High hydrostatic pressure influences the in vitro response to xenobiotics in Dicentrarchus labrax liver

2016 ◽  
Vol 173 ◽  
pp. 43-52 ◽  
Author(s):  
Benjamin Lemaire ◽  
Eric Mignolet ◽  
Cathy Debier ◽  
Pedro Buc Calderon ◽  
Jean Pierre Thomé ◽  
...  
Keyword(s):  
Hydrostatic Pressure ◽  
High Hydrostatic Pressure ◽  
Dicentrarchus Labrax ◽  
In Vitro Response

scholarly journals High hydrostatic pressure: a new way to improve in vitro developmental competence of porcine matured oocytes after vitrification

Reproduction ◽  
2007 ◽  
Vol 135 (1) ◽  
pp. 13-17 ◽  
Author(s):  
Y Du ◽  
C S Pribenszky ◽  
M Molnar ◽  
X Zhang ◽  
H Yang ◽  
...  
Keyword(s):  
Hydrostatic Pressure ◽  
High Hydrostatic Pressure ◽  
Developmental Competence

Verification of the Inactivation of Melanocytic Nevus in vitro Using a Newly Developed Portable High Hydrostatic Pressure Device

2016 ◽  
Vol 201 (3) ◽  
pp. 170-179 ◽  
Author(s):  
Naoki Morimoto ◽  
Chizuru Jinno ◽  
Atsushi Mahara ◽  
Natsuko Kakudo ◽  
Toshia Fujisato ◽  
...  
Keyword(s):  
Hydrostatic Pressure ◽  
High Hydrostatic Pressure ◽  
Physical Method ◽  
Melanocytic Nevus ◽  
Explant Culture ◽  
Cultured Fibroblasts ◽  
Giant Congenital Melanocytic Nevi ◽  
Tissue Specimens ◽  
Congenital Melanocytic Nevi

High hydrostatic pressure (HHP) technology is a physical method for inactivating tissue. We reported that nevus specimens were inactivated after HHP at 200 MPa and that the inactivated nevus could be used as autologous dermis for covering skin defects. In this study, we verified the inactivation of nevus specimens using a newly developed portable HHP device which will be used in a clinical trial. Nevus tissue specimens were obtained from 5 patients (mean age 7.2 years, range 1-19). We cultured fibroblasts and nevus cells from the tissue specimens and then evaluated their inactivation after HHP at 200 MPa by confirming the attachment of the suspensions and by the live/dead staining of the suspensions, through the dissociation of the cells on chamber slides and by the live/dead staining of the remaining cells. The cells were also quantitatively evaluated by WST-8 assay. We then confirmed the inactivation of the nevus specimens after HHP using explant culture. Our results indicated that fibroblasts and nevus cells were inactivated after HHP at 200 MPa, with the exception of a small percentage of green-colored cells, which reflected the remaining activity of the cellular esterases after HHP. No cells migrated from the nevus specimens after HHP at 200 MPa. We verified the inactivation of fibroblasts and nevus cells cultured from nevus specimens, and in the nevus samples themselves after pressurization at 200 MPa using this device. This device could be used in clinical trials for giant congenital melanocytic nevi and may thus become useful in various medical fields.

28 HIGH HYDROSTATIC PRESSURE IMPROVED DEVELOPMENTAL COMPETENCE OF PORCINE OOCYTES FOR HANDMADE CLONING

2008 ◽  
Vol 20 (1) ◽  
pp. 94 ◽  
Author(s):  
Y. Du ◽  
L. Lin ◽  
C. Pribenszky ◽  
M. Molnár ◽  
P. M. Kragh ◽  
...  
Keyword(s):  
Hydrostatic Pressure ◽  
High Hydrostatic Pressure ◽  
Nuclear Transfer ◽  
Technical Assistance ◽  
Developmental Competence ◽  
Control Groups ◽  
Donor Cells ◽  
And Control ◽  
Handmade Cloning

High hydrostatic pressure (HHP) has been introduced into the field of embryology recently, with the possible mechanism that a sublethal HHP could induce the synthesis of molecular chaperons to protect the embryos from further stresses. Improved cryotolerance has been achieved successfully in HHP-treated mouse (Pribenszky 2005 Anim. Reprod. Sci. 87, 143–150) and bovine (Pribenszky 2005 Reprod. Domest. Anim. 40, 338) embryos, and the semen of bull (Pribenszky 2007 Reprod. Fertil. Dev. 19, 181–182) and boar (Pribenszky 2005 Reprod. Fertil. Dev. 18, 162–163). The objective of the present study was to apply this new technique to in vitro-matured (IVM) porcine oocytes and further investigate its effect in the procedure of handmade cloning (HMC). After 40 h IVM, cumulus–oocyte complexes (COCs) were loaded in 0.5-mL straws by a 2-mL syringe, with HEPES-buffered TCM199 as the loading medium. COCs were then treated with 20 MPa (200 times greater than atmospheric pressure) for 60 min by a pressurizing device (Cryo-Innovation Inc., Budapest, Hungary), with an interval of 120 min between HHP treatment and subsequent HMC. Two different cell lines (from Day 40 fetuses of Yucatan and Danish Landrace breeds (LW1-2)) were used as donor cells for nuclear transfer. A total of 592 reconstructed embryos were produced from both HHP-treated and control groups and were in vitro cultured for 6 days to evaluate the developmental competence through to blastocyst formation. The effect of donor cells on blastocyst development was also investigated. SPSS 11.0 program (SPSS, Inc., Chicago, IL, USA) was used for statistical analysis; values with P < 0.05 were regarded as significant. Blastocyst rates of the different groups are shown in Table 1. Our results indicated that COCs treated with HHP had a much higher blastocyst rate than those untreated (P < 0.01) and this improvement was not affected by using different donor cells for nuclear transfer. In conclusion, the sublethal HHP treatment could improve the in vitro developmental competence of porcine IVM oocytes when they are used for HMC. Further in vivo experiments are required to investigate the long-term effect of HHP on embryo development. Table 1. Day 6 blastocyst rates of HHP-treated and control groups with different donor cells for nuclear transfer The authors thank Ruth Kristensen and Janne Adamsen for their help and excellent technical assistance.

Effects of high hydrostatic pressure on bacterial cytoskeleton FtsZ polymers in vivo and in vitro

Microbiology ◽  
2004 ◽  
Vol 150 (6) ◽  
pp. 1965-1972 ◽  
Author(s):  
Akihiro Ishii ◽  
Takako Sato ◽  
Masaaki Wachi ◽  
Kazuo Nagai ◽  
Chiaki Kato
Keyword(s):  
Cell Division ◽  
Hydrostatic Pressure ◽  
High Hydrostatic Pressure ◽  
Elevated Pressure ◽  
Immunofluorescent Staining ◽  
E Coli ◽  
Division Process ◽  
Indirect Immunofluorescent

Some rod-shaped bacteria, including Escherichia coli, exhibit cell filamentation without septum formation under high-hydrostatic-pressure conditions, indicating that the cell-division process is affected by hydrostatic pressure. The effects of elevated pressure on FtsZ-ring formation in E. coli cells were examined using indirect immunofluorescence microscopy. Elevated pressure of 40 MPa completely inhibited colony formation of E. coli cells under the cultivation conditions used, and the cells exhibited obviously filamentous shapes. In the elongated cells, normal cell-division processes appeared to be inhibited, because no FtsZ rings were observed by indirect immunofluorescent staining. In addition, it was observed that hydrostatic pressure dissociated the E. coli FtsZ polymers in vitro. These results suggest that high hydrostatic pressure directly affects cell survival and morphology through the dissociation of the cytoskeletal frameworks.

scholarly journals High Hydrostatic Pressure for Disinfection of Bone Grafts and Biomaterials: An Experimental Study

2009 ◽  
Vol 3 (1) ◽  
pp. 1-7 ◽  
Author(s):  
Hans Gollwitzer ◽  
Wolfram Mittelmeier ◽  
Monika Brendle ◽  
Patrick Weber ◽  
Thomas Miethke ◽  
...  
Keyword(s):  
Hydrostatic Pressure ◽  
Bacterial Growth ◽  
High Hydrostatic Pressure ◽  
Chronic Osteomyelitis ◽  
Bone Grafts ◽  
Biological Properties ◽  
Human Bone ◽  
Bacterial Inactivation ◽  
Bacterial Survival

Background:Autoclaving, heat, irradiation or chemical detergents are used to disinfect autografts, allografts and biomaterials for tissue reconstruction. These methods are often associated with deterioration of mechanical, physical, and biological properties of the bone grafts and synthetic implants. High hydrostatic pressure has been proposed as a novel method preserving biomechanical and biological properties of bone, tendon and cartilage. This is the first study to assess the inactivation of clinically relevant bacteria on biomaterials and human bone by high hydrostatic pressure.Methods:Bacterial suspensions ofStaphylococcus aureus,Pseudomonas aeruginosaandEnterococcus faecium, implants covered with infected blood, human bone infectedin vitro, and biopsies of patients with chronic osteomyelitis were subjected to different protocols of high hydrostatic pressure up to 600 MPa. Bacterial survival after high hydrostatic pressure treatment was determined and compared with bacterial growth in untreated controls.Results:S. aureusandP. aeruginosain suspension were completely inactivated by high hydrostatic pressure (> 5log levels), whereasE. faeciumshowed barotolerance up to 600 MPa. Blood and adherence to metal implants did not significantly alter inactivation of bacteria, and complete disinfection was achieved with barotolerant bacteria (S. aureusandP. aeruginosa). However, osteoarthritic bone demonstrated a non-homogeneous baroprotective effect, with single bone samples resistant to treatment resulting in unaltered bacterial growth, and complete disinfection of artificially infected bone specimens was achieved in 66% forS. aureus, 60% forP. aeruginosaand 0% forE. faecium. Human bone samples of patients with chronic osteomyelitis could be completely disinfected in 2 of 37 cases.Conclusion:High hydrostatic pressure offers new perspectives for disinfection of sensitive biomaterials and bone grafts, and contamination by blood did not significantly affect bacterial inactivation rates. However, a significant baroprotective effect was demonstrated in bone. Effectiveness is currently limited to colonization and / or infection with barosensitive micro-organisms.

scholarly journals The impact of high hydrostatic pressure (40 MPa and 60 MPa) on the apoptosis rates and functional activity of cryopreserved porcine mesenchymal stem cells

2018 ◽  
Vol 18 (1) ◽  
pp. 69-86
Author(s):  
Joanna Romanek ◽  
Jolanta Opiela ◽  
Zdzisław Smorąg
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Hydrostatic Pressure ◽  
Functional Activity ◽  
High Hydrostatic Pressure ◽  
Negative Impact ◽  
Caspase 8 ◽  
Significant Difference ◽  
The Impact

AbstractThe aim of the present study was to examine the influence of two varied high hydrostatic pressure (HHP) values on the apoptosis (assessing caspase-8, survivin, CAD, Bax, BclxL and BclxS) and functional activity (using cocultures with bovine embryos) of porcine mesenchymal stem cells (pBMSCs). pBMSCs were isolated from porcine bone marrow and cultured in vitro. Before cryopreservation and storage in liquid nitrogen, pBMSCs were subjected to HHP values of 40 MPa and 60 MPa for 1 h at 24°C. After thawing, the cells were analysed for caspase-8 activity and protein expression of survivin, CAD, Bax, BclxL and BclxS. To indirectly test the influence of HHP on the functional activity of pBMSCs, in vitro maturated bovine oocytes were fertilized in vitro, and the obtained embryos were cultured under 4 different conditions: 1. monoculture in SOF medium; 2. coculture with pBMSCs in SOF medium; 3. coculture with pBMSCs subjected to 40 MPa HHP in SOF medium and 4. coculture with pBMSCs subjected to 60 MPa HHP in SOF medium. The quality of the developed blastocysts was analysed by TUNEL assay. HHP did not induce apoptosis in pBMSCs, as no significant difference was noted in the expression of any of the analysed apoptosis- related proteins between pBMSCs subjected to HHP (40 MPa or 60 MPa) and control. The highest number of obtained blastocysts was observed when the embryos were cultured in SOF. A highly significant difference (P<0.005) was noted between embryos cultured in SOF and embryos cultured in the presence of pBMSCs subjected to 60 MPa HHP or untreated pBMSCs. A significant difference (P<0.05) was noted between embryos cultured in SOF and embryos cultured in the presence of pBMSCs subjected to 40 MPa HHP. In conclusion, HHP does not induce apoptosis in pBMSCs. The obtained results of the blastocysts cocultured in vitro with pBMSCs (HHP-treated and untreated cells) imply that coculture with pBMSCs has a negative impact on the developmental rates of blastocysts.

87 HIGH HYDROSTATIC PRESSURE TREATMENT IMPROVES THE QUALITY OF IN VITRO-PRODUCED OVINE BLASTOCYSTS

2010 ◽  
Vol 22 (1) ◽  
pp. 202 ◽  
Author(s):  
L. Bogliolo ◽  
F. Ariu ◽  
S. Uccheddu ◽  
A. Strina ◽  
I. Rosati ◽  
...  
Keyword(s):  
Hydrostatic Pressure ◽  
Beneficial Effect ◽  
High Hydrostatic Pressure ◽  
Cell Number ◽  
Embryo Cryopreservation ◽  
Hatching Rate ◽  
Pressure Treatment ◽  
Custom Made

High hydrostatic pressure (HHP) has been introduced into the field of embryology recently. It has been reported that a sublethal HHP could induce the synthesis of molecular chaperons to protect the embryos from other environmental stresses. The possible beneficial effect of HHP was examined on embryo vitrification, and improved cryotolerance has been achieved in HHP-treated mouse and bovine embryos. In the present study we evaluated, first, the behavior of in vitro-produced ovine blastocysts after treatment with 2 different HHP conditions. In the second part of the study, optimized pressure treatment was combined with a cryopreservation protocol. Day 7 in vitro-produced ovine blastocysts were loaded in 0.5-mL straws and pressure treated in a custom-made hydrostatic pressure chamber (Cryo-Innovation Ltd., Budapest, Hungary) according to the following conditions: -60 MPa, 70 min, 38°C; and -40 MPa, 70 min, 38°C. After HHP treatment, embryos were cultured for 24 h and their survival evaluated upon morphological appearance, re-expansion of the blastocoel, and hatching from the zona pellucida. Blastocysts were fixed and stained, observed with confocal microscope to evaluate blastocyst cell number, and mitotic and picnotic index. Untreated blastocysts were used as controls. On the basis of results obtained in this experiment, groups of blastocysts were exposed to 40 MPa for 70 min at 38°C and, following 2 h incubation after HHp treatment, were vitrified using cryotops. After warming, embryos were checked for re-expansion and hatching and finally fixed and stained as described for fresh embryos. Vitrified untreated blastocysts were used as controls. Results showed that the hatching rate of embryos treated at 40 MPa was significantly higher than that of 60 MPa-treated group (87.5 v. 0%; P < 0.01, chi-squared test) and similar to that of control embryos (85.3%) after 24 h culture. Blastocysts exposed at 40 MPa showed higher cell number compared with control embryos (161.3 ± 8.7 v. 123.9 ± 9.4, P < 0.01, ANOVA ± SD) and a lower picnotic index (PI: 1.3 ± 0.4% v. 3.8 ± 0.6%, P < 0.05). Similar re-expansion rate has been recorded between HHP treated and untreated (86.6 v. 89.6%) vitrified blastocysts as well as hatching rates (53.3 v. 48.3) and embryo cell number (131 ± 4.6 v. 139 ± 5.8). Conversely, HHP treatment significantly decreased picnotic index of vitrified treated embryos compared with control (2.3 ± 0.9 v. 4.8 ± 0.5, P < 0.05). We concluded that HHP treatment significantly improved the quality of in vitro-produced ovine blastocysts by increasing their cell number and reducing the proportion of picnosis. The HHP treatment exerted a positive effect in vitrified blastocysts, decreasing the number of picnotic nuclei. Further studies are needed to evaluate the possible beneficial effect at the molecular level of HHP treatment after vitrification and also to test if different interval times between HHP treatment and embryo cryopreservation could affect embryo response.

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