| Objective:Observe the microvessel volume density,platelet-endothelial cell adhesion molecule(CD31)expression and tissue vascular endothelial growth factor(VEGF)expression in New Zealand white rabbits carrying perforated vascular network arterialized venous flaps through histopathology.Effect of perforating vascular network on blood flow perfusion of arterialized venous flap.Methods:Twenty New Zealand white rabbits of 6 months old were selected,and the saphenous veins in the hind limbs formed arterialized vein flaps as the main trunk vessels.Using the principle of random allocation,the arterialized venous flap designed to carry no perforating vascular network on the medial leg of one hind limb was selected as the control group(Group A),and the arterialized venous flap designed to carry the perforated vascular network on the medial knee of the other hind limb was Experimental group(Group B).Tissue specimens were collected from25 points(four corners and the center)of 5 areas of the flaps of groups A and B on the 3rd and 7th days after surgery,and immediately immersed and fixed in formalin(10%).The tissue samples were HE stained and Immunohistochemical technique was used to observe and measure the microvessel volume density,CD31 and tissue VEGF expression levels of tissue samples under a microscope.SPSS22.0 software was used to test the obtained blood vessel density,CD31 and VEGF expression data by self-paired t test,P<0.05 was considered statistically significant.Result:1.Expression level of CD31 in flap pathological tissue(MVD):In this experiment,immunohistochemistry was used to measure the MVD value of 25 points in 5 areas of the tissue samples of A and B groups under200 times microscope,and the average value was taken.The results showed that on the third day after surgery,the average flap MVD value of group A was 10.94±1.67,the average flap MVD value of group B was10.28±0.77,and the flap MVD value of group A was higher than that of group B,The self-paired t test statistic P>0.05,it is still not possible to think that the MVD value measured by the CD31 marker on the third day after the flaps of the two groups is different;on the seventh day after the operation,the average MVD value of the flap in group A is 20.70±3.18The average MVD value of flaps in group B was 15.61±1.02,and the MVD value of flaps in group A was higher than that in group B.Self-paired t test statistic P<0.01,it was considered that CD31 marks on the7th day after flaps in both groups The difference in measured MVD values was statistically significant;the MVD of the flaps in both groups A and B showed an upward trend on the 3rd to 7th days after operation.The MVD value of the flap in group A increased by 14.1±1.65,and the flap MVD in group B The degree of value increase was 9.76±1.65.The MVD value of the flap in group A was higher than that in group B from 3 to 7days after surgery.The self-paired t test statistic P<0.01.The difference of the MVD rise value measured by the marker is statistically significant.2.VEGF expression in flap tissue:Immunohistochemical technique was used to detect the expression levels of VEGF at 25 points in 5 areas of flaps in groups A and B,and the average value was obtained.The results showed that the average VEGF IOD value of flaps in group A was12778.03±3097.71 on day 3 after operation,and the average VEGF IOD value of flaps in group B was 7862.53±2132.04.The VEGF IOD value was higher than the flap in group B.Self-paired t-test statistic P<0.01,it was considered that the difference in VEGF IOD value between the two groups of flaps on the third day after surgery was statistically significant;on the 7th day after surgery The average VEGF IOD value of the flap in group A was 5624.88±2791.58,and the average VEGF IOD value of the flap in group B was 2549.10±1346.77.The VEGF IOD value of the flap in group A was higher than that in group B on the 7th postoperative day.Self-paired t test statistic P<0.05,the difference of VEGF IOD value of the two groups of flaps on the 7th day after surgery was statistically significant;the VEGF IOD values of the two groups of flaps on the 3rd to7th days after surgery There was a downward trend.The average value of VEGF IOD of flaps in group A was 7153.15±4994.48,and the average value of VEGF IOD of flaps in group B was 5313.42±2441.51.The SPSS22.0 software self-paired t test statistics P>0.05 However,the difference in VEGF IOD value on the 7th day after flap operation between the two groups could not be considered statistically significant.3.HE staining results of flap tissue:A and B flaps were taken on the 3rd and 7th day after surgery to take tissue samples from 5 areas of the flap for HE staining pathological morphology analysis,and count and measure the 5 areas of the flap by Leica Q 500MC microscopic image analysis system.The volume density of the microvessels in the 25-point tissue specimen was taken as the average.The results showed that the average microvessel volume density of flaps in group A was 0.1056±0.0189mm~3/mm~3 on day 3 after operation,and the average microvessel volume density of flaps in group B was 0.2134±0.0407mm~3/mm~3.The microvessel volume density of the flap in group A was lower than that in group B,and the self-paired t test statistic P<0.01.The difference in microvessel volume density on the 3rd day after the flaps of the two groups was statistically significant.On the 7th postoperative day,the mean microvessel volume density of flaps in group A was 0.4104±0.0776mm~3/mm~3,and the mean mean microvessel volume density of flaps in group B was 0.4053±0.0248mm~3/mm~3.The microvessel volume density of the flap was slightly higher than that of the flaps in group B.The self-paired t test statistic P>0.05.There was no statistically significant difference in the microvessel volume density between the two groups of flaps on the 7th day after surgery.From 3 to 7 days after operation,the microvessel volume density of the flaps in both groups showed an upward trend.The mean value of the microvessel volume density of the flaps in group A was0.3048±0.0634mm~3/mm~3,and the flaps in group B was 0.1919±0.0297mm~3/mm~3,the degree of increase A group>B group,self-paired t test statistic P<0.01,the difference in the degree of increase in microvessel volume density between the three groups and 7 days after flap surgery was statistically significant.Conclusion:1.Observing histopathologically,the volume density of microvessels with perforated vascular network is higher than that of arterialized venous flaps without perforated vascular network,indicating that carrying perforated vascular network flaps can improve the blood flow perfusion volume of the flap;2.The expression levels of CD31 and VEGF in arterialized venous skin flaps with perforated vascular network measured by immunohistochemical technique were low,suggesting that arterialized venous skin flaps with perforated vascular network can reduce ischemia and hypoxia of arterialized venous skin flaps. |
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