| Objective: The model of the skin defect wounds on New Zealand white rabbits wasestablished in this study. Observing the healing time, wound healing rate and thequantity of wounds capillary vessel, and detect the expression of CD34, VEGF andbFGF in positive cells on granulation tissue of two kinds of concentrations ofautologous serum were used on the surface of wound. The study was investigated theeffects of autologous serum on the New Zealand white rabbits skin wound healing.Methods: Twenty-four New Zealand white rabbits model (male or female,weighing2.5to3.0kg) were made, and three wounds on the back skin were preparedby using40%(A) or60%(B) of autologous serum and0.9%Sodium chloride(C) tocover wound skin, debridement and dressing were done every other day. The efficacyof the treatment of the wound healing rate were respectively evaluated on4,8and12days after the surgering on the total of36wounds on the back of12New Zealandrabbits. For the other12models, the tissue samples on different locations of a marginof skin and granulation tissue were excisd and the sections were embedded withparaffin, and then detected by the routine HE stainning and immunohistochemicalafter4,8and12days after the surgering, respectively. The expression of CD34weretested by immunohistochemical method in order to observe capillary density andbiopsy with optical microscope.5view micro-vascular quantity at high magnificationwere counted and discrepancy of capillary quantity in group slices were contrasted.The expression of VEGF and BFGF in positive cells were detected byimmunohistochemical method and observated by using optical microscope. Fivediscontinuous Vision were randomly selected for each slice in the granulation tissueand skin layer by Axio Vision microscopic image acquisition system image and halfquantitative analysis were made through the medical image analysis system for theimmunohistochemical results. Results:1. The general observation for the wound:4days after the surgery, there is no swelling and exudates for the three groups ofwounds A, B and C. And granulation tissue hyperplasia is not obvious.8days afterthe surgery, groups of wounds A and B are moist but without infection. Granulationtissue was significantly increased and there is newborn epithelial on the edge of thewound. There is less granulation tissue on Group C wound. Some wounds are stillswelling and oozing.The granulation tissues were basically flat at the edge of woundand the entire wound were basically covered by epithelium on12days after surgeryfor Group B. Only a small residual wound is not healing.For Group A wound, thereare still exudates on part of the wound while granulation tissues were full and smooth.For Group C, the contraction of wounds were more obvious than before and woundcavity were shallow. Part of the wounds were still leaking but no obvious swellingand Bud-like tissues were obviously hyperplasia.2. Comparison on the wound healing time:The experimental results showed that the shortest wound healing time was about12.5days and the longest was about19.5days.For the wound of Group A, the healingtime is around13.67士0.77days.For the wound of group B, the healing time wasaround13.38士0.74days. For group C, the healing time was about17.45士1.03days.There was no statistical significance between group A and B and the differences ingroups A, B and group C are statistically significant (P <0.05)3. Comparison on the wound healing rate:After four days, the A, B, C healing rate was no significant difference (P>0.05).After8days and12days, For groups A and B, wound healing rate were higherthan that of the control group (P <0.05). And the wound healing rate of Group A wasslightly higher than that of Group B, but no significant difference (P>0.05)4. Observation on tissue sections by HE staining:4days after surgery, only a small amount of inflammatory cells were infiltratingand capillary and epithelial hyperplasia was not obvious for Group C. For Group A, there were not only more inflammatory cell infiltrating,but also epidermal cells,fibroblast proliferation and new capillary forming. For Group B, the content ofwound granulation tissue in each kind of inflammatory cells, epidermal cells,fibroblast proliferation and new capillary became richer. There was massive bloodcapillary forming and fibroblasts hyperplasia on8days after surgery for Group A.Collagen began to form. There was some epithelial hyperplasia. For Group B, thewound edge had more capillaries, fibroblasts and collagen and the epidermal cellproliferation was more obvious but than the other two groups. For Group C,therewere only a few fibroblasts, endothelial cells and epithelial cells hyperplasia.12daysafter surgery, The new skin cells had already covered the wound and been obviouslyhorny for A, B groups. In the wound tissue, the majority of fibroblasts were turnedinto fiber cells and a large number of collagen has been formed. For group A, theproliferations of wound fibroblast were still active and fiber cells were few, whereasCollagen was significantly increased.5.Comparison of capillary density:Through testing the positive indicators of the immunohistochemical CD34,thefollowing results were obtained by statistical methods.4days after the surgery, therewere no significant difference in the capillary density for the wound of group A, Band C (P>0.05).8days and12days after the surgery, the capillary number of GroupA, B were higher than that of the group C(P <0.05). There was no statisticallysignificant effect for the number of capillaries between Group A and group B,(P>0.05)6. Testing bFGF, VEGF by immunohistochemical methods:4days after the surgery, contents of bFGF and VEGF in each group weregradually increasing but there was no significant difference (P>0.05).8days after thesurgery, bFGF and the number of VEGF-positive cells in groups A and B weresignificantly higher than that of group C and there were significantly differences (P<0.05or0.01).12days after the surgery, compared with group C,bFGF and thenumber of VEGF-positive cells in groups A and B were significantly higher andthere were significant differences (P <0.05).Between Group A and group B, there was no significant difference (P>0.05).Conclusion:The40%and60%autologous serum can promote the proliferation of vascularendothelial cells, fibroblasts and epidermal stem cells on New Zealand white rabbitswound skin. At the same time, it can accelerate the formation of granulation tissue,wound epithelialization and wound healing. There was no significant differencebetween the effects of two concentrations of autologous serum. |
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