Platelet-Rich Plasma Combined With Yipanzhu Decoction Promote The Repair Of Tendon-Bone Junction In A Rabbit Model

Research Purposes: Based on the Traumatology of TCM,this study intends to place platelet-rich plasma of rabbit-self to damage model of rabbit Tendon-bone junction,guide and promote the release of various local cytokines and growth factors,and apply a Yanpanzhu decoction for treatment after surgery.This study is aimed at analyzing the combined effect of Yipanzhu decotion and platelet-rich plasma on the healing and regeneration of different tissues at the interface of the injury,which provides a new thinking for biological intervention after the injury of tibia’s ligament.Research Methods: A total of 144 mature New Zealand rabbits were given partial humeral resection to establish a damage model of rabbit tibia’s ligament.All the rabbits were divided into control group and Yipanzhu group(YPZ group): After operation,rabbits were given 5ml/kg of Yipanzhu(6g/kg)through intragastrical administration one times a day.Platelet-rich plasma group(PRP group): PRP was prepared by two centrifugation methods before operation.The prepared PRP was mixed with 10% calcium chloride solution + 200 U bovine thrombin at the same time before suturing the tendon and remaining tibia.Platelet-rich plasma combined with Yipanzhutang group(PRP+YPZ group): The surgical procedure and PRP intervention were the same as the PRP group.The application of Yipanzhu decoction was the same as that of the YPZ group.The iliac crest complex specimens were collected at 4 and 8 weeks postoperatively.q PCR was used to detect the expression of growth factors TGF-β1 m RNA,BMP-4 m RNA and Sox-9 m RNA;At 4 and 8 weeks postoperatively,histological H&E and Safranin/fast green staining were performed to observe the repair of the morphological structure of the callus junction;X-ray imaging was performed 8 weeks after surgery to measure the area and length of new bone at the epiphysis interface;At 8 weeks postoperatively,the specimens were scanned by synchronous radiation micro-CT for three-dimensional high-definition visualization,and the morphological parameters of the new bone microstructure were quantitatively evaluated;At 8 weeks postoperatively,the specimens were subjected to biomechanical tests to evaluate their mechanical properties such as tensile load and ultimate strength.Research Results:(1)q PCR detection: At 4 weeks postoperatively,the expression levels of TGF-β1 m RNA,BMP-4 m RNA and Sox-9 m RNA in the Tendon-bone junction of PRP group and PRP+YPZ group were significantly higher than those of the control group,and the difference was very significant(P< 0.01).There was no significant difference between YPZ group and control group(P>0.05),and the expression levels of TGF-β1 m RNA,BMP-4 m RNA and Sox-9 m RNA in the PRP group and PRP+YPZ group were the same.The difference was significantly higher than that of the YPZ group(P<0.01),but there was no significant difference between the PRP group and the PRP+YPZ group(P>0.05).At 8 weeks postoperatively,the expression levels of TGF-β1 m RNA,BMP-4 m RNA and Sox-9 m RNA in the YPZ group and PRP+YPZ group were significantly higher than those of the control group,the difference was very significant(P<0.01).There was no significant difference between the PRP group and the control group(P>0.05),and the expression levels of TGF-β1 m RNA,BMP-4 m RNA and Sox-9 m RNA in the YPZ group and the PRP+YPZ group were significantly different.There was no significant difference in expression level between the YPZ group and the PRP+YPZ group(P>0.05).(2)Histological examination: Compared with the control group,the growth and fusion of different tissues in the YPZ group,PRP group and PRP+YPZ group were more obvious.The 4th week after operation,the main part showed the increase of trabecular bone in the osteotomy surface of the tibial junction.The content and distribution of proteoglycan in the fibrocartilage layer and cartilage metaplasia area at the injury interface increased significantly in the last 8 weeks;and the growth and fusion of different tissues in the PRP+YPZ group were better than those in the YPZ group and PRP group.(3)X-ray detection: At 8 weeks after operation,the new bones gradually appeared at the osteotomy of the Tendon-bone junction,and the new bone mass of the three treatment groups was higher than that of the control group,and the area and length of the new bone in the PRP+YPZ group were significantly larger than the YPZ group and the PRP group,and there was no significant difference in the area and length of the new bone between the YPZ group and the PRP group.(4)Synchrotron radiation and micro-CT scanning: The bone volume fraction and trabecular bone number of the new bone in the YPZ group,PRP group and PRP+YPZ group were significantly higher than those in the control group.The trabecular thickness of the new bone in the PRP+YPZ group was significantly higher than that in the control group,and the new bone in the PRP+YPZ group.The Tb.Sp was significantly smaller than that of the control group;the bone volume fraction and trabecular bone number of the new bone in the PRP+YPZ group were significantly higher than those in the YPZ group.(5)Biomechanical test: In the YPZ group,PRP group and PRP+YPZ group,the tensile load,tensile energy and ultimate strength mechanical performance parameters of the tibia-bone complex specimens were superior to the control group,and the PRP+YPZ group was superior to YPZ group and PRP group.Research Conclusion: Platelet-rich plasma combined with Yipanzhu decoction can promote the repair of rabbit Tendon-bone junction injury,which is mainly to promote the growth and fusion of the tendon-bone interface,the formation of new bone and the remodeling of its fine structure,and enhance the healing in the middle and late healing stage.The bone interface of the fibrocartilage layer is regenerated and its mechanical properties are improved.