CKIP-1 Regulates The Mass Of The Different Parts Of Bone In The Growth And Development Of Mice

The bone metabolic disease,which is represented by osteoporosis,affects more and more people all over the world,and it has a great influence on the physical and mental health of the patients.With the change of the social life style,the age composition of the population of the bone metabolic disease is also showing a trend of younger age.Bone metabolism diseases are characterized by dynamic balance of bone formation and bone resorption.Therefore,the regulation of bone formation and bone resorption is an effective method to treat bone metabolic diseases.Although there are many related studies in this area,there is no systematic study on the changes of bone metabolism in different growth stages.In recent years CKIP-1 is a concern of the genetic factor,it is involved in a number of physiological processes in the body,which attracts the attention of us most is the negative regulatory effects of bone formation.According to the current research in bone CKIP-1 focus on at a particular point in time for a specific type of bone formation.So this study with the aid of ckip-1 gene knockout mouse model,by gross observation,radiological,histological and serum biochemistry test of different growth and development stages of ckip-1 gene knock in addition to different parts of mice bone were evaluated.Objective:1 To evaluate the effects of CKIP-1 gene on the whole bone and bone mass in different parts of bone in different stages of growth and development by gross observation,imaging and histology.2 To assess the effect of CKIP-1 gene on the whole bone metabolism of mice in different stages of growth and development by serum biochemistryMethods:1 Select littermate male WT(CKIP-1+/+)and KO(CKIP-1-/-)mice in 1,3 and 6 months for gross observation,and then draw the femur,the vertebrae and the mandible,then observe and analyze the condition of mouse overall development and bone growth and development.By using X-ray,micro-CT and tissue staining method,Determinate the differences of the bone imaging and bone related parameters of the femur,vertebrae and mandible of WT and KO mice in 1,3 and 6 months,and to observe the difference in morphology and histology.2 Collect the serum from WT and KO mice of 1,3 and 6 month time points,and evaluate the effect of CKIP-1 gene on bone formation and bone absorption index of mouse bone in different growth stages by Elisa.Results:1 General observation: the size of WT and KO mice showed a significant increase in the 1 to 3 months,while the body size was not significantly changed from 3 months to 6 months,but the body weight was increased.During the period of 1-6 months,the activities and the posture of WT and KO mice were normal and no significant difference.There was no significant difference in body weight of WT and KO mice aged 1 month.The weight of KO mice aged 3 and 6 months is larger than that of WT mice.The bone density of the mandible of KO mice was significantly more than that of WT mice from 1 month,and the difference was more obvious with the increase of the age of the mice.In 1 month,there was no significant difference between vertebrae and femur of KO and WT mice,but from 3 months to 6 months,it can be observed that the vertebrae and femur bone volume and form bone density of KO mice was higher than that of WT mice.2 X-ray scan: In 1 month,femur and vertebrae images of WT and KO mice showed no significant difference,and there is no significant difference between the two groups of the mandible,but more compact in KO groups.From 3 months,three kinds of bone imaging of KO mice showed obvious difference compared with WT mice,the radiopacity of KO mice bone was significantly higher than that of WT mice,and the morphological structure of the three kinds of bone in KO mice were more than WT mice.3 Micro-CT scan: we found that there was no significant difference in trabecular morphology between the three types of bone in the WT and KO mice in the group of 1 month,but the density of the KO mouse was slightly larger than that in the WT mouse.From 3 months,it was obvious that the bone of WT mice was less dense than that of KO mice,and the number and thickness of KO mice were significantly higher than that of WT mice.At the age of 1 month,either the volume fraction(BV/TV)and specific bone surface area(BS/BV)of femur and vertebrae of WT and KO mice bone,or the trabecular thickness(TB.Th),trabecular number(TB.N)and bone trabecular separation(TB.SP)showed no significant difference(P > 0.05).In the region of interest within the cancellous bone of KO mice mandibular,BV/TV,TB.Th and TB.N were higher than in WT mice(P < 0.05),and BS / BV and TB.SP were lower than WT mice(P < 0.05).In the group of 6 month,BV/TV,TB.Th and TB.N of femur,vertebrae and mandible of KO mice were higher than that of WT mice(P < 0.05),BS / BV and TB.SP were lower than WT mice(P < 0.05).4 HE staining: there was no significant difference between WT and KO mice bone microstructure in 1 and 3 months,but trabecular bone of mandibular of KO mice compared with WT mice arranged more closely The morphology of trabecular bone of three kinds of bones of KO mice in 3 and 6 months were still consistent with the WT group,but the number and size of trabecular bone of three kinds bone in KO mice were larger than that of WT mice.The arrangement of KO mouse bone trabecular was stronger than that of WT mice.5 Toluidine blue staining: there was no significant difference in microstructure of femur,vertebrae and mandible of WT and KO in 1 month.Mature trabecular number in femur,vertebrae and the mandible of KO mice in 3 month increased significantly,bone small beam has more thick,and the number of bone cells is more than the WT group,and the bone cells,osteoid and new bone around the cartilage of KO mice were also more than those in the WT mice.In 6 month,the microstructure of KO mice was more optimized,but the difference between the number of osteoblast,osteoblast around the cartilage and new bone of two groups was smaller than that in 1 month.6 Elisa: There were no significant differences in the indexes of WT and KO mice in 1 month.From 3 months,ALP,OCN,COL,PINP and PICP values in KO mice were higher than those in WT mice(P < 0.05),but the values of CTX in KO WT mice were not significantly different.The results of 6 month group and 3 month group was basically consistent,bone formation and metabolism index of KO mice were higher than those of WT mice(P < 0.05),the CTX value of KO and WT mice were not significantly different.Conclusions:1 During the whole process of growth and development,CKIP-1 gene had not lead to abnormal effects of its normal physiological function.2 The results of imaging and histology showed that the CKIP-1 gene had an effect on bone mass in the mandible at the early stage of 1 month,the bone structure was more optimized and the bone mass increased.In the 3 months of maturity,femur and lumbar vertebrae is beginning to reflect the difference,then continued to 6 months old.The bone mass change with time is more and more obvious,and it is consistent with the growth cycle of mice.3 The results of serum biochemical analysis showed that CKIP-1 gene had no significant effect on bone metabolism in 1month.The CKIP-1 gene promoted the process of bone formation in 3 and 6 months,but did not affect bone resorption.This regulator effect was more and more obvious with increasing of time.