmentation using a customized titanium mesh (CTM) that was preformed by trimming and bending the commercial titanium mesh on a virtually reconstructed 3D acrylic resin model using clinical, radiologic, histologic, and histomorphometric analyses. MATERIALS AND METHODS: This study was designed prospectively for patients who required vertical ridge augmentation using a staged approach before implant surgery. After installation of the CTM, grafting was performed using deproteinized porcine bone mineral covered with an absorbable membrane. Computed tomography was performed preoperatively and 6 months after simultaneous/staged guided bone regeneration to measure planned, reconstructed, and lacking bone volume, and the reconstruction rate was calculated based on these values. Clinical complications were also recorded, particularly the mesh exposure rate. At re-entry, the bone core was obtained using a trephine bur, and histologic and histomorphometric analyses were performed. RESULTS: A total of 10 sites in eight patients were used for the study analysis. The mean planned bone volume was 1.15 cm3 (range: 0.78 to 1.56 cm3), mean lacking bone volume was 0.13 cm3 (range: 0 to 0.59 cm3), and mean reconstructed bone volume was 1.02 cm3 (range: 0.56 to 1.43 cm3). The exposure rate was 30% (3 out of 10 sites). The reconstruction rate was over 80%, except for one case that showed suppuration. From histomorphometric analysis, 27.52% ± 16.87% of new bone, 7.62% ± 5.19% of residual graft, and 64.86% ± 23.76% of connective tissue were observed. The core biopsy samples demonstrated different pseudoperiosteum layer appearances based on the healing stage of the augmented sites. In the premature bone, the inner osteogenic layer consisted of multiple layers of osteoblast cells with adjacent large blood vessels. However, in the mature augmented site, there was no specific inner osteogenic layer, and the outer fibrous layer was dominant. CONCLUSIONS: The fabrication of CTM based on the application of the 3D-printing technique makes vertical ridge augmentation easier and can reduce complications and achieve target bone acquisition. In addition, it is expected that quantitative analysis of the pseudoperiosteum layer will be facilitated using the CTM.

Tannic-Acid-Enriched Poly(vinyl alcohol) Nanofibrous Membrane as a UV-Shie lding and Antibacterial Face Mask Filter Material.

Face masks are increasingly important in the battle against infectious diseases and air pollution. Nanofibrous membranes (NFMs) are promising filter layers for removing particulate matter (PM) without restricting air permeability. In this study, tannic-acid-enriched poly(vinyl alcohol) (PVA-TA) NFMs were fabricated by electrospinning PVA solutions containing large amounts of tannic acid (TA), a multifunctional polyphenol compound. We were able to prepare uniform electrospinning solution without coacervate formation by inhibiting the robust hydrogen bonding between PVA and TA. Notably, the NFM maintained its fibrous structure even under moist conditions after heat treatment without the use of a cross-linking agent. Further, the mechanical strength and thermal stability of the PVA NFM were improved by the introduction of TA. The functional PVA NFM with a high TA content showed excellent UV-shielding (UV-A: 95.7%, UV-B: 100%) and antibacterial activity against Escherichia coli (inhibition zone: 8.7 ± 1.2 mm) and Staphylococcus aureus (inhibition zone: 13.7 ± 0.6 mm). Moreover, the particle filtration efficiency of the PVA-TA NFM for PM0.6 particles was 97.7% at 32 L min-1 and 99.5% at 85 L min-1, indicating excellent filtration performance and a low pressure drop. Therefore, the TA-enriched PVA NFM is a promising mask filter layer material with excellent UV-blocking and antibacterial properties and has the potential for various practical applications.

Evaluation of a biodegradable polyurethane patch for repair of diaphragmatic hernia in a rat model: A pilot study.

INTRODUCTION: Congenital diaphragmatic hernia (CDH) repair is an area of active research. Large defects requiring patches have a hernia recurrence rate of up to 50%. We designed a biodegradable polyurethane (PU)-based elastic patch that matches the mechanical properties of native diaphragm muscle. We compared the PU patch to a non-biodegradable Gore-Tex™ (polytetrafluoroethylene) patch. METHODS: The biodegradable polyurethane was synthesized from polycaprolactone, hexadiisocyanate and putrescine, and then processed into fibrous PU patches by electrospinning. Rats underwent 4 mm diaphragmatic hernia (DH) creation via laparotomy followed by immediate repair with Gore-Tex™ (n = 6) or PU (n = 6) patches. Six rats underwent sham laparotomy without DH creation/repair. Diaphragm function was evaluated by fluoroscopy at 1 and 4 weeks. At 4 weeks, animals underwent gross inspection for recurrence and histologic evaluation for inflammatory reaction to the patch materials. RESULTS: There were no hernia recurrences in either cohort. Gore-Tex™ had limited diaphragm rise compared to sham at 4 weeks (1.3 mm vs 2.9 mm, p = 0.003), but no difference was found between PU and sham (1.7 mm vs 2.9 mm, p = 0.09). There were no differences between PU and Gore-Tex™ at any time point. Both patches formed an inflammatory capsule, with similar thicknesses between cohorts on the abdominal (Gore-Tex™ 0.07 mm vs. PU 0.13 mm, p = 0.39) and thoracic (Gore-Tex™ 0.3 mm vs. PU 0.6 mm, p = 0.09) sides. CONCLUSION: The biodegradable PU patch allowed for similar diaphragmatic excursion compared to control animals. There were similar inflammatory responses to both patches. Further work is needed to evaluate long-term functional outcomes and further optimize the properties of the novel PU patch in vitro and in vivo. LEVEL OF EVIDENCE: Level II, Prospective Comparative Study.

Poly(vinyl alcohol) nanofibrous membranes via green electrospinning and tannin coating for selective removal of Pb(II) ion.

The conventional adsorbent fabrication methods involve complicated processes and may cause secondary contaminations. Therefore, an effective eco-friendly method is required for the fabrication of heavy metal adsorbents using inexpensive and eco-friendly materials without secondary pollution during their process. In this study, nanofibrous membranes (NFMs) were fabricated via green electrospinning of poly(vinyl alcohol) (PVA), a hydrophilic polymer, and their water resistance was improved through simple heat treatment without using additional additives. Then, nanofibrous heavy metal adsorbents were prepared by dip-coating the NFMs in an aqueous solution of tannic acid (TA), a natural polyphenol. First, the adsorption/desorption behavior of TA on PVA NFMs during the TA coating process was investigated. In addition, the effects of TA coating on the mechanical properties and heavy metal adsorption characteristics of the PVA NFMs were analyzed. The TA coating significantly increased the mechanical strength, heat resistance, and heavy metal (Pb(II)) adsorption capacity of the PVA NFM. The Pb2+ adsorption amount of TA-coated PVA NFMs exhibited about 5-7 times higher than those of other heavy metal ions, indicating excellent selectivity for Pb2+. In addition, the TA-coated PVA NFMs retained >70% of its initial adsorption capacity even after four cycles of adsorption/desorption, indicating its reusability.

Rigiflex lithography-based nanodot arrays for localized surface plasmon resonance biosensors.

We present a facile and robust means of fabricating metallic nanodot arrays for localized surface plasmon resonance (LSPR) biosensors through the strategic coupling of a polymeric template prepared with rigiflex lithography and a subsequent metallization via electrodeposition. Rigiflex lithography provides the capability to realize large-scale nanosized features as well as process flexibility during contact molding. In addition, the electrodeposition process enables wet-based nanoscale metallization with high pattern fidelity and geometric controllability. Generated metallic nanodot arrays can be used as a general platform for LSPR biosensors via the sequential binding of chemicals and biomolecules. Extinction spectra of the corresponding LSPR signal are measured with UV-vis-NIR spectroscopy, from which the pattern size and shape dependence of LSPR are readily confirmed. The feasibility of a very sensitive biosensor is demonstrated by the targeted binding of human immunoglobulin G, yielding subnanomolar detection capability with high selectivity.

Lithographically-featured photonic microparticles of colloidal assemblies.

We have described a new and promising strategy for the fabrication of composite and porous photonic crystal microparticles that combines the self-assembly of colloidal particles with photolithography techniques. We fabricated silica/SU-8 composite microparticles with photonic bandgaps via four steps: (1) deposition of the silica colloidal crystals on the photoresist, (2) embedding of the colloidal crystals in the photoresist, (3) UV exposure through a photomask and subsequent development, and (4) release of the microparticles from the substrate. Embedding was performed above the glass transition temperature (T(g)) of uncrosslinked SU-8. At such temperatures, capillary forces on the silica particles facilitate the migration of colloidal crystals in the SU-8 matrix. Particle migration ceased when the top colloidal crystal layer was trapped at the interface between air and SU-8. In addition, we also prepared porous microparticles with an inverse opaline structure by dissolving the embedded silica particles from the composite structures. The porous microparticles showed enhanced reflectivity at the bandgap position due to the large refractive index contrast. The bandgap position of the microparticles was controlled by the size of the silica particles, which determined the lattice constant. Bilayered composite and porous microparticles with two distinct photonic bandgaps were also prepared by sequential deposition of colloidal crystals composed of two differently sized silica particles.

Whole-genome de novo sequencing of wood rot fungus Fomitopsis palustris (ATCC62978) with both a cellulolytic and ligninolytic enzyme system.

Fomitopsis palustris is a model brown rot fungus causing destructive wood decay based on the cellulase system. Endoglucanase secreted by F. palustris hydrolyzes cellulose in both the crystalline and amorphous form. In this study, whole-genome sequencing was conducted to identify genes related to F. palustris cellulose degradation and their functions. We determined the 43-Mb complete draft genome of F. palustris (ATCC 62978), comprising 14,592 predicted gene models. Gene annotation provided crucial information about the location and function of protein-encoding genes. Three types of endoglucanases were expressed: endo-1,3-beta-glucanase, endo-1,4-beta-d-glucanase, and endoglucanase. In addition, various ligninolytic enzymes such as laccase, aromatic compound dioxygenase, and aryl alcohol dehydrogenase were expressed in F. palustris (ATCC 62978). Colony polymerase chain reaction (PCR) indicated that the endo-1,4-beta-d-glucanase gene comprises 732bp. Optimization of the expression conditions of endoglucanase by real-time PCR revealed that endoglucanase was highly expressed after 7days in all conditions, which was secreted during the secondary metabolism. Studies for large-scale cellulase production from this fungus and investigation of its ligninolytic system will promote its extensive use in various applications. The genomic information determined herein provides a basis for molecular genetics studies to understand the genome functions of F. palustris (ATCC 62978).

Polydiacetylene/Anti-HBs Complexes for Visible and Fluorescent Detection of Hepatitis†B Surface Antigen on a Nitrocellulose Membrane.

The immunochromatographic assay (ICA) using a nitrocellulose (NC) membrane offers several advantages. This technique is a rapid and straightforward method in contrast to other immunoassays. Polydiacetylene (PDA) vesicles have unique optical properties, displaying red color and red fluorescence at the same time. In this system, red-phase PDA vesicles are used as a fluorescent dye as well as a surface for immobilized hepatitis B surface antibody (HBsAb). PDA has a remarkable stability compared with other fluorescent dyes. In this study, the most suitable PDA/HBsAb complexes are introduced for detecting hepatitis B surface antigen (HBsAg). Then, the PDA/HBsAb complexes affixed antibody is attached to NC membrane, which has two lines to confirm detection of HBsAg. The main advantage of this system is that the detection of HBsAg can be observed in both visible and fluorescent images due to the optical properties of polydiacetylene. Detection of HBsAg is observed up to 0.1 ng mL-1 by fluorescent analysis and confirmed by red line on the NC membrane up to 1 ng mL-1 (HBsAg) using the naked eye. Consequently, these results show that PDA/HBsAb complexes were successfully applied to ICA for the diagnosis of hepatitis B.

Shape changing thin films powered by DNA hybridization.

Active materials that respond to physical and chemical stimuli can be used to build dynamic micromachines that lie at the interface between biological systems and engineered devices. In principle, the specific hybridization of DNA can be used to form a library of independent, chemically driven actuators for use in such microrobotic applications and could lead to device capabilities that are not possible with polymer- or metal-layer-based approaches. Here, we report shape changing films that are powered by DNA strand exchange reactions with two different domains that can respond to distinct chemical signals. The films are formed from DNA-grafted gold nanoparticles using a layer-by-layer deposition process. Films consisting of an active and a passive layer show rapid, reversible curling in response to stimulus DNA strands added to solution. Films consisting of two independently addressable active layers display a complex suite of repeatable transformations, involving eight mechanochemical states and incorporating self-righting behaviour.

Degradation and polymerization of monolignols by Abortiporus biennis, and induction of its degradation with a reducing agent.

This study was carried out to better understand the characteristic modification mechanisms of monolignols by enzyme system of Abortiporus biennis and to induce the degradation of monolignols. Degradation and polymerization of monolignols were simultaneously induced by A. biennis. Whole cells of A. biennis degraded coniferyl alcohol to vanillin and coniferyl aldehyde, and degraded sinapyl alcohol to 2,6-dimethoxybenzene- 1,4-diol, with the production of dimers. The molecular weight of monolignols treated with A. biennis increased drastically. The activities of lignin degrading enzymes were monitored for 24 h to determine whether there was any correlation between monolignol biomodification and ligninolytic enzymes. We concluded that complex enzyme systems were involved in the degradation and polymerization of monolignols. To degrade monolignols, ascorbic acid was added to the culture medium as a reducing agent. In the presence of ascorbic acid, the molecular weight was less increased in the case of coniferyl alcohol, while that of sinapyl alcohol was similar to that of the control. Furthermore, the addition of ascorbic acid led to the production of various degraded compounds: syringaldehyde and acid compounds. Accordingly, these results demonstrated that ascorbic acid prevented the rapid polymerization of monolignols, thus stabilizing radicals generated by enzymes of A. biennis. Thereafter, A. biennis catalyzed the oxidation of stable monolignols. As a result, ascorbic acid facilitated predominantly monolignols degradation by A. biennis through the stabilization of radicals. These findings showed outstanding ability of A. biennis to modify the lignin compounds rapidly and usefully.

Compartment fabrication of magneto-responsive Janus microrod particles.

Monodispersed magneto-responsive microrod particles of variable magnetic/non-magnetic ratios and chemical compositions are created by compartment fabrication in a single poly(dimethylsiloxane) (PDMS) mold with periodic hole arrays. By labeling the two ends with green and red fluorescent dyes separately, we show that the particles can flip freely and reversibly in a confined geometry under the magnetic field, thereby displaying different patterned colors at the air-water interface.

Modulation of biomechanical properties of hyaluronic acid hydrogels by crosslinking agents.

Modulation of both mechanical properties and biocompatibilities of hyaluronic acid (HA) hydrogels is very importance for their applications in biomaterials. Pure HA solution was converted into a hydrogel by using butanediol diglycidyl ether (BDDE) as a crosslinking agent. Mechanical properties of the HA hydrogels have been evaluated by adding up different amount of BDDEs. While the mechanical properties of the obtained HA hydrogels were evaluated by measuring their crosslinking degrees, elastic modulus and viscosity, their in vitro biocompatibilities were done by measuring the degrees of anti-inflammatory reactions, cell viabilities and cytotoxicity. The degrees of anti-inflammatory reactions were determined by measuring the amount of nitric oxides (NOs) released from lipopolysaccharide(LPS)(+)-induced macrophages; cell viability was evaluated by observing differences in the behaviors of fibroblasts covered with the HA hydrogels, compared with those covered with the films of Teflon and Latex. Cytotoxicity of the HA hydrogels was also evaluated by measuring the degrees of viability of the cells exposed on the extracts of the HA hydrogels over those of Teflon, Latex and pure HA solutions by the assays of thiazoly blue tetrazolium bromide (MTT), neutral reds, and bromodeoxyuridine (BrdU). The results showed that employment of BDDEs beyond critical amounts showed lower biocompatibility of the crosslinked HA hydrogels but higher crosslinking degrees and mechanical properties, indicating the importance of controlling the HA concentrations, BDDE amounts and their reaction times for the synthesis of the crosslinked HA hydrogels for their clinical applications as biomaterials.

Gene expression profile in mesenchymal stem cells derived from dental tissues and bone marrow.

PURPOSE: The aim of this study is to compare the gene expression profile in mesenchymal stem cells derived from dental tissues and bone marrow for characterization of dental stem cells. METHODS: We employed GeneChip analysis to the expression levels of approximately 32,321 kinds of transcripts in 5 samples of bone-marrow-derived mesenchymal stem cells (BMSCs) (n=1), periodontal ligament stem cells (PDLSCs) (n=2), and dental pulp stem cells (DPSCs) (n=2). Each cell was sorted by a FACS Vantage Sorter using immunocytochemical staining of the early mesenchymal stem cell surface marker STRO-1 before the microarray analysis. RESULTS: We identified 379 up-regulated and 133 down-regulated transcripts in BMSCs, 68 up-regulated and 64 down-regulated transcripts in PDLSCs, and 218 up-regulated and 231 down-regulated transcripts in DPSCs. In addition, anatomical structure development and anatomical structure morphogenesis gene ontology (GO) terms were over-represented in all three different mesenchymal stem cells and GO terms related to blood vessels, and neurons were over-represented only in DPSCs. CONCLUSIONS: This study demonstrated the genome-wide gene expression patterns of STRO-1(+) mesenchymal stem cells derived from dental tissues and bone marrow. The differences among the expression profiles of BMSCs, PDLSCs, and DPSCs were shown, and 999 candidate genes were found to be definitely up- or down-regulated. In addition, GOstat analyses of regulated gene products provided over-represented GO classes. These data provide a first step for discovering molecules key to the characteristics of dental stem cells.