Application of Recombinant Human Fibroblast Growth Factor-5 in Promoting Fracture Healing
A technology of human fibroblasts and growth factors, applied in the field of medicine, can solve the problems of not shortening the fracture healing process, preventing delayed fracture healing or fracture nonunion, etc., and achieves the effect of simple operation
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Embodiment 1
[0031] Example 1. FGF5 changes in a trend during the fracture healing process
[0032] 1. Establish a mouse model of open femur fracture: In this experiment, 20-30 g male Kunming mice were anesthetized by intraperitoneal injection of 1.2% tribromoethanol. The femur was cut neatly, and the two broken ends were connected with a 1ml sterile syringe needle for fixation, and an open fracture model was successfully established. Experimental animals were divided into blank control group (Control group), which stripped muscle and tissue without cutting femur; fracture model group (Model group), which stripped muscle and tissue and cut femur.
[0033] 2. Observation results: 24 hours, 1 week and 2 weeks after modeling, X-ray scanning was used to detect the healing degree of the fracture ends. The results showed that obviously enlarged granulation tissue could be observed to wrap the fracture ends after 2 weeks of fracture healing, which was basically completed. Osseointegration of stu...
Embodiment 2
[0034] Example 2. The role of rhbFGF5 in promoting bone fracture healing in mice
[0035] 1. Establishment of an open mouse femur fracture model: the method is the same as that described in Example 1. Divided into the model group (Model) stripped muscle and tissue and cut the femur; rhbFGF5 group, stripped muscle and tissue and cut the femur, when the needle connects the broken end, will contain rhbFGF5 (2μg / mm 2 ) collagen sponge fixed at the broken end. At 1 week, 2 weeks, and 3 weeks after operation, the fracture ends were taken for Micro-CT scanning and H&E staining to observe the morphological changes of the fracture ends. RT-PCR was used to detect the changes of mRNA levels of fracture healing markers such as BMP2, ALP and COL1 in fracture ends.
[0036] 2. Observation results:
[0037]2.1 The promoting effect of rhbFGF5 on callus formation in mouse femoral fracture model
[0038] Micro-CT technology was used to detect the healing of the fracture ends of the animals ...
Embodiment 3
[0047] Example 3. Effect of rhbFGF5 on mouse bone marrow mesenchymal stem cells
[0048] 1. The effect of rhbFGF5 on the proliferation and migration of mouse bone marrow mesenchymal stem cells
[0049] 1.1 Cell culture and experimental grouping:
[0050] 1.1.1 CCK-8 assay for cell viability
[0051] Culture mouse bone marrow mesenchymal stem cell line (BMSC) in a 96-well plate, add different concentrations of rhbFGF5 (1, 5, 10, 12.5, 25, 50, 100, 200ng / ml), at 37°C, 5% CO 2 The cells were collected after 24 hours of incubation, and the cell viability was detected by the CCK-8 method.
[0052] 1.1.2 Cell scratch test
[0053] Mouse BMSCs were cultured in six-well plates and divided into control group and rhbFGF5 group. Blank medium was added to the control group, and medium containing 5 ng / ml rhbFGF5 was added to the rhbFGF5 group. At 37°C, 5% CO 2 After 24 hours of incubation in the incubator, the scratch test was performed.
[0054] 1.2 Observation results:
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