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Abstract A common element of internal thoracic artery harvesting techniques is a distal vascular clamp placement at the end of the procedure, not only to avoid bleeding, but also to increase the internal hydrostatic pressure, diameter and flow. The logic indicates that the placement of this clamp at the beginning of the dissection will allow the artery to benefit earlier from these advantages. After more than five years of experience, we present a modification in the classical technique of skeletonized harvesting of the internal thoracic artery, consisting of artery distal occlusion at the beginning of the procedure. Some of its advantages are discussed.
Subject(s)
Humans , Mammary Arteries/surgery , Tissue and Organ Harvesting , DissectionABSTRACT
ABSTRACT: The effect of high hydrostatic pressure (HHP) application on whey protein concentrate was evaluated both before (pre-treatment - PT) and during (hydrolysis assisted - HA) hydrolysis processes. A factorial design 22 with 3 central points was used with pressure (100, 250, 400 MPa) and time (5, 20 and 35 minutes) as independent variables. The hydrolysis was evaluated and monitored by soluble protein, aromatic amino acid contents and RP-HPLC. ABTS and ORAC tests were used to evaluate the in vitro antioxidant capacity. The reduction of soluble protein content was approximately 20% for conventional hydrolysis and for all PT treatments up to 4 h of reaction, while HHP assisted hydrolysis at 100 MPa showed a 35% protein reduction after 35 minutes of reaction. In addition, pressurization favored peptic hydrolysis of β-lactoglobulin by up to 98% and also improved the in vitro antioxidant capacity of the hydrolysates, which increased from 34.25 to 60.89 μmoles TE g-1 of protein in the best treatment. The results suggest that the use of HHP assisted hydrolysis favored the peptic hydrolysis, with a reduction in hydrolysis time and increased antioxidant activity.
RESUMO: Neste estudo, o efeito da aplicação de alta pressão hidrostática (HHP) sobre o concentrado proteico de soro de leite foi avaliado antes (pré-tratamento - PT) e durante os processos de hidrólise (assistida por hidrólise - HA). Utilizou-se o delineamento fatorial 22 com três pontos centrais, onde as variáveis independentes foram pressão (100, 250, 400 MPa) e tempo (5, 20 e 35 minutos). A hidrólise foi avaliada pelo conteúdo de proteínas solúveis e aminoácidos aromáticos, além do perfil peptídico por RP-HPLC. As análises de ABTS e ORAC foram utilizadas para avaliar a capacidade antioxidante in vitro. A redução do teor de proteína solúvel foi de aproximadamente 20% para a hidrólise convencional e para todos os pontos de PT até 4h de reação, enquanto a hidrólise assistida por HHP a 100 MPa mostrou uma redução de 35% de proteína em 35 minutos de reação. Além disso, a pressurização favoreceu a hidrólise péptica da β-lactoglobulina em até 98% e também melhorou a capacidade antioxidante in vitro dos hidrolisados, que aumentaram de 34,25 para 60,89 μmoles de TE g-1 de proteína no melhor tratamento. Os resultados sugerem que o uso da hidrólise assistida por HHP favoreceu a hidrólise péptica, com redução no tempo de hidrólise e aumento da atividade antioxidante.
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BACKGROUND: Cartilage tissue repair is an important field of tissue engineering. How to use engineering technology to effectively differentiate seed cells into chondrocytes is a focus and difficulty in the field of tissue engineering. At present, it is difficult to make seed cells differentiate into mature and stable chondrocytes by simple orientation-inducing culture. Thereafter, the authors preliminarily studied the induced directional differentiation using intermittent hydrostatic pressure stimulation based on the characteristics of ATDC5 chondrocytes, in addition to the use of effective culture solution. OBJECTIVE: To investigate the effects of intermittent hydrostatic pressure on the earlyterm chondrocyte differentiation of ATDC5 cells. METHODS: ATDC5 cell lines were cultured in multilayer. Cells adhered well with multiple-layer formation after 3 days, and were then sealed to maintain sterility. Intermittent hydrostatic pressure was applied to the cultures (10 MPa, 1 Hz, 4 h/d). Cells cultured with no intermittent hydrostatic pressure served as control group. Morphological changes of the cells were observed under microscope at 4, 7, 11, 14, and 17 days. Expression levels of Aggrecan, COL-2 and SOX-9 mRNA were detected by real-time PCR. RESULTS AND CONCLUSION: After application of intermittent hydrostatic pressure, ATDC5 cells aggregated and appeared with obvious patchy changes. The mRNA expression levels of Aggrecan and COL-2 were significantly increased. SOX-9 mRNA expression level showed no significant difference compared with the control group, but presented with fall-rise pattern. Intermittent hydrostatic pressure influences the mRNA expression related to chondrocyte differentiation and promotes the secretion of chondrogenic matrix. This method is contributive to the mature cartilage differentiation.
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BACKGROUND: Compressive stress can change the morphology and activity of cells, but whether the morphology and activity of nucleus pulposus cells change under hydrostatic pressure still needs further study. OBJECTIVE: To study the morphology and activity of human nucleus pulposus cells in vitro. METHODS: The human nucleus pulposus cells were separated, cultured and passed on for three generations, and pressurized for 2, 4 and 6 hours under the hydrostatic pressure of 0.3, 1, and 3 MPa. Then, the morphological changes and growth of the cells before and after pressurization were observed by inverted phase contrast microscope. Transmission electron microscope was used to observe the ultrastructural changes and differences of the cells. Cell counting kit-8 was used to detect the proliferation activity, morphology and activity of the cells under different hydrostatic pressures. RESULTS AND CONCLUSION: (1) Cell culture and passage: The growth curves of the first, second and third generations of human nucleus pulposus cells were S-shaped, and the cells proliferated fastest in a straight line from 3 to 7 days. The protuberances of the 5th and 6th generation cells were long shuttle shaped, grew slowly and degenerated. (2) Cell morphology: the human nucleus pulposus cells were shrunk under hydrostatic pressures of 0.3, 1, 3 MPa. At 0.3 and 1 MPa, the cells became slightly smaller and the morphology was basically complete; at 3 MPa, the cells were most obviously shrunk and the morphology was incomplete. The results showed that when the human nucleus pulposus cells were pressurized for 2, 4 and 6 hours under 0.3, 1 and 3 MPa hydrostatic pressures, the change of cell morphology was the most obvious under 3 MPa hydrostatic pressure, but there was no obvious change under the same hydrostatic pressure for different time. (3) Cell viability: Under 0.3 MPa hydrostatic pressure, the proliferation rate of human nucleus pulposus cells first increased and then decreased with the increase of time, and the cell proliferation rate reached the peak at 4 hours. Under 1 and 3MPa hydrostatic pressures, the proliferation rate of the cells gradually decreased with the increase of time, and the cell proliferation rate under 1 MPa hydrostatic pressure was significantly higher than that under 3 MPa hydrostatic pressure at the same action time (P < 0.05). These findings indicate that proper hydrostatic pressure stimulation helps to promote the proliferation of human nucleus pulposus cells, and long-term improperly high hydrostatic pressure stimulation can reduce the proliferation rate of human nucleus pulposus cells, leading to the occurrence of intervertebral disc degeneration.
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BACKGROUND: The relationship between nuclear factor (NF)-kB signaling pathway and intervertebral disc degeneration is a hotspot in the field of orthopedics. In-depth investigation on various signaling pathways in the intervertebral disc contributes to understanding the mechanism of intervertebral disc degeneration. OBJECTIVE: To investigate the regulation of the serum containing Yishen Huoxue Tongluo Recipe on the NF-kB signal pathway of human intervertebral disc nucleus pulposus cells under different hydrostatic pressures, attempting to explore the possible therapeutic mechanism and target of Yishen Huoxue Tongluo Recipe in the treatment of degenerative diseases of the intervertebral disc from the perspective of molecular biology. METHODS: Passage 3 Human intervertebral disc nucleus pulposus cells were divided into eight groups and were cultured in the drug-containing serum of Yishen Huoxue Tongluo Recipe. The cells were intervened for 2, 4, and 6 hours under different hydrostatic loading conditions (0.3,1, and 3 MPa). The morphology and growth of nucleus pulposus cells were observed by an inverted phase contrast microscope. Ultrastructural changes of intervertebral disc nucleus pulposus cells were observed by a transmission electron microscopy. Cell counting kit-8 method was used to detect the proliferation activity of nucleus pulposus cells. Annexin V-FITC/Propidium Iodide apoptotic kit was used to double-stain nucleus pulposus cells to detect the cell apoptotic rate. Western blot method was used to detect the changes of NF-kB p65, Collagen II, ADAMTS-4, MMP-13 and Caspase-3 in nucleus pulposus cells. RESULTS AND CONCLUSION: (1) Under the same pressure and time, the morphology and growth of nucleus pulposus cells in the pressure+drug-containing serum groups were better than those in the normal pressure group and the simple pressure groups. Among them, the 0.3 and 1 MPa pressure+drug-containing serum groups had more intact cell morphology and better cell growth than the 3 MPa pressure+ drug-containing serum group. (2) The proliferative activity of nucleus pulposus cells was higher in the pressure+drug-containing serum group, and there was a significant difference between the 0.3 MPa pressure+drug-containing serum group and the 0.3 MPa simple pressure group (P < 0.05). (3) The apoptotic rate of nucleus pulposus cells in the pressure+drug-containing serum group was lower than that in the normal pressure group and simple pressure intervention group (P < 0.05). (4) The expression of Collagen II and Caspase-3 increased in the pressure+drug-containing serum group, while the expression of NF-kappa B p65, ADAMTS-4 and MMP-13 decreased in the pressure+drug-containing serum group (P < 0.05). To conclude, Yishen Huoxue Tongluo Recipe can increase cell activity, reduce cell apoptosis and effectively delay the degeneration of nucleus pulposus cells. Its mechanism is likely to promote the expression of Collagen II and Caspase-3 through the NF-kB signaling pathway of nucleus pulposus cells of intervertebral disc, and inhibit the expression of NF-kB p65, ADAMTS-4 and MMP-13.
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Background : Cardiovascular surgery involves the use of several artificial materials as graft vessels. Although artificial blood vessels of medium and large diameters currently present a satisfactory patency and durability, those of small diameter remain inferior to one's own vessels to prevent issues such as early thrombosis and vascular stenosis. The present study aimed to investigate the functionality of decellularized tissues that hold structures and growth factors derived from a living body. Methods : Mini pigs were used for the study. The bovine-derived decellularized blood vessels were transplanted into the pigs' carotid artery, and no anticoagulant or antiplatelet drugs were used after the surgery. The blood vessels were dissected and evaluated for patency and tissue staining. Result : The patency of the blood vessels was confirmed in all cases ; however, a thrombus was confirmed in one transplanted vessel. Pathological findings showed maintenance of the blood vessel structure, presenting no issues with collagen or elastin. Conclusion : This study demonstrated that biologically derived decellularized blood vessels are highly functional and present an intact luminal basement membrane, even without antiplatelet therapy. This study suggested that decellularized blood vessels can potentially help in the development of medical devices with higher functionality than that of the existing materials.
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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.
Subject(s)
Humans , 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 , Hydrostatic Pressure , In Vitro Techniques , Mesenchymal Stem Cells , Osteocalcin , Real-Time Polymerase Chain Reaction , RNA, MessengerABSTRACT
ABSTRACT The effect of pressure on the fluorescence of tryptophan in the presence of metal ions was studied by fluorescence spectrometry. It was found that at 60 MPa, the fluorescence intensity of M/Trp mixtures (M represented metal ions) increased compared to that at atmosphere pressure. The relative fluorescence efficiency of M/Trp mixtures increased with pressure. When the M/Trp ratio was above 10:1, the relative fluorescence efficiency in decreasing order was Cu2+/Trp mixtures, Ni2+/Trp mixtures and Mg2+ (K+)/Trp mixtures. When the ratio was below 10:1, the decreasing order was Cu2+/Trp mixtures and Ni2+ (Mg2+, K+)/Trp mixtures. The relative fluorescence efficiency increased with the concentration of Cu2+ and Ni2+. The variation was relate to the quenching of tryptophan fluorescence in the presence of metal ions. A red shift was also observed, but the red shift was independent of metal ions.
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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.
Subject(s)
Humans , Asthma , Baths , Bronchoconstriction , Bronchodilator Agents , Cough , Hydrostatic Pressure , Immersion , Information Services , Inhalation , Japan , Nebulizers and Vaporizers , Observational Study , Outpatients , Premedication , Respiratory Sounds , ThoraxABSTRACT
BACKGROUND: Effusive-constrictive pericarditis (ECP) is traditionally diagnosed by using the expensive and invasive technique of direct pressure measurements in the pericardial space and the right atrium. The aim of this study was to assess the diagnostic role of echocardiography in tuberculous ECP. METHODS: Intrapericardial and right atrial pressures were measured pre- and post-pericardiocentesis, and right ventricular and left ventricular pressures were measured post-pericardiocentesis in patients with tuberculous pericardial effusions. Echocardiography was performed post-pericardiocentesis. Traditional, pressure-based diagnostic criteria were compared with post-pericardiocentesis systolic discordance and echocardiographic evidence of constriction. RESULTS: Thirty-two patients with tuberculous pericardial disease were included. Sixteen had ventricular discordance (invasively measured), 16 had ECP as measured by intrapericardial and right atrial invasive pressure measurements and 17 had ECP determined echocardiographically. The sensitivity and specificity of pressure-guided measurements (compared with discordance) for the diagnosis of ECP were both 56%. The positive and negative predictive values were both 56%. The sensitivity of echocardiography (compared with discordance) for the diagnosis of ECP was 81% and the specificity 75%, while the positive and the negative predictive values were 76% and 80%, respectively. CONCLUSION: Echocardiography shows a better diagnostic performance than invasive, pressure-based measurements for the diagnosis of ECP when both these techniques are compared with the gold standard of invasively measured systolic discordance.
Subject(s)
Humans , Atrial Pressure , Constriction , Diagnosis , Echocardiography , Heart Atria , Pericardial Effusion , Pericarditis , Pericardium , Sensitivity and Specificity , Tuberculosis , Ventricular PressureABSTRACT
Objective: To explore the application of high hydrostatic pressure in the development of melanoma vaccine.Methods:The high hydrostatic pressure,liquid nitrogen freezing and thawing were used to break the murine B16 melanoma cells and then the cell suspension was mixed with Freund's adjuvant to prepare vaccine for immunizing the mice.Results:After immu-nization,the murine B16 melanoma cells were injected intravenously and subcutaneously.The immune results of the vaccines were evaluated,by comparing survival time of the mice, subcutaneous tumor volume, DTH experiments and the fluorescence imaging of tumors.Conclusion:Compared with the method of liquid nitrogen freeze-thaw broken,high hydrostatic pressure crushing cells has more advantages in the development of tumor vaccine.
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Ibusuki hot spring sand bath (SB) (Sunamushi) has traditionally been used for the relief of musculoskeletal pain. It is specified by piling up heavy (40-60 kg) and hot (50°C) sands on the lied body heated by the hot spring water gushed at the seashore of Ibusuki. In this study, remarkable circulatory activation and metabolic improvements probably due to thermal vasodilation and higher hydrostatic pressure is examined. Subjects: The subjects examined were 20 healthy males (34.3±10.5 yrs) who accepted informed consents. Methods: The subjects were thin bathrobe and kept rest for 30 min in the supine position. BP, HR and sublingual temperature measurements and venous blood sampling from the indwelling catheter was done. Blood counts, blood gas pressure and plasma chemistry were examined. Then sand bath carried out for 10 min and 30 min rest under keeping sufficient warmth by blankets. Results: Systolic blood pressure was significantly increased though diastolic blood pressure was significantly reduced. HR and sublingual temperature were significantly increased by +22 bpm and +1.1°C, respectively, just after 10 min SB. Venous blood pO2 and pH were significantly increased by +18.3 mm Torr and +0.03pH, and pCO2 was significantly reduced by -5.8 mm Torr. Lactate and pyruvate were significantly reduced after 10 min and 30 min after sand bath suggesting the improved peripheral oxidative metabolism. Conclusion: Increase in blood pressure and heart rate indicating cardiac acceleration was considered to be induced by hydrostatic pressure with heavy sands and thermal vasodilation. Improved peripheral circulation and oxidative metabolism were also suggested by increased pO2, decreased pCO2 and decreased lactate and pyruvate level. Sufficient O2 supply and removal of wasted substances due to activated circulation was considered to be the basic mechanism of the effects of sand bath.
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Ibusuki hotspring sand bath (SB)(Sunamushi) is a special thermal therapy using heated sands by natural hotspring gushed at the seashore of Ibusuki city. Heated heavy sands (50°C, 40-60kg) was piled on the lied body. It has traditionally been used here for 250 years to relieve muculoskeletal and neuralgic pain, and still accepts 260 thousands visitors a year. In the present study, cardiovascular and metabolic effects by SB was studied from the viewpoint of accelerated circulation. Subjects and Methods: General physical parameters (BP, HR, sublingual temperature) and plasma chemistry were examined in 20 healthy males(36 ± 10yrs). The subjects wore thin bathrobe and a venous catheter for blood sampling were set in the forearm. They kept rest in the supine position for 30min and subjected 10min SB at the municipal SB institute with hotspring piping under the sands. In another 28 healthy subjects (44.3 ± 2.4yrs), cardiac outputs and plasma catecholamines (CA) and renin activity (PRA) were measured. In 6 subjects intracardiac study by Swan-Ganz catheterization were performed. Results: Diastolic pressure were significantly decreased by 6mmHg, and heart rate and sublingual temperature were significantly increased by +20bpm and +1.1°C, respectively, after 10min SB. Venous blood pO2 and pH was significantly increased by 20mm Torr and 0.03pH, and pCO2 was significantly reduced by 5mm Torr. Lactate, pyruvate and L/P ratio were significantly reduced suggesting improved oxidative metabolism of peripheral tissues. Plasma CAs and PRA were elevated after SB. All of these results gradually returned to the resting level after 30min. Cardiac output (CO) measured by dye dilution or thermo-dilution method was significantly increased from 5.6l/min to 10.5 l/min after 10min SB, and reduced to 8.1 l/min by removing piled heavy sands. Calculated total peripheral resistance (TPR) was significantly decreaased suggesting thermal vasodilation. Although mean right atrial pressure and pulmonary arterial pressure were increased during SB, they were immediately decreased by removing piled sands. Discussion: All of these results indicate that the basic effects of SB are derived from strong hydrostatic pressure of piled heavy sands and thermal vasodilation. Increased CO due to accelerated venous return and reduced afterload (TPR) will induce sufficient oxygen supply and increased discharge of wasted matters from peripheral tissues. These data seem to be compatible with the clinical effects of SB to relieve musculoskeletal pain and fatigue. Conclusion: Significant clinical effects is induced by increased CO due to the increased hydrostatic pressure of piled sands and thermal vasodilation.
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Ibusuki hotspring sand bath (SB)(Sunamushi) is a special thermal therapy using heated sands by natural hotspring gushed at the seashore of Ibusuki city. Heated heavy sands (50°C, 40-60kg) was piled on the lied body. It has traditionally been used here for 250 years to relieve muculoskeletal and neuralgic pain, and still accepts 260 thousands visitors a year. In the present study, cardiovascular and metabolic effects by SB was studied from the viewpoint of accelerated circulation.<BR><b>Subjects and Methods: </b>General physical parameters (BP, HR, sublingual temperature) and plasma chemistry were examined in 20 healthy males(36 ± 10yrs). The subjects wore thin bathrobe and a venous catheter for blood sampling were set in the forearm. They kept rest in the supine position for 30min and subjected 10min SB at the municipal SB institute with hotspring piping under the sands. In another 28 healthy subjects (44.3 ± 2.4yrs), cardiac outputs and plasma catecholamines (CA) and renin activity (PRA) were measured. In 6 subjects intracardiac study by Swan-Ganz catheterization were performed. <BR><b>Results: </b>Diastolic pressure were significantly decreased by 6mmHg, and heart rate and sublingual temperature were significantly increased by +20bpm and +1.1°C, respectively, after 10min SB. Venous blood pO<sub>2</sub> and pH was significantly increased by 20mm Torr and 0.03pH, and pCO<sub>2</sub> was significantly reduced by 5mm Torr. Lactate, pyruvate and L/P ratio were significantly reduced suggesting improved oxidative metabolism of peripheral tissues. Plasma CAs and PRA were elevated after SB. All of these results gradually returned to the resting level after 30min. Cardiac output (CO) measured by dye dilution or thermo-dilution method was significantly increased from 5.6l/min to 10.5 l/min after 10min SB, and reduced to 8.1 l/min by removing piled heavy sands. Calculated total peripheral resistance (TPR) was significantly decreaased suggesting thermal vasodilation. Although mean right atrial pressure and pulmonary arterial pressure were increased during SB, they were immediately decreased by removing piled sands.<BR><b>Discussion: </b>All of these results indicate that the basic effects of SB are derived from strong hydrostatic pressure of piled heavy sands and thermal vasodilation. Increased CO due to accelerated venous return and reduced afterload (TPR) will induce sufficient oxygen supply and increased discharge of wasted matters from peripheral tissues. These data seem to be compatible with the clinical effects of SB to relieve musculoskeletal pain and fatigue.<BR><b>Conclusion: </b>Significant clinical effects is induced by increased CO due to the increased hydrostatic pressure of piled sands and thermal vasodilation.
ABSTRACT
Ibusuki hot spring sand bath (SB) (Sunamushi) has traditionally been used for the relief of musculoskeletal pain. It is specified by piling up heavy (40-60 kg) and hot (50°C) sands on the lied body heated by the hot spring water gushed at the seashore of Ibusuki. In this study, remarkable circulatory activation and metabolic improvements probably due to thermal vasodilation and higher hydrostatic pressure is examined.<BR><b>Subjects:</b> The subjects examined were 20 healthy males (34.3±10.5 yrs) who accepted informed consents. <BR><b>Methods:</b> The subjects were thin bathrobe and kept rest for 30 min in the supine position. BP, HR and sublingual temperature measurements and venous blood sampling from the indwelling catheter was done. Blood counts, blood gas pressure and plasma chemistry were examined. Then sand bath carried out for 10 min and 30 min rest under keeping sufficient warmth by blankets.<BR><b>Results:</b> Systolic blood pressure was significantly increased though diastolic blood pressure was significantly reduced. HR and sublingual temperature were significantly increased by +22 bpm and +1.1°C, respectively, just after 10 min SB. Venous blood pO<sub>2</sub> and pH were significantly increased by +18.3 mm Torr and +0.03pH, and pCO<sub>2</sub> was significantly reduced by -5.8 mm Torr. Lactate and pyruvate were significantly reduced after 10 min and 30 min after sand bath suggesting the improved peripheral oxidative metabolism. <BR><b>Conclusion:</b> Increase in blood pressure and heart rate indicating cardiac acceleration was considered to be induced by hydrostatic pressure with heavy sands and thermal vasodilation.Improved peripheral circulation and oxidative metabolism were also suggested by increased pO<sub>2</sub>, decreased pCO<sub>2</sub> and decreased lactate and pyruvate level. Sufficient O<sub>2</sub> supply and removal of wasted substances due to activated circulation was considered to be the basic mechanism of the effects of sand bath.
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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.
Subject(s)
Bone and Bones , Cartilage , Cartilage, Articular , Chondrocytes , Cues , DNA , Fluorescent Antibody Technique , Hydrostatic Pressure , Intercellular Signaling Peptides and Proteins , Mesenchymal Stem Cells , Osteoblasts , Stem Cells , Syringes , Tissue EngineeringABSTRACT
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.
Subject(s)
Bone and Bones , Cartilage , Cartilage, Articular , Chondrocytes , Cues , DNA , Fluorescent Antibody Technique , Hydrostatic Pressure , Intercellular Signaling Peptides and Proteins , Mesenchymal Stem Cells , Osteoblasts , Stem Cells , Syringes , Tissue EngineeringABSTRACT
High hydrostatic pressure (HHP) has been investigated and industrially applied to extend shelf life of meat-based products. Traditional ham packaged under microaerophilic conditions may sometimes present high lactic acid bacteria population during refrigerated storage, which limits shelf life due to development of unpleasant odor and greenish and sticky appearance. This study aimed at evaluating the shelf life of turkey ham pressurized at 400 MPa for 15 min and stored at 4, 8 and 12 ºC, in comparison to the non pressurized product. The lactic acid bacteria population up to 10(7) CFU/g of product was set as the criteria to determine the limiting shelf life According to such parameter the pressurized sample achieved a commercial viability within 75 days when stored at 4 ºC while the control lasted only 45 days. Predictive microbiology using Gompertz and Baranyi and Roberts models fitted well both for the pressurized and control samples. The results indicated that the high hydrostatic pressure treatment greatly increased the turkey ham commercial viability in comparison to the usual length, by slowing down the growth of microorganisms in the product.
Subject(s)
Humans , Lactic Acid/analysis , Lactic Acid/isolation & purification , Food Preservation/methods , Food Analysis , Food Microbiology , Foods Modified by Air Incorporation , Meat Products/analysis , Food Samples , Hydrostatic Pressure , Methods , TurkeysABSTRACT
Objective To investigate the effect of hydrostatic pressure and estrogen on the proliferation, F-actin cytoskeleton, osteogenic and chondrogenic differentiation of rat bone marrow mesenchymal stem cells (BMSCs), and to test whether combined stimulation can exert the fortified stimulating effort on BMSCs． Methods BMSCs were separated by using the whole bone marrow culture method and purified by differential adherence method. BMSCs surface markers were detected by flow cytometer. BMSCs were randomly assigned to six groups：blank control group (Group C), 1 nmol/L 17β-Estradiol treatment group (Group E), 1 nmol/L tamoxifen treatment group (Group T), 90 kPa pressure treatment group for 1 h (Group P); 17β-Estradiol pretreatment for 12 h and 90 kPa pressure group for 1 h (Group P+E); and tamoxifen pretreatmet for 12 h and 90 kPa pressure group for 1 h (Group P+T). Cell cycle was measured by flow cytometry. Fluorescent staining under laser scanning confocal microscope observation was observed for F-actin cytoskeleton expression and re-assembly. After osteogenic differentiation for 7 d and 14 d, calcified nodules were detected with alizarin red staining. Further, the osteogenic markers including Col I, ON, OPN and BSP were analyzed by real-time PCR. Following chondrogenesis of BMSCs for 14 d and 28 d, proteoglycan contents were detected with toluidine blue staining, and chondrogenic markers including Sox9, Aggrecan and ColⅡwere evaluated by real time PCR. ANOVAs followed by the Dunnett t tests were adopted for comparisons among subgroups. All the experimental data were analyzed by SPSS 16.0 software. Results Both hydrostatic pressure (90 kPa, 1 h) and 1 nmol/L17β-estradiol could increase the proliferation of BMSCs and F-actin activation, but no bio-cooperation effects appeared. Calcified nodules were observed after 14 d osteogenic induction. Real-time PCR showed the estrogen enhanced osteogenetic gene (Col I, ON, OPN and BSP) expression in 7 d and 14 d. Combined effects of pressure and estrogen showed synergistic improving effects on early osteogenetic differentiation, but oppositional effects on advanced osteogenetic differentiation. Toluidine blue staining was positive after 28 d chondrogenic induction. With the hydrostatic pressure loading regime, the mRNA expression of chondrogenic genes (Sox9, Aggrecan and ColⅡ) was increased significantly, but with oppositional effects from estrogen on advanced chondrogenic differentiation. Conclusions The superposition effects of mechanical stimulation and estrogen acting only enhanced the differentiation of BMSCs in the early osteogenetic differentiation, but no effect was found in the proliferation and F-actin activation. Hydrostatic pressure and estrogen show antagonistic action in advanced osteogenetic differentiation and chondrogenic differentiation. Estrogen promotes osteogenetic differentiation, while hydrostatic pressure can enhance chondrogenic differentiation of BMSCs.
ABSTRACT
PURPOSE: This in-vitro study aimed to confirm the clinical potential of a newly suggested bone substitute fabricated by a bio-plotting system. MATERIALS AND METHODS: A 3-dimensional, PCL (polycaprolactone)-based bone substitute was fabricated by a bio-plotting system, which enables us to control the shape and porosity. Mesenchymal stem cells(MSCs) from a New Zealand white rabbits were seeded to the bone substitute. Through 2-week experiment, morphological and various biological responses were assessed depending on the use of osteogenic media and/or mechanical stimuli. RESULTS: The viability of cells was confirmed. Other findings in relation to proliferation and differentiation were also confirmed to be biocompatible. Specifically, the activity of ALP was increased in the groups using osteogenic media along the time. And the expressions collagen type I was more affected by intermittent hydrostatic pressure rather than by osteogenic media. But those of Cbfa-1 was affected by osteogenic media and mechanical stimuli, as well. CONCLUSION: The potential of a newly fabricated and suggested material was confirmed. However, further long term and animal study are recommanded for the clinical application.