Bone grafting is widely used to bridge major bone defects or to promote bone union. Natural calcium carbonate (CC) has been used as a bone substitute material and used to scaffold for bone morphogenetic protein (BMP). The aims of this study is to evaluate the biocompatibility of cuttlebone (CB) and hydroxyapatite from CB (CBHA). Each material was shaped into disks (5 mm in diameter and 2 mm in thickness). To test biocompatibility, the disks were implanted into the dorsal subcutaneous tissue in mice. Fibrous capsule thickness around each disk was evaluated histologically at 2 and 4 weeks after implantation. Concerning biocompatibility, fibrous capsule thickness of CBHA was significantly thinner than that of CB and CHA (p < 0.05) at 2 and 4 weeks after implantation. Based on the clinical and histological results, CBHA would be a safe material for use inside the body and has more effective osteoconduction than CB.
Kim, Jun-Dae;Kim, Hey-Jin;Koun, Soonil;Ham, Hyung-Jin;Kim, Myoung-Jin;Rhee, Myungchull;Huh, Tae-Lin
Molecules and Cells
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v.37
no.5
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pp.406-411
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2014
The initial step of atrioventricular (AV) valve development involves the deposition of extracellular matrix (ECM) components of the endocardial cushion and the endocardialmesenchymal transition. While the appropriately regulated expression of the major ECM components, Versican and Hyaluronan, that form the endocardial cushion is important for heart valve development, the underlying mechanism that regulates ECM gene expression remains unclear. We found that zebrafish crip2 expression is restricted to a subset of cells in the AV canal (AVC) endocardium at 55 hours post-fertilization (hpf). Knockdown of crip2 induced a heart-looping defect in zebrafish embryos, although the development of cardiac chambers appeared to be normal. In the AVC of Crip2-deficient embryos, the expression of both versican a and hyaluronan synthase 2 (has2) was highly upregulated, but the expression of bone morphogenetic protein 4 (bmp4) and T-box 2b (tbx2b) in the myocardium and of notch1b in the endocardium in the AVC did not change. Taken together, these results indicate that crip2 plays an important role in AV valve development by downregulating the expression of ECM components in the endocardial cushion.
Shin, Sun-Hye;Lee, Sangkyu;Bae, Jong-Sup;Jee, Jun-Goo;Cha, Hee-Jae;Lee, You Mie
Molecules and Cells
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v.37
no.4
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pp.330-336
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2014
Thymosin beta4 (TB4) has multiple functions in cellular response in processes as diverse as embryonic organ development and the pathogeneses of disease, especially those associated with cardiac coronary vessels. However, the specific roles played by TB4 during heart valve development in vertebrates are largely unknown. Here, we identified a novel function of TB4 in endothelial-mesenchymal transformation (EMT) in cardiac valve endocardial cushions in zebrafish. The expressions of thymosin family members in developing zebrafish embryos were determined by whole mount in situ hybridization. Of the thymosin family members only zTB4 was expressed in the developing heart region. Cardiac valve development at 48 h post fertilization was defected in zebrafish TB4 (zTB4) morpholino-injected embryos (morphants). In zTB4 morphants, abnormal linear heart tube development was observed. The expressions of bone morphogenetic protein (BMP) 4, notch1b, and hyaluronic acid synthase (HAS) 2 genes were also markedly reduced in atrio-ventricular canal (AVC). Endocardial cells in the AVC region were stained with anti-Zn5 antibody reactive against Dm-grasp (an EMT marker) to observe EMT in developing cardiac valves in zTB4 morphants. EMT marker expression in valve endothelial cells was confirmed after transfection with TB4 siRNA in the presence of transforming growth factor ${\beta}$ ($TGF{\beta}$) by RT-PCR and immunofluorescent assay. Zn5-positive endocardial AVC cells were not observed in zTB4 morphants, and knockdown of TB4 suppressed TGF-${\beta}$-induced EMT in ovine valve endothelial cells. Taken together, our results demonstrate that TB4 plays a pivotal role in cardiac valve formation by increasing EMT.
Objectives : Mangifera indica leaves are well known for having a variety of benefits, including anti-inflammatory, anti-tumor, diabetic retinopathy and diabetic vasculosis. However, the effects of Mangifera indica leaves on hair loss inhibition have not been studied. In this study, we investigated to find out the activity of Mangifera indica leaves on hair loss. Methods : 2,2'-azino-bis-3-ethylbenzothiazoline-6-sulphonic acid(ABTS) analysis was performed to confirm the antioxidant efficacy of the water extract of Mangifera indica leaves (WEML). To examine the effect of WEML on cell viability in dermal papillar (DP) cells, 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetra Zolium (MTS) analysis was performed. The changes in the mRNA expression level of the hair loss and hair growth-related genes in dermal papilla cells by WEML treatment were confirmed by quantitative RT-PCR. Results : In dermal papilla (DP) cells, ABTS analysis and MTS analysis of WEML showed antioxidant efficacy and low cytotoxicity. As a result of gene expression analysis through Quantitative RT-PCR, no changes in hair growth-related genes BMP6 and CTNNB1 was confirmed. but inhibitory activity of WEML on hair loss-related genes EGR1, SGK, DKK1, SRD5A1 and SRD5A2 was confirmed. Conclusion : We confirmed that WEML has excellent antioxidant efficacy and a inhibitory activity of hair loss-related genes including 5α-reductase genes. These results suggest that Mangifera indica leaves have a potential activity as a hair loss treatment for hair loss and hair growth. Biochemical or molecular biological research on hair loss is needed.
The present study investigated the effects of thermal pre-treatment on the enhancement of anaerobic biodegradability of waste activated sludge at varied TS concentration levels. The activated sludges were thermally oxidized for 30 minutes at $80{\sim}200^{\circ}C$ with varied TS concentrations (2%, 4% and 6%). and then, sludge characteristics, solubilization efficiency and methane production yield of thermally pre-treated sludges were analyzed. The higher the temperature in the thermal pre-treatment, the higher the concentration levels of dissolved matters such as $SCOD_{Cr}$, $NH_4{^+}$ and VFAs, which indicates that the thermal pre-treatment facilitates the hydrolysis and acid fermentation. Furthermore, the solubilization efficiency was increased in proportion to the temperature rise at all TS concentrations and was reached at 68.9%, 55.6% and 53.1%, respectively, at $200^{\circ}C$. In the BMP test of the pre-treated sludges, higher methane production yields were observed as 0.313. 0.314 and $0.299m^3\;CH_4/kg\;VS_{add}$ at the condition of TS 2% ($160^{\circ}C$), 4% ($160^{\circ}C$) and 6% ($180^{\circ}C$), respectively, and degradation rate was increased by 84%, 79% and 65% compared with non-pretreated waste activated sludge. These findings suggest the effectiveness of thermal pre-treatment of waste activated sludge for anaerobic biodegradable process.
Purpose: This study was performed to investigate the release behavior of bioactive materials as a BMP-2 embedding on the porous titanium implant. Methods: Porous Ti implant samples were fabricated by sintering of spherical Ti powders in a high vacuum furnace. Specimens diameter and height were 4mm and 10mm. Embedding materials were used to stamp ink. Sectional images, porosity and release behavior of porous Ti implants were evaluated by scanning electron microscope(SEM), mercury porosimeter and UV-Vis-NIR spectrophotometer. Results: Internal pore structure was formed fully open pore. Average pore size and porosity were $8.993{\mu}m$ and 8.918%. Embedding materials were released continually and slowly. Conclusion: Porous Ti implant was fabricated successfully by sintering method. Particles are necking strongly each other and others portions were vacancy. Therefore bioactive materials will be able to embedding to porous Ti implants. If the development of the fusion implant of the bioactive material will be able to have the chance to several patients.
The treatment characteristics of leachate produced from pretreatment facilities like composting and feeding were investigated in a mesophilic anaerobic treatment. Experiments were performed in two phase which were acidification and methane fermentation. The acidification step was optimized for OLR from 1 to $4.5kg\;COD/kg\;VS{\cdot}day$ without adding NaOH. As experiment dates became longer, the solubilization ratio of particles increased up to 30% over 70 days. TVA was generated up to maximum 9,970mg HAc/L at OLR of $2kg\;COD/kg\;VS{\cdot}day$. But TVA was generated to minimum 6,519mg HAc/L at OLR of $4.5kg\;COD/kg\;VS{\cdot}day$. The acidification ratio was analyzed from 10.9% to 3.8% at OLR of $2kg\;COD/kg\;VS{\cdot}day$ and $4.5kg\;COD/kg\;VS{\cdot}day$ respectively. After 55 days, salt contents in the acid fermenter were accumulated and stabilized at the concentration of 3,150mg/L. Sodium ion($Na^+$) concentration was stabilized at 1,300mg/L. At methane fermentation step, biogas was generated up to 750ml and 937.5ml at the feeding volume of 20ml and 25ml respectively for acid fermented liquid during 25 days. About 80% of total biogas was generated during early 15 days and 95% were generated during 18 days respectively. After 25 days of the BMP test, acetic acid was removed approximately 97% and 98%, in case of those two experimental conditions.
For the clinical application, it is needed to keep characteristics of stem cells after storage for a long time. In the present study, we examined stem cell properties of human cord-derived stem cells (HUC) after cryopreservation. Cells were isolated from human umbilical cord and cultured in vitro. At passage 2 or 3, HUC were suspended at a concentration of $1.0{\times}10^6/m{\ell}$ in cryomedium consisting of DMSO and FBS. After freezing at $-80^{\circ}C$ overnight, HUC were cryopreserved at $-196^{\circ}C$ nitrogen gas. After 6 months, HUC were thawed and cultured in vitro. Assessment for the stem cell properties was made upon the morphology, population doubling time, and expression profiles of genes and various proteins. Cryopreserved HUC showed more than 70% viability and maintained fibroblast-like morphology similar to HUC before cryopreservation. Throughout the culture, they underwent average 42.8 doublings and produced $6.75{\times}{10^{18}}$ cells. RT-PCR analyses showed that cryopreserved HUC expressed Oct-4, nanog, SCF, NCAM, nestin, GATA-4, BMP4, and HLA-1 genes. They did not express Brachyury and HLA-DR genes. Immunocytochemical studies showed that cryopreserved HUC reacted with antibodies against SSEA-3, -4, Thy-1, vimentin, fibronectin, HCAM, ICAM, HLA-1 proteins. They did not react with antibody against HLA-DR protein. Theses genes and proteins expression patterns of cryopresserved HUC were similar to those of HUC before cryopreservation. These results suggest that cryopreserved HUC could retain proliferative potential and they expressed various genes and proteins similar to HUC before cryopreservation. Thus, cryopreservation might be useful for HUC for future research and clinical application.
Kim, Min-Kyeong;Kwon, Soon-Ik;Jung, Goo-Bok;Hong, Seong-Chang;Chae, Mi-Jin;Yun, Sun-Gang;So, Kyu-Ho
Korean Journal of Environmental Agriculture
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v.32
no.4
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pp.355-358
/
2013
BACKGROUND: Water-born pollution loads by agricultural non-point source (NPS) pollution are expected to become intensified due to ongoing precipitation change. Therefore, it is essential to develop a best management practice (BMP) that is suitable to agricultural environments in Korea. This study aimed to develop an environmental-friendly BMP to reduce NPS pollution load by agricultural activities. An eco-friendly way, small drainage pond, was suggested in this study to avoid direct drainage of agricultural runoffs and eventually reduce the amount of pollutants discharged into the surrounding aqua-environment. METHODS AND RESULTS: A small pond ($12m^2$) was constructed at the corner of a rice paddy field ($1,715m^2$) located in Suwon, Korea. Water was allowed to drain only via a small drainage pond. Sampling was repeatedly made at two locations, one from an entrance and the other from an exit of a pond, during the rice cultivation period (May to October, 2012). Generally, sampling was made only when runoff water drained through a pond, such as during and/or after rain (irrigation). The water quality analysis showed that all quality parameters (SS, $COD_{Mn}$, T-N, and T-P) were improved as water passed through the pond. The amount of runoff water was reduced by 96~100%. Suspended solids and COD concentrations was reduced by 79.3% and 45.6%, respectively. In case of T-N and T-P concentrations, the reduction rates were 52.2% and 60.5%, respectively and the amount of T-N and T-P were reduced by 16.3~73.0% and 15.4~70.1%, respectively. CONCLUSION(S): Our data implies that agricultural NPS pollution from rice paddy fields can be effectively managed when an appropriate drainage water management practice is imposed. In this paper, it was suggested that an installation of a small drainage pond can be effective to prevent not only the nutrient loss from rice fields but also pollutant discharge to surrounding water environments.
Background: Kalkitoxin (KT) is an active lipopeptide isolated from the cyanobacterium Lyngbya majuscula found in the bed of the coral reef. Although KT suppresses cell division and inflammation, KT's mechanism of action in vascular smooth muscle cells (VSMCs) is unidentified. Therefore, our main aim was to investigate the impact of KT on vascular calcification for the treatment of cardiovascular disease. Objectives: Using diverse calcification media, we studied the effect of KT on VSMC calcification and the underlying mechanism of this effect. Methods: VSMC was isolated from the 6 weeks ICR mice. Then VSMCs were treated with different concentrations of KT to check the cell viability. Alizarin red and von Kossa staining were carried out to examine the calcium deposition on VSMC. Thoracic aorta of 6 weeks mice were taken and treated with different concentrations of KT, and H and E staining was performed. Real-time polymerase chain reaction and western blot were performed to examine KT's effect on VSMC mineralization. Calcium deposition on VSMC was examined with a calcium deposition quantification kit. Results: Calcium deposition, Alizarin red, and von Kossa staining revealed that KT reduced inorganic phosphate-induced calcification phenotypes. KT also reduced Ca++-induced calcification by inhibiting genes that regulate osteoblast differentiation, such as runtrelated transcription factor 2 (RUNX-2), SMAD family member 4, osterix, collagen 1α, and osteopontin. Also, KT repressed Ca2+-induced bone morphogenetic protein 2, RUNX-2, collagen 1α, osteoprotegerin, and smooth muscle actin protein expression. Likewise, Alizarin red and von Kossa staining showed that KT markedly decreased the calcification of ex vivo ring formation in the mouse thoracic aorta. Conclusions: This experiment demonstrated that KT decreases vascular calcification and may be developed as a new therapeutic treatment for vascular calcification and arteriosclerosis.
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