BACKGROUND: Articular cartilage damage is still a troublesome problem. Hence, several researches have been performed for cartilage repair. The aim of this study was to evaluate the chondrogenicity of demineralized bone matrix (DBM) scaffolds under cyclic hydrostatic pressure (CHP) in vitro. METHODS: In this study, CHP was applied to human bone marrow mesenchymal stem cells (hBMSCs) seeded on DBM scaffolds at a pressure of 5 MPa with a frequency of 0.5 Hz and 4 h per day for 1 week. Changes in chondrogenic and osteogenic gene expressions were analyzed by quantifying mRNA signal level of Sox9, collagen type I, collagen type II, aggrecan (ACAN), Osteocalcin, and Runx2. Histological analysis was carried out by hematoxylin and eosin, and Alcian blue staining. Moreover, DMMB and immunofluorescence staining were used for glycosaminoglycan (GAG) and collagen type II detection, respectively. RESULTS: Real-time PCR demonstrated that applying CHP to hBMSCs in DBM scaffolds increased mRNA levels by 1.3-fold, 1.2-fold, and 1.7-fold (p < 0.005) for Sox9, Col2, and ACAN, respectively by day 21, whereas it decreased mRNA levels by 0.7-fold and 0.8-fold (p < 0.05) for Runx2 and osteocalcin, respectively. Additionally, in the presence of TGF-β1 growth factor (10 ng/ml), CHP further increased mRNA levels for the mentioned genes (Sox9, Col2, and ACAN) by 1.4-fold, 1.3-fold and 2.5-fold (p < 0.005), respectively. Furthermore, in histological assessment, it was observed that the extracellular matrix contained GAG and type II collagen in scaffolds under CHP and CHP with TGF-β1, respectively. CONCLUSION: The osteo-inductive DBM scaffolds showed chondrogenic characteristics under hydrostatic pressure. Our study can be a fundamental study for the use of DBM in articular cartilage defects in vivo and lead to production of novel scaffolds with two different characteristics to regenerate both bone and cartilage simultaneously.
Aggrecans ; Alcian Blue ; Bone Marrow ; Bone Matrix ; Cartilage ; Cartilage, Articular ; Collagen Type I ; Collagen Type II ; Eosine Yellowish-(YS) ; Extracellular Matrix ; Fluorescent Antibody Technique ; Gene Expression ; Hematoxylin ; Humans ; Hydrostatic Pressure ; In Vitro Techniques ; Mesenchymal Stromal Cells ; Osteocalcin ; Real-Time Polymerase Chain Reaction ; RNA, Messenger

BACKGROUND: Adipose tissue accumulation in specific body compartments has been associated with diabetes, hypertension and dyslipidemia. Perirenal fat (PRF) may lead to have direct lipotoxic effects on renal function and intrarenal hydrostatic pressure. This study was undertaken to explore the association of PRF with cardiovascular risk factors and different stages of chronic kidney disease (CKD). METHODS: We studied 103 patients with CKD of different stages (1 to 5). PRF was measured by B-mode renal ultrasonography in the distal third between the cortex and the hepatic border and/or spleen. RESULTS: The PRF thickness was greater in CKD patients with impaired fasting glucose than in those with normal glucose levels (1.10 ± 0.40 cm vs. 0.85 ± 0.39 cm, P < 0.01). Patients in CKD stages 4 and 5 (glomerular filtration rate [GFR] < 30 mL/min/1.73 m²) had the highest PRF thickness. Serum triglyceride levels correlated positively with the PRF thickness; the PRF thickness was greater in patients with triglyceride levels ≥ 150 mg/dL (1.09 ± 0.40 cm vs. 0.86 ± 0.36 cm, P < 0.01). In patients with a GFR < 60 mL/min/1.73 m², uric acid levels correlated positively with the PRF thickness (P < 0.05). CONCLUSION: In CKD patients, the PRF thickness correlated significantly with metabolic risk factors that could affect kidney function.
Adipose Tissue ; Dyslipidemias ; Fasting ; Filtration ; Glucose ; Humans ; Hydrostatic Pressure ; Hypertension ; Kidney ; Renal Insufficiency ; Renal Insufficiency, Chronic ; Risk Factors ; Spleen ; Triglycerides ; Ultrasonography ; Uric Acid

PURPOSE: To compare the choroidal thickness of a branch retinal vein occlusion (BRVO) lesion and that of other areas in the eyes. METHODS: Patients who visited the Ophthalmologic Clinic of Inje University Sanggye Paik Hospital for BRVO between March 2015 and October 2015 were reviewed retrospectively. We performed basic ophthalmologic exam and enhanced depth imaging optical coherence tomography in 48 eyes of 24 patients with BRVO. The choroidal thickness was compared in a total of 4 places, the branch retinal vein occlusion lesion, the symmetric site in the same eye, and the equivalent sites in the fellow eye by paired t-test. All measurements were performed by 2 independent observers. RESULTS: Choroidal thickness had strong inter-observer correlation. Choroidal thickness of the BRVO lesion was significantly thicker than that in the symmetric site of same eye, the equivalent site of lesion, and the equivalent site of the symmetric site to lesion in the fellow eye. CONCLUSIONS: Choroidal thickness in acute BRVO lesions was thicker than choroidal thickness in other areas of the eyes. It is thought that both hydrostatic pressure and the effects of vascular endothelial growth factor influence choroidal thickness in the acute phase of BRVO.
Choroid* ; Humans ; Hydrostatic Pressure ; Retinal Vein Occlusion* ; Retinal Vein* ; Retinaldehyde* ; Retrospective Studies ; Tomography, Optical Coherence ; Vascular Endothelial Growth Factor A

BACKGROUNDGlaucoma is a progressive optic neuropathy characterized by degeneration of neurons due to loss of retinal ganglion cells (RGCs). High intraocular pressure (HIOP), the main risk factor, causes the optic nerve damage. However, the precise mechanism of HIOP-induced RGC death is not yet completely understood. This study was conducted to determine apoptosis of RGC-5 cells induced by elevated hydrostatic pressures, explore whether laminin is associated with apoptosis under pressure, whether laminin can protect RGCs from apoptosis and affirm the mechanism that regulates the process of RGCs survival.

METHODSRGC-5 cells were exposed to 0, 20, 40, and 60 mmHg in a pressurized incubator for 6, 12, and 24 h, respectively. The effect of elevated hydrostatic pressure on RGC-5 cells was measured by Annexin V-fluorescein isothiocyanate/propidium iodide staining, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay, and Western blotting of cleaved caspase-3 protein. Location and expression of laminin were detected by immunofluorescence. The expression of β1-integrin, phosphorylation of focal adhesion kinase (FAK) and protein kinase B (PKB, or AKT) were investigated with real-time polymerase chain reaction and Western blotting analysis.

RESULTSElevated hydrostatic pressure induced apoptosis in cultured RGC-5 cells. Pressure with 40 mmHg for 24 h induced a maximum apoptosis. Laminin was declined in RGC-5 cells after exposing to 40 mmHg for 24 h. After pretreating with laminin, RGC-5 cells survived from elevated pressure. Furthermore, β1-integrin and phosphorylation of FAK and AKT were increased compared to 40 mmHg group.

CONCLUSIONSThe data show apoptosis tendency of RGC-5 cells with elevated hydrostatic pressure. Laminin can protect RGC-5 cells against high pressure via β1-integrin/FAK/AKT signaling pathway. These results suggest that the decreased laminin of RGC-5 cells might be responsible for apoptosis induced by elevated hydrostatic pressure, and laminin or activating β1-integrin/FAK/AKT pathway might be potential treatments to prevent RGC loss in glaucomatous optic neuropathy.

Apoptosis ; Cells, Cultured ; Focal Adhesion Protein-Tyrosine Kinases ; physiology ; Humans ; Hydrostatic Pressure ; Integrin beta1 ; physiology ; Intraocular Pressure ; Laminin ; physiology ; Proto-Oncogene Proteins c-akt ; physiology ; Retinal Ganglion Cells ; physiology ; Up-Regulation

BACKGROUND: The influence of bathing in asthma patients is not yet fully known. OBJECTIVE: We conducted an observational study to investigate changes in symptoms and their degree by bathing in asthmatic patients. METHODS: A questionnaire focusing on ever experienced bathing-induced symptom changes and their degree, as well as contributing factors, was designed and administered to asthmatic patients in the outpatient department of our institute between January 2012 and November 2013. RESULTS: Two hundred fifteen cases were recruited. In 60 cases (27.9%), asthmatic symptoms appeared, including 20 cases of chest discomfort (33.3%), 19 cases of cough (31.7%), and 21 cases of wheezing (35.0%). The triggering factors included vapor inhalation (32 cases, 53.3%), hydrostatic pressure on the thorax due to body immersion in the bathtub (26 cases, 43.3%), and sudden change of air temperature (16 cases, 26.7%). Thirty-eight cases (17.7%) experienced improvement in active asthmatic symptoms by bathing. Vapor inhalation was the most common contributing factor (34 cases, 89.5%), followed by warming of the whole body (13 cases, 34.2%). There was no relationship between asthma severity and the appearance of bathing-induced symptoms or improvement of active asthmatic symptoms by bathing. CONCLUSION: The effects of bathing in asthmatic patients widely differed from patient to patient and their etiology includes several factors. For those who suffer from bathing-induced asthma symptoms, preventive methods, such as premedication with bronchodilators before bathing, should be established. This study is registered in the University Hospital Medical Information Network (UMIN) clinical trials registry in Japan with the registration number UMIN000015641.
Asthma ; Baths ; Bronchoconstriction ; Bronchodilator Agents ; Cough ; Humans ; Hydrostatic Pressure ; Immersion ; Information Services ; Inhalation ; Japan ; Nebulizers and Vaporizers ; Observational Study ; Outpatients ; Premedication ; Respiratory Sounds ; Thorax

PURPOSE: This study investigated the potential of dual differentiation of stem cells into osteo- and chodrogenesis depending on scaffold type even in the same environment. MATERIALS AND METHODS: For the part of the cartilage tissue section, MSCs were suspended in alginate solution and bead droplets were made using 23G syringe. For the bone tissue section, PCL/HA scaffolds were made using the bio-plotting system followed by seeding mesenchymal stem cells (MSCs) onto the scaffolds. Scaffolds with MSCs were cultured in cocktail media containing osteogenic and chondrogenic growth factors for up to 21 days. To provide mechanical environments which articular cartilage experiences in-vivo, intermittent hydrostatic pressure (IHP) was engaged. Various cellular responses were assessed: the quantitative analysis of DNA contents, GAG contents, ALP activities and immunofluorescence. RESULTS: We found that IHP promoted MSCs differentiation into the targeted cell types. That is, MSCs in alginate scaffolds were able to be differentiated into chondrocytes, while those onto PCL/HA scaffolds were able to be differentiated into osteoblasts. CONCLUSION: Depending on the scaffold characteristics MSCs can be differentiated into bone cells or chondrocytes. This technique can provide a cue for the treatment of osteochondral defects utilizing tissue engineering.
Bone and Bones ; Cartilage ; Cartilage, Articular ; Chondrocytes ; Cues ; DNA ; Fluorescent Antibody Technique ; Hydrostatic Pressure* ; Intercellular Signaling Peptides and Proteins ; Mesenchymal Stromal Cells* ; Osteoblasts ; Stem Cells ; Syringes ; Tissue Engineering

PURPOSE: This study investigated the potential of dual differentiation of stem cells into osteo- and chodrogenesis depending on scaffold type even in the same environment. MATERIALS AND METHODS: For the part of the cartilage tissue section, MSCs were suspended in alginate solution and bead droplets were made using 23G syringe. For the bone tissue section, PCL/HA scaffolds were made using the bio-plotting system followed by seeding mesenchymal stem cells (MSCs) onto the scaffolds. Scaffolds with MSCs were cultured in cocktail media containing osteogenic and chondrogenic growth factors for up to 21 days. To provide mechanical environments which articular cartilage experiences in-vivo, intermittent hydrostatic pressure (IHP) was engaged. Various cellular responses were assessed: the quantitative analysis of DNA contents, GAG contents, ALP activities and immunofluorescence. RESULTS: We found that IHP promoted MSCs differentiation into the targeted cell types. That is, MSCs in alginate scaffolds were able to be differentiated into chondrocytes, while those onto PCL/HA scaffolds were able to be differentiated into osteoblasts. CONCLUSION: Depending on the scaffold characteristics MSCs can be differentiated into bone cells or chondrocytes. This technique can provide a cue for the treatment of osteochondral defects utilizing tissue engineering.
Bone and Bones ; Cartilage ; Cartilage, Articular ; Chondrocytes ; Cues ; DNA ; Fluorescent Antibody Technique ; Hydrostatic Pressure* ; Intercellular Signaling Peptides and Proteins ; Mesenchymal Stromal Cells* ; Osteoblasts ; Stem Cells ; Syringes ; Tissue Engineering

BACKGROUND/AIMS: In Japan, it is customary to take a daily bath during which the body is immersed in water to the neck. During full-body immersion, hydrostatic pressure is thought to compress the chest and abdomen, which might influence esophageal motor function and intra-gastric pressure. However, whether water immersion has a significant influence on esophageal motor function or intragastric pressure has not been shown. The aim of this study was to clarify the influence of full-body water immersion on esophageal motor function and intragastric pressure. METHODS: Nine healthy male volunteers (mean age 40.1 +/- 2.8 years) were enrolled in this study. Esophageal motor function and intragastric pressure were investigated using a high-resolution 36-channel manometry device. RESULTS: All subjects completed the study protocol. Intragastric pressure increased significantly from 4.2 +/- 1.1 to 20.6 +/- 1.4 mmHg with full-body water immersion, while the lower esophageal high pressure zone (LEHPZ) value also increased from 20.5 +/- 2.2 to 40.4 +/- 3.6 mmHg, with the latter being observed regardless of dietary condition. In addition, peak esophageal peristaltic pressure was higher when immersed as compared to standing out of water. CONCLUSIONS: Esophageal motor function and intragastric pressure were altered by full-body water immersion. Furthermore, the pressure gradient between LEHPZ and intragastric pressures was maintained at a high level, and esophageal peristaltic pressure was elevated with immersion.
Abdomen ; Baths ; Esophageal Sphincter, Lower ; Gastroesophageal Reflux ; Humans ; Hydrostatic Pressure ; Immersion ; Japan ; Male ; Manometry ; Neck ; Peristalsis ; Thorax ; Water

PURPOSE: To compare the choroidal thickness in central serous chorioretiopathy (CSC) patients and normal controls using spectral domain optical coherence tomography (SD-OCT). METHODS: The authors compared the choroidal thickness in eyes with CSC, fellow eyes and in normal eyes. In addition, the authors attempted to determine any correlation between choroidal thickness and other factors such as age, height of serous retinal detachment, and spherical equivalent. Choroidal thickness was measured using a perpendicular line from the outer margin of the subfoveal retinal pigment epithelium to the inner surface of the sclera. RESULTS: Twenty-five eyes of 25 CSC patients, 17 fellow eyes and 29 age-matched normal eyes were examined and categorized as group 1, group 2 and group 3, respectively. Subfoveal choroidal thickness was 370.64 +/- 58.06 microm in group 1, 301.85 +/- 47.83 microm in group 2, and 261.84 +/- 48.22 microm in group 3. The choroidal thickness in group 1 was significantly greater than those in group 2 and group 3, and the choroidal thickness in group 2 was significantly greater than that in group 3 (p = 0.001, p < 0.001, p = 0.004, respectively), where the choroidal thickness showed a negative correlation with age (p = 0.015). CONCLUSIONS: The choroidal thickness was greater in eyes with CSC and in their fellow eyes compared to that in normal eyes. The results suggest that CSC may be caused by choroidal vascular hyperpermeability and increased hydrostatic pressure in the choroid.
Central Serous Chorioretinopathy ; Choroid ; Eye ; Humans ; Hydrostatic Pressure ; Retinal Detachment ; Retinal Pigment Epithelium ; Tomography, Optical Coherence

To explore the effects of the physiological range of hydrostatic pressure on human bladder smooth muscle cells (HBSMCs) cultured in vitro, we used a hydrostatic compression device designed in our laboratory into the experiments, which were grouped by varied hydrostatic pressure gradients. Cellular morphology was observed with HE staining; cytoskeleton F-actin, cell cycle, both proliferating cell nuclear antigen (PCNA) and matrix metalloproteinase 7 (MMP-7) were detected respectively with immunofluorescence, flow cytometry and RT-PCR. We found that the proliferation, cytoskeleton and cycle distribution of HBSMCs were not obviously different among the groups of different hydrostatic pressure; however, the mRNA expression of MMP-7 exhibited a trend of first increasing and then declining as the pressure gradually rises. Thus the physiological range of hydrostatic pressure may not have significant influence on proliferation, morphology, skeleton, and cell cycle of HBSMCs, but it may have great effect on the expression of MMP-7.
Cells, Cultured ; Humans ; Hydrostatic Pressure ; Matrix Metalloproteinase 7 ; genetics ; metabolism ; Myocytes, Smooth Muscle ; cytology ; physiology ; Proliferating Cell Nuclear Antigen ; genetics ; metabolism ; RNA, Messenger ; genetics ; metabolism ; Urinary Bladder ; cytology