Biomaterials, artificial cells, and immobilization biotechnology : official journal of the International Society for Artificial Cells and Immobilization Biotechnology最新文献

For the determination of stroma content in SFH, two analytical methods were developed, the HPLC-UV for phospholipids and the enzyme immunoassay for blood group antigen. These analytical methods were applied to evaluate three representative SFH preparations. The 36,000 x g centrifugation method was shown to contain higher amount of stroma components. On the other hand, the BMM-40 nm and the ultrafiltration method with a 100 kd membrane was shown to remove more than 99.7% of phospholipids and almost all of blood group antigen from hemoglobin solutions. These results demonstrated that these analytical methods were very useful in evaluating any highly purified hemoglobin solutions.

Over the past 10 years, the use of perfluorochemical emulsions (PFCE) and carbogen or oxygen breathing has been explored as an adjuvant to radiation therapy and/or chemotherapy in the treatment of solid tumors. The rationale for the use of PFCE and oxygen breathing in this therapeutic setting is that solid tumor masses contain areas of hypoxia which are therapeutically resistant. Since x-rays and many chemotherapeutic agents require oxygen to be maximally cytotoxic and most normal tissues are well-oxygenated, the additional oxygen put in circulation by the PFCE should not increase the normal tissue toxicities produced by the various therapies. The largest body of preclinical work and all of the clinical studies in cancer conducted with PFCE, thus far, have been done with Fluosol-DA, 20%. Oxygen microelectrode studies have confirmed increased oxygenation in previously hypoxic tumor regions after the administration of Fluosol-DA and carbogen breathing. The preclinical studies have shown very positive effects with single dose and fractionated radiation in several rodent solid tumor models. Many widely used anticancer drugs including antitumor alkylating agents and adriamycin are enhanced by PFCE and carbogen breathing for longer time periods (6 h). More recently, several experimental concentrated PFCE preparations have become available and work with these is actively under way in several laboratories. Clinical studies with radiation and four or five chemotherapeutic drugs as single agents have indicated that Fluosol-DA followed by oxygen breathing can be administered safely in a variety of cancer therapeutic settings. Further clinical studies with Fluosol-DA are planned.

To preserve isolated kidney normothermically, PHP containing UW components were evaluated as perfusates. Kidneys were flushed out by Lactate Ringer solution immediately after isolation from mongrel dogs, and then connected to the perfusion circuit which consists of a preservation box, a reservoir of perfusate, a membrane oxygenator and a drive unit. PHP containing 140 mEq/l of Na+ and 4 mEq/l of K+ (PHP(E)), UW solution (UW) and UW components added PHP(E) were prepared and adjusted at pH 7.4 prior to use. Temperature and perfusion pressure were controlled at 37 degrees C and 100 mmHg, respectively. During 12 hour perfusion, remarkable changes in pH were seen in UW group and PHP group while higher oxygen consumption was noted in PHP(E)+UW group than that in PHP(E) group. The histological findings showed moderate damages of tubular epithelial cells and maintaining normal glomerular structure in PHP(E)+UW while severe damage of both tubulus in UW group were seen. There was no edematous degeneration in both UW and PHP(E)+UW groups, however, it was seen in PHP(E) alone. It was suggested that components of UW solution have positive effect on normothermic machine perfusion with PHP(E) solution.

The effects of stroma-free hemoglobin (SFHgb) on the coronary circulation remain unclear. An intact canine model utilizing intracoronary adenosine to abolish the confounding effect of autoregulation was used to study maximal myocardial oxygen delivery during progressive hemodilution with polymerized bovine SFHgb. The circumflex coronary artery was instrumented with a flow probe, hydraulic constrictor, and proximal and distal catheters for adenosine infusion and distal pressure measurement, respectively. This preparation was used to generate diastolic coronary pressure-flow relations during maximal vasodilation. Maximal coronary conductance and maximal myocardial oxygen delivery were determined in two groups of 7 dogs each following hemodilution, first with 6% hetastarch (Control), followed by further hemodilution with ultra-pure, polymerized, bovine SFHgb. After hemodilution with SFHgb, maximal coronary flow increased slightly without evidence of coronary vasoconstriction. Since hemodilution with this material increases oxygen carrying capacity, maximal oxygen delivery is greater than Control, despite the very low canine hematocrit. These findings suggest: 1) SFHgb can provide adequate oxygen delivery to the myocardium despite extreme degrees of hemodilution, and 2) in this intact model, there is no evidence of adverse coronary vasomotion.

The effect of diaspirin cross-linked hemoglobin (DCLHb) on mean arterial pressure (MAP) and heart rate (HR) was compared to Ringer's lactate (RL) and shed blood in a 70% lethal model of hemorrhage (35 cc/kg blood loss) in conscious rats. All animals resuscitated with DCLHb regardless of dose (17.5 and 35 cc/kg) and concentration (7% and 10% solution) exhibited complete restoration of MAP and HR which was maintained for at least 5 hrs. Hemodynamic responses in DCLHb-treated animals were not significantly different from 35 cc/kg blood-treated animals. In RL (105 cc/kg) treated animals the MAP was restored to 60-70% of baseline. 24 hr survival in animals resuscitated with fluids ranged between 88-100% and was not significantly different between treatment groups.

Perfluorocarbons (PFCs) deliver oxygen due to their physical characteristics and the convective delivery to tissues; this has been amply demonstrated in numerous animal models and in clinical trials. PFC emulsions have also been used for organ perfusion and augmentation of oxygenation to cell cultures. By reason of their small particle size, PFC emulsions penetrate collateral capillaries of an ischemic microcirculation, both supplying oxygenation and possibly restoring flexibility of acidotically stiffened erythrocytes by reinstituting aerobic metabolism. There is also evidence that PFCs can augment tissue oxygenation by increasing oxygen solubility in the plasma phase of blood; this improves diffusion gradients between plasma and tissues even at relatively low doses of PFC. This hypothesis is supported by the ability of PFCs to increase oxygen tensions in tissues such as blood, brain, liver, kidney, pancreas, skeletal muscle, retina etc. PFCs can also on occasions increase critical oxygen extraction coefficients and should therefore be effective in supporting oxidative metabolism in states of extreme cardio-respiratory insufficiency. This theory represents a new concept in oxygen delivery and may define a new class of therapeutic agents that can improve tissue oxygen supply in various pathological states involving low oxygen delivery and ischemia.

Egg Yolk Phospholipid(EYP) has been used extensively as the primary surfactant in parenteral fat emulsions for many years. The simplicity, functionality and physiologic tolerance of EYP has contributed greatly to its success in the intravenous emulsion arena. The mechanism of stabilization in triglyceride emulsions is well understood; however, this is not the case with perfluorocarbon emulsions. Interfacial models, as well as emulsion stability studies, have been conducted utilizing EYP of varied composition in order to derive a structure/function relationship. Our studies indicate that minor components, total unsaturation, acyl chain length and presence of charged species have significant impact on the functional properties of EYP and the subsequent stability of the emulsion product. These findings contribute to our ability to design and manipulate natural surfactants with superior properties for use in medical applications of perfluorocarbon emulsions.

A double exchange transfusion-double carbon clearance method was evaluated for assessing reticuloendothelial (RE) function following exchange transfusion with hemoglobin solutions. Fifty percent of estimated blood volume (3% body weight) was withdrawn from anesthetized Sprague-Dawley rats and isovolumically replaced with shed blood (SB, control), lysed shed blood (LB, pos. control), human stroma-free hemoglobin solution (SFH), or polyhemoglobin solution (PHS). Thirty minutes after the exchange transfusion, colloidal carbon was injected intravenously and its vascular clearance followed for 1 hour. Then, the 50% exchange transfusion was repeated and the second carbon clearance measured. The intravascular carbon clearance constants, K, and clearance half-times, T1/2, were calculated and compared. No apparent differences in RE function were seen among the groups after the initial exchange transfusion. However, following the second exchange transfusion significant (P less than 0.05) slowing of carbon clearance was observed in lysed blood treated animals. The RE function of SFH or PHS treated animals were not different (P less than 0.05) from that of SB animals. A double exchange transfusion-double carbon clearance method seems to reveal changes in RE function that are not apparent after a single exchange transfusion and clearance test.