| Objective: By observing the change regularity of the thickness of fibra-peplos, quantity of micrangium, content of collagen protein and the minuteness architecture of tge fibroblast. To illuminating the influence of the process from fibroblast to myofibroblast, the influence of the fibroblast synthesis and secrete collagen fibers after chitosan interfere in the fibra-peplos around the expander. To reveal the mechanism of action of chitosan interfere in fibra-peplos and to explore a new feasible method to solve the question about the availability of slap depress cause of the fibra-peplos after the skin~soft tissue dilation.Methods:1 experimental animal15 weight approximation experimental New-Zealand albion rabbit were weighted and ordered.2 groupingPeeling a enough lacune on the infermediate piece of the back ,3cm from the back bone of each New-Zealand albion rabbit. Then embedding a 20ml skin-tissue expander in each lacune, left hand side is always experiment group and right hand side is always control group. 3 the methods of embedding expander and affusionMake 2% chitosan 0.5ml afflux into the lacune, and make it apply uniformly, then make a 20ml skin-soft tissue expander embedd into the lacune after the lacune is decoherenced enough and is not found active bleeding in the experiment group. Make a 20ml skin-soft tissue expander embedd into the lacune with routine method. Starting affusion and expansion after the wound healing and the stitches are taken out. Make affusion without narcotism, 3~5 days make one time ,according to the specific information. The capacity of every time affusion is on the basis of the tensile force of the flap, limitation of every time affusion is that flap's press reaction time shouldn't morn then 4~5s. Make the capacity of every time affusion to the same New-Zealand albion rabbit equal. Stopping affusion when the capacity of affusion to every 20ml skin-soft tissue expander is 40ml.4 draw the materialsBy means of intraperitoneal injection 10% chloral hydrate (8ml/kg), then draw the materials of flap around the expander in the same location of the experiment group and the control group with general anaesthesia. At the same time cut 4cm×4cm flap with fibra-peplos,and take it onto a glass which is dropped liquid paraffin. Takeing a picture after it naturally retract 5 minutes, and then calculating the shrinkage with topography analyza method. Cut 0.1cm×0.3cm piece of the flap example immerse into neutral glutaral then take it to transmission electron microscope observation. Tile the rest flap example on a asepsis gauze, suture it's four angles with suture silk to fix it onto the gauze. Put the flap example with the asepsis gauze into neutral formalin, and then take it to make paraffin section to part into HE dyeing, Masson dyeing and immunity histochemistry CD34 dyeing.5 observation index(1) shrinkage: every flap with fibra-peplos is 4cm×4cm before contract. Take a picture after it naturally retract 5 minutes ,then calculating the shrinkage with topography analyza method[1].Observe the change of experiment group and control group. (2) Observe routine HE dyeing microtome section, observe the histology change of experiment group and control group with light microscope and measure the thickness of fibra-peplos example with micro-ruler.Observe Masson dyeing microtome section with microscope, comper the content of collagen fibers in experiment group and control group.By quantitative analyzing the average gradation of the dyeing example with pathology image analysis system, quantificating the content of collagen fibers in two sets. Observe CD34 dyeing microtome section with microscope, compare the quantitative change of micrangium in fibra-peplos in experiment group and control group. (3) Observe the change of cell organ and microfilament with electron microscope, contrasting the capability of fibroblast excrete collgen in experiment group and control group, contrast the degree of the fibroblast transform tomyofibroblast in two sets.6 statistical treatmentMake statistical treatment with SPASS 11.5 statistical treatment software.Consequence:1 shinkageShinkage of experiment group is 43.371±7.312%,shinkage of control group is 5.637±6.613%,comper two sets (p<0.05)2 observation with light microscope2.1 observation of HE dyeing microtome section and determination of the thickness of fibra-peplosIt is thus clear that copious fibrablast and neogenesis micrangium scatter in the campus visualis not only experiment group but also control group. The number of immature fibrablast in experiment group more than it in control group. Immature fibrablast's cell body is comparatively large, there are ecphymas in it's two side,it's endochylema is a little basophilia so it is alittle blue. But the number of maturation period fibroblast in control group more than the number in experimentgroup. It is thus clear that there are many collagen fibers around maturation period fibrablast after the fibrablast excrete procollagen,so the maturation period fibrablast is long fusiform, it's endochylema is less than immature fibrablast and it's caryon is anachromasis than immature fibrablast. Stochastically selecting 15 pieces of campus visualis of HE dyeing microtome section amplified 100 times of experiment group and control group, and then measure the thickness of fibra-peplos with mico-ruler. Result: the thickness of fibra-peplos of experiment group is 516.000±128.491μm, the thickness of fibra-peplos of control group is 833.000±227.379μm(P<0.05).2.2 observation of Masson dyeing microtome section and topography quantitative analysisBy observing the Masson dyeing microtome section, it is thus clear that there are sporatic fibroblasts which caryon is gray-black, orbicular-ovate in the gray part of the fibrablast which is occupy 4/6~5/6 fibroblast's thickness and the rest 1/6~2/6 is green ,there are fibrablasts which caryon are long fusiform, gray-black, there is a layer intra-lining alike endothelial cells in it's inner face. The fibra-peplos of control group is all green, the fibrablast of it are long fusiform and there are fusiform cell alike endothelial cell in it. By topography quantitative analysis, the meangradation of experiment group is 170.383±14.313, the meangradation of control group is 159.762±10.623 (P<0.05).2.3 Obvservation of CD34 dyeing microtome section and the quantity of micrangiumThe micrangium endothelial cells are coloured cinnamomeous in CD34 dyeing microtome section , it scatter in the campus visualis. By random selecting 15pieces campus visualis of CD34 dyeing microtome section of not only experiment group but also control group with 100 times light microscope, artificially count the quant of micrangium. Result: the quant of micrangium in each campus of experiment group is 8.200±2.150,the quant of micrangium in each campus of control group is 7.900±1.729(P>0.05).3 Observation of transmission electron microscopeIn experiment group, rough endoplasmic reticulum in the fibrablast ie dilat, it's quant obviously decrease, the phenomenon of shed particle is obviously, part or all of the crista of bioblast coalesce or disappear, common Grolgi apparatus is rare, microfilament is rare. In control group, rough endoplasmic reticulum in the fibrablast is dilat, the phenomenon of shed particle is rare, the crista of bioblast is integrity, microfilament is more, the quant of ribosome isn't obviously decrease.Conclusion:The experiment confirm that chitosan can make the thickness of fibra-peplos around expender attenuation, the content of collagen fibers depression, the contractility of fibra-peplos decrease by repressing fibrablast secret collagen, delaying fibrablast transform to fibrocyte, repressing fibrablast transform to myofibrablast. So chitosan can effectively lessen the shrinkage of the expanding flap, at the same time can't effect the role of increasing the blood of the flap. Providing dependable experiment evidences for clinic to explore a new effective method that not only remaining the trophism of fibra-peplos but also avoiding the increase of the recovery rate of the flap because of the fibro-peplos.
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