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PubMed1992

Adaptation of rabbit cortical collecting duct to in vitro acid incubation.

Cortical collecting ducts (CCDs) isolated from acid-loaded rabbits and perfused in vitro absorb HCO3-, whereas CCDs from normal animals secrete HCO3-. We have previously shown that CCDs incubated in vitro for 3 h at pH 6.9 show a reduction in net (baseline and stimulated) HCO3- secretion. In this study we ascertained the minimum duration of an acidic stimulus necessary to induce adaptive changes in stimulated HCO3- secretion (determined in the absence of basolateral Cl-) and the roles of protein synthesis and cytoskeletal function in this process. CCDs incubated in acid (pH 6.8, HCO3- 6 mM) for 1 h followed by incubation at pH 7.4 (HCO3- 25 mM) for 2 h showed a 41% reduction in stimulated HCO3- secretion (P < 0.001), similar to that observed after 3 h of incubation at pH 6.8. However, this incubation protocol failed to enhance stimulated HCO3- absorption (determined in the absence of luminal Cl-). Addition of 10 microM anisomycin, a reversible inhibitor of protein synthesis, throughout the entire period of incubation (1 h at pH 6.8 plus 2 h at pH 7.4) blocked adaptive reduction in HCO3- secretion, as did exposure to anisomycin only during the initial 1 h of acid incubation. In contrast, anisomycin application during the 2-h incubation at pH 7.4 failed to block this adaptation of HCO3- secretion. Application of 4 microM actinomycin D, an inhibitor of DNA transcription, during the acid incubation also prevented the adaptive response, as did application during the total or during the 2-h pH 7.4 incubation period of 0.2 microM cytochalasin D, an inhibitor of actin filament function.(ABSTRACT TRUNCATED AT 250 WORDS)

Absorption↗
PubMed1992

Time-related morphological changes of porcine ovarian granulosa cells incubated with urea-EDTA solution.

Morphological profiles of porcine granulosa cells incubated with 10 mM Tris-HCl containing 1 M urea and 5 mM EDTA (urea-EDTA solution) were investigated. The percentages of granulosa cells incorporating the dye, trypan blue on incubation with urea-EDTA solution did not change during the initial 30 min. Thereafter, granulosa cells gradually took up the dye throughout the incubation. The amount of protein released from granulosa cells increased dramatically during initial 15 min of incubation, but decreased during the following 15-30 min of incubation. Thereafter, the amount of proteins released from granulosa cells increased gradually again. The releasing profile of 51Cr-bounded substances in granulosa cells increased markedly during the initial 15 min of incubation, and decreased during the next 15 min of incubation. Subsequently the amount of 51Cr released was enhanced. The min of incubation. Subsequently the amount of 51Cr released was enhanced. The plasma membranes of granulosa cells remained intact at 30 min of incubation, although chromatin clusters of granulosa cells disappeared. Thereafter, a number of cells showed signs of degeneration, including broken plasma membrane and cytolysis. The present study revealed that urea-EDTA solution is useful in extracting materials from porcine granulosa cells. The majority of the materials extracted from granulosa cells during the initial 30 min of incubation with urea-EDTA solution is considered to be from the cell surfaces and/or intercellular matrix.

Animals↗
PubMed1999

Preterm infant thermal care: differing thermal environments produced by air versus skin servo-control incubators.

OBJECTIVE: Incubator thermal environments produced by skin versus air servo-control were compared. STUDY DESIGN: Infant abdominal skin and incubator air temperatures were recorded from 18 infants in skin servo-control and 14 infants in air servo-control (26- to 29-week gestational age, 14 +/- 2 days postnatal age) for 24 hours. Differences in incubator and infant temperature, neutral thermal environment (NTE) maintenance, and infant and incubator circadian rhythm were examined using analysis of variance and scatterplots. RESULTS: Skin servo-control resulted in more variable air temperature, yet more stable infant temperature, and more time within the NTE. Circadian rhythm of both infant and incubator temperature differed by control mode and the relationship between incubator and infant temperature rhythms was a function of control mode. CONCLUSION: The differences between incubator control modes extend beyond temperature stability and maintenance of NTE. Circadian rhythm of incubator and infant temperatures is influenced by incubator control.

Body Temperature Regulation↗
PubMed2003

Accelerating embryonic growth during incubation following prolonged egg storage. 1. Embryonic livability.

The hypothesis was proposed that shorter incubation periods and faster growth rates for long-stored eggs would improve embryonic survival and poult hatchling quality. Increased incubation temperatures were tested for their efficacy in improving embryonic livability in fertilized eggs stored for 15 d prior to setting in the incubator compared to controls stored for only 3 d. Two temperature treatments were applied. In experiment 1, a 37.8 degrees C set point for dry bulb temperature was used to accelerate development for the initial 2 wk compared to the controls at 37.5 degrees C. Following treatment, the accelerated embryos were returned to the same machine as the controls. In experiment 2, higher temperature exposure was only for the initial week of incubation. The temperature and storage treatments were in a completely random 2 x 2 factorial arrangement of treatments. At the completion of 28 d of incubation, survival rates of all treatments were determined by opening all nonhatching eggs to differentiate truly fertilized eggs from unfertilized. Hatchability was determined by dividing the total number of poults on a hatching tray by the number of fertilized eggs on a tray. Incubator trays were the experimental unit. Tissues were sampled in both experiments to verify treatment effects on growth and metabolism. Hatching times were observed at 4-h intervals during the actual hatching process beginning at 25 d of incubation. It was concluded that delayed growth and depressed metabolism of fertilized turkey eggs stored for 15 d can be compensated for by exposure to higher incubation temperatures for the initial 1 or 2 wk of incubation.

Animals↗
PubMed1983

The effects of incubator carbon dioxide level on turkey hatchability.

Four experiments involving CO2 level in single-stage incubators were conducted at a commercial turkey hatchery. Carbon dioxide was injected into incubators to a level of .3% during the first 10 days of incubation. Noninjected control incubators were maintained at a .1% CO2. Eggs from the injected incubators had a higher hatchability and lower early, late, and malpositioned embryonic mortality. No difference in hatchability was found between .3 and .5% CO2 concentrations for the first 10 days of incubation. Carbon dioxide injection for the first 5 days of incubation decreased hatchability when compared to CO2 injection for the first 10 days. Carbon dioxide injection for the first 14 days of incubation resulted in no difference in hatchability from injection for the first 10 days. The artificial addition of CO2 to single-stage turkey incubation systems may be of economic importance in commercial hatcheries because of its effect on hatchability.

Animals↗
PubMed2003

Study of extremely low frequency electromagnetic fields in infant incubators.

OBJECTIVES: The aim of the work was to present the results of measurements of extremely low frequency electromagnetic fields (ELF EMF), namely the magnetic flux density, inside infant incubators, and to compare these results with the data published by other authors who point out to a possible association between leukemia or other diseases observed in newborns kept in incubators after the birth and the ELF EMF exposure in the incubator. The measured magnetic flux densities were compared with the reference values for this frequency range indicated in the European Union (EU) recommendations. MATERIALS AND METHODS: The repeated measurements in incubators were made with a calibrated magnetometer EFA 300 in the frequency range of 5-30 kHz. Effective values of magnetic flux densities of ELF EMF were determined taking account of the reference values. RESULTS: The results of many repeated measurements showing the values of magnetic flux density in modern incubators with plastic supporting frame, were compared with those obtained in old type incubators with iron skeleton. A power frequency of 50 Hz was detected in the incubator and the ELF EMF values were by over two orders lower than the EU reference values. CONCLUSIONS: The paper emphasizes the need to take a special care of newborns kept in incubators even if only the sub-reference values are detected. The EU reference values are intended for the adult human population. A baby in an incubator has much smaller dimensions, higher electric conductivity and maybe trigger another mechanism of response to ELF EMF than that indicated in this paper.

Electromagnetic Fields↗
PubMed1979

[Studies on noise stress caused by infant incubators (author's transl)].

The following acoustic effects of incubators were investigated: Noise level and vibration measurements inside incubators. Noise emanation into the vicinity of incubators. Deadening of incubator noise by the hood. The noise intensity inside the incubators was also registered under conditions of intensive care using sound emitting therapeutic and monitoring equipment. The results show that the noise level of incubator motors can be tolerated. This applies to well-serviced incubators only, however. The hood muffles outside noise, particularly in the range of greatest hearing acuity. But there is no protection against noisy intensive-care systems within the incubator. The infants own noise production is considered to contribute substantially to the noise within an incubator.

Humans↗
PubMed1996

Changes in pituitary somatotroph and lactotroph distribution in laying and incubating turkey hens.

Turkey hens can rapidly shift from a laying condition to one characterized by ovarian regression, incubation behavior, and hyperprolactinemia. Although remarkable changes occur in hormonal profiles as turkey hens pass from a laying to an incubating state, studies have not been undertaken to examine histochemical alterations of functionally relevant pituicytes in the adenohypophysis. The objective of this study was to compare the immunocytochemical changes in pituitary lactotrophs and somatotrophs in incubating turkey hens with those of egg laying hens. Based upon nest visiting and egg production records, laying and incubating hens were selected for sampling blood, pituitaries, and ovaries. Plasma prolactin (PRL) and growth hormone (GH) concentrations were determined. Sagittal pituitary sections of laying and incubating hens were immunostained using antibodies against turkey growth hormone or synthetic chicken PRL peptide. Somatotrophs were found predominantly in the caudal lobe while lactotrophs occurred only in the cephalic lobe of adenohypophysis in laying hens. In incubating hens, somatotrophs in the ventral half of the caudal lobe were replaced by lactotrophs. The sagittal area which immunostained for PRL was significantly greater while the area that immunostained for GH was less in the adenohypophysis of incubating turkey hens. Some of the lactotrophs were hypertrophied in incubating hens. The lactotrophic recruitment and hypertrophy provide a cellular basis for the hyperprolactinemia in incubating turkey hens.

Animals↗
PubMed2006

Role of phospholipid transfer protein on the plasma distribution of amphotericin B following the incubation of different amphotericin B formulations.

PURPOSE: The purpose of this study was to investigate the role of phospholipid transfer protein (PLTP) on the plasma distribution of amphotericin B (AmpB) following incubation with different AmpB formulations in human plasmas with varying lipid profiles. METHODS: In a first set of experiments, plasma distribution profiles of AmpB were determined following the incubation of Fungizone and lipid-based formulations (Abelcet and AmBisome) at a concentration of 20 microg AmpB/mL for 5-120 min at 37 degrees C in the plasma obtained from six different individuals (total cholesterol concentrations range between 62 and 332 mg/dL). In a second set of experiments, Abelcet, and AmBisome at a concentration of 20 microg AmpB/mL were incubated for 5 min at 37 degrees C in human plasma (total cholesterol = 163 mg/dL) that had been pretreated with an antibody raised up against PLTP (1:400 v/v dilution from stock solution) for 20 min at 37 degrees C. Following incubation, the human plasma was separated into its lipoprotein and lipoprotein-deficient fractions by density gradient ultracentrifugation and analyzed for AmpB content by high-performance liquid chromatography. RESULTS: The majority of AmpB was covered in the lipoprotein-deficient plasma and high-density lipoprotein (HDL) fractions following incubation of Fungizone in human plasma. The majority of AmpB (48.7-87.2%) was recovered in the HDL fraction following incubation of Abelcet and AmBisome in human plasma. The presence of the PLTP antibody resulted in a 20% decrease in the percentage AmpB recovered in the HDL fraction following the incubation of Abelcet. However, the plasma distribution of AmpB remained unchanged following the incubation of AmBisome in plasma containing the PLTP antibody. CONCLUSIONS: Taken together, these findings suggest indirect evidence that PLTP may play an important role in the plasma distribution profile of AmpB following the incubation of Abelcet and may be one of the factors responsible for the preferential association of AmpB with HDL when administered as Abelcet.

Amphotericin B↗
PubMed1981

Studies on the change of electrophoretic mobility and the decay of catalytic activity of brain-type creatine kinase isoenzyme (CK-BB) following incubation at 37 degrees C.

Incubation of CK-BB in serum (1:3, v/v) at 37 degrees C for 3 h caused a change of its electrophoretic mobility and decay of its catalytic activity. Similar effects were observed following incubation in water. Incubation in saline somehow preserved the electrophoretic mobility but not the catalytic activity. No effect was noted when incubated at 4 degrees C for 3 h. Further study on the rate of decay revealed that the decay in albumin solution (1:50, v/v) is quite similar to that in serum. More dramatic decay was noted when incubated in water and less when incubated in saline. It was further shown that the higher the incubation temperature (4 degrees C, 25 degrees C or 37 degrees C) the faster the decay. The rate of decay of CK-MM was much slower in all conditions of incubation. Determination of isoenzyme activities by means of an immunoprecipitation method again demonstrated that CK-BB lost a great deal of its catalytic activity following incubation at 37 degrees C for 1 h, and hence falsification of the isoenzyme pattern.

Antigen-Antibody Complex↗
PubMed1991

Influence of sepsis in rats on muscle protein turnover in vivo and in tissue incubated under different in vitro conditions.

We studied the influence of sepsis on muscle protein synthesis and degradation in vivo and in muscles, incubated flaccid or at resting length. Sepsis was induced in rats by cecal ligation and puncture (CLP). Control rats were sham-operated. A flooding dose of 14C-phenylalanine was used to determine muscle protein synthesis rate in vivo, and protein breakdown was calculated from the difference between protein synthesis and growth rates. Protein synthesis rate in vitro was assessed by determining incorporation of 14C-phenylalanine into protein in incubated extensor digitorum longus (EDL) and soleus (SOL) muscles. Total and myofibrillar protein breakdown rates were determined from release into incubation medium of tyrosine and 3-methylhistidine (3-MH), respectively. Muscle protein synthesis rate in vivo was reduced by 35%, similar to the reduction observed in muscles incubated flaccid or at resting length. The calculated protein breakdown rate in vivo was increased by 31% in septic rats. In incubated muscles, the increase in total protein breakdown (ie, tyrosine release) during sepsis was almost identical in muscles incubated flaccid or at resting length, ie, 83% to 88% in EDL and 47% to 49% in SOL. Myofibrillar protein degradation in vitro (ie, 3-MH release) was increased approximately 10-fold in EDL muscles incubated flaccid or at resting length, but was not significantly affected by sepsis in SOL. Results suggest that sepsis-induced changes in protein synthesis observed in muscles incubated either flaccid or at resting length reflect changes in vivo. Changes in protein breakdown were qualitatively similar in vivo and in vitro, but results in incubated muscles may overestimate the increase in muscle proteolysis caused by sepsis.

Adenosine Triphosphate↗
PubMed1996

Pulsatile secretion of prolactin in laying and incubating turkey hens.

Incubation behavior in the turkey hen is associated with a large increase in prolactin secretion. Previous research using hourly sampling of incubating hens has shown that prolactin levels fluctuate widely throughout a 24-hr period, suggestive of pulsatile secretion. This study compared the prolactin secretory patterns of laying and incubating turkeys to determine if prolactin is secreted episodically and if the high prolactin levels characteristic of the incubating hen may result, at least in part, from a change in the amplitude or frequency of secretory pulses. Blood samples were collected from cannulated, unrestrained laying and incubating hens at 10-min intervals for up to 24 hr. Data were analyzed with the PULSAR program to determine baseline prolactin levels and to establish the magnitude, frequency, and duration of episodic secretory peaks. The results revealed tha prolactin is secreted in a pulsatile pattern in both laying and incubating turkey hens. Incubating hens had ninefold higher mean and baseline plasma prolactin levels than laying hens. The prolactin pulses were of approximately 12-fold greater amplitude in incubating hens than in laying hens, but the duration and frequency of pulses were the same in both groups. Therefore, the high prolactin levels required for incubation do not appear to result from an increase in the frequency of lactotroph stimulation, but rather from an increase in the prolactin secretion rate.

Animals↗
PubMed2005

Sperm binding, in vitro fertilization, and in vitro embryonic development of bovine oocytes fertilized with spermatozoa incubated with norepinephrine.

The final stages of sperm maturation, fertilization, and early embryonic development occur within the oviduct and are essential for successful reproduction in mammals. Norepinephrine was previously identified in native bovine oviductal fluid and its in vitro effects on bull sperm capacitation and the acrosome reaction have been determined. It was unknown how physiological concentrations of norepinephrine influence sperm binding, fertilization, and embryo development. Therefore, the objective of this study was to determine if pre-incubating bovine spermatozoa with physiological concentrations of norepinephrine prior to insemination of bovine oocytes would improve sperm-oocyte binding, fertilization, and embryonic development in vitro. Norepinephrine, in concentrations representing those measured in bovine oviductal fluid, was used to treat bovine spermatozoa prior to insemination. Spermatozoa incubated in norepinephrine were used to inseminate bovine oocytes matured in vitro, and oocytes were evaluated for sperm binding and fertilization. Additional experiments were conducted to evaluate how early in the co-incubation period oocytes were fertilized by spermatozoa pre-incubated with norepinephrine, and to test the developmental competence of those oocytes fertilized with norepinephrine-treated sperm. Sperm binding to the zona pellucida was reduced by pre-incubation with norepinephrine. Rates of fertilization and embryo development did not increase as a result of pre-incubating spermatozoa with norepinephrine, but as early as 4h after insemination, spermatozoa treated with 20 ng/ml norepinephrine fertilized more oocytes than spermatozoa incubated in medium alone. Interestingly, this concentration of norepinephrine was found to capacitate spermatozoa in previous studies. These data suggest that oocytes fertilized by spermatozoa incubated in 20 ng/ml norepinephrine fertilize earlier in vitro than sperm pre-incubated in medium alone, and provide additional support for the role of norepinephrine in sperm capacitation and the acrosome reaction.

Animals↗
PubMed2006

Metabolic response to cooling temperatures in chicken embryos and hatchlings after cold incubation.

We asked to what extent cold exposure during embryonic growth, and the accompanying hypometabolism, may interfere with the normal development of thermogenesis. White Leghorn chicken eggs were incubated in control conditions (38 degrees C) or at 36 or 35 degrees C. Embryos incubated at a lower temperature (34 degrees C) failed to hatch. The cold-incubated embryos had lower oxygen consumption (VO2) and body weight (W) throughout incubation, and hatching was delayed by about, respectively, 1 and 2 days. The W-VO2 relationship of the cold-incubated embryos was as in controls, indicating that cold-induced hypometabolism was at the expense of the growth, not the maintenance, component of VO2. At embryonic day E11, the metabolic response to changes in ambient temperature (T) over the 30-39 degrees C range was typically poikilothermic, with Q10 = 1.8-1.9, and similar among all sets of embryos. Toward the end of incubation (E20), the thermogenic responses of the cold-incubated embryos were significantly lower than in controls. This difference occurred also in the few-hour old hatchlings (H1), even though, at this time, W was similar among groups. Exposure to cold during only the last 3 days of incubation (from E18 to H1), i.e. during the developmental onset of the endothermic mechanisms, did not lower the thermogenic capacity of the hatchlings. In conclusion, sustained cold-induced hypometabolism throughout incubation blunted the rate of embryonic growth and the development of thermogenesis. This latter phenomenon could be an example of epigenetic regulation, i.e. of environmental factors exerting a long-term effect on gene expression.

Adaptation, Physiological↗
PubMed1988

A short incubation time in insulin binding experiments leads to disappearance of negative cooperativity in H35 hepatoma cells.

The effects of different periods of incubation (8 min vs 20 min) on insulin binding kinetics were examined in a H35 hepatoma cell line. Scatchard plots from cells incubated for 8 min were linear (r = 0.987 +/- 0.006), in contrast to curvilinear Scatchard plots from cells incubated for 20 min. Hill plots showed a slope of 1.006 +/- 0.024 for the 8 min incubation, whereas the slope was 0.827 +/- 0.0026 (p less than 0.0005) for the 20 min incubation. TCA precipitation of the medium showed minimal insulin degradation products at 8 min with a significant increase at 20 min (1.38 +/- 0.11% vs. 3.06 +/- 0.37%, p less than 0.0005). Internalized insulin was also significantly increased at 20 min as compared to 8 min incubation (48.9 +/- 5.6% vs. 32.4 +/- 3.0%, p less than 0.0005) These data indicate that after 8 min of incubation no appreciable cooperativity of insulin binding was present, while negative cooperativity was present after 20 min of incubation. As significantly more insulin degradation has taken place after prolonged incubation these data support the hypothesis that insulin degradation leads to negative cooperativity of insulin receptors.

Animals↗
PubMed2001

Nitric oxide dependent and independent effects of in vitro incubation or endotoxin on vascular reactivity in rat aorta.

We sought to delineate contributions of nitric oxide (NO) and other mechanisms to impairment of contraction and endothelium-dependent relaxation following prolonged in vitro incubation, endotoxin and interleukin-1 exposure in isolated rat aorta. Responses from freshly-dissected (control) rings +/-endothelium were compared with those from rings incubated in sterile, antibiotic containing medium +/- E. Coli endotoxin (LPS, 100 microg/ml) +/- interleukin-1 (IL-1, 40 ng/ml) at 37 degrees C for 20-24 h. In some experiments, medium included dexamethasone (DEX, 1 microg/ml), cycloheximide (10 microg/ml), or N(G)-nitro-L-arginine (NNLA, 10(-4)M). After incubation, medium nitrite was measured. Incubation alone, without addition of inflammatory mediators, impaired contraction in an agonist-specific manner, by both NO-dependent and NO-independent mechanisms. Either LPS or IL-1 diminished contraction further, in a similarly heterogeneous manner. For example, contractions were changed in LPS-incubated endothelium-intact rings (vs. fresh controls) by -85%, +115%, -15%, -96%, and -37% for phenylephrine (PE), serotonin, prostaglandin F2alpha, angiotensin II, and U46619, respectively. NO synthase inhibition with NNLA either following, or during LPS incubation only partially normalized subsequent PE contractions, an effect which was smaller than that of DEX. Nitrite accumulation was inversely proportional to PE response, even though NO was not the sole mediator of LPS-impaired contraction. LPS and IL-1 nearly abolished ACh-induced relaxation, which was only mildly impaired by incubation alone. We conclude that prolonged incubation impaired vasoconstriction via both NO synthase induction and NO-independent mechanisms. LPS or IL-1 incubation impaired vasoconstriction further, primarily by NO-independent mechanisms. Moreover, vasoconstrictor responses following LPS varied with the agonist's ability to modulate endothelial NO release. These results are in accord with the failure of NO synthase inhibition to fully restore systemic vascular resistance indices in experimental endotoxemia or in hyperdynamic septic patients.

Animals↗
PubMed1999

Comparison of the longevity of motility of stallion spermatozoa incubated at 38 degrees C in different capacitating media and containers.

This study was designed to compare the effects of different media and containers on longevity of motility of spermatozoa during in vitro incubation at 38 degrees C in either air or 5% CO2 atmosphere. Three ejaculates were collected from each of 4 stallions. The media tested were skim milk-glucose, modified Krebs/Ringer and Hank's salts solution for incubation in an air atmosphere, and modified Krebs/Ringer and Brackett and Oliphant (BO) defined medium for incubation in a 5% CO2 atmosphere. All samples were incubated in 5-mL borosilicate glass tubes filled with 3 mL of extended spermatozoa, 5-mL borosilicate tubes filled with 6 mL (topped) of extended spermatozoa, 35-mm Petri dishes filled with 3 mL of extended spermatozoa, and 35-mm Petri dishes with 200-microL microdroplets of extended spermatozoa under sterile mineral oil. For all treatments, individual samples were removed at 2, 4, 6 and 12 h of incubation to determine the percentage of motile cells. Overall, spermatozoa incubated in Petri dishes in both 3-mL and microdroplet treatments had significantly higher motility than those incubated in glass tubes (P<0.01). At 6 and 12 h of incubation in Petri dishes, progressive motility was significantly higher for spermatozoa extended in the Hank's salts solution than in the other media. Both the medium and container used significantly affected the longevity of motility of spermatozoa incubated at 38 degrees C.

Animals↗
PubMed1991

Increased in-vitro incubation time of endothelial cells on fibronectin-treated ePTFE increases cell retention in blood flow.

Endothelial cell (EC) seeding is postulated as a mechanism of improving patency of small calibre vascular grafts. However, the majority of seeded cells are lost within hours following restoration of blood flow. We postulated that incubating EC in-vitro on a graft will improve adherence and resistance to the sheer stresses of pulsatile blood flow. Fibronectin-treated ePTFE (5 cm x 4 mm ID) seeded with Indium-111-labelled autologous canine EC (1.5 x 10(5) cells/cm2) were incubated for four different time periods; 90 min, 24 h, 72 h and 6 days. Incubated grafts were subjected to blood flow of 75 ml/min for 6 h, in a canine ex-vivo arteriovenous shunt circuit. EC retention during perfusion was studied by measuring gamma activity emitted by the grafts. Cell morphology of non-perfused control groups and perfused groups was compared using scanning electron microscopy (SEM). SEM of control grafts showed progressive EC spreading on the ePTFE surface for up to 72 h incubation. Gamma activity was significantly higher at 6 h perfusion in grafts incubated for 72 h (82 +/- 4%) and 24 h (63 +/- 6%) vs. 90 min (34 +/- 13%, p less than 0.05), and between grafts incubated for 72 h vs. 6 days (55 +/- 7%, p less than 0.05). Perfused grafts incubated for 72 h showed unaltered EC morphology on SEM, few cells remained on 90 min incubated grafts. We conclude that incubating EC on fibronectin-treated ePTFE for 72 h in-vitro after seeding improves cell retention during blood flow.

Animals↗