An Experimental Study On The Treatment Of Knee Joint's Synovitis Induced By Trauma With NanHu Gelatin

Objective: Distilling effective components respectively from traditional Chinese medicine of NanSheTeng, HuZhang, MoYao; Making them into NaHu Gelatin with carbomer and reagents else according to a certain proportion. To establish the model of knee's Synovitis by trauma through stimulating rabbit's hindlimbs with acusector .To discuss the function and the mechanism of prevention and treatment of NanHu Gelatin through animal experiment with New Zealand rabbits.Methods: Distilling effective components respectively from NanSheTeng and HuZhang with the method of using heated ethanol; Distilling naphtha from MoYao with method of vapor distillation. Making them into NaHu Gelatin with carbomer and other reagents according to a certain proportion. Observing the penetrating cutis effect of effective components of NanHu Gelatin through the trial of absorbability penetrating through skins. Rabbits were bound with nips, feather was fade locally with 8% sulfreted sodium. Let them in anaesthesia with 3% sodium pentobarbital injecting from the vein besides ears (The injecting time is less than 2 minutes) . The rabbit's hindlimbs were sticked vertically at the midpoint of quadriceps femoris and gastrocnemius muscles, connecting electroacupuncture apparatus, frequency is 50 Hz, intension is 1.5. Causing their knee joints do passive flexion and extension without intermission, twice a day, seven days and the time last 40±5 minutes each time. 48 rabbits were divided into 4 groups: group A prevented with NanHu Gelatin; group B treated with NanHu Gelatin; group C covered with FuTaLin latex; group D left modeluncovered. Observing the change of rabbit s knee joint circumference, the change of joint surface's temperature and the time of claudication after being sticked. The rabbits' knee joints of hindlimbs were scanned with ECT after experiment beginning nine days to observe the inflammation of joints . Using injection syringe inject 1 ml saline into knee joint's cavity from incisure of its inside or outside , drawing out the joint fluid to test the level of SOD, MDA and IL-1. Using oprating scalpe cutdown in the front of knee joint and its whirlbone inferior boarder, cut off the anterior ligament of whirlbone, stripping the Fat pad under whirlbone, the Synovium is a coat diaphragm which is achromatic color attaching on the surface of it. The Synovium and Fat pad were fixed with 10% neutrality formaldehyde solution, desiccationed twice with purified acetone, dipped wax, embedded, and done pathological section routinely to observe the form and constitution of Synovium. Observing the expression of NF- k Bp65, NF- k Bp50 in the tissue of Synovitis by using the method of immunohistochemistry Stain to discuss the effecting mechanism of NanHu Gelatin.Results: NanHu Gelatin can be smeared equably on the skin of nude mouse, and be agglutinated tightly with skin. It's effective components can be permeated skin in a short time.The change of knee joint circumference has no statistical difference between the model of hindlimb and the normal hindlimb in group D (P>0.05) , the joint surface temperature and the worth of ECT have statistical difference between them (P<0.05) , the level of SOD, MDA and IL-1 in joint fluid has statistical difference between them (P<0.01) . There are mass infiltrated inflammation cells, Vascular proliferation and distension, hyperplasia and thicken synovial villi in the model of hindlimb from pathological section. There are mass brown particles male substance which are expressed by NF- k Bp65, NF- k Bp50 ,which are appeared in macrophage , fibroblast, and lymphocyte of the lining coat and the second coat of synovitis. But there are no male substance appeared in the synovium tissue of the normal hindlimb. Group A compare with group D, the temperature of knee surface , the worth of ECT, SOD , MDA and IL— 1 in joint fluid, all have statistical difference (P<0.05, P<0.01) .The inflammation of synovium tissue is not obvious in group Athrough pathological section, a few inflammation cells infiltrated occasionally. However there are mass infiltrated inflammation cells, Vascular proliferation and distension, hyperplasia and thicken synovial villi in the model of hindlimb from pathological section. There are a little male substance appeared in synovium tissue of group A, much appeared in that of group D by using the method of immunohistochemistry Stain. . Group B compare with group D ,the temperature of knee surface , the worth of ECT , SOD ,MDA and IL— 1 in joint fluid ,all have statistical difference (P<0.05, P<0.01). There are a few neutrophilic leukocytosis and no obvious inflammation in synovium of group B, while typical inflammation was appeared in that of group D. There are a little male substance appeared in synovium tissue of group B, much appeared in that of group D by using the method of immunohistochemistry Stain. Group C compare with group D ,the temperature of knee suaface , the worth of ECT , SOD > MDA and IL— 1 in joint fluid ,all have statistical difference (P<0.05, P<0.01). There are a few neutrophilic leukocytosis and no obvious inflammation in synovium of group C, while typical inflammation was appeared in that of group D. There are a little male substance appeared in synovium tissue of group C, much appeared in that of group D by using the method of immunohistochemistry Stain. Group B compare with group C ,the temperature of knee surface , the worth of ECT , SOD , MDA and IL— 1 in joint fluid ,all have no statistical difference (P>0.05) . There are a few neutrophilic leukocytosis in synovium between group B and group C, and on typical inflammation. There are little male substance which was expressed by NF- k Bp65^ NF- k. Bp50 in synovium of both group B and group C. The rate of male substance has no statistical difference between group B and group C (P>0.05) .Conclusion: NanHu Gelatin can be spread well and be permeated skin rapidly, It has powerful viscosity and good biologic compatibility. The animal model of knee Synovitis by trauma is successful.lt can be tested its reliability from the result of the knee joint circumference, the joint surface's temperature , the time of claudication, the worth of ECT, the level of SOD, MDA and IL— 1, and the result of pathologicalsection and immunohistochemistry Stain. The result from comparison between group A and group D shows that NanHu Gelatin can decrease the temperature of join surface, shortening the time of claudication, adding the content of SOD in joint fluid , reducing the level of MDA and IL—1, relieving the inflammation, decreasing the expression of male substance of NF- k Bp65> NF- k Bp50 in synovium. From above, it can be draw a conclusion that NanHu Gelatin has the function of preventing Synovitis by trauma. The result from comparison between group B and group D shows that NanHu Gelatin acquired good therapeutic efficacy in treating Synovitis induced by trauma. The possible mechanism of it's prevention and cure of synovitis which induced by trauma is increasing oxygen-derived free radidicals and restraining the produce of inflammation medium (such as IL-1) .In this way, it decrease the expression of nuclear factor and interdict the enlargement of inflammation. The therapeutic efficacy is more protrusion and more economical and more compared with FuTaLin emulsion. It is worthy of being developed and being used in clinic.