The Effects Of The TSP-1 On Regeneration And Scarring In Skin Wound

Objective After the skin was wounded, he wound will heal in a period, this is a long elaborate and complicated process, the ending of the wound healing is either scar healing or regeneration, the delivering of cytokines, and lots energy and lots nutrition is needed in this process, so that we can see the new blood vessel in the surface of the wound is essential, as an angiogenesis inhibitors,TSP-1 could be very important in this process, a research found that the expression of TSP-lup-regulate in the early of the wound healing and the rat fetus’ skin, and the scar of the rabbits’ ear come to be flat and soft after inject the adenovirus that could secrete TSP-1, so we supposed that the TSP-1 could do some things to the wound healing and scarring, however the dynamic expression, the role and the function in the process was not confirmed. This thesis here is aimed to detect the expression of TSP-1 in the different period of the wound healing, and do some observation about the use of the TPS-1 and the inhibitor of the TSP-1 in the process of the wound healing.Methods 50 8weeks (2.0-2.5kg) Japanese white rabbits raised in the cages separately for one week, divide them into two groups randomly, twenty-five in one group, one group is the group TSP-1, the other one is group Anti-TSP-1, remove their hair on the ear of the abdomen side with the 5% sodium sulphide, after 3 days,we built the animal models(The area of the wound is 10.0mm×10.0mm), intravenous anesthesia with the 2%pentobarbital sodium through the ear vein, we cut one wound on the ear of the abdomen side, the wound is from the tip side of the ear to the end side of the ear with the depth is from surface of the ear to the deep to the cartilage, from the day the animal models were built, we injected TSP-10.1ml (5μg/m) and the antibody of TSP-1 0.1ml (20μg/ml) around the wound on the left ear of the rabbits in the group TSP-1 and the group Anti-TSP-1, we injected once two days until the wound re-epithelize,at the same time we inject the normal saline 0.1ml around the wound on the right ear of the rabbits in the two groups as group NS. At the 4th day,8th day,12th day,30th day after animal models were built, we observe the change of the wound healing, take photos and take them down, then we pick out five rabbits in every group randomly, and cut down the wound as the sample to do the hematoxylin eosin staining and immunohistochemical staining, and we observe the change of the expression of TSP-1 and the different expression of the Ⅰ and Ⅲ collagen in the wound between the experimental group and the control group. We use the microscope to detect the differences between deep side and the shallow side of the wound. We calculate the optical density average value in five visual field of the TSP-1 and the collagen Ⅰ and collagen Ⅲ with the Image-Pro-plus 6.0, and then we use the SSSP19.0 to analyze data, so that we can compare the differences between the experimental group and the control group.Result1. The wound 3.0mm apart from the shallow side of the wound lost the blood scab and re-epithelialized at the 8th day after wound, no scar appeared, at the 30th day after wound, the skin of the shallow side of the wound was flat, the color of the skin was similar to the normal skin; and the deep side of the wound at lost the blood scab and re-epithelialized at the 12th day after wound at the 30th day after wound the skin of the deep side of the wound was still dark red, the middle of the deep side was highlight out of the normal skin, and it was tough, that’s scarring.2. The wound of the group TSP-1 was healing and re-epithelialized more quickly than that of group Anti-TSP-1 and group NS, the skin of the group TSP-1 was much flatter, the shallow side of the wound in group TSP-1 re-epithelialized at 8th day after wound, that of group NS was the 9th day after wound, that of group Anti-TSP-1 was 12th. The skin’s color of the shallow side in group TSP-lwas more similar to the normal skin and the deep side has less scar and the scar was flatter and softer than that of group NS and group Anti-TSP-1.3. The expression of TSP-1 in the deep side of the wound of group NS began to raise at the 4th day, it reached the top at 8th day, at the 30th day, it came to the normal, the shallow side reached the top at 4th day, and it decreased to the normal at the 12th day.4. The expression of type Ⅰ collagen in the deep side of the wound is higher than that in the shallow side of the wound of every group at every moment after wound (P<0.05), and the expression of type Ⅲ collagen in the deep side of the wound is lower than that in the shallow side of the wound of every group at every moment after wound(P<0.05).5. The expression of type Ⅰ collagen in the shallow side of the wound of group TSP-1 reached the top at the 8th day after wound, and began to decrease at 12th day, it was higher than that of the group NS and group Anti-TSP-1 at every moment (P<0.05); the expression of type Ⅰ collagen in the deep side of the wound of group TSP-1 was increasing gradually, and it was always higher than that of the group NS and group Anti-TSP-1 (P<0.05); the expression of type Ⅲ collagen in the wound of group TSP-1 was higher than that of group NS and group Anti-TSP-1 at every moment (P<0.05), in the shallow side of the wound, it always increased gradually, and in the deep side of the wound, it reached the top at 8th day, and after this, it began to decrease.6. The expression of type Ⅰ collagen in deep side of the wound of group Anti-TSP-1 reached the top at 12th day after wound, and it decreased to normal at 30th day, it always increased gradually after wound in the shallow side of the wound, and the expression of it was always lower than that of group NS and group TSP-1 (P<0.05), the expression of type Ⅲ collagen in the wound of group Anti-TSP-1 was always lower than that of group NS and group TSP-1 at every moment (P<0.05), and it reached the top at 12th day after wound in the deep side of the wound, it always increased gradually after wound in the shallow side.7. The proportion of type Ⅲ/Ⅰ collagen in the group TSP-1 was higher than that of group NS and group Anti-TSP-1 (P<0.05), and the proportion of type Ⅲ/Ⅰ collagen in the shallow side of the wound was higher than that in the deep side of every group (P<0.05).Conclusions:1. we realized the rabbit ear regeneration-scaring wound model, the shallow side of the wound achieved regeneration, and the deep side of the wound formed scar.2. TSP-1 promoted the expression of the type Ⅰ and type Ⅲ collagen, and it increased the proportion of the type Ⅲ/Ⅰ collagen.3. The injection of the TSP-1 improved the healing of the shallow side of the wound, and it also inhibited the forming of the scar in the deep side of the wound.