Surface Protection of Exposed Biological Tissues
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example 1
[0059]A reproducible and standardized rat and rabbit model was adopted. Forty eight female MRI mice weighing about 25-30 g were used to induce the adhesions and forty two for further tests. The animals were kept under standardized conditions and had free access to pellet and tap water.
[0060]Anesthesia was induced by ketamine 150 mg / kg (Ketalar, Parke Davis) and zylazine 7.5 mg / kg (Rompun, Bayer Sverige AB) intramuscular injection. After disinfection, a 25 mm long midline laparotomy was performed. Both peritoneal surfaces of the lateral abdominal wall were exposed, and 2×15 mm long sharp incisions were performed at the same distance from the midline, including the muscles. The wounds were immediately closed with 2×4 single sutures at equal distances by using 5.0 polypropylene (Prolene, Ethicon, Johnson & Johnson). The midline laparotomy was closed in two layers with a continuous 5.0 polypropylene suture. At the evaluation time an overdose of anesthetic was adminis...
example 2
Phagocytosis and Particle Ingestion Index.
[0066]The time course of the phagocyte function was tested in vitro on peritoneal resident macrophages from mice after 0.5 h, 1 h, 2 h, 3 h, 4 h, 5 h, 6 h, 8 h, 10 h, 12 h, 16 h, and 24 h incubation with poly-L-lysine+poly-L-glutamate (40 μg / ml) and / or fluorescent particles (1 μm).
[0067]Macrophage samples were taken by abdominal lavage with 10 ml ice cold DMEM-solution. The samples in medium were immediately centrifuged at 1200 rpm for 10 min. The cells were resuspended in DMEM containing 10% FBS and penicillin / streptomycin and then plated on 48 wells cell culture plates; 5×105 cells in each well. After 1.5 h non-adherent cells were washed away, particles (100 / cell) together with test drugs (poly-L-lysine+poly-L-glutamate) were added in a dose of 40 μg / ml to 12×5 wells, and particles only were added to the remaining 12×5 wells. More-over negative controls were performed at each time point. The cells were incubated (37° C., 5% CO2) and detach...
example 3
Transmission Electron Microscopy.
[0073]Peritoneal macrophages were harvested from two healthy non-treated animals as described above and plated on cell culture plates (Thermanox, Naperville, Ill., USA). The cells were washed away after 1.5 h and poly-L-lysine+poly-L-glutamate (40 μg / ml) in supplemented DMEM solution were added in sequence followed by a 24 h incubation. The incubation medium was removed and the cells were fixed in 2.5% phosphate buffered glutaraldehyde was followed by rinsing in Milloning's phosphate solution. Samples were postfixed in 1% osmium tetroxide and subsequently dehydrated with graded series of ethanol, which was followed by embedding in Araldite 502 kit. Vertical sections were obtained with a diamond knife and stained with uranyl acetate and lead citrate in a LKB Ultrastainer. Samples were examined in a JEOL 1200 EX transmission electron microscope (TEM).
[0074]Electron microscopy verified that macrophages phagocyte the test compound particles, resulted in ...
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