| Objective:under the guide of "paying equal attention to the bone and tend theory of TCM(Traditional Chinese Medicine),through establishment the model of reconstruction rabbit anterior cruciate ligament to study the influence of using the nourishing liver and kidney Chinese medicine radix dipsaci.To explore the tendon to bone healing mechanism by using Chinese medicine radix dipsaci,as to provide theoretical basis for its application in clinical.Methods:The 40 healthy adult male New Zealand white rabbits were used to simulate the left knee reconstruction model of anterior cruciate ligament rupture,and selecting 32 successful model for the experiment,all of them were randomly divided into four groups by using the random number table.The model were divided into blank control group,low dose radix dipsaci group,medium dose radix dipsaci model group and high dose radix dipsaci model group,each group of eight.Under the guide of "paying equal attention to the bone and tend theory of TCM,using physiological saline,low,medium and high dose of nourishing liver and kidney medicine radix dipsaci to lavage,each group executed 4 rabbits in 4 weeks,8 weeks,before putting them to death,observing the general situation of all rabbits,taking out all the rabbits ear margin venous blood to analyze biochemical examination analysis of blood calcium and blood concentration of alkaline phosphatase;Micro-CT scans all the rabbit knee joint,calculation and analysis of bone tunnel area and the average CT attenuation value around the bone tunnel;And the tissues surrounding the femoral and tibial tunnel in model;Using vernier calipers measures the femoral medial tunnel diameter,and calculates its expansion rate;Each group executed 1 model to make histology slice and then took it for HE staining as well as observed the formation and distribution of Sharpey sample fiber by light microscopy to understand the change of tendon tissue component and cell differentiation in bone and tend healing;Each group took three model to make the specimen with the tendon transplantation in femoral and tibial side.Finally,using specimen to conduct biomechanical test,observing the reconstruction ligament's biggest limit tensile load when it totally torn or emerged;Then Comparing with blank control group and statistical analysis.Results:All the experimental rabbit made model of anterior cruciate ligament rupture construction in their left knee.While in the process of building model,1 rabbit died of anesthesia,3 rabbit die of failure,the rest were finished building successfully.After building model,in experimental process there did not appear a rabbit death.Overall,in 4 and 8 weeks after building model,the low,medium and high dose of radix dipsaci model groups in general situation,gross observation,the femoral medial tunnel expansion rate,concentration of blood alkaline phosphatase,blood calcium,bone tunnel area,the average CT attenuation value around the bone tunnel and ultimate tensile load were better than that of blank control model group.Among the results,high dose of radix dipsaci model group was the best,second was the medium dose of radix dipsaci model group,blank control model group was the relatively worst;And in HE staining histological observation of 4 weeks and 8 weeks,the Sharpey collagen fibers as well as fibroblasts' growth and ossification of low,medium and high dose radix dipsaci model groups were better than blank control model,high doses of radix dipsaci model group was the best,by contrast;After building model in 4 and 8 weeks,using the collected data to carry on statistical analysis of four groups specimens,the results showed:(1)After anterior cruciate ligament reconstruction of rabbit left knee in 4 weeks,four groups compared blood calcium concentration with each other.The result showed statistically significant differences(F=214.823,P=0.000<0.05);Four groups of blood concentration of alkaline phosphatase had statistically significant differences(F=231.660,P=0.000<0.05);Four groups of bone tunnel area had statistically significant differences(F=213.030,P=0.000<0.05),except the pairwise comparison between low doses of radix dipsaci and the blank control group(P=0.210>0.05);Four groups of the average CT attenuation value around tibial tunnel had statistically significant comparative differences(F=128.852,P=0.000<0.05);Comparing with the four groups of femoral medial tunnel expansion rate,the difference was statistically significant(F=43.371,P=0.000<0.05),except the pairwise comparison between low doses of radix dipsaci group and the blank control group(P=0.219> 0.05);Comparing with the four groups ultimate biggest tensile load in 4 weeks,the differences had statistically significant(F=109.802,P=0.000<0.05).(2)After anterior cruciate ligament reconstruction of rabbit left knee in 8 weeks,four groups compared blood calcium concentration with each other.The result showed statistically significant differences(F=69.489,P=0.000<0.05);Four groups of blood concentration of alkaline phosphatase had statistically significant differences(F=65.796,P=0.000<0.05);Four groups of bone tunnel area had statistically significant differences(F=74.589,P=0.000<0.05);Four groups of the average CT attenuation value around tibial tunnel had statistically significant comparative differences(F =75.548,P=0.000<0.05);Comparing with the four groups of femoral medial tunnel expansion rate,the difference was statistically significant(F=73.222,P=0.000<0.05);Comparing with the four groups ultimate biggest tensile load in 8 weeks,the differences had statistically significant(F= 77.134,P=0.000<0.05).Conclusion:After knee anterior cruciate ligament reconstruction,traditional Chinese medicine radix dipsaci has a positive role in tendon to bone healing.To promote osteogenesis,increase the number of trabecular bone,reduce trabecular interspaces,increase bone density,promote fibroblasts and Sharpey collagen fibrous proliferation in area of tendon to bone junction,reinforce tendon bone healing strength are of great significance in such aspects.And there is a certain positive correlation with concentration of radix dipsaci. |
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