Treatment and composition for wound healing
a technology of composition and wound healing, applied in the direction of blood coagulation/fibrinolysis factor, protease inhibitor, animal/human protein, etc., can solve the problems of inability to heal venous stasis, poor understanding of the molecular mechanisms of impaired wound healing, and failure to heal the venous stasis of most clinical trials using growth factors/cytokines
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example 1
[0078] APC Promotion of Endothelial Wound Repair
[0079] Activated protein C (APC) was tested for its ability to promote repair of endothelial wounding using a modification of an in vitro assay, as described previously (19). Briefly, confluent microvascular endothelial cells (FSE) from neonatal foreskins were cultured for 5 days in 24-well culture plates in growth medium (Biorich plus 50 .mu.g / ml heparin, 50 .mu.g / ml endothelial cell growth supplement and 5% human serum). The endothelial monolayers were wounded by a single stroke across the diameter of the well with a pipette tip. The media and dislodged cells were then aspirated, and the plates rinsed with Hanks buffer. Fresh growth medium was added to the plates along with APC at various concentrations or the potent tumour-promoting angiogenic factor, phorbol myristate acetate (PMA) (10 ng / ml) and the cells were incubated at 37 degrees C. After 24 hr, the width of the wound was visualised microscopically and results at different dos...
example 2
[0080] APC Promotion of Angiogenesis
[0081] In view of the ability of APC to activate gelatinase A and promote endothelial wounding, APC was investigated as to whether it could promote angiogenesis. APC was added to the chicken embryo chorio-allantoic membrane (CAM) assay using gelatin sponges (Gelfoam). Sponges were cut to approximately 2 mm.times.2 mm. Five .mu.g APC in phosphate buffered saline (PBS) or PBS alone was added to gelatin sponges which were subsequently placed on the 9 day old CAM, as previously described (34). The CAMs were inspected daily and on day 14 were photographed and fixed for histological sectioning. Macroscopically, on day 14, the APC-treated gelatin sponges were surrounded by blood vessels that grew radially inwards towards the sponge in a "spoke-wheel" pattern (data not shown). In contrast, gelatin sponges treated with PBS had no surrounding vascular formation. Histological sections showed that APC-treated sponges were infiltrated with many new blood vesse...
example 3
[0082] APC Promotion of Wound Healing
[0083] In view of APC's ability to stimulate endothelial migration and enhance re-epithelialisation, fibroblast invasion and angiogenesis, APC was examined for a capacity to improve wound healing in a rat model. Sprague-Dawley rats were anaesthetised and four full-thickness wounds were excised, using a 8 mm punch biopsy, on the back of the rat, exposing the underlying dorsolateral skeletal muscle fascia. Hemostasis was achieved by even compression with sterile gauze. APC was diluted in isotonic, sterile, pyrogen-free saline solution and each excision was treated with a 50 .mu.l topical application of sterile, pyrogen-free saline solution or saline containing 20 .mu.g APC. The wounds were left open with no dressing and rats caged one per cage. Wound closure was assessed visually and after 40 hr, 4 days and 7 days. At each timepoint the wounds were digitally photographed using a Nikon Coolpix 950, with a distance calibration scale in the frame. The...
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