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Restricted dextran transport in the sheep lung lymph preparation.

The ease with which plasma proteins and other large solutes enter sheep lung lymph had led previous investigators to conclude that such large molecules probably move through a pathway that has transport properties similar to pores 100 nm or larger in radius. To test this hypothesis, the transport of some very large solutes into lung lymph was measured. Dextran fractions having Stokes-Einstein radii of 10--150 nm were prepared and labeled with fluorescein isothiocyanate (FITC). Five sheep were given 2--2.5 g FITC-dextran 24--36 h prior to cannulation of the efferent duct of the caudal mediastinal lymph node. Analysis of paired plasma and lymph samples for fluorescence as a function of molecular size gave the following results. FITC-dextran 10 nm in radius appeared in sheep lung lymph at 0.30 +/- 0.06 of its plasma concentration. Larger FITC-dextran molecules appeared at still lower concentrations until, above 35 nm, fluorescence could no longer be detected in lymph. The restriction to transport of FITC-dextran was found to be inconsistent with values predicted by a pore pathway 100 nm or larger. These data are compatible with a pore pathway 22--35 nm in radius.

Animals↗

Passage of uncharged dextrans from blood to lung lymph in awake sheep.

To examine how molecular size alone influences the passage of macromolecules from the pulmonary microcirculation into lymph collected from the caudal mediastinal lymph node of the sheep, we infused polydisperse uncharged [3H]dextrans intravenously at a constant rate over a period of 7.5 h in nine awake sheep with lung lymph fistulas. Lymph and plasma were collected during hours 5.5-7.5 of the infusions, and the [3H]dextrans were separated by molecular sieve chromatography into fractions that ranged from 1.6 to 8.4 nm in effective molecular (Stokes-Einstein) radius. Lymph-to-plasma (L/P) ratios for [3H]dextrans were near 1.0 at 1.6-nm radius, decreased with increasing molecular size, and approached zero at radii above 5.0 nm. We confirmed that these L/P ratios represented steady-state values by extending the duration of the infusion to approximately 30 h in two of the nine sheep and finding that the L/P ratios remained unchanged. These results were consistent with molecular sieving through a homoporous membrane with cylindrical pores of 5.0-nm radius. We also found that the L/P ratio for albumin [0.76 +/- 0.13 (SE)] in five of the same sheep was much higher than that for the [3H]dextran fraction of the same effective molecular radius [0.11 +/- 0.02 (SE)]. These results suggest that the movement of macromolecules from the pulmonary microcirculation into pulmonary lymph collected from the caudal mediastinal node of the sheep is influenced by both molecular size and molecular charge and that, compared with uncharged dextrans, the steady-state passage of anionic endogenous proteins from plasma to lymph is enhanced.

Animals↗

Dextran sulfate inhibits PMN-dependent hydrostatic pulmonary edema induced by tumor necrosis factor.

We tested the hypothesis that neutrophil sequestration is required for the development of tumor necrosis factor- (TNF) induced neutrophil- (PMN) dependent pulmonary edema. TNF (3.2 X 10(5) U/kg ip) was injected into guinea pigs 18 h before lung isolation. After isolation, the lung was perfused with a phosphate-buffered Ringer solution. Dextran sulfate (mol wt 500,000) prevented the changes in pulmonary capillary pressure (Ppc; 8.5 +/- 0.8 vs. 12.8 +/- 0.8 cmH2O), lung weight gain (dW; +0.240 +/- 0.135 vs. +1.951 +/- 0.311 g), and pulmonary edema formation or wet-to-dry wt ratio [(W - D)/D; 6.6 +/- 0.2 vs. 8.3 +/- 0.5] at 60 min induced by PMN infusion into a TNF-pretreated lung. The unsulfated form of dextran had no protective effect [Ppc, dW, and (W - D)/D at 60 min: 11.9 +/- 0.9 cmH2O, +1.650 +/- 0.255 g, and 7.3 +/- 0.2, respectively], whereas the use of another anionic compound, heparin, inhibited the TNF + PMN response [Ppc, dW, and (W - D)/D at 60 min: 5.6 +/- 0.4 cmH2O, +0.168 +/- 0.0.052 g, and 6.4 +/- 0.2, respectively]. Isolated lungs showed increased PMN myeloperoxidase (MPO) activity compared with control in TNF-treated lungs at baseline and 60 min after PMN infusion. Dextran sulfate, dextran, and heparin inhibited the increase in MPO activity. The data indicate that inhibition of PMN sequestration alone is not sufficient for the inhibition of PMN-mediated TNF-induced hydrostatic pulmonary edema and that a charge-dependent mechanism mediates the protective effect of dextran sulfate.

Animals↗

Fluorescein-labeled dextran concentration is increased in BAL fluid after ANTU-induced edema in rats.

Several methodologies have been developed to assess alveolocapillary membrane permeability in acute lung injury. The purpose of this study was to determine the reliability of FITC-dextran compared with radioactive tracers to assess lung permeability alterations. After intraperitoneal administration of alpha-naphthylthiourea (ANTU, 50 mg/kg) or DMSO-ANTU vehicle, the animals were euthanized and their lungs were studied in an isolated-lung preparation. FITC-dextran or radiolabeled tracers were added to the perfusate. At 2 h the bronchoalveolar lavage (BAL) fluid from the ANTU group showed a significantly greater amount of fluorescence in the supernatant after centrifugation of BAL fluid compared with the DMSO group. Consistent results were observed with the radioactive tracers: there was an increase in extravascular albumin space and extravascular lung water compared with the control group. No cleavage of the FITC from the dextran molecule was evident by chromatography comparing samples recovered from the BAL fluid to the pure FITC-dextran molecule. In conclusion, measurement of FITC-dextran in the supernatant of BAL fluid after intravascular administration is a reliable method of assessing lung permeability changes in vivo and ex vivo.

Animals↗

Dextran restores albumin-inhibited surface activity of pulmonary surfactant extract.

We examined the effect of dextran (molecular weight 71,000) in counteracting the surfactant inhibitory action of plasma albumin. The surface adsorption time of 0.5 mg/ml modified natural surfactant (MNS; porcine lung extract consisting of phospholipids and hydrophobic surfactant proteins) with 7.5 mg/ml albumin decreased from 681 to 143 s by addition of dextran at a concentration of 10 mg/ml (P < 0.01). The minimum surface tension of 2.0 mg/ml MNS with 30 mg/ml albumin decreased from over 21 mN/m to below 3 mN/m when dextran was added at a concentration of 10 mg/ml (P < 0.01). Surfactant-deficient newborn rabbits given 10 ml/kg of a liquid containing 2.0 mg/ml MNS with 30 mg/ml albumin had a mean tidal volume </=5 ml/kg after 5 min of mechanical ventilation, but, in those animals given the liquid containing 10 mg/ml dextran also, the volume was >13 ml/kg (P < 0.05). Although the underlying mechanism remains to be elucidated, we conclude that dextran restores the albumin-inhibited surface activity of MNS.

Adsorption↗

Iron dextran causes renal iron deposition but not renal dysfunction in angiotensin II-treated and untreated rats.

BACKGROUND: Angiotensin II infusion into rats causes iron deposition in the kidney, which may augment the pro-proteinuric effects of this octapeptide. We have investigated whether administration of iron mimics the renal damage induced by angiotensin II. METHODS: Rats were treated with iron dextran at a total dose of 960 mg/kg either with or without angiotensin II treatment at a dose of 0.7 mg/kg/day for 7 days. Protein expression of ferritin and heme oxygenase-1, an oxidative stress-sensitive gene, was determined by Western blot analysis and immunohistochemistry. RESULTS: Administration of iron dextran did not significantly increase proteinuria or decrease creatinine clearance in the rats with or without angiotensin II treatment. Prussian blue staining showed that iron deposition was observed mainly in the glomerular and medullar regions in the iron dextran-treated rats, but in the tubular epithelial cells in angiotensin II-infused rats. Administration of iron dextran upregulated ferritin, but not heme oxygenase-1. CONCLUSION: Iron dextran did not enhance or cause the renal dysfunction in the angiotensin II-treated or untreated rats, respectively. The distribution of deposited iron and presumably the type of iron compound administered may be important determinants of the development of renal injury.

Angiotensin II↗

Influence of dextrans on cardiac preservation in an extracorporeal rat heart model.

Despite advances in preservation techniques for thoracic organs, the ischemic tolerance of the donor heart is still limited. Recently, a beneficial effect of oncotic substances such as dextran was shown in lung transplantation. Clinically, only in the University of Wisconsin (UW) solution oncotic substances for the prevention of cellular edema are used. Since little is known about the perspective value of dextrans in cardiac preservation, we investigated dextrans with different molecular weights added to the St. Thomas Hospital solution in an experimental working rat heart Langendorff model for functional and histological aspects. By comparison of various dextrans with molecular weights of 40,000, 70,000 and 160,000 daltons, best results were achieved by the addition of 5% dextran with the highest molecular weight.

Animals↗

Delayed hypersensitivity reactions in rats and their response to clinical dextran.

Four colonies of rats were tested for their ability to produce adjuvant-induced arthritis, oxazolone contact hypersensitivity and the dextran anaphylactoid reaction. Tuck Wistar and Hooded Lister rats, both of which respond to dextran, showed disseminated inflammatory lesions after adjuvant and exhibited oxazolone sensitisation, regardless of the age of the animal. Spontaneously hypertensive rats, which at all ages respond to dextran, also responded to adjuvant and oxazolone but only when they were young; as they grew older, this response to these two agents diminished. Wistar NELP rats, which at all ages do not respond to dextran, responded to oxazolone; sensitivity to adjuvant, however, was maximal only in young animals. The link between the ability of rats to resist the dextran anaphylactoid reaction and their failure to respond to adjuvant with disseminated inflammatory lesions has not been confirmed.

Anaphylaxis↗

Dextran and antidextran antibodies in the sera of patients with liver diseases.

Dextran and antidextran antibodies were examined by enzyme immunoassay (EIA) and immunodiffusion in sera of 108 patients with various liver diseases. In EIA, IgG class antidextran antibody was detected in 16 patients (14.8%), mostly with chronic liver diseases such as liver cirrhosis (26.9%) and chronic hepatitis (22.2%). IgM class antibody was detected in 10 patients (9.2%). Results obtained by EIA inhibition revealed that dextran antigen was present mostly in sera of patients with acute liver diseases such as fulminant hepatitis (75.0%) and acute hepatitis (17.2%). Identity of the dextran antigen in the liver diseases serum and the dextran preparation recognized by an antibody-containing serum was demonstrated. These results suggest that the damaged hepatocytes in the process of the liver disease may release the dextran antigen into the patient's circulation which is responsible for the formation of antidextran antibodies by the patients with liver diseases.

Acute Disease↗

Effect of a dextran derivative associated with TGF-beta1 or FGF-2 on dermal fibroblast behaviour in dermal equivalents.

Dextran derivatives that mimic the action of heparin have been shown to protect heparin-binding growth factors, such as Transforming Growth Factor-beta1 (TGF-beta1) and Fibroblast Growth Factor-2 (FGF-2). The aim of this study was to investigate the effect of LS21 DMCBSu, a dextran derivative which contains methylcarboxylate, benzylamide and sulfate groups, both by itself and when combined with TGF-beta1 and FGF-2, on the behaviour of fibroblasts. Two systems were assessed: a monolayer culture and three-dimensional collagenous matrices (dermal equivalent). Polymeric biomaterial LS21 DMCBSu and LS21 DMCBSu associated with either TGF-beta1 or FGF-2, were added to the monolayer culture on day 3. After 7 days of culture the number of cells was determined. Two treatments were carried out on the dermal equivalents: 9 days of treatment from day 0 to day 9 of culture and 9 days of treatment from day 21 to day 30 of culture for the premature and the mature dermal equivalents respectively. In the monolayer culture, the bioactive polymer produced a slight increase in fibroblast growth (10% with 10 microg/ml of LS21 DMCBSu) and promoted the stimulating effect of the growth factors on cell growth. In the premature dermal equivalents growth was stimulated by 20% when 10 microg/ml LS21 DMCBSu was added. The dextran derivative mixed with TGF-beta1 slightly inhibited the growth effect of the growth factor in the dermal equivalents. The functionalized dextran with FGF-2 enhanced the stimulating effect of the growth factor in the premature dermal equivalent. A significant increase in cell growth was observed with the fibroblasts treated with the FGF-2 LS21 DMCBSu mixture and FGF-2 (51% and 40%, respectively). However, none of the described treatments affected the cell growth in the mature dermal equivalent. Furthermore, the dextran derivative had no effect on dermal contraction under these experimental conditions (3D culture).

Cell Differentiation↗

Determination of iron sucrose (Venofer) or iron dextran (DexFerrum) removal by hemodialysis: an in-vitro study.

BACKGROUND: Intravenous iron is typically administered during the hemodialysis (HD) procedure. HD patients may be prescribed high-flux (HF) or high-efficiency (HE) dialysis membranes. The extent of iron sucrose and iron dextran removal by HD using HF or HE membranes and by ultrafiltration rate (UFR) is unknown. METHODS: Two in vitro HD systems were designed and constructed to determine the dialyzabiltiy of iron from a simulated blood system (SBS) containing 100 mg iron sucrose or iron dextran (system A) or 1000 mg iron sucrose (system B). Both in vitro systems utilized a 6-L closed-loop SBS system that was subject to 4 different HD conditions conducted over 4 hours: HE membrane + 0 ml/hr UFR; HE membrane + 500 ml/hr UFR; HF membrane + 0 ml/hr UFR; HF membrane + 500 ml/hr UFR. Blood flow and dialysate flow rates were 500 ml/min and 800 ml/min, respectively. The dialysate compartment was a 192-L open system for system A and a 6-L closed-loop system for system B. Samples from the SBS and dialysate compartments were taken at various time points and iron elimination rate and HD clearance was determined. Iron removal from the SBS > 15% was considered clinically significant. RESULTS: The greatest percentage removal from the SBS was 13.5% and -0.03% utilizing system A and B, respectively. Iron sucrose and iron dextran dialysate concentration was below the lower limits of assay (< 2 ppm) for system A. Dialysate recovery of iron was negligible: 0-5.4 mg system A and 5.47-23.59 mg for system B. Dialyzer type or UFR did not affect iron removal. CONCLUSION: HF or HE dialysis membranes do not remove clinically significant amounts of iron sucrose or dextran formulations over a 4-hour HD session. This effect remained constant even controlling for UFR up to 500 ml/hour. Therefore, iron sucrose and iron dextran are not dialyzed by HE or HF dialysis membranes irrespective of UFR.

Ferric Compounds↗

Vascular to alveolar leak of iron dextran (120 kD) in the immature ventilated rabbit lung.

Rabbit fetuses were delivered by hysterotomy on day 27 or 28 of gestation. Immediately after birth, the animals were tracheotomized and received by intravenous injection 0.2 mu Ci radiolabeled albumin and 11 mg iron dextran in 0.2 ml saline. The newborn rabbits then were ventilated artificially with a tidal vol of 12 ml/kg for 5-20 min. One group of nonventilated animals served as controls. At the end of the experiment, one lung was lavaged via the airways and the other was fixed for histologic examination. The recovery of labeled albumin and iron dextran in the lavage fluid was quantified. Iron dextran complexes were easily identified in the lung sections by staining with Prussian blue. Iron dextran accumulated in the airspaces of animals delivered on day 27 (about 4% of the injected dose during 10-20 min of ventilation). The albumin leakage was slightly higher than that of the dextran, a result consistent with different mol wt of the markers. The vol density of leaking alveoli in histologic sections increased with time, from 0 at birth to a mean value of 0.36 after 20 min of ventilation. The leakage starts as a focal event, gradually involving more and more terminal airspaces. In the histologic sections, there was no indication of a significant leakage at the bronchiolar level, although the epithelium of terminal and preterminal airways was clearly injured in all ventilated animals.

Albumins↗

Percutaneous penetration of fluorescein isothiocyanate-dextrans and the mechanism for enhancement effect of enhancers on the intercellular penetration.

To identify the mechanism involved in the enhancement effect of enhancers on the intercellular penetration of large polar molecules, the skin penetration of fluorescein isothiocyanate (FITC)-dextrans (average molecular weight; 4400, 9400, and 69000 Da) and the lipid removal from the intercellular spaces by enhancers were studied using hairless rat skin. Pretreatment of hairless rat skin with enhancers such as n-octanol (20%), laurocapram (2%), isopropylmyristate (IPM, 20%), oleic acid (5%) and cineol (2%), which are water-immiscible, significantly enhanced the flux of FITC-dextrans, while pretreatment with water-miscible enhancers, i.e. dimethyl sulfoxide (DMSO, 5%) and N-methyl-2-pyrrolidone (NMP) did not increase the flux compared with the control. The penetration of FITC-dextrans was approximately size dependent. n-Octanol, laurocapram, IPM and oleic acid dramatically removed ceramides which are the intercellular lipids, whereas NMP and DMSO partly extracted the sphingolipids. A linear relationship was observed between the flux and removal of ceramides (p < 0.01), indicating that the removal of intercellular lipids would cause dramatic dilations between adherent cornified cells and enhance the penetration through the intercellular pathways. When the penetration of FITC-dextrans through Wistar rat skin was compared with that via hairless rat skin, the steady state flux of FITC-dextrans through Wistar rat skin pretreated with water-immiscible enhancers was 1.2- to 4.9-fold higher, suggesting that the penetration of large polar molecules through follicles may play at least some role in the percutaneous absorption.

Animals↗

Exponential increases in pulmonary intravascular and extravascular fluid volumes by overperfusion with dextran 70 in anesthetized dogs.

In the lung overperfused by dextran 70, it is not known how intravascular blood volume and extravascular water volume increase as a function of pulmonary microvascular pressure. To determine their characteristics, we produced an overperfusion state (high pressure and high blood flow) by infusing dextran 70 (100 ml/kg over 30 min) into 15 anesthetized dogs. Using the thermal and dye dilution technique, we simultaneously estimated the pulmonary blood volume (PBV) and the extravascular lung thermal volume (EVLTV), and correlated mathematically these variables with pulmonary artery end-diastolic pressure (PAEDP). Here, we defined PAEDP as the pulmonary microvascular pressure. From the values determined before, during and after dextran 70 infusion, we obtained the following exponential relations. PBV(ml/kg) = 30[1-0.902(e-0.021PAEDP(mmHg)) EVLTV(ml/kg) = 5.15(e0.027PAEDP (mmHg)) From these mathematical relations, we conclude that: (1) the pulmonary blood volume increases rapidly at low pressure and slowly at high pressure; and (2) the pulmonary extravascular water volume increases slowly at low pressure and then increases rapidly at high pressure. In addition, this equation indicates that the critical pressure from which the pulmonary extravascular water volume exceeds the upper limit of its normal volume is about 17 mmHg in PAEDP. Infusion of dextran 70 increased plasma macromolecular osmotic pressure from 20.8 +/- 0.6 (mean +/- SD) mmHg before infusion to 43.7 +/- 1.4 mmHg after infusion. Therefore, dextran 70 does not change the critical microvascular pressure against pulmonary edema.

Animals↗

Dextran and adhesions in guinea-pigs.

This prospective, randomized, 'blind' study with guinea-pigs was performed to assess the possible benefit of 6% dextran 70 (molecular weight 70 000) in the prevention of post-operative intra-abdominal adhesions and recurrent adhesions after adhesiolysis. In 50 guinea-pigs lesions for inducing adhesions were applied at the end of the uterine horn. On the right side a strip lesion was made and on the left side an end-to-end anastomosis was performed after section. Before closing the peritoneum 20 ml 6% dextran 70 (N = 25) or saline (N = 25) were introduced into the peritoneal cavity. A second laparotomy 4 weeks later showed no differences in adhesion formation in the animals treated with 6% dextran 70 and saline. In the animals with adhesions adhesiolysis was performed and 6% dextran 70 or saline was left in the peritoneal cavity. Again no beneficial effect of dextran was seen. The end-to-end procedure appeared to be far more suitable for producing adhesions than was the strip lesion.

Abdomen↗

The effect of topical application of dextran on the gingiva of the beagle dog.

Dextrans derived from Leuconostoc mesenteroides were placed on clinically healthy gingiva of Beagle dogs once a day for 21 days. Control gingival tissues received saline. Both healthy controls and dextran-treated tissues were brushed daily. Inflamed control tissues were obtained by allowing plaque to accumulate for 21 days. Tissues receiving daily application of dextran solutions developed chronic gingival inflammation but displayed no clinical signs of gingivitis. Healthy control gingival tissues showed no clinical signs of gingivitis and minor histologic inflammatory changes. Tissues exposed to dental plaque showed the typical clinical and histological inflammatory changes of gingivitis. Thus dextran, a substance similar to the extracellular polysaccharide found in dental plaque, was able to penetrate the sulcular epithelium, enter healthy gingival connective tissue and cause chronic inflammation. This connective tissue inflammation occurred without inducing any of the clinical signs of gingivitis. Therefore, it is concluded that dextran produced one component of the gingivitis response, chronic histologic inflammation, independent of another major component of the disease, clinical inflammation.

Administration, Topical↗

Dextran-induced inflammation and its effect on keratinized gingival epithelium in monkeys.

The cell population present during dextran-induced inflammation and its effect upon induced keratinization of the sulcular epithelium was investigated in two young adult male Rhesus monkeys. Keratinization of the sulcus epithelium was induced by a combined regimen of scaling, an intravenous injection of achromycin and daily rubber cup prophylaxes. After keratinization was confirmed by means of biopsies, inflammation was induced either by injecting 200 microliters of a 5% dextran saline solution or by applying the solution topically on the marginal gingiva for 2 weeks. Clinical grade dextran, molecular weight 70,000, was used. Physiologic saline solution, either injected or topical, was also used. At the same time, the daily prophylaxes were continued. After the 2 weeks, gingival biopsies were taken from each tooth treated with the different regimens. One-half of each biopsy was routinely processed and stained with hematoxylin and eosin or Rhodamine B, while the other half was processed for and stained with alcoholic and aqueous PAS to detect dextran in tissues. Histologic evaluation was carried out in three areas: a crestal zone, a cervical zone and an oral gingival zone. An Inflammatory Index (II) was determined and the width and length of keratin were measured. Dextran, either topical or injected, produced mainly a chronic inflammatory response characterized by lymphocytes (30-35%), monocytes-macrophages (5-10%), plasma cells (10%), polymorphonuclear leukocytes (PMNs) (15%) and unidentified cells (35%). Conversely, the physiologic saline-induced inflammation showed PMNs (75%), lymphocytes (5%) and unidentified cells (20%). The II for injected areas was significantly higher than for those topically treated or for nontreated controls. However, the increased II did not affect the degree of keratinization achieved.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Fetal risks with dextrans during delivery.

Epidural analgesia for caesarean section is increasingly used and is gradually replacing general anaesthesia. Hypotension is one of the main risks: preloading of the maternal circulation is used to prevent maternal hypotension and its consequences. For this, various colloid and crystalloid solutions are used. We report a case of maternal anaphylactoid reaction with apparent death in a neonate after dextran administration to the mother. After 100ml of a dextran 40 solution administered intravenously, immediately before an epidural blockade, the mother fainted and developed urticaria and mild respiratory disturbances, without hypotension. At that point dextran infusion was stopped. An apparently dead neonate was rapidly delivered. Immediate and vigorous cardiopulmonary resuscitation was successful. Clonismus appeared 12h later, followed by 3 general epileptic fits treated by phenytoin infusion and subsequently oral phenobarbital. No aetiology was found. After 2 months of treatment, barbiturates were stopped following clinical and electroencephalogram (EEG) improvement. Several similar cases of neonatal disorders resulting from preventive dextran administration during delivery were studied in a national pharmacovigilance survey in France. There were 32 cases reported with moderate maternal anaphylactoid reaction associated with severe acute fetal distress; it is probably advisable to take a cautious approach and avoid preventive fluid preload by dextran administration. Gelatins or crystalloid solutions should be preferred, with intravenous vasopressive amine administered promptly and repeated if necessary should significant maternal hypotension occur during epidural anaesthesia.

Anaphylaxis↗