Establishment of a neovascular bed in a collagen-impregnated polyurethane sponge.

A technique for promoting vascularization of a polyurethane sponge is demonstrated in the present study. Collagen-impregnated polyurethane sponges (Hypol, 2002) Foamable Hydrophilic Prepolymer (FHP) were implanted in the femoral fossa of rats for 1 day to 6 weeks. The ligated femoral artery/vein was pulled through the sponges to facilitate more complete neovascularization. Light-microscopic evaluation of the implanted sponges revealed that significant vascularization had occurred by the seventh day of implantation, and was maximal by the fourth to sixth week. Sponges containing collagen had a more thorough vascularization process than sponges without collagen, perhaps due to a more uniform pore size as demonstrated by scanning EM. Time course studies suggested that the artery/vein pull-through enhanced the development of the neovascularization process in the center of the sponges. We conclude that significant vascular tissue in-growth can be developed in polyurethane sponges and that both collagen and centrally placed blood vessels help promote the vascularization process. Potential applications could extend to a variety of bioartificial systems including endocrine or hepatic transplantation, soft-tissue prosthetic materials, bone grafts, or drug delivery systems. Further studies would be useful in providing additional information on the factors promoting neovascularization, and on the potential applications of this methodology using the present or similar biomaterials.

Circulatory isolation of the liver for studying energy metabolism in trauma and sepsis.

An isolated level model was used in 25-30 kg pigs to study the uptake and/or release of various energy substrates across the liver and splanchnic bed during trauma and sepsis. This was accomplished by bypassing the hepatic portion of the inferior vena cava (IVC) using a polytetrafluoroethylene (PTFE) (IMPRA, Inc.) graphite-impregnated graft, from above the renal veins to the right atrial junction. The IVC was then ligated between the distal anastomosis and the liver. Sampling was done from inlying catheters placed in the main portal vein, hepatic vena cava, and an artery. Total hepatic blood flow was determined by the dye dilution technique. Our data showed the same pattern of metabolic derangements in both the traumatized and the septic-starved animals, but differed significantly from the normal fed controls. The correlation between glucogenesis and other energy functions of the liver and the proteolysis in skeletal muscle seen in trauma and sepsis is demonstrated. This model is functional and is applicable to the study of various kinds of shock and/or liver failure.

Trypan blue dye uptake and lactate dehydrogenase in adult rat hepatocytes--freshly isolated cells, cell suspensions, and primary monolayer cultures.

Leakage of lactate dehydrogenase and staining by the vital dye trypan blue were investigated in adult rat hepatocytes at the time of isolation, in suspensions up to 3 h and in primary monolayer cultures up to 3 d. These two parameters of plasma membrane integrity were found to correlate closely in hepatocyte suspensions, but to a lesser degree in monolayer cultures. Functional activity was demonstrated in culture by glucose consumption and lactic acid production. There was a balance of total lactate dehydrogenase (LDH) activity over time for both hepatocyte suspensions and cultures. Loss of LDH activity in the cell fraction was accompanied by a corresponding increase in enzyme activity in the media fraction. Lactate dehydrogenase activity per dye-excluding hepatocyte was calculated to be 9.2 +/- 1.5 X 10(-6) IU assayed at 37 degrees C for 25 preparations of isolated hepatocytes. The results suggest that leakage of cytoplasmic enzyme and vital dye staining are of comparable sensitivity in evaluating hepatocyte preparations. Measurement of LDH leakage offers a less subjective alternative to cell counting procedures and is applicable to both attached and suspended cells.

Benzylacyclouridine enhances 5-fluorouracil cytotoxicity against human prostate cancer cell lines.

At a nontoxic growth inhibitory concentration benzyloxyacyclouridine (BAU), a potent and specific inhibitor of uridine phosphorylase (UrdPase), enhanced 5-fluorouracil (5-FU) cytotoxic activity against human prostate cancer PC-3 and DU-145 cell lines. The BAU/5-FU combination exhibited greater antitumor activity in vivo using PC-3 human xenografts compared to 5-FU alone, with no associated increase in animal host toxicity. The mechanism(s) responsible for the enhanced in vitro and in vivo activity of this combination may involve enhanced formation of the 5-FU nucleotide metabolites FdUMP, FdUTP, and FUTP resulting in enhanced inhibition of thymidylate synthase (TS) and increased incorporation of fluoropyrimidine metabolites into tumoral RNA and DNA.

Inhibition of growth factor mitogenicity and growth of tumor cell xenografts by a sulfonated distamycin A derivative.

Interference with growth factor-receptor interactions may have particular relevance in efforts to intervene clinically in both autocrine and paracrine aspects of malignancy. Suramin is a synthetic anticancer agent that works, in part, by blocking the binding of growth factors to their receptors. While initial clinical trials have been encouraging, its use in clinical applications is associated with significant toxicities. Suradista is a novel sulfonated distamycin derivative that is also effective at complexing and inactivating growth factors and cytokines while remaining relatively nontoxic. The goal of this study was to compare the antineoplastic properties of suramin and Suradista. To achieve this, the effects of these compounds on growth factor induced mitogenesis in normal mouse fibroblasts and human umbilical vein endothelial cells were examined, as well as their ability to inhibit the growth of NIH/3T3 cells that had been transformed by the introduction of a fibroblast growth factor (FGF) 1 coding region (residues 1-154) fused to the signal peptide of the hst/KS3 gene (sp-hst/KS3:FGF1-154). In each case, Suradista was more effective than suramin in inhibiting mitogenesis in normal cells, as well as the growth of the transformed cells. Furthermore, Suradista was also shown to be as effective as suramin at inhibiting the growth of sp-hst/KS3:FGF1-154-transformed NIH/3T3 xenografts grown in athymic nude mice when given at only 50% the dosage used for suramin (50 mg/kg for Suradista versus 100 mg/kg for suramin). In summary, these results indicate that novel compounds acting like suramin may be developed as effective antineoplastic agents and may also prove to be of clinical benefit.

Predictive sensitivity of human cancer cells in vivo using semipermeable polysulfone fibers.

An in vivo experimental model was developed to predict efficiently and accurately chemosensitivity of human tumors. Human cancer cells either from cultured cell lines or from patients' tumors were injected directly into semipermeable polysulfone fibers subsequently implanted into immunocompetent rats. Results suggest utility of this novel model system for predicting tumor sensitivity to a wide range of anticancer agents and for potentially guiding the treatment of cancer patients in the clinical setting.