The Study Of Curing Burn By Transplanting Fibrin Membrane With Auto Epidermal Cells On Raw Surface

Objective 1.Auto-epidermal cells transplanting is a very good therapeutic method in curing for serious patients who burned in a large area.But in the progress of transplanting,the epidermal cells suspension cannot be limited in raw surface of cutaneous deficiency because of its fluidity,so the maximal curing effect cannot be played.In order to solve this problem,we want to use fibrin membrane as the matrix of epidermal cells.2.To find out the best density of inoculating epidermal cell on fibrin membrane,we inoculate epidermal cells on fibrin membrane and observe their growth.So we put the epidermal cells and the fibrin membrane together,and provide a foundation for curing burned patients with cutaneous deficiency.3.In order to study the feasibility of using the fibrin membrane of epidermal cells cure the burn raw surface in large area.In this study,with the help of our previous empirical study,we want to put the auto-epidermal cells and fibrin membrane together,and to build the fibrin membrane with epidermal cells,then we put this special membrane on the rat's burn raw surface a large area.Methods 1.The thrombin and blood plasm are needed in the preparation of fibrin membrane.We used the injected water to dilute thrombin to 1 mh 10 IU,1 ml.50 IU and 1 ml:100 IU.Then we made three kinds of the fibrin membrane using the thrombin and blood plasm in equal-volume,and to measure its thickness by the micrometer.We can get the average value.About measure the physics performance of fibrin membrane,we used the same thickness sample with 1.5 cm width,2.0 cm length and each kind do five samples to measure the index.To calculate by the equation S=P/(A×B) and L=(L₂-L₁)/L₁.We test the fibrin membrane's biocompatibility and degradation use the methods of direct observation and histology. 2.We use trypsin to digest the epidermis,and finish the isolation of rat's epidermal cells.We count cells by the slice of cell counting,detect vigour of cell by trypan blue dyeing.We identify the cells by the way of immunehistochemistry.We observe the epidermal cells morphologic change and their growth status by light microscope. Through the comparison about the rate of epidermal cells adherence,we find out the suitable density of inoculating epidermal cells.Also,we put the primary culture epidermal cells into the six shadow mask that having the fibrin membrane,then put the six shadow mask into incubator where under condition of thirty-seven degrees centi-degree,five percent carbon dioxide,and observe the morphologic change of the epidermal cells.3.We respectively found the cutaneous deficiency model in the two group rats.Group A as control group,we just routinely did bandaging after the cutaneous deficiency was done.In group B,we transplanted the fibrin membrane of epidermal cells to the raw surface.After the operation,we observed the two group rat's mental status and the condition of the cutaneous deficiency raw surface.We also record the time when the rat's raw surface is closed,and calculated the shrink rate of the rat's raw surface.At the last,we compared occurrence of scar in the two groups, and to evaluate therapeutic effects of curing by transplanting the fibrin membrane of epidermal cells.Results 1.We get the best fibrin membrane by 1 mh 50 IU thrombin,that has good physical property.Also the fibrin membrane's biocompatibility and degradation are very nice.2.The epidermis is digested by the trypsin which concentration is zero point two five percent is good.The quantity and vigour of the epidermal cells we get is very well.The epidermal cells are inoculated into culture flask.In the beginning, the cells are round,single or agglomerate floating in the nutrient medium;after four to six hours,the epidermal cells begin to sink to the bottom of culture flask;and twenty-four hours later,the cells begin to adherence bottom.At that time,we can see the shape of the epidermal cells.The cells are polygon,cell nucleus are round,and the appearance of cells are transparent,the growth appearance as the stone for road proving.The dying of epidermal cells about antikeratin antibody is positive,but anti-vimentin antibody is negative,that proving the cells we isolated are right. Through comparing the rate of the adherence about epidermal cells,we get the answer:the epidermal cells which on fibrin membrane grow better,and the proper density is 5×10⁴/cm².When we put the epidermal cells on fibrin membrane,at the beginning,the cells are round,single or agglomerate floating in the nutrient medium. And twenty-four hours later,the cells begin to adherence onto fibrin membrane. Forty-eight hours later,epidermal cells changes to big,and spread out.After seventy-two hours,some cells are to died,we can see the round cells floating in the nutrient medium.3.The two groups rat's mental status is well after the operation.The raw surface of the rats in group B healed very well;there were no to form a scab.In the process of wound healing of the rats in group A,we can see that a scab was formed in the next day after the operation,and in the end there is a scar there.There is one rat in each group found wound be infected.We managed them actively and made them better.About the time of wound healing,the rat in group B was shorter than group A.After calculating the shrink rate of the rat's raw surface,the rate of group A is larger than the rate of group B.We can see the way of wound healing in group A was cicatricle,but in the group B,there is no scar can be seen.Conclusion 1.The preparation of fibrin membrane needs to split fibrin by thrombin with an appropriate concentration.There is good biocompatibility and degradation about the fibrin membrane.2.We find out that we can obtain enough epidermal cells which quantity and vigor is good by trypsin digestion.The epidermal cells can grow normally on the fibrin membrane.We put them together to get the fibrin membrane with epidermal cells.3.The proper density of epidermal cells on fibrin membrane is 5×10⁴/cm².4.Using transplanting the fibrin membrane with auto-epidermal cells to the burn cutaneous deficiency in a large area,that can prevent scarring,lessen wound contraction,accelerate the speed of cutaneous deficiency healing.From now on,that transplanting the fibrin membrane with auto-epidermal cells can be a therapeutic method for the burn patients in a large area.