Isolation of interstitial fluid from skeletal muscle and subcutis in mice using a wick method

2004 ◽  
Vol 287 (5) ◽  
pp. H2085-H2090 ◽  
Author(s):  
Carl Erik Markhus ◽  
Helge Wiig
Keyword(s):  
Skeletal Muscle ◽  
Plasma Proteins ◽  
Interstitial Fluid ◽  
Colloid Osmotic Pressure ◽  
Tissue Fluid ◽  
Protein Extravasation ◽  
Plasma Protein Extravasation ◽  
Wick Technique ◽  
Region Plasma ◽  
Gain Access

Until recent years, mice were sparsely used in physiological experiments, and therefore, data on the basic cardiovascular parameters of mice are lacking. Our aim was to gain access to interstitial fluid and thereby study transcapillary fluid dynamics in this species. Using a modified wick method, we were able to isolate interstitial fluid from subcutis and skeletal muscle in mice. Three-stranded, dry, nylon wicks were inserted post mortem in an attempt to avoid local inflammation and thus eliminate protein extravasation and wick contamination. Colloid osmotic pressure (COP) was measured with a colloid osmometer for submicroliter samples and averaged (means ± SE) 18.7 ± 0.4 in plasma, 9.1 ± 0.4 in subcutis, and 12.3 ± 0.5 mmHg in muscle. HPLC of plasma and wick fluid showed similar patterns except for some minor peaks eluting in the <40-kDa region. Plasma protein extravasation as determined by 125I-labeled human serum albumin showed that contamination of wick fluid by plasma proteins was negligible (<2%). Capillary hyperfiltration induced by intravenous infusion of saline (10% of body wt) was reflected in tissue fluid isolated by wicks as shown by the average postinfusion COP values of 14.5 ± 0.6, 6.8 ± 0.3, and 7.7 ± 0.4 mmHg in plasma, subcutis, and muscle, respectively. We conclude that the wick technique can be easily adapted for use in mice and may represent a reliable method to isolate interstitial fluid and study transcapillary fluid flux in this species.

Isolation of pulmonary interstitial fluid in rabbits by a modified wick technique

2001 ◽  
Vol 280 (5) ◽  
pp. L1057-L1065 ◽  
Author(s):  
Daniela Negrini ◽  
Alberto Passi ◽  
Katia Bertin ◽  
Federica Bosi ◽  
Helge Wiig
Keyword(s):  
Osmotic Pressure ◽  
Plasma Proteins ◽  
Regional Differences ◽  
Protein Concentration ◽  
Interstitial Fluid ◽  
Lung Parenchyma ◽  
Colloid Osmotic Pressure ◽  
Rabbit Lung ◽  
Pressure Plasma ◽  
Wick Technique

Interstitial fluid protein concentration (Cprotein) values in perivascular and peribronchial lung tissues were never simultaneously measured in mammals; in this study, perivascular and peribronchial interstitial fluids were collected from rabbits under control conditions and rabbits with hydraulic edema or lesional edema. Postmortem dry wicks were implanted in the perivascular and peribronchial tissues; after 20 min, the wicks were withdrawn and the interstitial fluid was collected to measure Cprotein and colloid osmotic pressure. Plasma, perivascular, and peribronchial Cproteinvalues averaged 6.4 ± 0.7 (SD), 3.7 ± 0.5, and 2.4 ± 0.7 g/dl, respectively, in control rabbits; 4.8 ± 0.7, 2.5 ± 0.6, and 2.4 ± 0.4 g/dl, respectively, in rabbits with hydraulic edema; and 5.1 ± 0.3, 4.3 ± 0.4 and 3.3 ± 0.6 g/dl, respectively, in rabbits with lesional edema. Contamination of plasma proteins from microvascular lesions during wick insertion was 14% of plasma Cprotein. In control animals, pulmonary interstitial Cprotein was lower than previous estimates from pre- and postnodal pulmonary lymph; furthermore, although the interstitium constitutes a continuum within the lung parenchyma, regional differences in tissue content seem to exist in the rabbit lung.

Isolation of interstitial fluid from rat mammary tumors by a centrifugation method

2003 ◽  
Vol 284 (1) ◽  
pp. H416-H424 ◽  
Author(s):  
Helge Wiig ◽  
Knut Aukland ◽  
Olav Tenstad
Keyword(s):  
Interstitial Fluid ◽  
Tumor Tissue ◽  
Venous Blood ◽  
Lymphatic Drainage ◽  
Colloid Osmotic Pressure ◽  
Low Molecular Weight ◽  
Tissue Fluid ◽  
Arterial Plasma ◽  
Lower Ratio ◽  
Superficial Tumor

Access to interstitial fluid is of fundamental importance to understand tumor transcapillary fluid balance, including the distribution of probes and therapeutic agents. Tumors were induced by gavage of 9,10-dimethyl-1,2-benzanthracene to rats, and fluid was isolated after anesthesia by exposing tissue to consecutive centrifugations from 27 to 6,800 g. The observed51Cr-EDTA (extracellular tracer) tissue fluid-to-plasma ratio obtained from whole tumor or from superficial tumor tissue by centrifugation at 27–424 g was not significantly different from 1.0 (0.92–0.99), suggesting an extracellular origin only. However, fluid collected from excised central tumor parts had a significantly lower ratio (0.66–0.77) for all imposed G forces, suggesting dilution by fluid deriving from a space unavailable for51Cr-EDTA. The colloid osmotic pressure in tumor fluid was generally higher than in fluid isolated from the subcutis, attributable to less selective capillaries and impaired lymphatic drainage in tumors. HPLC analysis of tumor fluid showed that low-molecular-weight macromolecules not present in arterial plasma were present in tumor fluid obtained by centrifugation and in venous blood draining the tumor, most likely representing proteins derived from tumor cells. We conclude that low-speed centrifugation may be a simple and reliable method to isolate interstitial fluid from tumors.

scholarly journals The Colloid Osmotic Pressure Across the Glycocalyx: Role of Interstitial Fluid Sub-Compartments in Trans-Vascular Fluid Exchange in Skeletal Muscle

2021 ◽  
Vol 9 ◽  
Author(s):  
Fitzroy E. Curry ◽  
C. Charles Michel
Keyword(s):  
Skeletal Muscle ◽  
Steady State ◽  
Osmotic Pressure ◽  
Capillary Pressure ◽  
Interstitial Fluid ◽  
Protein Composition ◽  
Colloid Osmotic Pressure ◽  
Endothelial Glycocalyx ◽  
Fluid Exchange ◽  
Small Pore

The primary purpose of these investigations is to integrate our growing knowledge about the endothelial glycocalyx as a permeability and osmotic barrier into models of trans-vascular fluid exchange in whole organs. We describe changes in the colloid osmotic pressure (COP) difference for plasma proteins across the glycocalyx after an increase or decrease in capillary pressure. The composition of the fluid under the glycocalyx changes in step with capillary pressure whereas the composition of the interstitial fluid takes many hours to adjust to a change in vascular pressure. We use models where the fluid under the glycocalyx mixes with sub-compartments of the interstitial fluid (ISF) whose volumes are defined from the ultrastructure of the inter-endothelial cleft and the histology of the tissue surrounding the capillaries. The initial protein composition in the sub-compartments is that during steady state filtration in the presence of a large pore pathway in parallel with the “small pore” glycocalyx pathway. Changes in the composition depend on the volume of the sub-compartment and the balance of convective and diffusive transport into and out of each sub-compartment. In skeletal muscle the simplest model assumes that the fluid under the glycocalyx mixes directly with a tissue sub-compartment with a volume less than 20% of the total skeletal muscle interstitial fluid volume. The model places limits on trans-vascular flows during transient filtration and reabsorption over periods of 30–60 min. The key assumption in this model is compromised when the resistance to diffusion between the base of the glycocalyx and the tissue sub-compartment accounts for more than 1% of the total resistance to diffusion across the endothelial barrier. It is well established that, in the steady state, there can be no reabsorption in tissue such as skeletal muscle. Our approach extends this idea to demonstrate that transient changes in vascular pressure favoring initial reabsorption from the interstitial fluid of skeletal muscle result in much less fluid exchange than is commonly assumed. Our approach should enable critical evaluations of the empirical models of trans-vascular fluid exchange being used in the clinic that do not account for the hydrostatic and COPs across the glycocalyx.

Transcapillary fluid shifts in tissues of the head and neck during and after simulated microgravity

1991 ◽  
Vol 71 (6) ◽  
pp. 2469-2475 ◽  
Author(s):  
S. E. Parazynski ◽  
A. R. Hargens ◽  
B. Tucker ◽  
M. Aratow ◽  
J. Styf ◽  
...  
Keyword(s):  
Time Course ◽  
Interstitial Fluid ◽  
Baseline Period ◽  
Colloid Osmotic Pressure ◽  
Tissue Fluid ◽  
Fluid Filtration ◽  
Lower Lip ◽  
Facial Edema ◽  
Capillary Pressures ◽  
Male Subjects

To understand the mechanism, magnitude, and time course of facial puffiness that occurs in microgravity, seven male subjects were tilted 6 degrees head-down for 8 h, and all four Starling transcapillary pressures were directly measured before, during, and after tilt. Head-down tilt (HDT) caused facial edema and a significant elevation of microvascular pressures measured in the lower lip: capillary pressures increased from 27.7 +/- 1.5 mmHg (mean +/- SE) pre-HDT to 33.9 +/- 1.7 mmHg by the end of tilt. Subcutaneous and intramuscular interstitial fluid pressures in the neck also increased as a result of HDT, whereas interstitial fluid colloid osmotic pressures remained unchanged. Plasma colloid osmotic pressure dropped significantly by 4 h of HDT (21.5 +/- 1.5 mmHg pre-HDT to 18.2 +/- 1.9 mmHg), suggesting a transition from fluid filtration to absorption in capillary beds between the heart and feet during HDT. After 4 h of seated recovery from HDT, microvascular pressures in the lip (capillary and venule pressures) remained significantly elevated by 5–8 mmHg above baseline values. During HDT, urine output was 126.5 ml/h compared with 46.7 ml/h during the control baseline period. These results suggest that facial edema resulting from HDT is caused primarily by elevated capillary pressures and decreased plasma colloid osmotic pressures. The negativity of interstitial fluid pressures above heart level also has implications for maintenance of tissue fluid balance in upright posture.

Interaction of transcapillary Starling forces in the isolated dog forelimb

1977 ◽  
Vol 233 (1) ◽  
pp. H136-H140 ◽  
Author(s):  
R. A. Brace ◽  
A. C. Guyton
Keyword(s):  
Osmotic Pressure ◽  
Plasma Proteins ◽  
Interstitial Fluid ◽  
Fluid Pressure ◽  
Colloid Osmotic Pressure ◽  
Interstitial Fluid Pressure ◽  
Average Increase ◽  
Average Decrease ◽  
Average Value ◽  
Intact Limb

Three of the four Starling forces were measured in the intact dog forelimb after anesthetization and all four of the Starling forces were measured in the same forelimb which was surgically isolated yet innervated. In the isolated forelimb, isogravimetric capillary pressure (Pci) averaged 15.6 mmHg; colloid osmotic pressure of the plasma proteins (IIp) averaged 19.9 mmHg; mean interstitial fluid pressure (Pif) was +0.4 mmHg, and the average value of interstitial colloid osmotic pressure (IIif) was 4.9 mmHg. Thus the net imbalance in the Starling forces, i.e., (Pci - Pif) - (IIp - IIif), averaged 0.3 mmHg. Furthermore, the value of IIif was consistently decreased after isolation (average decrease of 1.2 mmHg) while Pif was always increased following isolation (average increase of 4.3 mmHg). In addition, it was found that if the forelimb was denervated during isolation, then Pif was increased by an average of 2 mmHg above Pif in the innervated, isolated forelimb. In summary, these studies show that the differences between the intact and isolated forelimb are that Pci averages 10-11 mmHg in the intact forelimb and 15-16 mmHg in the isolated innervated forelimb while interstitial fluid pressure is negative in the intact limb and positive in the isolated limb.

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