| OBJECTIVE:To investigate the feasibility of accelerated bone repair and reconstruction of fracture combined with traumatic brain injury,and to study the effect of serum of rats with fracture combined with traumatic brain injury on the proliferation and differentiation of osteoblasts,and provide a theoretical basis for the treatment of orthopedic diseases with brain injury and fracture serum.The next step is to apply the clinical foundation.METHODS:Twenty Sprague Dawley(SD)rats with fracture and brain injury were prepared.The serum was prepared in stages.Twenty healthy rats were randomly selected to prepare negative control serum.All rats were collected by abdominal aorta and serum was labeled.refrigeration.96 rats with model femur fractures were randomly divided into three groups:A,B,and C,namely,treatment group,negative control group and blank group,with 32 rats in each group.Group A was treated with intramuscular injection of serum,group B was injected with negative control serum,and group C was injected with normal saline.Three groups were intramuscularly injected at the corresponding time points.Each group died 8 rats at 1 w,2 w,3 w,and 4 w postoperatively.Radiographic X-ray projection and dual-energy X-ray bone density were performed.Detection and histomorphological observations were performed to compare the three groups of bone repair and reconstruction.The osteoblasts were cultured and identified,and the effects of ccc-8 method and alkaline phosphatase activity on the proliferation and differentiation of osteoblasts in the combined trauma group,brain trauma and simple fracture group were compared.result:1.Radiological and histomorphological observation of the serum of rats with fracture combined with traumatic brain injury in rats1.1 X-ray filmThe results of postoperative X-ray results showed that the bone repair and reconstruction process in group A was significantly faster than that in group B and C.One week after operation,a small amount of callus was seen in group A.The fracture line was slightly blurred and local swelling was obviously relieved.The fracture lines in group B and C were clear,local soft tissue was swollen,and no obvious osteophyte formation was observed.At the second week after operation,the osteophytes of group A increased and the fracture line was blurred.The fracture lines of group B and C were slightly blurred,and a small amount of callus was formed.In the third week,the osteophytes in group A decreased,and the fracture line was further blurred.In group B and C,the fracture line was blurred and the callus increased.At the 4th week,the osteophytes in group A were further reduced,and the fracture line disappeared.The fracture lines in group B and C were blurred,and the callus continued to increase.At 2 weeks after operation,the volume of callus in group A was significantly higher than that in group B and C,and the difference was statistically significant(p=0.000).At 3 weeks after operation,there was no significant difference in the A,B,and C groups(P=0.669).At 4 weeks after surgery,the volume of callus in group A was smaller than that in group B and C;the difference was statistically significant(p=0.000).1.2 Bone mineral density(BMD)At the first week,there was no significant difference in bone mineral density between the three g:roups(p=0.882).From the second week,the bone mineral density of group A was higher than that of group B and C,and the difference was statistically significant(p=0.000).1.3.Hematoxylin-eosin(H E)stainingAt the second week after operation,the cartilage callus and calcified tissue were seen in the fracture of group A,There was no obvious regularity in the arrangement of trabecular bone.In the same period,the bones of group B and C grew sparsely,and a large number of cartilage bones were seen at the fracture end,and hypertrophic chondrocytes were seen.At the third week after operation,there were a large number of hard osteophytes in the fractures of group A.The trabecular bones increased and arranged regularly,and obvious lamellar bone structure appeared.In the same period,groups B and C only found immature woven bones,and the trabecular bones increased,and the arrangement gradually became regular.At the 4th week after surgery,most of the braided bone of group A was changed into lamellar bone,and a little cartilage was found in the outer epiphysis.In the B and C groups,the woven bone and the lamellar bone began to fuse,and the trabecular bone was thickened.2.The effect of different serum on the proliferation and differentiation of osteoblasts.2.1 The effect of three groups of serum on the proliferation of osteoblasts2.1.1 Effect of serum between groups on osteoblast proliferationThe statistical differences between the three groups of serum prepared at each time point were significant(P<0.01).For the serum prepared at 12h,4d and 1w,LSD test showed that the OD value of the fracture combined with traumatic brain injury group was higher than that of the fracture group and brain trauma group(p<0.05).There was no significant difference between the fracture group and the brain trauma group(p>0.05).).For the serum prepared by 2w and 3w,the OD value of the fracture combined with traumatic brain injury group was significantly higher than that of the fracture group and brain trauma group(p<0.01);the fracture group was higher than the brain trauma group,and there was statistical difference(p<0.05).2.1.2 Effect of serum prepared at different time points in the group on osteoblast proliferationThe OD values of serum prepared at each time point in the three groups were significantly different(P<0.01).There was no significant difference in the OD values between the 1st and the 3w serum in the combined trauma group(p>0.05).The multiple comparisons of the OD values at each time point were statistically significant(P<0.01).The proliferation of osteoblasts was gradually enhanced,and the serum of the 2nd was the strongest,and then began to decline.The OD of the serum in the fracture and brain injury group was higher than that in the fracture group and the brain trauma group.The serum OD value of the fracture group gradually increased with the preparation time,and reached the peak at the 1st,then decreased gradually.The OD values at different time points in the group were compared,and there was no significant difference between the 12h and 2w,3w(p>0.05).There was a statistically significant difference(p<0.01)from the comparison of 4d and 1w.4d was statistically different(p<0.05)compared with serum OD values prepared at other time points.There was a statistically significant difference(p<0.01)between 1w and 2w and 3w.There was no statistical difference between 2w and 3w(p>0.05).The serum of the brain injury group gradually enhanced the proliferation of osteoblasts.The serum proliferation activity reached the maximum at the 1st week,and then decreased gradually.There was no statistical difference between the 12h and 3w in the group(p>0.05),and the other two points had the contrast.Statistical difference(p<0.05)2.2 The effect of three groups of serum on osteoblast differentiation.2.2.1 Effect of serum between groups on osteoblast differentiationThere was no significant difference in serum OD between the three groups(P>0.05).There were significant differences in serum OD values at 12h,4d,2w and 3w(P<0.05).Multiple comparisons of serum OD values at 12h showed that the fracture group was significantly higher than the combined trauma group and brain trauma group,the difference was statistically significant(p<0.01);the combined trauma group was higher than the brain trauma group,and there was significant statistical difference(p<0.01).Multiple comparisons of serum OD values in 4d showed that the fracture group was higher than the brain trauma group and had statistical difference(p<0.05).The combined trauma group was higher than the brain trauma group and lower than the fracture group,but it was not statistically significant(p>0.05).The 2w serum OD values were significantly higher than those in the fractaure group and the brain trauma group(p<0.01).There was no significant difference between the fracture group and the brain trauma group(p>0.05)..Multiple comparisons of 3w serum OD values showed that the combined trauma group was significantly higher than the fracture group and brain trauma group,the difference was statistically significant(p<0.01);the fracture group was higher than the brain trauma group and had statistical significance(p<0.01).2.2.2 Effect of serum in the group on osteoblast differentiationThe effect of serum preparation on the differentiation of osteoblasts was stronger in the time of serum preparation.There was no significant difference in the effect of serum from different time in the fracture group on osteoblast differentiation.The effect of brain trauma group on osteoblast differentiation was slightly enhanced.Peaked at 1w,then gradually decreasedThe OD values of serum prepared at different time points in the combined trauma group and brain trauma group were statistically different(p<0.05).The serum prepared at different time points in the combined trauma group showed that the serum OD value of the 4th day was less than the 1W serum.OD values,but no statistical difference(p>0.05);serum OD values in each period were statistically different(p<0.05).There was no significant difference in serum OD between the fracture group at different time points(p>0.05).The OD values of serum in the traumatic brain injury group showed that there was no significant difference between the 12h and 3w,4d and 2w,1w and 21w(p>0.05);the serum OD values in the remaining time periods were statistically different(p>0,05)<0.05).Conclusion1.Improved the operation method of rat femur fracture model,and successfully established SD rat fracture combined with brain trauma model.2.To determine the feasibility of serum accelerated bone repair and reconstruction in rats with intramuscular fracture combined with traumatic brain injury.3.The serum of rats with traumatic brain injury has a significant effect on the proliferation and differentiation of osteoblasts.When the serum concentration of rats with traumatic brain injury was 10%,the enhancement of osteoblast proliferation was the strongest.From the time point,the serum of 2w preparation had the strongest proliferation,then began to weaken;the effect on osteogenic differentiation was 2w.The peak was reached and the 3w did not decrease significantly.4.Accelerated bone repair and reconstruction after combined traumatic brain injury may be due to the increased permeability of the blood-brain barrier after brain trauma,a large number of osteogenic factors enter the blood through the blood-brain barrier;it may also be that the body after the combined brain trauma has produced some kind of Hormones greatly promote the formation of osteogenic factors and promote bone repair and reconstruction.5.This study applied SD rat fracture combined with brain trauma serum to simulate the accelerated healing process of fracture combined with traumatic brain injury,provided theoretical basis for the treatment of orthopedic diseases,and laid the foundation for its clinical application. |
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