Design and validation of a biomechanical bioreactor for cartilage tissue culture

Specific tissues, such as cartilage, undergo mechanical solicitation under their normal performance in human body. In this sense, it seems necessary that proper tissue engineering strategies of these tissues should incorporate mechanical solicitations during cell culture, in order to properly evaluate the influence of the mechanical stimulus. This work reports on a user-friendly bioreactor suitable for applying controlled mechanical stimulation—amplitude and frequency—to three-dimensional scaffolds. Its design and main components are described, as well as its operation characteristics. The modular design allows easy cleaning and operating under laminar hood. Different protocols for the sterilization of the hermetic enclosure are tested and ensure lack of observable contaminations, complying with the requirements to be used for cell culture. The cell viability study was performed with KUM5 cells.

[1]  A. Huang,et al.  Long-term dynamic loading improves the mechanical properties of chondrogenic mesenchymal stem cell-laden hydrogel. , 2010, European cells & materials.

[2]  Zhen Li,et al.  Chondrogenesis of human bone marrow mesenchymal stem cells in fibrin-polyurethane composites is modulated by frequency and amplitude of dynamic compression and shear stress. , 2010, Tissue engineering. Part A.

[3]  D. Carter,et al.  Articular cartilage functional histomorphology and mechanobiology: a research perspective. , 2003, Bone.

[4]  Robert L Mauck,et al.  Mechanics and mechanobiology of mesenchymal stem cell-based engineered cartilage. , 2010, Journal of biomechanics.

[5]  B. A. Byers,et al.  The beneficial effect of delayed compressive loading on tissue-engineered cartilage constructs cultured with TGF-beta3. , 2007, Osteoarthritis and cartilage.

[6]  Keita Ito,et al.  Effect of TGF beta1, BMP-2 and hydraulic pressure on chondrogenic differentiation of bovine bone marrow mesenchymal stromal cells. , 2009, Biorheology.

[7]  Pauline M Doran,et al.  Chondrogenic differentiation of human adipose-derived stem cells in polyglycolic acid mesh scaffolds under dynamic culture conditions. , 2010, Biomaterials.

[8]  Rui L Reis,et al.  Effect of flow perfusion conditions in the chondrogenic differentiation of bone marrow stromal cells cultured onto starch based biodegradable scaffolds. , 2011, Acta biomaterialia.

[9]  K. Jepsen,et al.  Cyclic hydrostatic pressure enhances the chondrogenic phenotype of human mesenchymal progenitor cells differentiated in vitro , 2003, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[10]  P. Eggli,et al.  Chondrocyte biosynthesis correlates with local tissue strain in statically compressed adult articular cartilage , 1997, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[11]  G. Vunjak‐Novakovic,et al.  Gas exchange is essential for bioreactor cultivation of tissue engineered cartilage. , 1999, Biotechnology and bioengineering.

[12]  Stuart B Goodman,et al.  Effects of hydrostatic pressure and transforming growth factor-beta 3 on adult human mesenchymal stem cell chondrogenesis in vitro. , 2006, Tissue engineering.

[13]  Juan Alberto Panadero,et al.  Mechanical fatigue performance of PCL-chondroprogenitor constructs after cell culture under bioreactor mechanical stimulus. , 2016, Journal of biomedical materials research. Part B, Applied biomaterials.

[14]  Y. Toyama,et al.  Hyaline cartilage formation and enchondral ossification modeled with KUM5 and OP9 chondroblasts , 2007, Journal of cellular biochemistry.

[15]  R. Kandel,et al.  Cyclic compressive mechanical stimulation induces sequential catabolic and anabolic gene changes in chondrocytes resulting in increased extracellular matrix accumulation. , 2006, Matrix biology : journal of the International Society for Matrix Biology.

[16]  Mandi J. Lopez,et al.  In Vitro Mesenchymal Trilineage Differentiation and Extracellular Matrix Production by Adipose and Bone Marrow Derived Adult Equine Multipotent Stromal Cells on a Collagen Scaffold , 2013, Stem Cell Reviews and Reports.

[17]  D. Kelly,et al.  Hydrostatic pressure acts to stabilise a chondrogenic phenotype in porcine joint tissue derived stem cells. , 2012, European cells & materials.

[18]  Christopher R Jacobs,et al.  The role of mechanical signals in regulating chondrogenesis and osteogenesis of mesenchymal stem cells. , 2010, Birth defects research. Part C, Embryo today : reviews.

[19]  A. Braccini,et al.  Dynamic compression of cartilage constructs engineered from expanded human articular chondrocytes. , 2003, Biochemical and biophysical research communications.

[20]  Stephen D. Thorpe,et al.  THE EXTERNAL MECHANICAL ENVIRONMENT CAN OVERRIDE THE INFLUENCE OF LOCAL SUBSTRATE IN DETERMINING STEM CELL FATE , 2012 .

[21]  J. Steinmeyer,et al.  The proteoglycan metabolism of mature bovine articular cartilage explants superimposed to continuously applied cyclic mechanical loading. , 1997, Biochemical and biophysical research communications.

[22]  S. Thorpe,et al.  Dynamic compression can inhibit chondrogenesis of mesenchymal stem cells. , 2008, Biochemical and biophysical research communications.

[23]  S. Thorpe,et al.  European Society of Biomechanics S.M. Perren Award 2012: the external mechanical environment can override the influence of local substrate in determining stem cell fate. , 2012, Journal of biomechanics.

[24]  S. Bryant,et al.  The role of hydrogel structure and dynamic loading on chondrocyte gene expression and matrix formation. , 2008, Journal of biomechanics.

[25]  L. Bian,et al.  Dynamic compressive loading enhances cartilage matrix synthesis and distribution and suppresses hypertrophy in hMSC-laden hyaluronic acid hydrogels. , 2012, Tissue engineering. Part A.

[26]  J. A. Panadero,et al.  Fatigue prediction in fibrin poly-ε-caprolactone macroporous scaffolds. , 2013, Journal of the mechanical behavior of biomedical materials.