Different Concentrations Of Nimesulide On The Intervention Of Wound Healing

Wound healing is one of the most common encountered situation in clinical. And scar conformation, prevention and treatment of wound healing has been a problem. The mechanism, development and prognosis of the scar conformation is a very complex process, affected by many cytokines. Among them the key factor COX (Cyclooxygenase, COX) has two isozymes including cyclooxygenase 2 (COX-2). It is a key enzyme in the biosynthesis of prostaglandin (Prostaglandins, PGs), which play an important role in the metabolism of membrane phospholipid, and it is an important rate-limiting enzyme in prostaglandin synthesis. Previous study found that COX-2 play an important role in angiogenesis and migration of fibroblasts cell, suggesting that the formation of pathological scar may be interrelating with the expression of COX-2. But there is no related research on the relationship of COX-2 inhibitors and pathological scar conformation. We study COX-2 signaling pathway through vitro and vivo experiments and try to clarify the impact of COX-2 pathway to inflammatory cells in the skin wound, angiogenesis and evaluate the wound healing, in order to reduce scar formation, promote healing and provide experimental basis for diagnosis and treatment methods.Methods1. Vitro experiment: Carry on cell culture with human fibroblasts from normal connective tissue. Observe the migration of fibroblasts using optical microscopy, phase contrast microscopy.2. Qualitative analyze the relationship between the concentration of COX-2 inhibitors and the ability of fibroblast migration, using scratch test in vitro fibroblast cell culture.3. Form surgical wound model using strain Kunming mouse, and intervene the skin wound healing process using COX-2 selective inhibitor nimesulide with the control group and 3 experimental groups with different concentrations of nimesulide. 4. Observe and evaluate the wound healing process by observing angiogenesis, scar tissue of the skin, the skin stretched, immunohistochemical methods, inflammatory cells, the growth of fibroblasts et cetera. And using digital software analyze the healing process and scars. Using statistical methods analyze the results.Results1.Cell culture experiments showed that COX-2 inhibitor had a significant effect on inhibiting the migration of fibroblasts, and the inhibit level were positively correlated to the concentration of COX-2 inhibitor, the experimental group using 25μmol / L nimesulide has better results of cell growing comparing to other groups: cells can evenly migrate into the scratches gap and did not appear order chaos.2. Vivo experiments show: Through the intervention of COX-2 signaling pathways using nimesulide, the skin wound healing process has been significant impacted. Using appropriate concentration of nimesulide can inhibit wound inflammation and control the migration of fibroblasts in the right level. Comparing the scar area of experimental group and the control group, we found statistical significance between the experimental groups and the control group on the incidence of hypertrophic scars, pigmentation, scar size and other aspects. The 25μmol / L nimesulide group received the best healing results.3. Using Immunohistochemistry method to analyze the healing wound showed: In vitro applications, Nimesulide could significantly inhibit the expression of COX-2 in inflammatory cell, and the inhibition is positive correlation to the drug concentration.4. Statistical analysis indicated that there was statistically difference between the experimental group and control group, and statistical differences were significant between most of the experimental groups.Conclusion1. Using Nimesulide in vitro applications on mouse surgical wound can intervene the expression level of COX-2 regulation of the inflammatory cells, COX-2 expression level has a negative correlation with the drug concentration.2. Conducting COX-2 inhibitor Nimesulide in mouse surgical wound can intervene the growth of new blood vessels, level the migration of fibroblasts, control the activity of inflammatory cells. Application of appropriate Nimesulide (COX-2 selective inhibitor) has a positive result on surgical wound in mice, proper concentration of COX-2 selective inhibitor Nimesulide can reduce scar formation, it is worth further study in trauma, Cosmetic Surgery, Burn Surgery and other applications.