A systematic study for optimal cell seeding and culture conditions in a perfusion mode bone-tissue bioreactor

被引:25
作者
Beskardes, Isil G. [1 ]
Aydin, Gizem [2 ]
Bektas, Seyma [2 ]
Cengiz, Alper [2 ]
Gumusderelioglu, Menemse [1 ,2 ]
机构
[1] Hacettepe Univ, Dept Chem Engn, TR-06800 Ankara, Turkey
[2] Hacettepe Univ, Dept Bioengn, TR-06800 Ankara, Turkey
关键词
Perfusion bioreactor; Dynamic cell seeding; Chitosan; Superporous hydrogels; Bone tissue engineering; MARROW STROMAL CELLS; MATRIX DEPOSITION; IN-VITRO; SCAFFOLDS; FLOW; DIFFERENTIATION; OSTEOBLASTS; CONSTRUCTS;
D O I
10.1016/j.bej.2018.01.006
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
Bioreactors have an important role in tissue engineering to improve cell seeding and cell culturing conditions for the production of clinically relevant bone grafts. In our study, we systematically investigated the effects of static and dynamic approaches on cell seeding and in vitro culturing of a scaffold-containing perfusion bioreactor system. For this purpose, MC3T3-E1 pre-osteoblasts were seeded on chitosan-hydroxyapatite scaffolds under different flow rates and directions. We showed that static seeding with a five-day preculture ensured higher mitochondrial activity during perfusion culture. Bidirectional perfusion culture at 0.1 and 0.5 mL/min were applied for 21 days after static seeding and 5 days of static preculture to evaluate the effects of flow rate on the viability, morphology and osteogenic differentiation of cells. We found out that 0.1 mLimin flow rate significantly increased the mitochondrial activity of the cells, whereas osteogenic differentiation could be enhanced by 0.5 mL/min flow rate. In the last part of the study, in combination with static culturing bidirectional perfusion at 0.1 mLimin was applied for cell seeding and culturing. According to the results obtained from our study, perfusion seeding followed by static culture neither increased the mitochondria! activity nor enhanced the expression of osteogenic genes of cells except for osteopontin gene. However, static seeding combined with perfusion culture enhanced cell viability and osteogenic differentiation. (C) 2018 Elsevier B.V. All rights reserved.
引用
收藏
页码:100 / 111
页数:12
相关论文
共 28 条
[1]   Flow perfusion improves seeding of tissue engineering scaffolds with different architectures [J].
Alvarez-Barreto, Jose F. ;
Linehan, Shawna M. ;
Shambaugh, Robert L. ;
Sikavitsas, Vassilios I. .
ANNALS OF BIOMEDICAL ENGINEERING, 2007, 35 (03) :429-442
[2]   Osteoblast adhesion on biomaterials [J].
Anselme, K .
BIOMATERIALS, 2000, 21 (07) :667-681
[3]   Fluid flow increases mineralized matrix deposition in 3D perfusion culture of marrow stromal osteloblasts in a dose-dependent manner [J].
Bancroft, GN ;
Sikavitsast, VI ;
van den Dolder, J ;
Sheffield, TL ;
Ambrose, CG ;
Jansen, JA ;
Mikos, AG .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2002, 99 (20) :12600-12605
[4]   Bone tissue engineering with scaffold-supported perfusion co-cultures of human stem cell-derived osteoblasts and cell line-derived osteoclasts [J].
Beskardes, Isil G. ;
Hayden, Rebecca S. ;
Glettig, Dean L. ;
Kaplan, David L. ;
Gumusderelioglu, Menemse .
PROCESS BIOCHEMISTRY, 2017, 59 :303-311
[5]   Microwave-assisted fabrication of chitosan-hydroxyapatite superporous hydrogel composites as bone scaffolds [J].
Beskardes, Isil Gercek ;
Demirtas, Tugrul Tolga ;
Durukan, Muge Dagli ;
Gumusderelioglu, Menemse .
JOURNAL OF TISSUE ENGINEERING AND REGENERATIVE MEDICINE, 2015, 9 (11) :1233-1246
[6]   Effects of medium perfusion rate on cell-seeded three-dimensional bone constructs in vitro [J].
Cartmell, SH ;
Porter, BD ;
García, AJ ;
Guldberg, RE .
TISSUE ENGINEERING, 2003, 9 (06) :1197-1203
[7]   3D Culture of Osteoblast-Like Cells by Unidirectional or Oscillatory Flow for Bone Tissue Engineering [J].
Du, Dajiang ;
Furukawa, Katsuko S. ;
Ushida, Takashi .
BIOTECHNOLOGY AND BIOENGINEERING, 2009, 102 (06) :1670-1678
[8]   A Single Short Session of Media Perfusion Induces Osteogenesis in hBMSCs Cultured in Porous Scaffolds, Dependent on Cell Differentiation Stage [J].
Filipowska, Joanna ;
Reilly, Gwendolen C. ;
Osyczka, Anna M. .
BIOTECHNOLOGY AND BIOENGINEERING, 2016, 113 (08) :1814-1824
[9]  
Gardel LS, 2014, TISSUE ENG PART B-RE, V20, P126, DOI [10.1089/ten.teb.2013.0010, 10.1089/ten.TEB.2013.0010]
[10]   Novel markers of osteogenic and adipogenic differentiation of human bone marrow stromal cells identified using a quantitative proteomics approach [J].
Graneli, Cecilia ;
Thorfve, Anna ;
Ruetschi, Ulla ;
Brisby, Helena ;
Thomsen, Peter ;
Lindahl, Anders ;
Karlsson, Camilla .
STEM CELL RESEARCH, 2014, 12 (01) :153-165