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Method for preparing basic fibroblast growth factor sustained-release carrier

A technology of fibroblasts and slow-release carrier, which can be applied to medical preparations containing active ingredients, pharmaceutical formulas, peptide/protein components, etc. wait for the question

Inactive Publication Date: 2011-11-02
舒泰经贸(广州)有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The bFGF-PLGA slow-release carrier disclosed in Chinese patent 02133476.5 can only be used in joint space, fracture or intravenous administration, and has shortcomings in the application of large wound healing, and the application range is relatively narrow, and because it is a synthetic polymer material, it has Hydrophobic, so there is a disadvantage of poor tissue biocompatibility during use
The bFGF-collagen sponge carrier disclosed in Chinese patent 200710010844.4, the collagen sponge has good water absorption and high biocompatibility, but as the carrier of bFGF alone, the microfibers of the collagen sponge have only a low affinity with bFGF, and the physical adsorption method Loading bFGF, because of its porosity, once in contact with body fluids or water, the sponge will swell and degrade too quickly, which is not conducive to the maintenance of bFGF activity, and cannot effectively release bFGF slowly

Method used

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  • Method for preparing basic fibroblast growth factor sustained-release carrier
  • Method for preparing basic fibroblast growth factor sustained-release carrier

Examples

Experimental program
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Effect test

Embodiment 1

[0019] 1. Dissolve 100mg of PLGA (PLA:PLG=50:50) in dichloromethane as the oil phase, dissolve 70ug of bFGF sample in double distilled water as the water phase, then add the bFGF solution into the oil phase and homogenize Then colostrum is formed. Add 40ml of 1% PVA solution, through the action of high-speed milk homogenizer (20000rpm, 2min) to form W / O / W, remove the organic solvent by rotating under reduced pressure with a rotary evaporator, centrifuge to precipitate the microspheres and freeze-dry to obtain the encapsulation PLGA Microspheres of bFGF.

[0020] 2. Add type I collagen to 0.15% acetic acid solution, stir well, and prepare collagen solution.

[0021] 3. Add the microspheres prepared in step 1 to the collagen solution in step 2, stir evenly, add to the mold (or a 96-well plate), pre-freeze at -70°C for 24 hours, and then perform conventional freeze-drying. 80% ethanol was used to solidify for 25 minutes, and then conventional freeze-drying was performed again t...

Embodiment 2

[0023] 1. Dissolve 140mg of PLGA (PLA:PLG=50:50) in dichloromethane as the oil phase, dissolve 90ug of bFGF sample in double distilled water as the water phase, then add the bFGF solution into the oil phase and homogenize to form colostrum. Add 40ml of 1% PVA solution, through the action of high-speed milk homogenizer (20000rpm, 2min) to form W / O / W, remove the organic solvent by rotating under reduced pressure with a rotary evaporator, centrifuge to precipitate the microspheres and freeze-dry to obtain the encapsulation PLGA Microspheres of bFGF.

[0024] 2. Add type I collagen to 0.2% acetic acid solution, stir well, and prepare collagen solution.

[0025] 3. Add the microspheres prepared in step 1 to the collagen solution in step 2, stir evenly, add to the mold (or a 96-well plate), pre-freeze at -70°C for 12 hours, and then perform conventional freeze-drying. The collagen sponge carrier embedded with bFGF-PLGA microspheres was obtained by solidifying with 75% ethanol for ...

Embodiment 3

[0027] 1. Dissolve 155mg of PLGA (PLA:PLG=50:50) in dichloromethane as the oil phase, dissolve 100ug of bFGF sample in double distilled water as the water phase, then add the bFGF solution into the oil phase and homogenize to form colostrum. Add 40ml of 1% PVA solution, through the action of high-speed milk homogenizer (20000rpm, 2min) to form W / O / W, remove the organic solvent by rotating under reduced pressure with a rotary evaporator, centrifuge to precipitate the microspheres and freeze-dry to obtain the encapsulation PLGA Microspheres of bFGF.

[0028] 2. Add type I collagen to 0.15% acetic acid solution, stir well, and prepare collagen solution.

[0029] 3. Add the microspheres prepared in step 1 to the collagen solution in step 2, stir evenly, add to the mold (or a 96-well plate), pre-freeze at -70°C for 18 hours, and then perform conventional freeze-drying. The collagen sponge carrier embedded with bFGF-PLGA microspheres was obtained by solidifying with 80% ethanol for ...

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Abstract

The invention discloses a method for preparing a basic fibroblast growth factor (bFGF) sustained-release carrier, which comprises the following steps of: 1, preparing bFGF-poly(lactide-co-glycolide) (PLGA) sustained-release microspheres encapsulating 0.00001 to 0.001 percent of bFGF; 2, dissolving type I collagen in an acetic acid solution to obtain a collagen solution; 3, mixing and stirring the bFGF-PLGA sustained-release microspheres and the collagen solution, performing freeze drying, solidifying, and performing freeze drying again to obtain a bFGF-PLGA collagen sustained-release carrier; and 4, performing in-vitro release experiments and Balb / c3T3 cell proliferation promotion experiments, wherein experimental results show that the sustained-release carrier has good biocompatibility, can slowly control the release of the bFGF and effectively promote Balb / c3T3 cell growth, and is an ideal bFGF sustained-release carrier.

Description

technical field [0001] The invention relates to a preparation method for constructing a slow-release carrier of basic fibroblast growth factor by using type I collagen and lactic acid-glycolic acid copolymer (PLGA) microspheres. Background technique [0002] 1. Basic fibroblast growth factor is a biologically active substance that can widely promote the proliferation of cells derived from mesoderm and neuroectoderm, such as epithelial cells, nerve cells, and vascular endothelial cells. Exogenous bFGF can also promote the release of endogenous bFGF and other growth factors, upregulate the activity of growth factors and the expression of growth factor receptors, and play biological roles. In recent years, it has been widely used in clinical medicine and cosmetics, and has shown great application potential in the fields of burns, trauma, nerve injury and vascular injury repair. However, basic fibroblasts have a short half-life in vivo, low local drug concentration, are sensiti...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K47/42A61K47/34A61K9/00A61K38/18A61P17/02
Inventor 罗丽华万宇陈凡徐华吴容英
Owner 舒泰经贸(广州)有限公司
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