| Objective: To study the expressions of Bone morphogenetic protein2(BMP2)and Transforming growth factor-form1(TGF-β1)in peripheral blood of rabbits during tibial Bone transport and their correlation with the amount of Bone callus during Bone regeneration,and to explore the feasibility of regulating the migration rate with BMP2 and TGF-form1 as monitoring indicators。Methods: Rauch F and other methods were used to slightly improve the tibial bone migration and prolongation of rabbits.The 36 animals needed for the tibial bone transport and prolongation animal model experiment were all provided by our school’s experimental animal center,and they were mature male New Zealand white rabbits at the age of 6 months.The experimental modeling was divided into two parts: the first part randomly selected 20 healthy mature male New Zealand white rabbits and gave them bone transfer molding treatment.They were divided into 4 groups of ABCD,group A as the control group,and BCD group as the experimental group with different distraction speed,5 rabbits in each group.After the latency period of 5 days,each group was distractied respectively by 0mm/24 h,0.5mm/24 h,1mm/12 h,1.5 mm/8h.BCD group of rabbit tibia eventually extend 30 mm,the distraction time of the group A with group C as the control point(the intermediate group of BCD group),respectively in group A preoperative,latency period 5 days(5 days after operative),11 days after operative,17 days after operative,23 days after operative,29 days after operative,35 days after operative and BCD group of preoperative,latency period 5 days(5 days after operative),distraction period a total of five times(distraction length is 6 mm,12 mm,18 mm,24 mm,30 mm)these seven point detection of peripheral blood serum BMP2 respectively TGF-β 1 concentration and X-ray films were compared with bone mineral density(BMD)values 。In the second part.The second part,the other 24 mature New Zealand white rabbit,and consists of four groups of the ABCD,6 in each group.respectively in group A preoperative,latency period 5days(5 days after operative),11 days after operative,17 days after operative,23 days after operative,29 days after operative,35 days after operative and BCD group of preoperative,latency period 5 days(5 days after operative),distraction period a total of five times(distraction length is 6 mm,12 mm,18 mm,24 mm,30 mm)these seven point,a rabbit bone was randomly selected from each group to remove the gap tissue,and wax blocks were made after formaldehyde fixation for HE staining and masson staining analysis to evaluate the callus formation in the distraction gap.Results: the tibia distraction model of the experimental group and the control group was successfully established,and the results were all analyzed.1.Comparing the serum BMP2 concentration among the four groups of ABCD,there was no statistical difference in the serum BMP2 concentration before and during the latency period(P > 0.05).At the time point of moving 6mm,there was a statistical difference in the serum BMP2 concentration between groups A and B,A and C(P < 0.05);at the time point of moving 12 mm,there was a statistical difference in the serum BMP2 concentration between groups A and B(P < 0.05);at the time points of moving 18 mm,moving 24 mm and moving 30 mm,there was a statistical difference in the serum BMP2 concentration between groups.There was no statistical difference in degree(P >0.05).Compared with the serum BMP2 concentration in the four groups of ABCD,there was no significant difference in the serum BMP2 concentration in group A at each time point(P > 0.05);there was a significant difference in the serum BMP2 concentration in group B between the 12 mm and 6mm distraction length(P < 0.05);there was a significant difference in the serum BMP2 concentration in group C between the 12 mm distraction length and the preoperative concentration(P < 0.05);and there was a significant difference in the serum BMP2 concentration in group D between the 24 mm distraction length and the 6mm distraction length(P < 0.05);and there was a significant difference in the serum BMP2 concentration in group D between preoperative and the30 mm distraction length(P < 0.05)。The main effect of different distraction speed on serum TGF-beta 1concentration was not statistically significant(P > 0.05).The time factor had statistical significance(P < 0.05)on serum TGF-beta 1 concentration,in which the difference between latency and preoperative serum TGF-beta 1concentration was statistically significant(P < 0.05);the difference between6 mm and preoperative serum TGF-beta 1 concentration was statistically significant(P < 0.05);the difference between 12 mm and preoperative serum TGF-beta 1 concentration was statistically significant(P < 0.05).2.In X-ray image analysis,In the control group,a small amount of high density shadow was observed in the bone defect area 17 days after operation,that is,a small amount of new bone formation.On the 23 rd day after operation,a high density shadow was seen in most of the bone transfer space,i.e.,a large number of callus formed to fill the bone defect area.The bone-distraction space was basically filled 35 days after operation.In group B,a small amount of high density shadow,i.e.,a small amount of new bone formation,could be seen under the periosteum when the removal length was 6 mm(17 days after operation).At12 mm(29 days after operation),there were many high density shadows in the bone-distraction space,which extended from the broken ends of the two bones to the center in the bone-distraction area.At 30 mm(65 days after the operation),the new bone extended from the broken ends of both sides to the center,but still not connected.The new bone structure at the osteotomy end was remodeled,showing a marrow cavity-like structure,but not penetrated.In group C,a small amount of high-density shadow appeared at the osteotomy end of the bone-distraction space at 12 mm(17 days after operation),and the new bone in the bone-distraction space was gradually increased with the extension of bone-distraction distraction.Up to 30 mm(35 days postoperatively),new bone extending from the osteotomy end to the center of the gap on both sides remained unconnected;new callus shadow was more and more uniform in Group C compared with X-ray films of the same length of removal in Group B.In group D,a small amount of high density shadows appeared in the bone-distraction space of 18 mm(17 days after operation),mainly new callus under periosteum.On the 25 th day after operation,the callus extending from the osteotomy ends on both sides to the center still did not reach the connection,and the amount of new callus in the whole removal space was significantly reduced compared with the control group,group B and group C.There was no significant difference in BMD between the four ABCD groups at the time point of 6mm distraction(P > 0.05);there was significant difference between groups A and C,groups A and D,groups B and C at the time point of 12 mm distraction(P < 0.05);there was significant difference between groups A and D,groups B and D at the time point of 18 mm distraction(P <0.05);there was significant difference between groups A and D,groups B and D,groups C and D at the time point of 24 mm distraction.The difference was statistically significant(P < 0.05);there was no significant difference in bone mineral density between the four ABCD groups at the time point of 30 mm of distraction(P > 0.05).There was no significant difference in bone mineral content between the four ABCD groups at the time point of 6 mm of distraction(P > 0.05);there was significant difference between groups A and D,B and D at the time point of12 mm of distraction(P < 0.05);there was significant difference between groups A and B,B and C at the time point of 18 mm of distraction(P < 0.05);there was no significant difference in bone mineral content between the four ABCD groups at the time point of 24 mm of distraction and at the time point of 30 mm of distraction.Statistical significance(P > 0.05).3.In H&E staining,compared with callus near the osteotomy end in the ABCD group,the control group distracted 30mm(25 days after operation)to see a larger amount of new callus but arranged in disorder;Group B distracted30mm(65 days after operation)to see that the bone transfer area was mainly filled with fibrous tissue,with a little callus;Group C distracted 30mm(35 days after operation)to see a larger amount of new callus in the bone transfer area and arranged in the same direction;Group D distracted 30mm(25 days postoperatively)some new callus could be seen to fill the bone transfer area,and the callus was arranged in the same direction basically.4.In Mason staining,mineralized bone tissue was found near the osteotomy end of the bone defect gap and cartilage tissue was found in the middle of the bone defect in group A 35 days after operation.In group B,a small amount of mineralized bone and some adipose tissue could be seen near the osteotomy end in the distraction gap of 30 mm,and some mineralized bone could be seen in the middle of the distraction gap,and the new bone tissues were regularly arranged in the same direction of distraction.In group C,some mineralized bone could be seen near the osteotomy end in the distraction gap of 30 mm,and the callus amount was more than that in group B at the same site,and arranged regularly in the same direction.The callus amount of new bone tissue in the middle of the bone distraction gap was less than that in group B,and the callus distribution in the whole distraction gap was more uniform in group C.A small amount of callus could be seen near the osteotomy end and the middle of the distraction gap of 30 mm in group D,but the callus amount was less than that in group B and C.Conclusion: 1.The concentration changes of serum BMP2 and TGF-β 1have the potential to regulate the transport speed.2.The results showed that the optimal transport speed of rabbit tibia was 1 mm / d.3.The time of callus formation was similar in different moving speed groups,which was 17 days after operation;but it would affect the amount of new callus,which decreased with the acceleration of different transport speed under the same length of distraction.4.Serum BMP2 can promote the formation of new callus;TGF-β 1plays an important role in the early stage of transfer.5.In the process of bone transportation,the external mechanical environment can affect the spatial arrangement of new bone and blood vessels. |
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