Effects Of Vascular Endothelial Growth Factor In Bone Repairing With Setting Broken Bone Mixture

Objective:To investigate correlative experimental study and mechanis of vascular endothelial growth factor during the homologous spongy bone repairing defect with setting broken bone mixture(nature copper xuduan gusuibu ground beetle) of invigorator the circulation of blood.But the Chinese drug go to correlate virus test in smaller white mouse. Methods:96 Kunming smaller white mouse which weight are about 20g devide into two groups.Which go on acute test and deuto～test with setting broken bone mixture,every group is 46.acute test group devid into the biggest dose group(21.5g/kg),middle dose group (10g/kg),smaller dose group(4.64g/kg) and the smallest dose group.deuto～test group devid into the bigger dose group(20g/kg),middle dose group(10g/kg) and the smallest dose group (5g/kg).100 common adult family rabbit weigh from 2.0kg to 2.5kg without limitation of sex,masculinity and females werw feed in two cages.90 of them were randomly divided into 5 groups.And then each group 18 was divided into a test team and a controled team,every group is 9.The other ten rabbits offer homologous spongy bone.They were killed by pouring 5ml of air into the vein of each rabbit's ear.Spongy bone of bilaterally ilium was taken to make 2×2×2mm sponge bone particle.Rabbits in both teams were local anesthesia with 1% dolicaine,and then 1/3 of their middle part of radius were exposed in asepsis environment. Then a 15mm standard bone defect model with a bone grinding machine was made,the bone marrows were enclosed by bone wax and the left and right radical bone defect was repaired by above sponge bone particles.The test team was fed with Chiense drug mixture forage and the controled team was fed with common forage nor Chiense drug mixture.In order to prevent rabbits from infection,they were injected with 0.15g/kg of cefazolin sodium into muscle for 4 days in a row.After they were killed separately in the 1st,2nd,4th,6th,8th week,After their radius were x～rayed.Ttree rabbits specimen were taken for polymerase chain reaction and other three were taken from for histological observation,situ hybridization as well as electron microscope observation.Results:Virus test consult:acute virus test is 46 smaller white rouses without anomal action and without one case dead after one week.deuto～acute virus test is that the biggest dose test group wite rouses find obvious fatty change,increase transaminase and creatinine than in the control group in lobes liver histological observation,the group with medine is no obvious difference to the other group except high dose group and creatinine,the middle dose test group wite rouses find minimal fatty change,increase transaminase and creatinine than in the control group in lobes liver histological observation,the group with medine is no obvious difference to the other group except high dose group and creatinine.the smaller dose test group wite rouses find without fatty change,increase transaminase and creatinine than in the control group in lobes liver histological observation,the group with medine is no obvious difference to the other group except high dose group but creatinine.X-Ray observation:There was no obvious change in the test and group team at first week.The porosis were obvious increase from two to eight weeks.there were a lot of blurred porosis on the bone defect,and the bone defect vanished In the eighth week,the marrom cavity is formed.The porosis was less in controled team than in the test team at the same time.Histological observation:There was no obvious change in the test and group team at first week.there are part of the osteocyte necrosed and some nuclei of osteocyte concentrated in implanted bone;and even vanished.there are less inflammation in the test team than in the controlled group team.There were a lot of cartilage cells formed and blood vessels formed growing into implanted bone from two to eight weeks,marrow cavity are formed in the eight weeks.There was a few cartilage cells and blood vessels formed during second and fourth week.During sixth and eighth week,there were an increase of porosis and blood vessels,but the marrow cavity had not formed yet.The electron microscope observation:There were destruction on part of the bone trabecula in the test and controlled group and with a little inflammatory cell in the first week.There was a lots of preosteoblast and a few osteoblast cells in the test team and the in the second week.team, There was more capillaries around preosteoblast and collagen fiber after bone matrix being Decalcification in the second week.controled team.,There were more capillaries,osteoblast and bone trabecula in the test team than in the controled team in the fourth week.There were more some formation of bone matrix capillaries,bone trabecula,chromocenter and increasing of the endoplasmic reticulum in the the test team than in the controled team from six weeks to eighth week.Immunity chemical staining observation:Expression of VEGF is weak positive, positive,stronger positive,positive,weak positive in the test group at the one week,two week,four week,six week,eight week.but Expression of VEGF is weak positive,weak positive,positive,weak positive,weak positive in the control group at the one week,two week,four week,six week,eight week.Situ hybridization observation:Expression of VEGF was negative in grafting bone specimen of bone defeact after one week in the test team,.It was weakly positive in grafting bone specimen of bone defeact after two week in the test team and was strongly positive in grafting bone specimen of bone defeact after 4 weeks in the test team.It was weakly positive in the sixth and eighth week.However,Expression of VEGF was negative in grafting bone specimen of bone defeact after one week in the controled groups and it was positive after 4 weeks.And it was weakly positive after 2,6,8 weeks. RT～PCR result:Under the condition of 94℃,30 second,55～65℃,30 second,30 Cycles 72℃,1 min/kbp,Target gene of VEGF in the controled team were:2547copies in the first week,4972 copies in the second week,1230 copies in the fourth week,411 copies in the sixth weeks,and 856 copies in the eighth week.target gene of VEGF were1105 copies in the first week,1386 copies in the second week,1048 copies in the fourth weeks,824 copies in the sixth week and 883 copies in the eighth week.Result of t analysis statistics was P＜0.05 in the vascular endothelial growth factor Conclusion:1.The medial lethal dose of setting broken bone mixture exceed 21.5g/kg to Kunming mice in acute toxicological experiment.2. Setting broken bone mixture can significantly shorten the time to repair bone defect.3. Setting broken bone mixture can significantly increase the expression of vascular endothelial growth factor with homologous spongy bone to repair bone defects.