In Vitro Antifungal Activity of Epigallocatechin 3-O-Gallate against Clinical Isolates of Dermatophytes

Previously, we reported that epigallocatechin 3-O-gallate (EGCg) has growth-inhibitory effect on clinical isolates of Candida species. In this study, we investigated the antifungal activity of EGCg and antifungal agents against thirty-five of dermatophytes clinically isolated by the international guidelines (M38-A2). All isolates exhibited good susceptibility to EGCg (MIC50, 2-4 microg/mL, MIC90, 4-8 microg/mL, and geometric mean (GM) MICs, 3.36-4 microg/mL) than those of fluconazole (MIC50, 2-16 microg/mL, MIC90, 4-32 microg/mL, and GM MICs, 3.45-25.8 microg/mL) and flucytosin (MIC50, MIC90, and GM MICs, >64 microg/mL), although they were less susceptible to other antifungal agents, such as amphotericin B, itraconazole, and miconazole. These activities of EGCg were approximately 4-fold higher than those of fluconazole, and were 4 to 16-fold higher than flucytosin. This result indicates that EGCg can inhibit pathogenic dermatophyte species. Therefore, we suggest that EGCg may be effectively used solely as a possible agent or combined with other antifungal agents for antifungal therapy in dermatophytosis.

The Effects of Growth Factors on Motility of Cultured Human Dermal Microvascular Endothelial Cell

Cell migration is essential for many important biological events, including embryonic development, wound healing, inflammatory response, and tumor metastasis. As a result of endothelial cell migration, angiogenesis is very important factor in embryogenesis, wound healing, tumor development and flap survival. Angiogenesis is dependent on endothelial cell proliferation, migration and motility is one of the most essential for many important biological events. The speed of cell migration is regulated by extension, attachment, detachment of cell membrane and adhesiveness of cell to extracellular matrix. Growth factors such as FGF, TGF, VEGF is well known to play a major roles in the migration of endothealial cells. This study was designed to compare the motilities of human dermal microvascular endothelial cell(HDMEC) in growth factors such as bFGF, TGF-beta1 and VEGF. The motility of cultured HDMEC was compared using a video-microscopy system that was developed in combination with a self-designed CO2 mini- incubator. To determine migration speed, cells were viewed with a 4 phase-contrast lens and video recored. Images were captured using a color CCD camera and saved in 8-bit full-color mode. Experimental groups were divided into four groups: group I(with a Control, HDMEC only), group II(HDMEC with bFGF), group III (HDMEC with TGF-beta1), group IV(HDMEC with VEGF). At the concentration of 1ng/ml(bFGF), 1ng/ml(TGF-beta1), and 10ng/ml(VEGF) as the most effective dose for cell migration through preliminary study, the speed of migration are 8.736+/-0.948micrometer/hr, 9.869+/-1.904micrometer/ hr, 10.293+/-1.612micrometer/hr, respectively. These data shows that groups with growth factor accelerate the HDMEC migration than a control group, and the VEGF is most effective growth factor in the HDMEC migration than bFGF and TGF-beta1.

The Effects of Recombinant Human BMP-7, Prepared from a COS-7 Expression System, on the Proliferation and Differentiation of Rat Newborn Calvarial Osteoblasts

A family of proteins, the bone morphogenetic proteins (BMPs), which promote osteoblast differentiation and bone mineralization, have recently been identified. One, BMP-7, has shown the ability to induce cartilage and bone formation processes. In this report, the possibility that other cell lines, to CHO cells, may also be available as host cells for the expression of hBMP-7 was validated. Recombinant human BMP (rhBMP) -7 was produced in COS-7 cells, as a processed mature disulfide-linked homodimer, with an apparent molecular weight of 36, 000. Examination of the expressions of the markers characteristic of osteoblast phenotypes showed that the rhBMP-7 specifically stimulated the inductions of alkaline phosphatase (ALP) (5-fold increase at 100 ng of rhBMP-7/ml), parathyroid hormone (PTH) -mediated intracellular cAMP production (4-fold increase at 100 ng of rhBMP-7/ml) and osteocalcin synthesis (5-fold increase at 100 ng of rhBMP-7/ml). In summary, the in vitro mineralization assay results provide evidence that the rhBMP-7 peptide, produced by COS-7 expression system, possesses intact biological activity. A similar pattern of biological activity was observed for the BMP-7 in COS-7 cells compared to the corresponding CHO cell expression system. Thus, these findings can be experimentally utilized for the production of rhBMPs for in vitro or in vivo studies.

Evaluation of the Cytotoxicity of Polyetherurethane (PU) Film Containing Zinc Diethyldithiocarbamate (ZDEC) on Various Cell Lines

A polyetherurethane (PU) film containing 0.1% zinc diethyldithiocarbamate (ZDEC) is the international standard reference material for testing the in vitro cytotoxicity of polymer based biomaterials. Nowadays, culturing L929 or BALB/3T3 cells in direct contact or in an extract dilution condition is the most frequently using method for evaluating the cytotoxicity from biomaterials and medical devices. However, the results often vary, because it is directly related to the cellular functions and the mechanism of the toxicity of the contacting cells. In this study, 13 cell lines originating from various tissues were used to detect the cytotoxic activities of a PU film containing 0.1% ZDEC (PU-ZDEC). The correlation between the reactivity zone size and the relative cytotoxicity by quantifying the released total protein from each cell in the direct contact testing method was investigated. Hepa-1c1c7 cells demonstrated the highest sensitivity in the reactivity zone size, while CHO/dhFr(-) cells were the most sensitive in terms of the relative cytotoxicity. A correlation between the two processes in each cell line was not found (r=-0.478). In the extract dilution method, which involved cultivating the cells in the medium with various ZDEC concentrations prepared by diluting the PU incubation, the cytotoxicity increased with increasing ZDEC concentration in all cell lines. The BALB/ 3T3 cells demonstrated the highest sensitivity in the extract dilution method. No correlation in a comparison of the relative cytotoxicity from the direct contact method with the extract dilution method in each cell line, was found (r=-0.445). In this experiment, Hepa-1c1c7, BALB/3T3, CHO/dhFr(-) and L-929 cells among the 13 types of cell lines were the sensitive cell lines according to the two methods. The preliminary results suggest that a comparison of at least one or more cytotoxicity testing methods and many cell lines is necessary for an in vitro cytotoxicity test of biomaterials.

Comparative study on motility of the cultured fetal and neonatal dermal fibroblasts in extracellular matrix

One of the differences between fetal and adult skin healing is the ability of fetal wounds heal without contraction and scar formation. Extracellular matrix (ECM) provides a substratum for cells adhesion, migration, and proliferation and can directly influence the form and function of cells. As motility is essential for many important biological events, including wound healing, inflammatory response, embryonic development, and tumor metastasis, this study was designed to compare the motilities cultured dermal fetal and neonatal fibroblasts in the extracellular matrix. The motility of cultured fetal and neonatal fibroblasts was compared using a video-microscopy system that was developed in combination with a self-designed CO2 mini-incubator. To determine migration speed, cells were viewed with a 4X phase-contrast lens and video recorded. Images were captured using a color CCD camera and saved in 8-bit full-color mode. We found that cultured fetal fibroblasts move faster than neonatal fibroblast on type I collagen (fetal fibroblast, 15.1 micrometer/hr; neonatal fibroblast, 13.7 micrometer/hr), and in fibronectin (fetal fibroblast, 13.2 micrometer/hr; neonatal fibroblast, 13.0 micrometer/hr) and hyaluronic acid (fetal fibroblast, 11 micrometer/hr; neonatal fibroblast, 9.8 micrometer/hr).

Rotation / 대한정형외과학회잡지

Rotation-plasty procedure was first done by Borggreve, in 1930 for the congenital short femur. Recently this procedure was used for the malignant bone tumor at the distal femur by Kotz and Salzer in 1982. In spite of its cosmetic problem of the distal stump, this procedure has great functional advantage of converting the above-knee amputation to the below-knee amputation. The authors applied this procedure in case of severe soft tissue crushing and large bone defect from the distal femur to the proximal tibia by the rail-road accident. Conventional below-knee prosthesis with thigh corset gave him normal ambulation. The inverted foot was good to control the prosthesis as a below-knee stump and heel functioned as a patella to support the body weight.

A Clinical Study of Popliteal Artery Injury / 대한정형외과학회잡지

With the increase of motor accident, the rate of popliteal artery injury is increased, As the injuries are critical, early diagnosis and prompt treatment can save the limb. 27 cases of popliteal artery injury patients were reviewed and the result was as follows ; l. Amputation rate was 11%, as 3 out of 27 cases failed. There was no correlation between ischemic time and the rate of amputation. 2. Complete resection of all injured portion of vessel and reconstruction of patency through interposed saphenous vein graft was often indicated to avoid tension at the anastomosis site. 3. The rate of fasciotomy was positively correlated with the ischemic time. 4. Medial approach is preferred to posterior approach, since it makes proximal extension of exploration possible, and decrease the incidence of joint contracture. 5. Complications noted are wound infection, systemic sepsis and venous stasis, but they could be treated with proper management. However, nerve deficit due to associated nerve injuries was resist to treatment.

The Clinical Study of Transient Synovitis of the Hip Joint in Children / 대한정형외과학회잡지

Transient synovitis of the hip joint in children was first described by Lovett and More in 1892and is the commonest cause of painful hip in children. It is characterized by pain and limp of acute onset, generally shart-lived, unassociated with structural changes in the femoral capital epiphysis demonstrable roentgenographically.. The pathology is of interest, not because it is a disabling condition, but because of the difficulties in differentiating it from more serious disease of the hip such as Legg-Calve-Perthes disease, tuberculosis, osteomyelitis and etc. The 76 cases so diagnosed at the Han-II hospital from 1974 to 1983 were reviewed and reported as follows; 1. There was probably associated with predisposing factors such as infection, and trauma. 2. The diagnostic criteria were clinical symptoms, laboratory and radiological findings. 3. The treatment was bed rest, skin traction on the affected leg, non-weight bearing, antibiotics, sedatives and in a few cases, INH was administrated for the prevention of the tuberculosis. 4. Four hips among them revealed later as Legg-Calve-Perthes disease and tuberculous arthritis of the hip joint.