Application of recombinant human basic fibroblast growth factor-5 in promotion of 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 fracture healing delay or fracture nonunion, etc., and achieves the effect of simple operation

Active Publication Date: 2018-07-27
HARBIN MEDICAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, simply promoting bone calcium deposition cannot shorten the fracture healing process, nor can it prevent the occurrence of complications such as delayed fracture healing or fracture nonunion.
At present, there is no treatment strategy that can promote fracture healing at the cellular and molecular levels.

Method used

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  • Application of recombinant human basic fibroblast growth factor-5 in promotion of fracture healing
  • Application of recombinant human basic fibroblast growth factor-5 in promotion of fracture healing
  • Application of recombinant human basic fibroblast growth factor-5 in promotion of fracture healing

Examples

Experimental program
Comparison scheme
Effect test

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:

[0055] CC...

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Abstract

The invention discloses application of recombinant human basic fibroblast growth factor-5 (rhbFGF5) in preparation of drugs for promoting fracture healing. A collagen sponge containing the rhbFGF5 isfixed in an open mice femur fracture broken end. A result shows that the rhbFGF5 remarkably promotes the generation of osteoblast, bone trabecular and callus of the fracture broken end, shortens fracture healing course and reduces fracture complications. In vitro level experiment results show that the rhbFGF5 can remarkably increase mRNA expression levels of fracture healing marker molecules suchas alkaline phosphatase (ALP), collagen I (COL1) and bone morphogenetic protein-2 (BMP2), promote proliferation, migration and differentiation of mesenchymal stem cell in bone marrow, and promote proliferation of mature osteoblast. The application of the rhbFGF5 provided by the invention provides a powerful theoretical basis for the fact that rhbFGF5 can be used for preparing drugs for treating fracture and provides a more effective treatment manner for promoting fracture healing in clinic. The rhbFGF5 has characteristics of being safe and effective and being easy to operate, and significancefor fracture healing in clinic.

Description

technical field [0001] The invention relates to a new application of recombinant human fibroblast growth factor-5, in particular to the application of recombinant human fibroblast growth factor-5 in promoting fracture healing. The invention belongs to the technical field of medicine. Background technique [0002] Fracture is a common exogenous trauma in clinical practice. As my country's population ages and the incidence of tumors increases year by year, the incidence of pathological fractures secondary to traumatic fractures, senile osteoporosis, or tumor metastasis is increasing sharply. . The process of fracture healing is to restore the biological functions of injured tissues and organs, but the process is complicated, takes a long time to heal, and has many complications, which not only seriously affects the quality of life of patients, but also causes serious economic burdens to society and families. . According to clinical statistics, the incidence of fracture nonun...

Claims

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Application Information

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IPC IPC(8): A61K38/18A61P19/08
CPCA61K38/1825
Inventor 杨磊杜智敏许超千王宁
Owner HARBIN MEDICAL UNIVERSITY
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