| Objective Perioperative blood oozing and postoperative wound blood oozing arecommon complications in the surgical procedure, so reducing intraoperative andpostoperative blood loss has being taken seriously attention in clinical practice. Drugand physical hemostasis are commonly used in clinic operation to stop bleeding,however, currently there are few studies focus on these two hemostasis methods oflong-term observation of tissue repair.This study aimed to compare the effects ofhemocoagulase atrox(HA) and electric cauterization(EC) hemostasis on blood oozingby using the carotid arteries adventitia removal rabbit model and explore the effects ofHA and EC on vascular modeling in rabbit carotid artery, so we can guide the clinicalapplication.Methods A total of New Zealand White Rabbits (NZW rabbits,3.0~3.5kg;20weeks ofage, half male and female, n=27) were randomly divided into3groups by the feedingtime:0d group(n=9)、14d group(n=9)、28d group(n=9). All of them were removed theadventitia to make the blood oozing model,9in each group were randomly divided into3subgroups, in which animals were respectively treated with control group(CON group,n=3), hemocoagulase atrox (HA group, n=3), and electric cauterization(EC group, n=3).After blunting dissection the adventitia of the left and right common carotid arteries andcausing capillary hemorrhage, CON group will use0.9%physiological normalsaline(NS), HA group will receive2U/ml HA for topical hemostatic, EC group willreceive EC (power=40w) for topical hemostatic. Groups of animals were euthanized by 0,14and28days after surgery which performed with an aseptic technique. Thespecimens were divided into3parts:①0.5cm samples embedded in paraffin wasobserved for arterial repair and remodeling of pathological changes by HE staining andMASSON tricolor collagen staining, and the expression of MCP-1, TGF-β1, VEGFprotein by immunohistochemical staining;②1cm samples was observed fordifferentiation of arterial vascular cell in different subgroups by transmission electronmicroscopy analysis;③1cm samples was observed for the expression of MCP-1, TGF-β1, VEGF mRNAby RT-PCR.Results1、All rabbits were successfully caused the bleeding model, and the extent ofarterial bleeding in each group were roughly equal. There were no significantdifferences (P>0.05) in general and surgical conditions of all groups’ rabbits.2、Hemostatic effects on vascular remodeling by NS、HA and EC after dissection theadventitiaThe adventitia was completely removed and the intima, membrane weremorphologically intact by HE and Masson staining. The structure of the smooth musclecell and the collagen fiber was loosely in membrane, and the endothelial cells wererarely observed and irregular in EC group. In14d groups, the adaptation process ofvascular resulted in activitely proliferation of adventitial cells, thickening of the mediaand neoadventitia, and the density of CFD increased significantly. In the HA group, theincrease in intimal, membrane and adventitial thickness was significantly reducedcompared to the EC group（P<0.05）, and the rises of RI and LS index in HA group wasrises compared to the EC group（P<0.05）. The HA grpoup’s measurements after28days showed better than the EC group（P<0.05）. And there were no statisticallydifference in the HA group compared with the CON group by14to28d.3、Hemostatic effects on proliferation and differentiation of arterial vascular cell in NS、HA and EC groups In0d group, the endothelial cell integrity and smooth muscle cells morphologicallynormal were observed both in HA group and CON group, and the endothelial cellsshowing fusiform, cytoplasmic vacuoles, the smooth muscle cells in medial is hydropicdegeneration in EC treated animals; In14d group, fibroblasts(F) cells were observed inthe neoadventitia and migrate to the intima, which showed the characteristics with alarge number of the filaments and rough endoplasmic reticulum; In CON and HAgroup by28d postoperative, myofibroblasts differentiate into fibroblasts inneoadventitia, However, myofibroblasts can also be observed in neoadventitia in ECtreated animals.4、Hemostatic effects of the expression on MCP-1, TGF-β1, VEGF in NS、HA and ECgroupsThere were no statistically difference in the TGF-β1, α-SMA and VEGF positive cellsand mRNA between CON and HA group after0、14and28days postoperative. and28days later the positive cells and mRNA were return to normal level in HA treatedanimals and control animals （P>0.05）; Furthermore, the expression of TGF-β1,α-SMA and VEGF positive cells and mRNA was detected at a elevated level in ECtreated animals after14d and28d compared to HA group（P<0.05）.Conclusion In the carotid arteries adventitia removal rabbit model, usinghemocoagulase atrox can effectively reduce vascular inflammation injury, and decreasevascular stenotic ratio and remodeling index, as the same results as the normal saline.Hmocoagulase atrox as a local hemostatic is safety, efficiently and closer to naturalconditions. |
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