-
Food Research International (Ottawa,... May 2020The objective of this work was to produce solid lipid microparticles using fully hydrogenated anhydrous milk fat (FHAMF) and to evaluate their physical stability during...
The objective of this work was to produce solid lipid microparticles using fully hydrogenated anhydrous milk fat (FHAMF) and to evaluate their physical stability during 90 days of storage at different temperatures. To obtain the lipid microparticles, the FHAMF was sprayed in a double fluid atomizer at 1 bar pressure, in a chilled chamber (2 °C). After atomization, the microparticles were divided into three batches and stored for 90 days at three different temperatures (5, 15 and 25 °C). During storage, samples were periodically removed (7, 15, 30, 60 and 90 days) for evaluation of particle size, melting behavior, morphology, and polymorphic habit. The microparticles presented spherical shaped, with a smooth surface and wide size variation. When stored at 5 °C, the microparticles showed the smaller size and smaller agglomeration, due to the lower liquid fat content in the system, which that makes it difficult the adhesion of one particle to another. The lipid microparticles presented β' crystals immediately after processing and at all temperatures during the storage. This study demonstrated the potential of FHAMF as an appropriate lipid phase for the production of lipid microparticles, and may contribute to further studies on the delivery of active compounds.
Topics: Animals; Dairy Products; Hydrogenation; Lipids; Milk; Nanoparticles; Particle Size; Preservation, Biological; Spray Drying; Temperature
PubMed: 32247456
DOI: 10.1016/j.foodres.2020.109009 -
Poultry Science Oct 2021The present study focuses on the effect of different levels of ergothioneine and thawing temperature on rooster semen cryopreservation. Semen was diluted in Lake...
The present study focuses on the effect of different levels of ergothioneine and thawing temperature on rooster semen cryopreservation. Semen was diluted in Lake extender containing ergothioneine at 5, 10, 15, and 20 µM and cryopreserved. Two thawing temperatures (37°C for 30 s and 60°C for 5 s) were consequently examined. Sperm motility parameter, membrane integrity, abnormal morphology, viability, apoptotic status, mitochondria activity, and lipid peroxidation were determined after freeze-thaw process. Ergothioneine levels of 5 and 10 µM led to higher (P < 0.05) total motility (66.58 ± 1.44 and 72.11±1.44, respectively) and average path velocity (VAP) (34.54 ± 0.89, 37.28 ± 0.89, respectively). Higher (P < 0.05) significant membrane integrity and mitochondria activity after freeze-thawing were observed in the groups supplemented with 10 µM ergothioneine (68.62 ± 1.24 and 69.12 ± 1.26, respectively). Also, 5 and 10 µM of ergothioneine led to the lowest significant percentage of apoptotic and dead sperm. The total motility and progressive motility resulted in significantly (P < 0.05) higher amount when sperm were thawed with 60°C (60.58 ± 0.91 and 24.76 ± 0.53, respectively) compared to thawed sperm in 37°C. The membrane integrity, viability and mitochondria activity led to significantly (P < 0.05) higher when sperm were thawed with 60°C (58.2 ± 0.78, 63.21 ± 0.80 and 56.85 ± 0.79, respectively). It could be concluded the addition of 5 and 10 µM ergothioneine in the semen extender and thawing temperature at 60˚C in 5 s can be an efficient strategy to preserve rooster cryopreserved semen quality.
Topics: Animals; Chickens; Cryopreservation; Cryoprotective Agents; Ergothioneine; Male; Semen; Semen Analysis; Semen Preservation; Sperm Motility; Spermatozoa; Temperature
PubMed: 34464932
DOI: 10.1016/j.psj.2021.101405 -
Reproductive Biology and Endocrinology... Mar 2020Sperm cryopreservation has been widely used in assisted reproductive technology (ART) and has resulted in millions of live births. Two principal approaches have been... (Review)
Review
Sperm cryopreservation has been widely used in assisted reproductive technology (ART) and has resulted in millions of live births. Two principal approaches have been adopted: conventional (slow) freezing and vitrification. As a traditional technique, slow freezing has been successfully employed and widely used at ART clinics whereas the latter, a process to solidify liquid into an amorphous or glassy state, may become a faster alternative method of sperm cryopreservation with significant benefits in regard to simple equipment and applicability to fertility centers. Sperm vitrification has its own limitations. Firstly, small volume of load is usually plunged to liquid nitrogen to achieve high cooling rate, which makes large volume sample cryopreservation less feasible. Secondly, direct contact with liquid nitrogen increases the potential risk of contamination. Recently, new carriers have been developed to facilitate improved control over the volume and speed, and new strategies have been implemented to minimize the contamination risk. In summary, although sperm vitrification has not yet been applied in routine sperm cryopreservation, its potential as a standard procedure is growing.
Topics: Cryopreservation; Freezing; Humans; Male; Reproducibility of Results; Reproductive Techniques, Assisted; Semen Preservation; Sperm Motility; Spermatozoa; Vitrification
PubMed: 32145746
DOI: 10.1186/s12958-020-00580-5 -
Lancet (London, England) Jul 2019
Topics: Animals; Museums; Physicians; Preservation, Biological; Zoology
PubMed: 31327361
DOI: 10.1016/S0140-6736(19)31569-7 -
Platelets Dec 2023Platelet Rich Plasma (PRP) is a biological treatment which, thanks to its enhanced growth factors content, is widely used in the field of regenerative medicine for its...
Platelet Rich Plasma (PRP) is a biological treatment which, thanks to its enhanced growth factors content, is widely used in the field of regenerative medicine for its reparative effects. Although it is usually used fresh immediately after preparation, its freezing for preservation for future usage could be key in increasing its versatility and new applications. To assess the suitability of freezing, after collecting PRP and platelet lysates (PL) from 6 patients, they were preserved for 1 or 3 months at temperatures of -20ºC and -80°C. Measurements were then made on platelet number and integrity, growth factor levels, biomechanical properties of the clot and its bioactivity on cultured cells. Fresh PRP and PL were used as controls. The results showed an increase in platelet size ( < .01) and clot elasticity ( < .01), as well as decrease in levels of PDGF ( < .05) and VEGF ( < .05), though the overall bioactivity was not affected as culture cells showed the same responsiveness to both frozen and fresh PRP and PL in terms of cell viability. Based on these results, it could be assumed that preservation of PRP by freezing is a feasible and suitable option for its further use.
Topics: Humans; Cryopreservation; Intercellular Signaling Peptides and Proteins; Cells, Cultured; Cell Culture Techniques; Platelet-Rich Plasma
PubMed: 37165543
DOI: 10.1080/09537104.2023.2210243 -
Postepy Biochemii Dec 2022Cryopreservation (banking) techniques have been known to nature for centuries. Many species of insects, amphibians, fish and even reptiles use natural cryopreservation...
Cryopreservation (banking) techniques have been known to nature for centuries. Many species of insects, amphibians, fish and even reptiles use natural cryopreservation methods to survive the harsh conditions of winter or to live in extremely cold temperatures. Cryopreservation and dreams of immortality have intrigued humanity for years. The first reports of observing the effects of freezing sperm (stored in snow) date back to 1776. In 1866, Montegazza was the first to suggest a vision completely unimaginable for the time: "a man dying on the battlefield can conceive an heir from sperm frozen and stored at home". The first, at that time still unsuccessful, reports of laboratory freezing of human sperm date back to the 1930s [1]. Finally, mankind "learned" cryopreservation in the middle of the twentieth century, when on October 15, 1949, the article "Revival of spermatozoa after vitrification and dehydration at low temperatures" appeared in print in the Nature journal, summarising the pioneering research of scientists from the National Institute for Medical Research, Mill Hill, London [2]. This concerned the freezing of fowl sperm in the presence of glycerol, ethylene glycol and propylene glycol in such a way that after thawing it was able to fertilise eggs effectively. The subsequent use of dimethyl sulfoxide (DMSO) revolutionised modern cryobiology [3-5]. Thus began the era of cryopreservation, without which today it is difficult to imagine the work of cell biology laboratories, modern animal breeding, or the development of modern medicine.
Topics: Animals; Male; Humans; Cryoprotective Agents; Semen; Semen Preservation; Sperm Motility; Cryopreservation
PubMed: 36649141
DOI: 10.18388/pb.2021_461 -
American Journal of Transplantation :... Feb 2022Ex vivo lung perfusion (EVLP) is a novel lung preservation strategy that facilitates the use of marginal allografts; however, it is more expensive than static cold...
Ex vivo lung perfusion (EVLP) is a novel lung preservation strategy that facilitates the use of marginal allografts; however, it is more expensive than static cold storage (SCS). To understand how preservation method might affect postoperative costs, we compared outcomes and index hospitalization costs among matched EVLP and SCS preserved lung transplant (LTx) recipients at a single, high-volume institution. A total of 22 EVLP and 66 matched SCS LTx recipients were included; SCS grafts were further stratified as either standard-criteria (SCD) or extended-criteria donors (ECD). Median total preservation time was 857, 409, and 438 min for EVLP, SCD, and ECD lungs, respectively (p < .0001). EVLP patients had similar perioperative outcomes and posttransplant survival compared to SCS SCD and ECD recipients. Excluding device-specific costs, total direct variable costs were similar among EVLP, SCD, and ECD recipients (median $200,404, vs. $154,709 vs. $168,334, p = .11). The median direct contribution margin was positive for EVLP recipients, and similar to that for SCD and ECD graft recipients (all p > .99). These findings demonstrate that the use of EVLP was profitable at an institutional level; however, further investigation is needed to better understand the financial implications of EVLP in facilitating donor pool expansion in an era of broader lung sharing.
Topics: Costs and Cost Analysis; Humans; Lung; Lung Transplantation; Organ Preservation; Perfusion; Tissue Donors
PubMed: 34379885
DOI: 10.1111/ajt.16794 -
International Journal of Molecular... Mar 2022Spermatozoa cryopreservation is an important technique to preserve fertility for males. This study aimed at exploring the stability of epigenetics information in human...
INTRODUCTION
Spermatozoa cryopreservation is an important technique to preserve fertility for males. This study aimed at exploring the stability of epigenetics information in human spermatozoa, manipulated by two different technologies, freezing and vitrification.
METHODS
Spermatozoa samples were distributed into three groups: 1. Fresh spermatozoa (control group), 2. Frozen spermatozoa, 3. Vitrified spermatozoa. Epigenetic differences of fresh and cryopreserved spermatozoa were evaluated using high-throughput RNA sequencing.
RESULTS
Differentially expressed genes (DEGs) in frozen (1103 genes) and vitrified (333 genes) spermatozoa were evaluated. The bioinformatical analysis identified 8 and 15 significant pathways in groups of frozen and vitrified spermatozoa, respectively. The majority of these pathways are most relevant to immune and infectious diseases. The DEGs of the fertilization process are not detected during vitrification. The freezing process induces more down-regulation of genes and is relevant to apoptosis changes and immune response.
CONCLUSION
Cryopreservation of human spermatozoa is an epigenetically safe method for male fertility preservation. Cryoprotectant-free vitrification can induce more minor biological changes in human spermatozoa, in comparison with conventional freezing.
Topics: Cryopreservation; Cryoprotective Agents; Freezing; Humans; Male; Semen Preservation; Sperm Motility; Spermatozoa; Vitrification
PubMed: 35328464
DOI: 10.3390/ijms23063047 -
International Journal of Molecular... Sep 2023Telomeres play pivotal roles in processes closely related to somatic senescence and aging, making them a compelling target for interventions aimed at combating aging and...
Telomeres play pivotal roles in processes closely related to somatic senescence and aging, making them a compelling target for interventions aimed at combating aging and age-related pathologies. Ginsenoside, a natural compound, has emerged as a potential remedy for promoting healthy aging, yet how it protects telomeres remains incompletely understood. Here, we show that treatment of F1 can effectively restore the level of TRF2, thereby preserving telomere integrity. This restoration leads to inhibition of the DNA damage response and improvements in mitochondrial function and, ultimately, delays in cellular senescence. Conversely, depletion of TRF2 causes mitochondrial dysfunction, accompanied by increased oxidative stress, autophagy inhibition, insufficient energy metabolism, and the onset of cellular senescence. These observations underscore the critical role of TRF2 in maintaining telomere integrity and direct association with the initiation of cellular senescence. We conduct a further analysis, suggesting F1 could bind in proximity to the TRF2 heterodimer interface, potentially enhancing dimerization stability. These findings suggest that F1 may be a promising natural remedy for anti-aging, and restoring TRF2 could potentially prevent telomere-dependent diseases commonly associated with the aging process.
Topics: Humans; Ginsenosides; Cellular Senescence; Preservation, Biological; Syndrome
PubMed: 37762556
DOI: 10.3390/ijms241814241 -
Frontiers in Immunology 2022Normothermic machine perfusion (NMP) is a technique of kidney preservation designed to restore cellular metabolism after cold ischemia. Kidneys are perfused with an... (Randomized Controlled Trial)
Randomized Controlled Trial
Normothermic machine perfusion (NMP) is a technique of kidney preservation designed to restore cellular metabolism after cold ischemia. Kidneys are perfused with an oxygenated banked red blood cell (RBC) based solution for 1h at 36°C. During NMP, RBCs can become damaged, releasing free heme into the perfusate. This can act as a damage-associated molecular pattern (DAMP) activating inflammatory signalling pathways. The aim of this study was to measure the levels of free heme during NMP, assess the effect on kidney function and determine any association with inflammatory and stress related gene expression. Levels of free heme were measured in perfusate samples from a series of donation after circulatory death (DCD) kidneys undergoing NMP as part of a randomised controlled trial (RCT). The age of RBCs and levels of free heme were correlated with perfusion parameters. Changes in gene expression were analysed in a series of kidneys declined for transplantation using the NanoString nCounter Organ Transplant Panel and qRT-PCR. Older units of RBCs were associated with higher levels of free heme and levels increased significantly during NMP (Pre 8.56 ± 7.19µM vs 26.29 ± 15.18µM, P<0.0001). There was no association with levels of free heme and perfusion parameters during NMP (P > 0.05). Transcriptional and qPCR analysis demonstrated the upregulation of differentially expressed genes associated with apoptosis (FOS and JUN), inflammatory cytokines (IL-6, SOCS3, ATF3), chemokines (CXCL8, CXCL2, CC3/L1) and oxidative stress (KLF4) after NMP. However, these did not correlate with levels of free heme (P >0.05). A significant amount of free heme can be detected in the perfusate before and after NMP particularly when older units of red cells are used. Although transcriptional analysis demonstrated significant upregulation of genes involved with apoptotic, inflammatory and oxidative pathways these were not associated with high levels of free heme.
Topics: Cold Ischemia; Heme; Humans; Kidney; Organ Preservation; Perfusion
PubMed: 35585981
DOI: 10.3389/fimmu.2022.849742