A multi-cytokines sequential slow-release microsphere-hydrogel composite system and its preparation method

A composite system and slow-release microsphere technology, which is applied in the field of multi-cytokine sequential slow-release microsphere-hydrogel composite system and its preparation field, can solve the problem that seed cells and cytokines cannot aggregate, proliferate, and affect the repair of defective cartilage. Issues such as speed and extent, and reduced efficacy of stem cell homing

Inactive Publication Date: 2017-12-08
HANGZHOU CITY XIAOSHAN DISTRICT TRADITIONAL CHINESE MEDICAL HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] After the seed cells are implanted into the cartilage to be repaired, due to the daily activities of the body and the flow of blood and joint fluid in the damaged tissue in the body, the seed cells and cytokines cannot effectively gather and proliferate for a long time, which seriously affects the repair of the defective cartilage. Normal repair speed and extent
Studies on the migration and homing of BMSCs cultured in vitro to the infarcted myocardium showed that the uptake rate of MSCs in the infarcted heart at each time period was lower than 1%, which greatly reduced the efficacy of stem cell homing.

Method used

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  • A multi-cytokines sequential slow-release microsphere-hydrogel composite system and its preparation method
  • A multi-cytokines sequential slow-release microsphere-hydrogel composite system and its preparation method
  • A multi-cytokines sequential slow-release microsphere-hydrogel composite system and its preparation method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] Example 1. Preparation of IGF-1-loaded PLGA Microspheres

[0048] The W / O / W emulsification-solvent evaporation method was used to prepare IGF-1-loaded PLGA microspheres. The specific process was as follows: 1 mg of rhIGF-1 freeze-dried powder was dissolved in 100 μl of deionized water to prepare an aqueous solution with a concentration of 10 mg / ml as The inner aqueous phase (W1 phase). Take 500mg of PLGA and 140mg of Span-80 dissolved in 10ml of dichloromethane, as the oil phase (O phase). Add the W1 phase to the O phase, and ultrasonically emulsify for 1 min with an ultrasonic cell pulverizer under ice bath to form the primary emulsion W1 / O. Slowly and uniformly add W1 / O primary emulsion into 100ml of 2% (w / v) PVA aqueous solution containing 150mg tween-80 (outer water phase W2) under continuous stirring, and stir magnetically at a constant speed of 3000rpm under ice bath for 10min to form W1 / O / W2 double layer complex emulsion. Then magnetically stir at a constant ...

Embodiment 2

[0049] Example 2. Preparation of TGF-ß3-loaded PLGA microspheres

[0050] The W / O / W emulsification-solvent evaporation method was used to prepare TGF-ß3-loaded PLGA microspheres. The specific process was as follows: 1 mg of rhTGF-ß3 lyophilized powder was dissolved in 100 μl of deionized water to prepare an aqueous solution with a concentration of 10 mg / ml as The inner aqueous phase (W1 phase). Take 500mg of PLGA and 140mg of Span-80 dissolved in 10ml of dichloromethane, as the oil phase (O phase). Add the W1 phase to the O phase, and ultrasonically emulsify for 1 min with an ultrasonic cell pulverizer under ice bath to form the primary emulsion W1 / O. Slowly and uniformly add W1 / O primary emulsion into 100ml of 2% (w / v) PVA aqueous solution containing 150mg tween-80 (outer water phase W2) under continuous stirring, and stir magnetically at a constant speed of 3000rpm under ice bath for 10min to form W1 / O / W2 double layer complex emulsion. Then magnetically stir at a constan...

Embodiment 3

[0051] Example 3. Preparation of PLGA-PEG-PLGA hydrogel

[0052] The PLGA-PEG-PLGA block copolymer hydrogel was prepared by the ring-opening polymerization method. The specific method is: weigh a certain amount of DLLA, GA and PEG1500 according to the ratio of DLLA:GA:PEG=6:1:3 and put them in the oven After drying in medium for 2 hours, mix it and add it to the vacuum polymerization reaction bottle, place it at room temperature for 2 hours to cool, then add the catalyst stannous isooctanoate (0.02%, W / W), repeat nitrogen and vacuum, remove trace moisture and oxygen, Vacuum seal the tube, heat to 150°C and react for 10 hours to obtain PLGA-PEG copolymer. Afterwards, the polymer was dissolved in dichloromethane, and ethanol was precipitated out to obtain a purified polymer, which was dried in vacuo to constant weight to obtain a viscous transparent product, which was stored at -20°C for future use.

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Abstract

The invention relates to a multi-cytokines sequential slow-release microsphere-hydrogel composite system and a preparation method thereof. A multi-cytokines sequential slow-release microsphere-hydrogel composite system, the composite system includes the following components: IGF-1 PLGA microspheres, TGF-ß3 PLGA microspheres, SDF-1 and PLGA-PEG-PLGA . Due to the adoption of the above-mentioned technical scheme, the present invention integrates SDF-1 and PLGA slow-release microspheres respectively wrapped with IGF-1 and TGF-ß3 into the PLGA-PEG-PLGA thermosensitive hydrogel, so that it can Consecutive and sustained slow release of 3 kinds of growth factors, and gather stem cells in the cartilage damage site to improve the repair effect of articular cartilage damage.

Description

technical field [0001] The invention relates to a multi-cytokines sequential slow-release microsphere-hydrogel composite system and a preparation method thereof. Background technique [0002] Growth factor (GF) is a kind of protein substances that can promote or inhibit cell proliferation, differentiation, migration and gene expression, and play an important role in regulating cell division, matrix synthesis and tissue differentiation. It is an indispensable part of engineering technology and plays an important role in the repair of damaged tissues. Studies have shown that many growth factors are involved in the repair process of articular cartilage, mainly including insulin-like growth factor (insulin-like growth factor, IGF) family, transforming growth factor (Transforming Growth Factor, TGF) superfamily and so on. Insulin-like growth factor-1 (insulin-like growth factor-Ⅰ, IGF-Ⅰ) can promote the proliferation and redifferentiation of chondrocytes, maintain the specific p...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K9/06A61K38/19A61K47/34A61P19/08A61L27/52A61L27/54
Inventor 黄忠名李俊华胡文跃全仁夫
Owner HANGZHOU CITY XIAOSHAN DISTRICT TRADITIONAL CHINESE MEDICAL HOSPITAL
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