Nanoscale topography of artificial substrates can greatly influence the fate of stem cells including adhesion, proliferation, and differentiation. Thus the design and manipulation of nanoscale stem cell culture platforms or scaffolds are of great importance as a strategy in stem cell and tissue engineering applications. In this report, we propose that a graphene oxide (GO) film is an efficient platform for modulating structure and function of human adipose-derived stem cells (hASCs). Using a self-assembly method, we successfully coated GO on glass for fabricating GO films. The hASCs grown on the GO films showed increased adhesion, indicated by a large number of focal adhesions, and higher correlation between the orientations of actin filaments and vinculin bands compared to hASCs grown on the glass (uncoated GO substrate). It was also found that the GO films showed the stronger affinity for hASCs than the glass. In addition, the GO film proved to be a suitable environment for the time-dependent viability of hASCs. The enhanced differentiation of hASCs included osteogenesis, adipogenesis, and epithelial genesis, while chondrogenic differentiation of hASCs was decreased, compared to tissue culture polystyrene as a control substrate. The data obtained here collectively demonstrates that the GO film is an efficient substratum for the adhesion, proliferation, and differentiation of hASCs.
We describe a scrub typhus patient with acute renal failure for whom a diagnosis was made based on serology as well as immunohistochemical (IHC) staining and an electron microscopic examination (EM) of a renal biopsy specimen. For our case, we demonstrated by IHC staining and EM that renal failure was caused by acute tubular necrosis due to a direct invasion of Orientia tsutsugamushi. CASE REPORTA healthy-looking, 33-year-old male patient received a drug treatment for a 2-week history of cough and sputum. One day before admission to our hospital, he complained of abdominal pain and nausea after he played football. He was suspected of having renal failure on examination at a local clinic and was transferred to Chosun University Hospital in Gwang-ju, South Korea. At the time of admission, his vital signs were a blood pressure of 120 over 80 mm Hg, a pulse rate of 72 times/min, a respiratory rate of 20 breaths/min, and a body temperature of 37.2°C. A urinalysis showed no gross hematuria, with a urine output of 50 ml/h. His mental status was alert. There was no skin rash or lymphadenopathy. The patient's complete blood count showed a white blood cell count of 11,630/mm 3 , a hemoglobin count of 14.5 g/dl, and a platelet count of 243,000/ mm 3 . An arterial blood gas analysis showed a pH of 7. , 120 meq/liter; blood urea nitrogen, 39.3 mg/dl; and creatinine, 5.0 mg/dl. The biochemistry results showed a total protein level of 6.73 g/dl, albumin at 3.78 g/dl, aspartate transaminase at 30 IU/liter, alanine aminotransferase at 18 IU/liter, creatine phosphokinase at 446 U/liter, lactate dehydrogenase at 565 U/liter, myoglobin at 168.4 ng/ml, fibrin degradation products at 1 mg/ml (range, 0 to ϳ5.0 mg/ml), and a D-dimer concentration of 143.6 ng/ml (range, 0 to ϳ255 ng/ml). On urinalysis, hematuria (score, ϩ3) and proteinuria (score, ϩ4) were shown to be present. A microscopic urinalysis revealed 10 to 19 red blood cells/high-power field (HPF), 2% dysmorphic red blood cells/HPF, 1 to 4 white blood cells/HPF, and no cast/HPF. A urinalysis using urine collected during 24 h detected selective proteinuria (albumin, 895 mg/day). Serologic tests were all negative for rheumatoid factor, antinuclear antibody, antineutrophil cytoplasmic antibody, and cold agglutinin, human immunodeficiency virus, hepatitis B virus, and hepatitis C virus antibodies, and the VDRL test was negative. Antistreptolysin O and complement levels were all normal.A self-employed person, our patient presented with no typical symptoms of scrub typhus, including rash, fever, and headache.
Most chronic tympanic membrane (TM) perforations require surgical interventions such as tympanoplasty because, unlike with acute perforations, it is very difficult for the perforations to heal spontaneously. The purpose of this study was to develop novel therapeutic techniques and scaffolds that release growth factors to treat chronic TM perforations. We evaluated the cell proliferation effects of the epidermal growth factor (EGF) and fibroblast growth factor (FGF) on in vitro cultures of TM cells using an MTT assay. They both showed similar efficacy, so we used EGF because of its lower cost. We then constructed an EGF-releasing chitosan patch scaffold (EGF-CPS) based on previous studies. We analyzed its toxicity and strength, and we studied it using scanning electron microscopy. EGF was released from the EGF-CPS for 8 weeks in an in vitro system. In animal studies, the EGF group, which was treated with EGF-CPS, showed healing in 56.5% of the animals (13/23), while the control group, which did not receive any treatment, revealed 20.8% healing (4/24) (p=0.04). Transmission electron microscopic studies of regenerated eardrums in the EGF group showed much greater preservation of histological features, and TMs of the EGF group were thinner than spontaneously healed TMs. In conclusion, this novel EGF-CPS can be used as a nonsurgical intervention technique for treatment of chronic TM perforations.
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