Osteogenic Differentiation of adipose mesenchymal stem cells with BMP-2 embedded microspheres in a rotating bed bioreactor

Stefanie Boehm,Cornelia Kasper,Marcelle Machluf,Yael Lupu

doi:10.1186/1753-6561-5-s8-p74

Stefanie Boehm, Cornelia Kasper + Show 2 more

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https://doi.org/10.1186/1753-6561-5-s8-p74

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Abstract

scaffolds with a diameter of 65 mm and a thickness of 3 mm were used. The PLGA microspheres were produced by solvent evaporation and at a size of about 50 µm. The BMP-2 concentration in the microspheres was about 0.05 µg per mg PLGA. The total amount of BMP2 in the dynamic cultivations was 2.5 µg in each bioreactor. The BMP-2 release and the concentration in the medium was measured with a BMP-2 ELISA (Quantikine, R&D Systems) The BMP-2 concentration in medium samples of bioreactor 4 decreases almost linear from 875 pg/ml to zero pg/ml in the first 16 days of the dynamic cultivation. Cells

Highlights

Bone tissue engineering aims at the generation of functional bone tissue for replacement of defect bone tissue in order to reestablish normal function
During 28 days of dynamic cultivation about 200 mg glucose were consumed in each bioreactor
The glucose consumption increased during the cultivation indicating a continuous cell growth on the ceramics

Summary

Introduction

Bone tissue engineering aims at the generation of functional bone tissue for replacement of defect bone tissue in order to reestablish normal function. For this purpose mesenchymal stem cells (MSCs) are widely used since they can be isolated from different sources and can be differentiated in vitro into the mesenchymal lineages [1,2]. The ceramic discs were loaded with PLGA microspheres releasing BMP-2. For this experiments a system was used which allows the parallel cultivation of four bioreactors. During the cultivation glucose and lactate concentrations were measured and after the experiments histological stainings were performed (DAPI, von Kossa and alizarine red). The concentration of alkaline phosphatase was measured in medium samples and mRNA of the cells was isolated to perform RT-PCR to investigate the expression of different bone markers

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