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The process of neovascularization was analyzed in vivo in different expanded polytetrafluoroethylene (e-PTFE) implants which are frequently used in cardio-thoracic and vascular surgery. We have used the model of the hamster dorsal skinfold chamber which allows quantitative analysis of the microcirculation by means of intravital fluorescence microscopy. Pieces of approximately 1 mm2 of the cardiovascular patch (CVP, fibril length: 30 microns; n = 21), surgical membrane (SM, fibril length: 1 micron; n = 16), and soft tissue patch (STP, fibril length: 22 microns; n = 12) were implanted into the skinfold chambers. On day 10 after implantation, the functional density of newly formed microvessels was significantly (P less than 0.05) higher in CVP (145.0 +/- 10.9 cm-1) as compared to SM (688 +/- 13.9 cm-1) and STP (86.9 +/- 21.2 cm-1). In addition, CVP revealed a larger zone of neovascularization (311.6 +/- 19.4 microns) and the tightest integration (dynamic breaking strength: 17.9 +/- 3.0 cN/mm2) into the perigraft tissue, while SM demonstrated only few microvessels and no integration (6.0 +/- 1.9 cN/mm2) into the perigraft. None of the three different PTFE-implants revealed transmural ingrowth of capillaries. The internodal distance of PTFE implants seems to be the most important factor for neovascularization. Surgical membrane used for the replacement of passive biological membranes demonstrated, as is its purpose, little neovascularization and no integration into the perigraft tissue.(ABSTRACT TRUNCATED AT 250 WORDS)

We present a new technique for specific ex situ fluorescent staining of islets of Langerhans with neutral red. Depending on the concentration of neutral red (1 microgram/ml), this technique allows the differentiation between exocrine and endocrine pancreatic tissue, according to differences in fluorescence intensity. Pancreatic islets can be stained not only in situ, but in particular ex situ after pancreas digestion. Since intravital staining of the pancreas in situ with neutral red is not applicable in human transplantation, the staining procedure ex situ may be a useful tool for islet purification by particle sorting, using a fluorescent-light-activated system; thus, a sufficient mass of purified islets for free transplantation may be isolated.

A new experimental skin-flap model is presented in which direct observations of blood flow in individual capillaries can be made from the time of flap creation throughout the entire evolution of the establishment of necrosis. After flap creation, one observes through the microscope that at 1 hour a large area of tissue is nonperfused as a result of the surgical trauma. This is followed by vasodilatation at 6 hours, resulting in an increase in the area of perfused tissue. At 24 hours, the vasodilatation persists, and the red cells that have entered the tissue during the vasodilatation (6 hours) accumulate in the capillaries, this being reflected by an increased area of nonperfused tissue. This increase continues to 72 hours, at which time the perfusion-nonperfusion interface becomes well defined and remains so throughout the 5-day experiment. Analyses of the relationship between early postoperative capillary perfusion and eventual necrosis are presented. Advantages and disadvantages of this model are listed.

Reperfusion injury following prolonged ischemia is thought to be caused primarily by microvascular failure. The aim of the present study was to investigate whether prophylactic isovolemic hemodilution with Dextran 60 (hct 30%) could improve microvascular perfusion after 4 h of pressure-induced ischemia in skeletal muscle. In 28 Syrian golden hamsters (6-8 weeks/60-80 g b. wt.) a dorsal skinfold chamber and permanent arterial and venous catheters were implanted under Nembutal anesthesia (50 mg/kg b. wt.). Following a recovery period of 48 h pressure-induced ischemia was applied to the skeletal muscle within the skinfold chamber by means of a transparent stamp. Quantitative analyses of microhemodynamics were performed in the awake animal prior to and 15 min, 1, 2, 4 and 24 h after ischemia using vital fluorescence microscopy. In non-treated animals, functional capillary density decreased after 4 h of ischemia to 30% of the initial values (P less than 0.001); after 24-h reperfusion only 50% of the initially perfused capillaries were reperfused (P less than 0.001). The heterogeneity of functional capillary density increased after ischemia to a maximum of 2.19 +/- 0.94 as compared to 0.48 +/- 0.11 prior to ischemia. Capillary RBC-velocity suffered a marked reduction in the early reperfusion phase and did not recover up to the 24-h observation time. In contrast, prophylactic isovolemic hemodilution was associated with only a small and reversible reduction of functional capillary density after 4-h ischemia. At 24-h reperfusion 90% of the initially perfused capillaries were reperfused. Capillary RBC-velocity was reduced in the early reperfusion phase, but returned to normal values within 24 h. Thus, prophylactic isovolemic hemodilution resulted in a marked reduction of microvascular reperfusion failure in skeletal muscle. A hematocrit lower than normal prior to ischemia provides better conditions for capillary reperfusion after prolonged ischemia.

The homozygous (hr/hr) hairless mouse ear was introduced in 1980 by Eriksson and coworkers as a model for in vivo studies of the skin microcirculation. Herein we expand on this work, presenting results of in vivo microvascular parameter measurements and morphologic studies in the intact ear. The in vivo measurements include microvascular diameter, RBC velocity, capillary density, and the frequency and amplitude of arteriolar vasomotion. In connection with the in vivo studies, a detailed anatomic description of the overall and vascular anatomy is given. Additionally, the preparation techniques for carrying out these in vivo and morphologic studies in the mouse ear are presented in detail.

Transplantation of isolated islets of Langerhans is frequently followed by early loss of islet function. Because whether this is caused by insufficient vascularization or graft rejection is unknown, angiogenesis and microvascularization of islet grafts were studied in vivo by means of intravital microscopy. After transplantation of syngeneic islets in hamster dorsal skin-fold chambers, 97% (n = 66) of the islets exhibited the first signs of angiogenesis at days 2-4, characterized by sinusoidal sacculations and capillary sprouts. After 10 days, angiogenesis was completed, consisting of a microvascular network similar to those of islets in situ: arterial supply, afferent and efferent capillary loops, and venular drainage. Functional density of microvessels was 700.1 +/- 127.0 cm-1, and erythrocyte velocity was 0.58 +/- 0.35 mm/s. Intracellular insulin was demonstrated immunohistochemically. Electron-microscopic studies revealed normal fine structure of the capillary wall. The model allows in vivo analysis of microvascular phenomena occurring in host-vs.-graft reaction after allogeneic and xenogeneic islet transplantation. Furthermore, it may be used to quantitatively assess immunosuppressive regimens.

In 147 patients suffering from arterial occlusive disease of the lower extremities 162 PTAs of the iliac, femoral and popliteal arteries were performed. Thirteen complications occurred in 12 patients, while surgical intervention was necessary in seven cases (4.3%). Hematoma at the site of the arterial puncture were the most frequent complications, requiring acute surgical intervention (n = 3). In two patients, acute vascular occlusion after PTA required bypass operation and thrombendarteriectomy, respectively. In one patient, presenting with an iliac stenosis, PTA was performed using a transaxillar approach and was complicated by an axillar hematoma due to the ruptur of the arteria subscapularis. Furthermore, one patient with an extended hematoma of the abdominal wall required surgical intervention. The results presented indicate that PTA of the lower extremities for treatment of arterial occlusive disease is a procedure with low risk. Complications, which require acute operative therapy, can be treated mostly successful by small surgical interventions.

Prolonged ischemia is known to cause severe damage in skeletal muscle and skin as a result of reperfusion failure. Isovolemic hemodilution has been suggested as a modality to reverse microcirculatory disorders by improving flow properties and flow conditions of the blood. The aim of the present study was to investigate whether prophylactic isovolemic hemodilution could improve tissue oxygenation after 4h of pressure induced ischemia in skeletal muscle. In 17 Syrian golden hamsters a dorsal skin fold chamber and two permanent arterial and venous catheters were implanted. Following a recovery period of 48h ischemia was induced for 4h by means of a transparent stamp compressing the tissue within the chamber. In 9 animals (control, hct 43%) measurements of tissue PO2 (platinum multiwire electrode) were performed prior to and 15 min, 2h and 24h after release of ischemia. In 8 animals isovolemic hemodilution with Dextran 60 (hct 29%) was carried out prior to ischemia and measurements of local tissue PO2 were performed as reported for the control group with an additional measurement 30 min after hemodilution. In control animals tissue PO2 decreased significantly (p less than 0.01) from 20.7 +/- 2.4 mmHg prior to ischemia to 8.8 +/- 3.1 mmHg after 15 min of reperfusion; after 24h tissue PO2 was 15.6 +/- 6.1 mmHg. In hemodiluted animals tissue PO2 increased due to hemodilution from 20.9 +/- 1.6 mmHg to 23.5 +/- 2.5 mmHg (p less than 0.05); after 15 min of reperfusion tissue PO2 was 19.8 +/- 6.8 mmHg and remained unchanged for 24h (20.0 +/- 2.5 mmHg).(ABSTRACT TRUNCATED AT 250 WORDS)

In order to study changes in functional capillary density and tissue PO2 in chronically ischemic skeletal muscle, a new model, using the Syrian golden hamster was developed. In the hamster dorsal skin fold, which receives its vascular supply from two cranial and two caudal feeding arteries, a double frame chamber was implanted and ischemia was induced in the cranial part by heat coagulation of the cranial arteries outside of the chamber. This technique allows for analysis of microvascular hemodynamics and local tissue PO2 prior to and during a prolonged period of ischemia in skin muscle. As result of ischemia the diameters of the arterioles increased (p less than 0.001) over the whole 11 day observation period. Functional capillary density decreased significantly (p less than 0.01) during the first 7 days, while capillary RBC-velocity was reduced throughout the 11 days of observation. RBC-velocity in collecting venules was diminished significantly throughout the postischemic observation period. The diameters of the collecting venules first increased upon ischemia (p less than 0.001) but were found decreased at 4, 7 and 11 days. Measurements of tissue PO2 demonstrated a marked decrease from a mean PO2 of 20.5 mmHg prior, to 9.5 mmHg following induction of ischemia. The model allows for induction of chronic ischemia and is suitable to study the effect of therapeutic measures on the microcirculation in chronically ischemic skeletal muscle in vivo.