Secured delivery of basic fibroblast growth factor using human serum albumin-based protein nanoparticles for enhanced wound healing and regeneration.

BACKGROUND: Basic fibroblast growth factor (bFGF) is one of the critical components accelerating angiogenesis and tissue regeneration by promoting the migration of dermal fibroblasts and endothelial cells associated with matrix formation and remodeling in wound healing process. However, clinical applications of bFGF are substantially limited by its unstable nature due to rapid decomposition under physiological microenvironment. RESULTS: In this study, we present the bFGF-loaded human serum albumin nanoparticles (HSA-bFGF NPs) as a means of enhanced stability and sustained release platform during tissue regeneration. Spherical shape of the HSA-bFGF NPs with uniform size distribution (polydispersity index < 0.2) is obtained via a simple desolvation and crosslinking process. The HSA-bFGF NPs securely load and release the intact soluble bFGF proteins, thereby significantly enhancing the proliferation and migration activity of human dermal fibroblasts. Myofibroblast-related genes and proteins were also significantly down-regulated, indicating decrease in risk of scar formation. Furthermore, wound healing is accelerated while achieving a highly organized extracellular matrix and enhanced angiogenesis in vivo. CONCLUSION: Consequently, the HSA-bFGF NPs are suggested not only as a delivery vehicle but also as a protein stabilizer for effective wound healing and tissue regeneration.

Improvement of osseointegration efficacy of titanium implant through plasma surface treatment.

A novel plasma treatment source for generating cylindrical plasma on the surface of titanium dental implants is developed herein. Using the titanium implant as an electrode and the packaging wall as a dielectric barrier, a dielectric barrier discharge (DBD) plasma was generated, allowing the implant to remain sterile. Numerical and experimental investigations were conducted to determine the optimal discharge conditions for eliminating hydrocarbon impurities, which are known to degrade the bioactivity of the implant. XPS measurement confirmed that plasma treatment reduced the amount of carbon impurities on the implant surface by approximately 60%. Additionally, in vitro experiments demonstrated that the surface treatment significantly improved cell adhesion, proliferation, and differentiation. Collectively, we proposed a plasma treatment source for dental implants that successfully removes carbon impurities and facilitate the osseointegration of SLA implants.

Enhanced osseointegration through direct energy deposition porous coating for cementless orthopedic implant fixation.

Direct energy deposition (DED) is a newly developed 3D metal printing technique that can be utilized on a porous surface coating of joint implants, however there is still a lack of studies on what advantages DED has over conventional techniques. We conducted a systematic mechanical and biological comparative study of porous coatings prepared using the DED method and other commercially available technologies including titanium plasma spray (TPS), and powder bed fusion (PBF). DED showed higher porosity surface (48.54%) than TPS (21.4%) and PBF (35.91%) with comparable fatigue cycle. At initial cell adhesion, cells on DED and PBF surface appeared to spread well with distinct actin stress fibers through immunofluorescence study. It means that the osteoblasts bind more strongly to the DED and PBF surface. Also, DED surface showed higher cell proliferation (1.27 times higher than TPS and PBF) and osteoblast cell activity (1.28 times higher than PBF) for 2 weeks culture in vitro test. In addition, DED surface showed better bone to implant contact and new bone formation than TPS in in vivo study. DED surface also showed consistently good osseointegration performance throughout the early and late period of osseointegration. Collectively, these results show that the DED coating method is an innovative technology that can be utilized to make cementless joint implants.

Exposure assessment for the abandoned metal mine area contaminated by arsenic.

Among the results of community health impact assessments completed in 2014, residents of the Indae abandoned metal mine area showed high average urinary concentrations of harmful arsenic (As), at 148.9 µg/L. The concentration of harmful As was derived as the sum of As(V), As(III), MMA, and DMA concentrations known to be toxic. In this area, mining hazard prevention work was not carried out and the pollution source was neglected, and the health effect of the residents due to arsenic exposure was concerned. We re-assessed As exposure levels and tried to identify exposure factors for residents of this area. Analysis of the soil, sediment, and river water to assess the association between the soil of the Indae abandoned metal mine area and the soil in residential areas confirmed a correlation between Pb and As concentrations in the soil. Since Pb and As behave similarly, the use of the stable Pb isotope ratio for assessment of the pollution source tracking was validated. In the 3-isotope plot (207/206Pb vs. 208/206Pb) of soil samples in this area, a stable Pb isotope ratio was located on the same trend line, which confirmed that the soil in the residential area was within the area of influence of the Indae abandoned metal mine. Therefore, we judged that the pollution source of As was the Indae abandoned metal mine. The results by As species were As (III) 1.45 µg/L, As (V) 0.74 µg/L, monomethylarsonic acid (MMA) 2.43 µg/L, dimethylarsinic acid (DMA) 27.63 µg/L, and arsenobetaine 88.62 µg/L. The urinary harmful As was 31.92 µg/L, much lower than the 148.9 µg/L reported in a 2014 survey, due to the implementation of a multi-regional water supply in November 2014 that restricted As exposure through drinking river water. However, concerns remain over chronic exposure to As because As in river water used for farming and in agricultural soil still exceeds environmental standards; thus, ongoing work to address hazards from former mining areas and continued environmental monitoring is necessary.

Difluorobenzothiadiazole and Selenophene-Based Conjugated Polymer Demonstrating an Effective Hole Mobility Exceeding 5 cm2 V-1 s-1 with Solid-State Electrolyte Dielectric.

We report synthesis of a new poly(4-(4,4-bis(2-ethylhexyl)-4 H-silolo[3,2- b:4,5- b']dithiophene-2-yl)-7-(4,4-bis(2-ethylhexyl)-6-(selenophene-2-yl)-4 H-silolo[3,2- b:4,5- b']dithiophene-2-yl)-5,6-difluorobenzo[ c][1,2,5]thiadiazole (PDFDSe) polymer based on planar 4,7-bis(4,4-bis(2-ethylhexyl)-4 H-silolo[3,2- b:4,5- b']dithiophen-2-yl)-5,6-difluorobenzo[ c][1,2,5]thiadiazole (DFD) moieties and selenophene linkages. The planar backboned PDFDSe polymer exhibits highest occupied molecular orbital and lowest unoccupied molecular orbital levels of -5.13 and -3.56 eV, respectively, and generates well-packed highly crystalline states in films with exclusive edge-on orientations. PDFDSe thin film was incorporated as a channel material in top-gate bottom-contact organic thin-film transistor with a solid-state electrolyte gate insulator (SEGI) composed of poly(vinylidene difluoride-trifluoroethylene)/poly(vinylidene fluoride- co-hexafluroropropylene)/1-ethyl-3-methylimidazolium bis(trifluoromethylsulfonyl)imide, which exhibited a remarkably high hole mobility up to µ = 20.3 cm2 V-1 s-1 corresponding to effective hole mobility exceeding 5 cm2 V-1 s-1 and a very low threshold voltage of -1 V. These device characteristics are associated with the high carrier density in the semiconducting channel region, induced by the high capacitance of the SEGI layer. The excellent carrier mobility from the PDFDSe/SEGI device demonstrates a great potential of semiconducting polymer thin-film transistors as electronic components in future electronic applications.

Structure-Property Relationships of Semiconducting Polymers for Flexible and Durable Polymer Field-Effect Transistors.

We report high-performance top-gate bottom-contact flexible polymer field-effect transistors (FETs) fabricated by flow-coating diketopyrrolopyrrole (DPP)-based and naphthalene diimide (NDI)-based polymers (P(DPP2DT-T2), P(DPP2DT-TT), P(DPP2DT-DTT), P(NDI2OD-T2), P(NDI2OD-F2T2), and P(NDI2OD-Se2)) as semiconducting channel materials. All of the polymers displayed good FET characteristics with on/off current ratios exceeding 107. The highest hole mobility of 1.51 cm2 V-1 s-1 and the highest electron mobility of 0.85 cm2 V-1 s-1 were obtained from the P(DPP2DT-T2) and P(NDI2OD-Se2) polymer FETs, respectively. The impacts of the polymer structures on the FET performance are well-explained by the interplay between the crystallinity, the tendency of the polymer backbone to adopt an edge-on orientation, and the interconnectivity of polymer fibrils in the film state. Additionally, we demonstrated that all of the flexible polymer-based FETs were highly resistant to tensile stress, with negligible changes in their carrier mobilities and on/off ratios after a bending test. Conclusively, these high-performance, flexible, and durable FETs demonstrate the potential of semiconducting conjugated polymers for use in flexible electronic applications.

Highly Sensitive Flexible NH3 Sensors Based on Printed Organic Transistors with Fluorinated Conjugated Polymers.

Understanding the sensing mechanism in organic chemical sensors is essential for improving the sensing performance such as detection limit, sensitivity, and other response/recovery time, selectivity, and reversibility for real applications. Here, we report a highly sensitive printed ammonia (NH3) gas sensor based on organic thin film transistors (OTFTs) with fluorinated difluorobenzothiadiazole-dithienosilole polymer (PDFDT). These sensors detected NH3 down to 1 ppm with high sensitivity (up to 56%) using bar-coated ultrathin (<4 nm) PDFDT layers without using any receptor additives. The sensing mechanism was confirmed by cyclic voltammetry, hydrogen/fluorine nuclear magnetic resonance, and UV/visible absorption spectroscopy. PDFDT-NH3 interactions comprise hydrogen bonds and electrostatic interactions between the PDFDT polymer backbone and NH3 gas molecules, thus lowering the highest occupied molecular orbital levels, leading to hole trapping in the OTFT sensors. Additionally, density functional theory calculations show that gaseous NH3 molecules are captured via cooperation of fluorine atoms and dithienosilole units in PDFDT. We verified that incorporation of functional groups that interact with a specific gas molecule in a conjugated polymer is a promising strategy for producing high-performance printed OTFT gas sensors.

Exposure factors of cadmium for residents in an abandoned metal mine area in Korea.

This study evaluated blood and urine cadmium (Cd) levels and human exposure factors for residents in an abandoned metal mine in Korea. We collected blood, urine, soil, water, and rice grain samples to analyze Cd concentrations and analyzed heavy metal concentration patterns in soil. We estimated the major exposure factor of Cd through non-carcinogenic risk assessment depending on exposure routes. The blood Cd concentration in the case group was 5.33 µg/L (geometric mean), significantly higher than that in the control group (1.63 µg/L, geometric mean). Urine Cd concentrations were also similar. The Cd concentrations in paddy soil (1.29 mg/kg) and rice grains (0.14 mg/kg) in the study area were higher than those in the control area (0.91 and 0.07 mg/kg, respectively). The analysis of heavy metal concentration in soil showed that the Cd levels in agricultural soil in the case group were attributable to the mine. The hazard quotient (HQ) of Cd by rice ingestion in the case group (1.25) was 2 times higher than that in the control group (0.6). We found that the HQ of rice ingestion contributed to more than 97 % of the total HQ, indicating that rice grains were the major exposure source. However, it is likely that the continuous intake of Cd-exposed crops led to chronic exposure among the residents in mine area.

Directing human embryonic stem cells towards functional endothelial cells easily and without purification.

Hemangioblasts or blood islands only arise in early development thereby the sources to obtain these bi-potential cells are limited. While previous studies have isolated both lineages in vitro through the hemangioblast, derivation efficiency was rather low due to cellular damage attributed by enzyme usage and fluorescent activated cell sorting (FACS). This study focused on avoiding the use of damaging factors in the derivation of endothelial cells (ECs). Single cell H9-human embryonic stem cells (hESCs) were obtained by using a mild dissociation protocol then human embryoid body (hEB) formation was performed under hemangioblast differentiation conditions. The hEBs were subjected to a two-stage cytokine treatment procedure. Subsequent culture of the adhesive cells in day 4 hEBs gave arise to a seemingly pure population of ECs. The hESC-derived ECs were characterized by identifying signature endothelial gene and protein markers as well as testing for in vitro functionality. Furthermore, in vivo functionality was also confirmed by transplanting the cells in hindlimb ischemic murine models. We demonstrate that the genetic change required for EC derivation precedes blast colony formation. Furthermore, cell damage was prevented by abating enzyme usage and FACS, resulting in a high yield of ECs upon adhesion. Under this method, confluent cultures of ECs were obtainable 4 days after hEB formation which is significantly faster than previous protocols.

Association of urinary 3-phenoxybenzoic acid levels with self-reported depression symptoms in a rural elderly population in Asan, South Korea.

OBJECTIVES: This study aimed to evaluate the association between presence of depression symptoms and the exposure level to insecticides among aged population in rural area, determined via measured levels of urinary 3-phenoxybenzoic acid (3-PBA), after controlling for socioeconomic confounding factors. METHODS: Using a cross-sectional study design, we randomly recruited participants for our study (161 male and 239 female) from rural areas of Asan, Chungnam, Korea. Environmental risk factor exposure was assessed using a questionnaire, and gas chromatography- mass spectrometry was used to analyze urinary 3-PBA levels. We used a logistic regression analysis to assess the association of urinary 3-PBA levels with the presence of self-reported depression symptoms. RESULTS: After controlling for creatinine levels, the median (interquartile range) concentration of 3-PBA was approximately 1.5 times (p<0.05) higher among female (1.54 [0.90 to 2.35]) µg/g) than among male (1.06 [0.64 to 1.81] µg/g). Our study found that among female participants, the unit increase in 3-PBA levels exhibited a likely positive association (odds ratio, 1.12; 95% confidence interval, 1.00 to 1.25) with an increased risk of presence of self-reported depression symptoms, after adjusting for socioeconomic insurance type, daily physical condition, marital status, smoking status, and age. CONCLUSIONS: Given our finding of a potential association between the presence of selfreported depression symptoms and 3-PBA levels, precautions should be considered to minimize exposure to insecticides and thus protect the health of aged residents in rural areas.

Mesenchymal stem cell-derived microparticles ameliorate peritubular capillary rarefaction via inhibition of endothelial-mesenchymal transition and decrease tubulointerstitial fibrosis in unilateral ureteral obstruction.

INTRODUCTION: Microparticles (MPs) derived from kidney-derived mesenchymal stem cells (KMSCs) have recently been reported to ameliorate rarefaction of peritubular capillaries (PTC) in ischemic kidneys via delivery of proangiogenic effectors. This study aimed to investigate whether KMSC-derived MPs show anti-fibrotic effects by ameliorating endothelial-to-mesenchymal transition (EndoMT) in human umbilical vein endothelial cells (HUVEC) in vitro and by preserving PTC in kidneys with unilateral ureteral obstruction (UUO) in vivo. METHODS: MPs isolated from the supernatants of KMSC were co-cultured with HUVEC to assess their in vitro biologic effects on endothelial cells. Mice were treated with MPs via the tail vein after UUO injury to assess their anti-fibrotic and PTC sparing effects. Renal tubulointerstitial damage and inflammatory cell infiltration were examined with Masson's trichrome, F4/80 and α-smooth muscle actin (α-SMA) staining and PTC rarefaction index was determined by CD31 staining. RESULTS: KMSC-derived MPs significantly ameliorated EndoMT and improved in vitro proliferation of TGF-ß1 treated HUVEC. In vivo administration of KMSC-derived MPs significantly inhibited EndoMT of PTC endothelial cells and improved PTC rarefaction in UUO kidneys. Furthermore, administration of KMSC-derived MPs inhibited inflammatory cell infiltration as well as tubulointerstitial fibrosis in UUO mice as demonstrated by decreased F4/80 and α-SMA-positive cells and Masson's trichrome staining, respectively. CONCLUSIONS: Our results suggest that KMSC-derived MPs ameliorate PTC rarefaction via inhibition of EndoMT and protect against progression of renal damage by inhibiting tubulointerstitial fibrosis.

Well-defined differentiation of hESC-derived hemangioblasts by embryoid body formation without enzymatic treatment.

Human hemangioblasts exist only during the early embryonic developmental stage thereby limiting the adult cellular source from which to obtain such cells for study. To overcome this, hemangioblast studies have focused on utilizing human embryonic stem cell (hESC) derivatives but current methods are cell-line dependent. Single cell dissociation of a hESC colony quickly led to cell death in most hESC lines due to enzyme treatment which, in turn, reduced induction potential and hemangioblast differentiation efficiency. Therefore, we sought to effectively improve the process of cell dissociation that is adaptable to various hESC lines and increase the initial induction potential of embryoid body (hEB). As a result, we determined an effective cell dissociation method through a comparison study involving various reagents which demonstrated successful dissociation regardless of cell line and enhanced hemangioblast differentiation efficiency.

Microparticles from kidney-derived mesenchymal stem cells act as carriers of proangiogenic signals and contribute to recovery from acute kidney injury.

We recently demonstrated the use of in vitro expanded kidney-derived mesenchymal stem cells (KMSC) protected peritubular capillary endothelial cells in acute renal ischemia-reperfusion injury. Herein, we isolated and characterized microparticles (MPs) from KMSC. We investigated their in vitro biologic effects on human endothelial cells and in vivo renoprotective effects in acute ischemia-reperfusion renal injury. MPs were isolated from the supernatants of KMSC cultured in anoxic conditions in serum-deprived media for 24 hours. KMSC-derived MPs demonstrated the presence of several adhesion molecules normally expressed on KMSC membranes, such as CD29, CD44, CD73, α4, 5, and 6 integrins. Quantitative real time PCR confirmed the presence of 3 splicing variants of VEGF-A (120, 164, 188), bFGF and IGF-1 in isolated MPs. MPs labeled with PKH26 red fluorescence dye were incorporated by cultured human umbilical vein endothelial cells (HUVEC) via surface molecules such as CD44, CD29, and α4, 5, and 6 integrins. MP dose dependently improved in vitro HUVEC proliferation and promoted endothelial tube formation on growth factor reduced Matrigel. Moreover, apoptosis of human microvascular endothelial cell was inhibited by MPs. Administration of KMSC-derived MPs into mice with acute renal ischemia was followed by selective engraftment in ischemic kidneys and significant improvement in renal function. This was achieved by improving proliferation, of peritubular capillary endothelial cell and amelioration of peritubular microvascular rarefaction. Our results support the hypothesis that KMSC-derived MPs may act as a source of proangiogenic signals and confer renoprotective effects in ischemic kidneys.

Optimum cutoff value of urinary cotinine distinguishing South Korean adult smokers from nonsmokers using data from the KNHANES (2008-2010).

INTRODUCTION: Cutoff values for distinguishing smokers from nonsmokers have generally been derived from studies in White populations. Even though recent population-based studies have demonstrated that cutoff values can differ by race and ethnicity, few studies have explored cutoff values among Asian populations. We established the cutoff values for urinary cotinine and cotinine:creatinine ratio (CCR) using a nationally representative South Korean adult population sample. METHODS: Data were obtained from the Korea National Health and Nutrition Examination Survey (KNHANES) 2008-2010 with pooled sampling weights. Adult participants (n = 11,629) aged ≥19 were included. Optimal cutoff values were determined by performing receiver operating characteristic curve analysis. RESULTS: The optimum cutoff values for urinary cotinine and CCR for Korean adults were found to be 164ng/mL and 1122ng/mg, respectively. The application of these urinary cotinine and CCR cutoff values for the female subpopulations aged 19-34, 35-54, and ≥55 years provided sensitivities ranging from 87.1% to 93.8% and from 82.9% to 94.9%, respectively, while maintaining specificity of ≥92%. For the 3 male age-based subpopulations, we obtained sensitivities and specificities for cotinine ranging from 93.1% to 94.5% and from 92.8% to 97.0%, respectively, and for CCR ranging from 90.0% to 96.9% and from 92.2% to 96.8%, respectively. CONCLUSIONS: Our cutoff values should allow researchers conducting environmental epidemiological or clinical studies in South Korea to distinguish adult smokers from nonsmokers effectively. However, different values may be applicable for subpopulations with different smoking prevalence rates or higher exposure levels to secondhand smoke.

Winter season temperature drops and sulfur dioxide levels affect on exacerbation of refractory asthma in South Korea: a time-trend controlled case-crossover study using soonchunhyang asthma cohort data.

OBJECTIVE: According to the American Thoracic Society, approximately 5% of all asthmatics have refractory asthma (RA); these patients fail to recover lost lung function even after long-term treatment with high doses of medications. METHODS: Using a time-trend controlled case-crossover study design, we calculated odds ratios to evaluate whether exposure to ambient air pollutants and certain meteorological conditions on the day of admission (Lag 0) and up to 3 days before admission (Lag1 through Lag 3) were associated with acute RA exacerbation. Results were obtained after controlling for the effects of seasonality, smoking, and allergen sensitivity; we stratified our data into four seasons with respect to the median temperature of each month and further stratified them according to self-reported smoking status and skin-prick test results. RESULTS: In our study, RA patients (n = 82), living in metropolitan city of Seoul and Kyunggi Province, accounted for 3.7% of all asthmatics (n = 2298) registered in our asthma cohort between 2005 and 2009. In winter, a 1°C decrease in ambient temperature and a 1 ppb increase in sulfur dioxide concentration on the day of Lag 1 were associated with 14.8% (95% confidence interval [CI]: 0.9-26.7) and 19.7% (95% CI: 3.3-38.7) increases in the risk of RA exacerbation among nonsmokers, respectively. Similar associations were obtained on the day of Lag 2. The association remained unchanged after excluding patients sensitive to Dermatophagoides farinae and Dermatophagoides pteronyssinus. CONCLUSIONS: Exposure to temperature drops and increased sulfur dioxide concentrations are positively associated with the occurrence of acute RA exacerbation during winter with 1 or 2 day lags.