Injectable PLGA (poly(lactic-co-glycolic acid)) porous composite microsphere preparation embedded with BMP-2 and TGF-beta1 containing microspheres as well as preparation method and application of composite microsphere preparation
A technology of porous composite microspheres and BMP-2, which is applied in the field of medicine to reduce systemic side effects and improve curative effect
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Embodiment 1
[0068] Embodiment 1: preparation contains the polylactic acid glycolic acid ( Poly(lactic-co-glycolicacid, PLGA) porous composite microspheres
[0069] A, prepare chitosan microspheres containing BMP-2 or TGF-β1:
[0070] 根据文献报道(KimSE,ParkJH,ChoYW,ChungH,JeongSY,LeeEB,etal.Porouschitosanscaffoldcontainingmicrospheresloadedwithtransforminggrowthfactor-beta1:implicationsforcartilagetissueengineering.Journalofcontrolledrelease:officialjournaloftheControlledReleaseSociety.2003;91:365-74),采用如下方法制备:称取120mg壳聚糖(分子量 100000, Zhejiang Jinke Biochemical Co., Ltd.), add 3mL 2% (V / V) acetic acid aqueous solution, vortex to dissolve, add 1mL 1% (W / V) BMP-2 (U.S. Peprotech company) aqueous solution, mix After uniformity, drop in 90mL n-octanol (containing 4% Span 80, V / V), stir at 1200 rpm for 30 minutes, then add dropwise 10mL 5% (W / V) sodium tripolyphosphate (Sodiumtripolyphosphate, TPP, U.S. Amresco company) for cross-linking reaction, after continuing to stir for 1 hour, the chitosan mic...
Embodiment 2
[0074] Example 2: Morphological observation of PLGA porous composite microspheres embedded with BMP-2 and TGF-β1 chitosan microspheres
[0075] Get a certain amount of BMP-2 or TGF-β1 chitosan microspheres prepared in Example 1, and the PLGA porous composite microspheres containing chitosan microspheres, place on the sample plate that is glued with double-sided adhesive , observed under a scanning electron microscope (ZESSMA10, Germany) after gold plating, and obtained chitosan microspheres ( figure 2 A, the scale is 20 μm) and PLGA porous composite microspheres containing BMP-2 and TGF-β1 chitosan microspheres ( figure 2 B, morphology with scale bar 100 μm).
[0076] Chitosan microspheres are round in shape and smooth in surface, such as figure 2 As shown in A. The shape of PLGA porous microspheres is round, and a large number of pores are randomly distributed on the surface, such as figure 2 Shown in B.
Embodiment 3
[0077] Embodiment 3: the particle size determination of the PLGA porous composite microspheres embedded with BMP-2 and TGF-β1 chitosan microspheres
[0078] The PLGA porous composite microspheres containing BMP-2 or TGF-β1 chitosan microspheres inlaid prepared in Example 1 were dispersed in deionized water, and the particle diameter was measured with Malvern Masersizer2000 ( image 3 A). Randomly select 3 microspheres from the scanning electron microscope photos of porous microspheres, measure their surface pore size (negligible if the pore size is less than 5 μm), take the average value, and the pore size distribution diagram is as follows image 3 Shown in B. Measure the particle diameter of the microspheres on the scanning electron microscope photos containing BMP-2 or TGF-β1 chitosan microspheres, get the average value ( image 3 C).
[0079] Such as image 3 As shown in A, the average particle size of the PLGA porous composite microspheres embedded with BMP-2 and TGF-...
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