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Gene transfer of naked plasmid encoding for rhVEGF in ischemic TRAM flap

Author: YeXiuZuo
Tutor: YuYuSheng
School: Zhejiang University
Course: Surgery (General Surgery)
Keywords: Recombinant gene Vascular endothelial growth factor Rats Rampant TRAM flap Microvessel density Fluorescein Survival area Immunohistochemistry Enzyme - linked immunosorbent assay
CLC: R622.1
Type: Master's thesis
Year: 2001
Downloads: 47
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Abstract

Research background and purpose of rampant TRAM flap (TRAM flap) is widely used in soft tissue repair and recycling, in particular, play an important role in breast reconstruction. Ischemic flap partial necrosis caused by the high incidence of problems to be solved. Many scholars through a variety of surgical methods, physical methods and drugs improve the blood supply of the TRAM flap to reduce necrosis occurred. Therapeutic angiogenesis in ischemic diseases research more VEGF pro-angiogenic effect is most concerned about the project. Gene therapy is a very promising treatment, has proven in animal myocardial ischemia and lower limb ischemia model in VEGF gene can promote angiogenesis, improve hemodynamics, increase the blood supply of the ischemic area. In the the flap transplantation experiments also proved that the human VEGF gene was injected into the blood vessels supplying the flap, the results of the flap survival area increases. But in general, gene therapy in ischemic skin flap is still small. The purpose of this study: (a) the establishment of stable ischemic TRAM flap model. (B) study of recombinant human VEGF gene direct subcutaneous injection of the TRAM flap survival and subcutaneous microvascular density, to provide experimental evidence for the gene therapy of ischemic skin flap. (C) study of recombinant human VEGF gene directly injected subcutaneously, VEGF expression levels flap survived and subcutaneous microvascular density. Research methods in 2001, Zhejiang University master's degree thesis, plasmid extraction: The purpose of transfected plasmid bacterial amplification, the the soda process a large number of extraction. TRAM flap model: the unilateral inferior epigastric artery pedicled TRAM flap bit rampant. 3, the mode of administration: 32 animals were randomly divided into four groups, each of eight four points, drug injection in flap District rectus pedicle on both sides. The experimental group was five days before the administration of each 500tyPCDNAVEGF; positive control group was administered per each lml saline administered 5 days before surgery 100ngVEGF blank control group; negative control group preoperative administration of each 500ngCDNA. A week after sodium fluorescein test flap blood supply, the photographic entered into the computer, the computer image processing software AUt. CAD calculated survival area. 4, PcDNAVEGF prior to injection, injection Bu 3,5,7,9-day serum specimens measured using ELLISA kit VEGF levels in the blood. 5 PCDNAVEGF lower abdomen after injection five days of the injection zone skin biopsy using streptavidin - peroxidase complex (SP) immunohistochemical determination of the expression of VEGF. 6, microvessel counts: groups flap HE staining slice subcutaneous tissue per mm 'number of microvascular results, TRAM flap model survival area: 3.61 Guests 1 knife 6cm'. Flap survival area groups flap survival area: saline group 4 * Guests 1.77 Cm 'PcDNA group flap survival area 4.22 Guests 1.80Cffi' pCDNAVEGF group flap survival area. gS with disabilities 2.64 Cm 'the VEGF groups flap survival area of ??7.31 Guests 1.22 cm'. The flap survival area PcDNAVEGF group was significantly higher than the saline group (P <0.01) and PCDNA group (P <0.01), the VEGF groups flap survival area was significantly higher than the saline group (P <0.05). 3, VEGF levels in the blood: injection PCDNAVEGF ago after injection, 1,3,5,7,9 a large concentration of VEGF in the blood without significant differences. 4 slices microvessel density: saline group 44.56i7. S7 root / mm ', PcDNA group 37.49i3.34 root / mm', pCDNAVEGF group 58.71 ± 8. The ZS root / mm 'VEGF group 14.29 65.61 Guests / mm'. 12001 Zhejiang University Master's degree thesis D PCDNAVEGF group and VEGF group microvessel density was significantly higher than the saline group (P <0.05), microvessel density PCDNAVEGFl group also was significantly higher than PcDNA. 15 rabbits infected group: PcDNAVEGF injection area skin tan antigen-antibody complex deposition in the the small blood D vessel endothelial cytoplasm and leaching Pat tube cytoplasm of endothelial cells, sometimes banded around the vessel lumen, without injection Area D is not found in similar situations. The J, The direct injected subcutaneously under PCDNAVEGF can rat abdominal skin expression of biologically active IVEGF. 12, directly the subcutaneous injection PCDNAVEGF promote subcutaneous angiogenesis. 13, direct injected subcutaneously PCDNAVEGF improve ischemic TRAM flap survival area, play a positive role in the treatment of pre-D anti-ischemic skin flap.

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