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PubMedDate not supplied

In vitro model for the evaluation of inferior vena cava filters: effect of experimental parameters on thrombus-capturing efficacy of the Vena Tech-LGM filter.

PURPOSE: To determine the experimental parameters in an in vitro model that influence the thrombus-capturing efficacy of the Vena Tech-LGM filter. MATERIALS AND METHODS: The Vena Tech-LGM filter was evaluated in an in vitro model of the vena cava with a computer-controlled flow system with a total of 5,200 thrombi. The influences of the following experimental parameters on the capture rate were analyzed with a multiple logistic regression model: type of testing (single, double, and multiple shot testing), thrombus diameter and length, IVC diameter and orientation, flow quality and quantity, flow velocity, and the length of the prepositioned thrombus. RESULTS: A significant influence on the capture rate could be demonstrated for the type of testing, the thrombus diameter and length, the IVC diameter, and with double shot testing for the length of the prepositioned thrombus and the IVC orientation. The flow quality and the peak velocity were not significant. Based on these results, a protocol for in vitro testing of IVC filters was designed. CONCLUSIONS: Experimental parameters influence the thrombus-capturing efficacy of the Vena Tech-LGM filter and should be taken into account when in vitro testing is performed.

Filtration↗
PubMed1989

Isolation of human sebaceous glands and cultivation of sebaceous gland-derived cells as an in vitro model.

An experimental technique is presented as an in vitro model for the study of human sebaceous gland-derived cells. Intact sebaceous glands were isolated from full-thickness human skin after incubation in dispase (2.4 U/ml) and in deoxyribonuclease (0.02%) by using microsurgical instruments under microscopical observation of the epidermal underface. Subsequently, the ducts of the glands were removed, the isolated gland lobules were seeded on a 3T3-cell feeder layer in Dulbecco's modified Eagle's medium and Ham's F 12 medium (3:1) supplemented with fetal calf serum (10%), L-glutamine, antibiotics, epidermal growth factor (10 ng/ml), hydrocortisone (0.4 microgram/ml), and cholera toxin (10(-9) M), and were then cultivated in a CO2-incubator at 37 degrees C. After 2-3 wk cell outgrowths resulting from the periphery of the gland lobules were obtained and dispersed cells were passaged for three subcultures with or without 3T3-cell feeder layer. The cultured cells preserved in vitro morphologic characteristics and differentiation patterns comparable to those described for normal human sebocytes in vivo, with a high rate of viable cells. Their labeling pattern with MoAb showed close similarities to the pattern reported for sebocytes in vivo but differences to the pattern of keratinocytes in vivo and in vitro. In their cytoplasm oil red and nile red stained droplets were detected, and the observed density and distribution evidenced in vitro lipogenesis. The technique presented here may provide a promising model for further experimental studies on sebaceous gland cell development and function.

Antibodies, Monoclonal↗
PubMed1999

An in vitro model for the study of the role of innervation in circumvallate papillae morphogenesis.

The following study was done to demonstrate the reliability of an in vitro model for use in the study of early events and the role of innervation in mouse circumvallate papillae development. Gestational day (gd)-11 fetuses were partially dissected to produce explants that included the mandibular, hyoid, third and fourth branchial arches and their ganglia. In ganglionectomized explants, the nodose ganglia and either the geniculate, petrosal or both ganglia were removed. Explants were cultivated in roller tube culture for 24, 48, 72, and 96 h of culture and examined for the presence of papillary structures. Innervation was verified by immunostaining for neural cell adhesion molecule (NCAM). In all control explants, circumvallate papillae had formed by 72 h in culture. These papillae were innervated by fibers originating in petrosal or nodose ganglia, although, in a small number, fibers from the geniculate also contributed. Circumvallate papillae also formed in some explants in which either the geniculate or petrosal ganglia had been removed. However, placodal structures failed to mature into papillary structures even by 96 h in explants in which both ganglia had been removed. Our results demonstrate that an in vitro model using branchial arch explants supports the morphogenesis of an epithelial placode through the formation of a definite papillary structure, the circumvallate papilla, with an integrated nerve. Our results also indicate that, whereas the initial stages in gustatory papillae formation, the formation of a placode, are nerve-independent, the maturation of the placodal structure to form a papilla requires the presence of an intact nerve.

Animals↗
PubMed1985

The use of bacteria producing the aminoglycoside inactivating enzyme ANT-(2") in an in-vitro model.

A series of transconjugants of Escherichia coli W3110 containing different plasmids coding for the aminoglycoside inactivating enzyme ANT-(2") were tested in an in-vitro model simulating the concentration time curves of aminoglycosides after intramuscular administration of 80 or 120 mg. There was good correlation of the MIC, which ranged from 2 to 32 mg/l, with the killing ability of the drugs. A second dose had no or only little effect on ANT-(2") producing strains. All strains, although of 'intermediate' sensitivity were found to be resistant in the in-vitro model. Only netilmicin could eliminate some of the strains but its activity on ANT-(2")-positive strains was not as good as on ANT-(2")-negative ones.

Aminoglycosides↗
PubMed1999

In vitro models of intracranial arteriovenous fistulas for the evaluation of new endovascular treatment materials.

BACKGROUND AND PURPOSE: The purpose of this study was to create and test an in vitro model of intracranial arteriovenous fistulas (AVFs) that simulates the geometry of human vasculature and allows realistic testing of devices used in endovascular therapy. METHODS: The models were derived from corrosion casts of the main cervicocranial arteries and veins obtained from two nonfixed human specimens. Wax copies of the casts were produced and combined to create complex models simulating various types of intracranial AVFs. Wax assemblies were embedded with liquid silicone solidified into transparent blocks containing, after wax evacuation, hollow reproductions of the original vascular trees. The models were connected to a pulsatile pump and their compatibility with various imaging techniques and endovascular treatment materials was evaluated. RESULTS: The models were compatible with digital subtraction angiography, CT, MR imaging, and transcranial Doppler sonography. They provided a realistic endovascular environment for the simulation of interventional neuroradiologic procedures. CONCLUSION: Anatomically accurate and reproducible in vitro models of intracranial AVFs provide a valuable method for evaluating new endovascular treatment materials and for teaching purposes.

Aged↗
PubMedDate not supplied

Evaluation of the placental environment with a new in vitro model of histocultures of early and term placentae: determination of cytokine and chemokine expression profiles.

We aimed to set up and validate a new in vitro model of placental histocultures, for the evaluation of cytokine and chemokine profiles of the placental environment, over a long culture period. Micro-explant cultures from 6 early and 6 term placentae were set up on collagen sponge gel supports at a liquid/air interface. At various times during culture, we analyzed tissue morphology and cell death by microscopy and quantified beta-hCG production and mRNA levels for beta-hCG and insulin-like 4 (INSL4). Levels of IL-6, LIF, TNF alpha, IL-10, IFN-gamma, IL-16 and RANTES in the medium were measured by ELISA on days 1, 4 and 7 of culture. SDF-1 mRNA expression was determined by real-time PCR at the same time points. Histocultures from early and term placentae remained viable until day 10. High levels of IL-6 and LIF production, low levels of TNF alpha, IL-10 and IFN-gamma production and significant SDF-1 expression were observed. These data indicate that placental histoculture is a suitable and reliable in vitro model for studying the placental environment.

Adult↗
PubMed2005

A homologous in vitro model to study interactions between alphaherpesviruses and trigeminal ganglion neurons.

A key aspect in the life cycle of alphaherpesviruses is their neurotropic behaviour. Sensory neurons of the trigeminal ganglion (TG) are important target cells for many alphaherpesviruses (including herpes simplex virus 1, pseudorabies virus (PRV), bovine herpesvirus 1) and constitute major sites for latent infections. The aim of this study was to develop an in vitro model that simulates the in vivo infection pattern of TG neurons by alphaherpesviruses. To this end, we developed a homologous in vitro two-chamber model using PRV and porcine TG neurons. TG of 4- to 6-week-old piglets were dissociated and cultured in the inner chamber of the in vitro model, which is separated from the outer chamber by a medium- and virus-impermeable silicon barrier. Outgrowth of axons from neuronal cell bodies in the inner chamber through the silicon barrier into the outer chamber could be observed after 2-3 weeks of cultivation. Subsequent addition of PRV to the outer chamber resulted in exclusive infection of the TG neurons by transport of virus through the axons, subsequently giving rise to productively infected TG neurons that transmitted virus to contacting neurons and non-neuronal cells in the inner chamber. Thus, we established a homologous in vitro model that mimics the natural route of alphaherpesvirus infection of TG neurons that can be used to study interactions between these viruses and this pathogenetically very important cell type.

Alphaherpesvirinae↗
PubMed2006

Optional inferior vena cava filter retrieval with retained thrombus: an in vitro model.

PURPOSE: Retrieval of an optional inferior vena cava (IVC) filter with retained thrombus may result in pulmonary emboli if the trapped thrombus is not removed along with the filter. An in vitro model was developed to determine the fate of trapped thrombus during filter removal. MATERIALS AND METHODS: An in vitro IVC flow model was created with 25-mm inner diameter tubing and a 50% glycerol/water solution. Three different optional filters-Recovery (Bard, Tempe, AZ), Günther-Tulip (Cook Inc., Bloomington IN), and OptEase (Cordis Endovascular/Johnson & Johnson, Warren, NJ)-were evaluated in the study. A known mass of mature thrombus (porcine, aged 1 wk) was trapped within the optional filters. The filters were then retrieved according to the manufacturers' protocol, and the mass of thrombus recovered with the filter was determined. For each filter, five iterations were performed with initial thrombus sizes less than 1 g (group A) and an additional five iterations with initial thrombus sizes greater than 1 g (group B). RESULTS: Thrombi from group A were statistically significantly smaller than those from group B (P < .0001). Retrieval of the Recovery filter resulted in an average of 25% (range, 0%-53%) and 4% (range, 0%-7%) of the clot being removed in group A and group B, respectively. Retrieval of the Günther-Tulip filter resulted in an average of 22% (group A) and 13% (group B) of the clot being removed. Retrieval of the OptEase filter resulted in an average of 43% (group A) and 0% (group B) of the clot being removed. CONCLUSIONS: In our in vitro model, we have established that the mass of thrombus retrieved with optional filters is only a fraction of the initial clot burden. Because of the risk of pulmonary emboli, care should be taken when IVC filters with large amounts of trapped thrombus are removed from patients.

Analysis of Variance↗
PubMed1977

Everted rat intestinal sacs as an in vitro model for assessing absorptivity of new drugs.

An in vitro model that utilizes everted rat intestinal sacs was evaluated for assessing the absorptivity of several analogs of potential drug substances prior to formulation work and clinical trials. This model not only is a useful qualitative tool for assessing absorptivity of structurally related compounds but also yields some insight into the process involved in drug absorption. Notwithstanding the complexities involved in the absorption processes, the data support the hypothesis that the absorption of organic electrolytes mainly takes place by the partitioning of the unionized species into the lipoidal membranes and then diffusion.

Animals↗
PubMed1993

A simplified in vitro model of oxidant injury using vascular endothelial cells.

Oxidant injury of the vascular endothelium is considered an early event in the pathogenesis of atherosclerosis. The model of oxidant injury is crucial to the investigation of antioxidants. In the present study, a convenient in vitro model of oxidant injury induced by hydrogen peroxide (H2O2) was developed using bovine pulmonary artery endothelial cells (PAEC). Viability of PAEC grown in 96-well culture plates was determined with methylthiazol tetrazolium (MTT) colorimetric assay. Cell membrane integrity was measured by lactate dehydrogenase (LDH) release from PAEC grown in 24-well plates. Malondialdehyde (MDA, a product of lipid peroxidation) in PAEC grown in 6-well plates was detected by a thiobarbituric acid fluorometric assay. Incubation of H2O2 with PAEC caused a dose-dependent decrease of cell viability, an increase of LDH release, and an elevation of MDA production. MTT assay was convenient, quantitative, non-radioactive, and suitable for testing a large number of samples. The fluorometric assay for measuring MDA production in endothelial cells used 6-well plates instead of 80-cm2 flasks employed by previous investigators. The use of multiwell culture plates in these assays made it possible for more samples to be tested in any single experiment. The three assays are reproducible with low intraplate and interplate coefficients of variation. This in vitro model is suitable for screening antioxidants and for studying pharmacodynamics at the cellular level.

Animals↗
PubMed1992

Effect of phytanic acid on cultured retinal pigment epithelium: an in vitro model for Refsum's disease.

Refsum's disease (heredopathia atactica polyneuritiformis) is an autosomal recessive retinitis pigmentosa syndrome caused by the excessive deposition of phytanic acid in ocular tissues. It is thought that phytanic acid causes retinal degeneration either by interfering with vitamin A metabolism in the retinal pigment epithelium or by altering photoreceptor cell membrane structure. Efforts to elucidate the molecular mechanism of phytanic acid's retinal toxicity have been hampered by the rarity of human pathological specimens and by the inability to reproduce the disease in living animal models. In this study, an in vitro model for Refsum's disease was established by exposing cultured human and bovine retinal pigment epithelial cells to phytanic acid bound to bovine serum albumin at concentrations comparable to levels found in affected humans. Ultrastructural studies show that these cells exhibit morphological changes consistent with those observed in pathological specimens from patients with Refsum's disease. Biochemical assays of retinoid metabolism by cell membranes from control cells and from cells exposed to 200 microM phytanic acid demonstrate that the ability to esterify retinol and to isomerize all-trans retinoids to 11-cis retinoids remains intact despite the deposition of large amounts of phytanic acid. The work described here is strong evidence against the hypothesis that phytanic acid inhibits vitamin A metabolism in the retinal pigment epithelium, and it demonstrates the potential use of cultured retinal pigment epithelial cells in modeling this and other degenerative diseases of the retina.

Animals↗
PubMed1996

[Free microsurgical superior musculocutaneous trapezius flap of the swine: an ideal training model for microvascular reconstructions and in vitro model for experimental microsurgery].

BACKGROUND: Until now there have been no published studies on musculocutaneous porcine free flaps with an uncompromised vascular pedicle, easy surgical access and anatomic orientation allowing unlimited postoperative movement, as German animal research regulations require. RESULTS: The authors present the porcine musculocutaneous superior trapezius flap. It is supplied by the transverse neck artery, a branch of the thyrocervical trunk of the subclavian artery as in human anatomy. CONCLUSION: The advantages of this flap are its easy surgical access, its uncompromised vascular anatomy, and the easy closure of the donor area. The authors demonstrated the uncompromised vascular anatomy in 40 pigs with body weights ranging between 24 and 68 kg. The pigs were able to move normally following surgery.

Animals↗
PubMed1983

In vitro model for the study of platelet-vessel wall interactions following a freeze- thaw injury.

Platelet-endothelial cell interactions are important for maintaining normal hemodynamics. The intact endothelial cell lining is considered nonthrombogenic, but following disruption of the lining, platelets bind to the subendothelium. There is also much conjecture concerning the affinity of platelets for damaged endothelial cells. A model is described for the study of platelet-aorta vessel wall interactions following freeze-thaw insult. Using this model, we found that control aortas (37 degrees C) perfused with platelet-rich plasma or gel-filtered platelets showed no generalized platelet-endothelial cell interactions, although some platelets did adhere to areas of exposed subendothelium. Following freeze-thaw insult (-15 degrees or -20 degrees C), the endothelial lining was grossly disrupted. The remaining endothelium was severely damaged, demonstrating holes and pits in the plasma membranes and separation of adjacent cell borders. Platelets readily adhered to the basal lamina but were rarely noted in sole contact with the damaged endothelium. Platelet binding did not result in morphologic changes, degranulation, or aggregation. Using transmission electron microscopy, we noted platelets in contact with amorphous material and microfibrils but not collagen fibers of the subendothelium. It is concluded that this model is suitable for the in vitro study of certain hemodynamic phenomena associated with blood vessel freeze-thaw injury. In addition, freeze-thaw damage in this in vitro model indicated that platelet-vessel wall interactions were limited to areas of exposed subendothelium.

Animals↗
PubMed2003

Optimization of Schwann cell adhesion in response to shear stress in an in vitro model for peripheral nerve tissue engineering.

The design of nerve guidance channels (NGCs) is evolving to produce a favorable environment for neural regeneration. We created an in vitro model to evaluate the interactions between three centrally important components of this altered host environment: (1). Schwann cells, (2). substrate, and (3). sustained mechanical stimulus in the form of shear stress with laminar fluid flow. Preconfluent Schwann cells were plated on slides coated either with laminin, poly-D-lysine, type IV collagen, or fibronectin. These slides were placed into custom-designed, parallel-plate, flow chambers and were administered laminar fluid flow at a rate of 15 mL/min for 2 h. Schwann cell adhesion assays demonstrated that laminin (mean, 86.1%; SEM, 4.47%) and fibronectin (mean, 81.7%; SEM, 3.24%) were statistically superior to collagen type IV (mean, 57.7%; SEM, 3.96%) and poly-D-lysine (mean, 58.0%; SEM, 4.97%) (p < 0.001). Fibronectin (mean, 12.20%; SEM, 0.374%) induced statistically greater Schwann cell proliferation than did laminin (mean, 8.14%; SEM, 0.682%) (p < 0.001). Therefore, we recommend that fibronectin should be used as an important component of NGCs with further in vivo studies. As mechanical stress is an integral part of the host environment, our study is the first to incorporate this factor into an in vitro model for peripheral nerve tissue engineering.

Animals↗
PubMed1997

A physiologic in vitro model of the inferior vena cava with a computer-controlled flow system for testing of inferior vena cava filters.

RATIONALE AND OBJECTIVES: The authors develop a physiologic in vitro model of the inferior vena cava (IVC) for testing of filters. METHODS: The model is driven by a centrifugal pump. A computer-controlled electromagnetic valve is used for generation of different flow patterns. Limitation of the pressure increase in case of IVC occlusion is achieved by a bypass circuit. A glycerin solution is used for perfusion. Artificial clots are made from polyacrylamide gel. Data acquisition includes continuous monitoring of flow and difference pressure over the filter and video recording of the testing events. RESULTS: The model can generate constant and pulsatile flows. The pressure increase can be limited to 70 mm Hg in case of occlusion. Calculation of the flow velocities in the IVC is possible. A classification of thrombus capturing is presented. The testing of most of the results are reproducible. CONCLUSIONS: The in vitro model simulates the physiologic conditions in the IVC. It can be used for comparative testing of different filters and the evaluation of new filter designs.

Blood Flow Velocity↗
PubMed2006

Development of an in vitro model for study of the efficacy of ischemic preconditioning in human skeletal muscle against ischemia-reperfusion injury.

Ischemia-reperfusion (I/R) injury causes skeletal muscle infarction and ischemic preconditioning (IPC) augments ischemic tolerance in animal models. To date, this has not been demonstrated in human skeletal muscle. This study aimed to develop an in vitro model to investigate the efficacy of simulated IPC in human skeletal muscle. Human skeletal muscle strips were equilibrated in oxygenated Krebs-Henseleit-HEPES buffer (37 degrees C). Aerobic and reperfusion phases were simulated by normoxic incubation and reoxygenation, respectively. Ischemia was simulated by hypoxic incubation. Energy store, cell viability, and cellular injury were assessed using ATP, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT), and lactate dehydrogenase (LDH) assays, respectively. Morphological integrity was assessed using electron microscopy. Studies were designed to test stability of the preparation (n = 5-11) under normoxic incubation over 24 h; the effect of 1, 2, 3, 4, or 6 h hypoxia followed by 2 h of reoxygenation; and the protective effect of hypoxic preconditioning (HPC; 5 min of hypoxia/5 min of reoxygenation) before 3 h of hypoxia/2 h of reoxygenation. Over 24 h of normoxic incubation, muscle strips remained physiologically intact as assessed by MTT, ATP, and LDH assays. After 3 h of hypoxia/2 h of reoxygenation, MTT reduction levels declined to 50.1 +/- 5.5% (P < 0.05). MTT reduction levels in HPC (82.3 +/- 10.8%) and normoxic control (81.3 +/- 10.2%) groups were similar and higher (P < 0.05) than the 3 h of hypoxia/2 h of reoxygenation group (45.2 +/- 5.8%). Ultrastructural morphology was preserved in normoxic and HPC groups but not in the hypoxia/reoxygenation group. This is the first study to characterize a stable in vitro model of human skeletal muscle and to demonstrate a protective effect of HPC in human skeletal muscle against hypoxia/reoxygenation-induced injury.

Adenosine Triphosphate↗
PubMed1985

Pharmacokinetic considerations for the setting of in-vitro models.

Nearly all in-vitro kinetic models hitherto employed only consider serum concentration curves of the antibiotic. Data thus obtained at best reflect the situation in septicaemia. However, they are not applicable for infections where infecting bacteria are not localized within the blood. This paper presents a method for calculating the time course of the non-protein bound drug in tissue water. The concept makes use of serum concentration curves, the extent of serum protein binding and the peak time of total drug concentration in the tissue. This is practicable since all data necessary for calculation are available experimentally. Using cefmenoxime, cefotaxime, latamoxef (moxalactam) and ceftriaxone as examples, the vast differences between the total concentration of the antibiotic in serum and the concentration of the non-protein bound antibiotic in the tissue water are demonstrated. Therefore, only the results of in-vitro experiments, which are based on time courses of the non-protein bound drug in the tissue water, are considered relevant for assessing therapeutic efficacy of an antibiotic.

Anti-Bacterial Agents↗
PubMed2002

Studies on dental erosion: An in vivo-in vitro model of endogenous dental erosion--its application to testing protection by fluoride gel application.

BACKGROUND: The objective in this study was to develop an in vivo-in vitro model of endogenous erosion, with a view to exploring the potential for some degree of its control by the use of topical fluoride gel application to teeth. METHODS: Six volunteers each wore a small clasp retained palatal acrylic appliance to which six sterilized enamel tiles were bonded. Three tiles on each appliance were coated extra-orally with either 1.23 per cent acidulated phosphate fluoride (APF) or 2.2 per cent sodium fluoride (NaF) neutral gel for four minutes prior to multiple periods of exposure to the simulated gastric acid, cumulating in 16, 36, 80 and 150 minutes of exposure. Impressions of the enamel tiles prior to and following acid exposure permitted dies to be prepared. These were sectioned through the exposed areas and examined under a stereomicroscope to assess maximum depths of erosion. RESULTS: The depth of erosive demineralization of enamel was found to be greatly reduced with increased frequency of APF gel application. The reduction in enamel loss was less following topical application of NaF gel. CONCLUSIONS: It was concluded that fluoride gels significantly reduced enamel erosion using this in vivo-in vitro model and therefore, if prescribed appropriately, should help reduce tooth tissue loss from endogenous erosion.

Acidulated Phosphate Fluoride↗