| Objective: To prepare Papaverine Hydrochloride gel and investigate its effect on survival of dorsal supernormal random flaps. Explore the local application of papaverine on the influence of the survival area of random skin flap and histological varieties ,and infirm that the effects on preventing and curing the putrescence of flap.In order to enhance the rate of the transplantation of skin flap and explore a new effective approach.Method: 1 Laboratory animals: 40 healthy Wistar rats(provide by Hebei medical university laboratory animal center,weighing 280±10g),male . 2 The group of laboratory animals: The rats were randomly divided into 4 groups: external group, the injection group, wet packing group and control group. All rats were operated abnormal random flap on back. external group: Treated with papaverine hydrochloride gel; the injection group: Injected Papaverine Hydrochloride3. 0 mg/ kg; wet packing group: Treated with Papaverine hydrochloride saturated solution; control group: Injected with normal saline, the dose reference injection group. 5 min after operation , 2 times a day(8 :00和14 :00).3 Skin flap formation: All the animals were anesthetized with 2%Sodium pentobarbital (35~40mg/kg) by intraperitoneal injection.When induce anesthesia successfully,depilate the back hair,Prone fixed,disinfected by iodine and alcohol.The line connected with the iliac crest on the center of rat's dorsal as the base line,the back of midline as the axis,made a random skin flap with a size of 7.0cm×1.0cm (including muscle lipid membrane,the length and breadth ratio was 7:1), the pedicel was on the rump.Close the incision at the original position by 3-0 Suture.4 Papaverine gel preparationFormula: Papaverine15%(W/V),B-CD7.5%(W/V), Isopropanol40 % (V/V),CMC-Na 6%(W /V),10%OA (V/V),distillated water45mL,Total 100 mL。Preparation process:To take B-CD dissolved in 50℃~ 60℃isopropyl alcohol - water (1:1) solution, and then adding papaverine, papaverine until completely dissolved, add transdermal absorption enhancers OA and CMC-Na, stirring until at room temperature that is too.5 The detection method and observation marker5.1 The general observation:Observe the survival situation of skin flap on the 7th day after operation.5.2 The observation of flap surviving rate:Flap necrosis adjust by its appearance,color and texture.The 7th day after operation,take photo of flap and enter into computer image,calculating the ratio of surviving flap(the proportion of the flap surviving area in total area)by using Image-Pro Plus.v6.0 software system.5.3 The observation through light microscope: Take 0.5cm×0.2cm skin tissue at the middle of the flap on the 7th day after operation in each group,4% formalin-fixed,Paraffin-embedded histotomy,stain of HE.5.3.1 Observe the varies of tissue structure.5.3.2 Capillary calculate.Method:All the vascular were included in.Five histotomis in every group and five high multiples visual fields were selected randomly from central and peripheral area,calculating the number of capillary and obtaining the mean value.5.3.3 Fibroblast calculate.Five histotomis in every group and five high multiples visual fields were selected randomly from central and peripheral area,calculating the number of fibroblast and obtaining the mean value.5.4 The observation through laser doppler flowmetry: The back flap in the rat central axis line, away from the pedicle 2, 4, 6 cm Department, in surgery before and after 4 h using laser Doppler flowmetry measurement of flap blood flow, take three parts of the blood flow averages were statistically analyzed.6 Analysis of results: Dealt with different kinds of results by using SPSS14.0 software. Results: 1 The general observation: The 7th day after operation, External group and the injection group:the near section of flap:Light red,good elasticity,and a little back hair grew up can be seen;middle section:The color of the center flap gradually darken.There were not scab formation on surface,no hair grew.The color was black in peripheral area and covered with a little scabs.The elasticity was diminished.Far section:The color was black,and covered with scabs,bad elasticity.There can be seen bleeding activity when lift up the flap at the original position,and the quantity is large;There were no hematocele under the flap,little effusion,and vessels were abundance.The flap color and elasticity of external group was better than the injection group. Wet packing group and control group:the near section of flap:The color was light red and gradually darken from near to far.A little hair grew at the pedicel of flap;Middle and far section:The color was black and covered with scabs,bad elasticity.There can be seen a little bleeding when lift up the flap at the original position,much inflammatory exudates under the flap,vessels relatively sparse.2 The observation of flap surviving rate:external group(83.6±11.3)> the injection group(67. 5±8.9)> wet packing group(44.3±12.5) and control group(36.1±10.5) (P<0.01)。Flap survival rate of wet packing group and the control group is no significant difference(p>0.05).3 The observation through light microscope3.1 The 7th day after operation, External group and the injection group: Cuticle atrophy was not obvious or partly damaged,cutis'structure had not obvious changing,neutrophils infiltration.There can be seen freshmen capillary in cutis, subcutaneous fibrous tissue hyperplasia,fiber array rules. Wet packing group and control group: Cuticle and cutis were completely necrosis,the structure was disordered, a mount of neutrophils infiltration, muscles fiber obviously necrotizing.3.2 Capillary calculate: The 7th day after operation, External group(7.0±0.71)and the injection group(6.0±0.85)﹥wet packing group(4.0±0.51)and control group(3.5±0.39). External group was higher than the other groups( P < 0. 01). 3.3 Fibroblast calculate: The 7th day after operation, External group(40.5±3.16)and the injection group(34.2±3.08)﹥wet packing group(15.3±1.65)and control group(10.1±3.11). External group was higher than the other groups( P < 0. 01).4 Flap blood flow monitoring: There is no obvious difference in the blood flow of all groups before operation ( P > 0. 05),,After treatment,the blood flow of the external application group was higher than other groups( P < 0. 01).Conclusions:1 Preparing of Papaverine hydrochloride gel is convenient and effeetive , Papaverine gel has a good transdermal absorption.2 Papaverine hydrochloride solution can not effectively penetrate the skin ,so it is difficult to play its pharmacological effects and promote flap survival.3 Papaverine hydrochloride gel can effeetively Permeate through skin and expand local vessels in local tissue, thus increase the blood flow and Promote survival of flaps. So Papaverine hydrochloride gel is veay useful to transplantation of flap in clinical.
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