| ObjectiveThe study aimed to investigate the mechanism of BuShenHuoXue Decoction (BSHXD) promoting fracture healing under the guidance of traditional Chinese medicine (TCM) theory, "kidney governing bones" and "extravasated blood gone, callus grew, bone formed". Based on reviewing the latest literature related to Mesenchymal stem cells (MSCs) and fracture healing, the clinical effects of distal radius fractures (DRFs) in the elderly treating with BSHXD were analyzed, and animals experiment were conducted to explored the methods of promoting fracture healing better, which would provide new ideas and theoretical basis for the treatment of bone defects, fracture delayed union or nonunion, osteonecrosis, osteoporosis after long-term immobilize, and other intractable bone diseases by TCM.Methods1. Literature SearchMSCs were the most important cells during fracture healing with immense potential of self-proliferation, multilineage differentiation, homing to the injury, repairing the injury. Since it’s potential differentiation to osteoblasts and chondrocytes, and promoted fracture healing, MSCs became a research hotspot. Chemokines were a superfamily of small molecule secreted protein, which could induce MSCs migration. The latest research found that chemokines and their receptors could induce MSCs homing to injury, inflammation, ischemia and et al, involved in repairing the injury. Therefore, induction and summarizing the chemokines which could induce migration of MSCs and understanding its preliminary mechanism were meaningful to explore ways promoting fracture healing.2. Experimental StudyThe SD rat femur fracture model was conducted and treated with BSHXD. Then the MSCs in the fracture site was isolated and cultured at 7thday,14thday and 21st day respectively, and identified by surface antigen Immunofluorescence staining and differentiation to osteoblasts and chondrocytes. At the third passage, Transwell assay was applied to examine the migration of MSCs influenced by BSHXD. The expression of CCR2, a chemokine receptor, was detected by Western Blot and qPCR.3. Clinical Research60 patients of DRFs in the elderly were divided into the treatment group and control group randomly. The treatment group was treated with BSHXD for 4 weeks, while control group accepted nothing special treatment after reduction of fractures and got stable fixation. X-ray was applied to conduct the Radius union scoring system (RUSS) and identify the healing time of fracture. Functional evaluation was performed using the Gartland-Werley scoring system and the PRWE (patient-rated wrist evaluation) scale after fracture healing.Results1. Literature SearchChemokines and their receptors’multi-channel interaction approach could affect each stage of MSCs migration by a single or multiple chemokine and their receptors, blocking the integration of chemokine ligand and receptor, which may result in binding or changing the direction of the moving MSCs. At the same time, we could find that a single chemokine receptor activator could significantly promote MSCs migration in some experimental studies. Therefore, to make out whether the chemokines and their receptors expressed in fracture microenvironment and how did they work could not only help us find out the key chemokine which induce the MSCs migrate to the fracture site, but also further clarify the regulation of endogenous MSCs migrate to the fracture site promoting fracture healing. At the same time, if we could grasp the rules and mechanisms controlling the expression of chemokines, we would bring a new prospect to the treatment of fractures, bone defects, and promote fracture healing by the use of BSHXD regulating the expression of chemokine and its receptor.2. Experimental StudyThe MSCs in the fracture site was authenticated by surface antigen Immunofluorescence staining and differentiation to osteoblasts and chondrocytes confirming that the cells were mesenchymal stem cells. BSHXD could promote migration of MSCs significantly at 7th,14th day but not 21st day, compared with the model group and control group. Similarly the expression of CCR2 was up-regulated significantly at 7th,14th day but not 21st day, detected by Western Blot and qPCR.3. Clinical ResearchCompared with the control group, the treatment group got higher scores in 2nd,4th,6th week significantly, but not in 8th week according to RUSS. In the case of healing time, Gartland-Werley scoring system and the PRWE scale, the treatment group got better results significantly compared with the control group.Conelusions1. Chemokines and their receptors could induce MSCs homing to the fracture sites participating in fracture healing. To study the expression of its mechanism was meaningful to explore ways to promote fracture healing.2. BSHXD could promote migration of MSCs during fracture healing, and its mechanism may be associated with the up-regulation of chemokine receptors CCR2.3. BSHXD could promote healing of DRFs in the elderly, shorten healing time of fracture and improve the function of wrist. |
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