| Objective: through studying the origin and development of the theory of "lesser yang governing bone" and the clinical application of xiaochaihu decoction modified recipe,we have further expanded and enriched its theoretical connotation.moreover,through modern scientific means,we have studied the application performance of xiaochaihu decoction modified recipe in improving the microenvironment of bone metabolism from the microscopic field,and verified its role in promoting the repair of bone defects with Mg-Zn-Ca amorphous alloy material,so as to provide a theoretical basis and experimental support for the clinical application of traditional chinese medicine collaborative medical metal material for the repair of bone defects.Methods: By the method of combining theoretical discussion with animal experiment,we combed the related ancient literature of traditional Chinese medicine,and from the perspective of meridian theory,specifically explained the ideological framework and connotation of "Shaoyang governing bone" in Huangdi’s Internal Classic from the following aspects: running of Shaoyang meridians,taking Shaoyang as the pivot,and the rise and development of gallbladder qi of Shaoyang,and systematically constructed the close relationship between "Shaoyang" and "bone".Taking advantage of the classic famous recipe Xiaochaihu Decoction’s efficacy of both regulating qi movement and harmonizing zang-fu organs,as well as the efficacy advantages of multiple active components and multiple target points in traditional Chinese medicine(TCM)prescriptions,we modified the original recipe into the modified recipe and conducted an in-depth study on its important role in regulating both ends by using the pivot in the bone repair process.In the animal experiment,20 SPF SD rats(Sprague-Dawley,SD)were used to establish the femoral bone defect model and the Mg-Zn-Ca amorphous alloy material was implanted.Ten rats randomly selected as the experimental group and the other 10 control groups were given the Xiaochaihu Decoction cut and the same dose of 0.9% normal saline by gavage for 12 weeks.After the samples were taken,they were separately scanned by Micro-CT to observe the osteogenesis around the material and its degradation.HE staining of hard tissue sections was used to evaluate the growth of bone tissue around the material and the binding of the “ bone-implant ” interface.The fluorescence labeling of calcein–xylenol orange– tetracycline hydrochloride was used to trace the formation rate and growth mode of new bone,and the tissues of important organs were collected for pathological observation and blood biochemical analysis to evaluate its biological safety.Result:As for the TCM mechanism,the theory of "Shaoyang governing bone" originates from the bone moving along the meridians of Foot-Shaoyang,and is based on the qi-promoting effect of liver and gallbladder rise and hair rise of Shaoyang on bone formation,but it is not limited to the bone diseases coverage along the meridians,etc.,thus providing a theoretical basis for the treatment of bone and joint diseases based on Shaoyang.The imaging test results showed that the Mg-Zn-Ca amorphous alloy had good osteogenic property and excellent biodegradability in vivo,and matrix collapse did not occur in the early stage of the material after administration of Chinese medicine,so it could provide early load-bearing for bone defect area.The staining of hard tissue sections showed that the Mg-Zn-Ca amorphous alloy underwent significant biodegradation and bone tissue regeneration was increased.The Elisa method used to detect the osteogenesis-related genes in serum showed that the expressions of COL-1,BMP2,Runx-2 and TGF-β in the Xiaochaihu Decoction +Mg-Zn-Ca amorphous alloy group were up-regulated compared with those in the control group,and COL-1 expression was particularly significant.The rats in the two groups were labeled with calcein–xylenol orange–tetracycline hydrochloride fluorescence,and the obvious fluorescence staining bands were observed around the implant materials,forming concentric rings or irregular shapes with the relative materials turning green,red and yellow from far to near.The spacing between different fluorescence bands could indirectly indicate the speed of new bone formation,and the fluorescence range could indirectly reflect the bone mass.The fluorescence band spacing and fluorescence range in the control group were narrow,while those in the experimental group were large,with wide band spacing.New bone formation was more active than that in the control group.Conclusion: The iconography,hard tissue section staining and osteogenesis-related gene expression levels of the Xiaochaihu Decoction cut +Mg-Zn-Ca amorphous alloy group are superior to those of the Mg-Zn-Ca amorphous alloy group.Besides,its good biological safety has been confirmed by hematological tests and histology of vital organs.Therefore,Xiaochaihu Decoction cut under the guidance of the theory of "Shaoyang governing bone" has the effect of promoting the repair of bone defects with Mg-Zn-Ca amorphous alloy. |
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