| Objective:Collect and centrifuge the rat venous blood to extract IPRF(injectable concentrated platelet fibrin).By comparing the difference in growth factor concentrations between IPRF and whole blood,determine whether centrifugation has successfully activated platelets;use IPRF for treatment Deep ? degree burn wounds in rats.Observe that during and after healing of rat burn wounds,IPRF has anti-inflammatory and anti-infective properties in local wound tissues,promotes tissue repair and healing,shortens healing time,improves healing quality,and reduces healing.The role of scars and other aspects to clarify the mechanism and principle of IPRF to promote tissue repair.Method:1.Select 50 female SD rats,Weighing 250g-300g,and divided into three groups:15 in experimental group A,15 in control group B,20 in blood supply group,Given intraperitoneal anesthesia in AB group rats,right 3cm*3cm area of hair was shaved on the back of the side,and a round iron block was used to burn the external flame of the alcohol lamp for 15 seconds to immediately contact the exposed skin on the back of the rat to create a 1.5cm diameter,Deep second degree burn wound.Wound iodine disinfection.2.Group C:4 rats were taken.After abdominal anesthesia,Blood was collected through the abdominal veins.About 8ml of each blood was collected using a pp centrifuge tube.6ml of the blood was centrifuged within 60 seconds after blood collection(700 rpm)./3min),the remaining 2ml is used for platelet count and VEGF concentration detection.After centrifugation,the blood in the blood collection tube is divided into two layers.The upper layer is about 2/5 of a light yellow layer and the lower layer is a red layer.The upper layer of liquid is extracted,which is the target product I.-PRF,a total of about 8ml IPRF can be prepared from the blood of 4 rats.3.ELISA to detect IPRF and VEGF content in whole blood,and manually count IPRF and platelet content in whole blood under a microscope.4.The burn wounds of rats were injected subcutaneously.One point in the center,the depth of the needle is about 2-3mm,and 0.1ml is injected at each point.5.Use saline to subcutaneously inject the burn wounds of group B rats.The injection method is the same as that of group A.6.Each rat after modeling Three rats were randomly selected in group C for 5 days.After abdominal blood collection,IPRF was prepared and injected into the wounds of group A rats.Group B was injected with normal saline.The injection method was the same as before.Rats were given every 5 days after modeling.The wounds of the two groups of AB rats were photographed to record the changes in the wound area.Three rats were randomly selected within the two groups of AB and sacrificed.Skin within 5 mm of the wound week was excised and examined in the laboratory.The rats were sacrificed after 20 days.The remaining 5 rats were used to calculate the average healing time and wound healing rate.7.The remaining rats in the two groups were sacrificed after the wound healing rate was greater than 90%.The skin tissue at the healing site was excised for laboratory testing 8.Tissues were removed during the experiment,and each body tissue was divided into two parts.One tissue was ground and homogenized,and the supernatant was used to detect IL-6,IL-8,and VEGF.Some tissues were embedded in paraffin.Sections,thickness 4-6 ?m,VEGF content detected by immunohistochemical staining And distribution,HE staining was used to detect the infiltration degree of tissue inflammatory cells,Masson staining was used to detect the collagen content and distribution,and the quantitative detection method was to use IMAGE J to calculate the optical density value after collecting microscopic images.Result:1.Platelet count results showed that the platelet count value in IPRF was more than 6 times higher than that of whole blood,and the results of ELISA showed that the measured VEGF concentration in blood was lower than the detection range of the reagent,which was defined as not detected;The VEGF concentration measured in IPRF is higher than the detection range of the reagent,which confirms that the preparation process completes the purpose of platelet concentration and successfully activates some platelets to produce VEGF.The difference is statistically significant(P<0.01).The results of two experiments confirm IPRF was successfully prepared.2.The wound tissues of the experimental group and the control group were homogenized.The supernatant was centrifuged and the VEGF content was measured by ELISA.The results showed that the VEGF content of the experimental group was from the second day of modeling.Compared with the control group,the VEGF concentration in the two groups increased during the experiment,the difference was statistically significant(P<0.01);immunohistochemical staining showed that the VEGF concentration in the experimental group was distributed in the vascular endothelium of the wound tissue,and the blood in the control group was rich Set,IMAGE j analysis of optical density values,the difference between the two groups of AB was statistically significant(P<0.01)3.The experimental and control groups were homogenized,and the supernatant was centrifuged to perform ELISA enzymes.The levels of IL4 and IL8 in the experimental group were increased by the immunoassay method.The results showed that the growth rate of the IL4 and IL8 concentration in the experimental group was lower than that in the group B,while the increase in the group AB was increased with time,and the growth rate of the group B was higher than that of the group A.The difference between them was statistically significant(P<0.01).4.Tissue sections were stained with HE.The results showed that the degree of infiltration of nucleated cells in group A was lower than that in group B.The difference between the two groups was statistically significant(P<0.01)5.Massion staining of tissue sections.IMAGE-j calculation results showed that the collagen content of the experimental group was higher than that of the control group.The collagen distribution in the experimental group was uniform and the structure was clear.The collagen content in the control group was less and the structure was disordered.The difference in protein content was statistically significant(P<0.01)6.Wound healing rate:The results showed that group A was higher than group B,and the results were statistically significant(both P<0.01).There was no significant difference in healing time between the two groups.Conclusion:1.IPRF can increase the VEGF content in burn wounds and promote vascular growth in tissues.2.IPRF can reduce the degree of inlflammatory cell infiltration in burn wounds in rats.3.IPRF reduces the inflammatory response in wound tissues and creates a more favorable healing environment.4.IPRF promotes the recovery of normal tissue structure.5.IPRF reduces burn collagen production and reduces scars after wound healing.6.IPRF improves burn wound healing rate in rats,shortens healing time,increases healing quality,and reduces healing.After scar formation. |
PDF Full Text Request