Comparison of chondrogensis in static and perfused bioreactor culture

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Pazzano, David¹; Mercier, Kathi A.^1,2; Moran, John M.^1,3; Fong, Stephen S.^1,3; DiBiasio, David D.³; Rulfs, Jill X.⁴; Kohles, Sean S.²; Bonassar, Lawrence J.¹

Comparison of chondrogensis in static and perfused bioreactor culture

Biotechnol Prog. September–October 2000;16(5):893-896

©2000 American Chemical Society and American Institute of Chemical Engineers

Affiliations

¹University of Massachusetts Medical School

²Department of Biomedical Engineering, Worcester Polytechnic Institute.

³Department of Chemical Engineering, Worcester Polytechnic Institute

⁴Department of Biology, Worcester Polytechnic Institute
Abstract

As a result of the low yield of cartilage from primary patient harvests and a high demand for autologous cartilage for reconstructive surgery and structural repair, primary explant cartilage must be augmented by tissue engineering techniques. In this study, chondrocytes seeded on PLLA/PGA scaffolds in static culture and a direct perfusion bioreactor were biochemically and histologically analyzed to determine the effects of fluid flow and media pH on matrix assembly. A gradual media pH change was maintained in the bioreactor within 7.4−6.96 over 2 weeks compared to a more rapid decrease from 7.4 to 6.58 in static culture over 3 days. Seeded scaffolds subjected to 1 μm/s flow demonstrated a 118% increase (p < 0.05) in DNA content, a 184% increase (p < 0.05) in GAG content, and a 155% (p< 0.05) increase in hydroxyproline content compared to static culture. Distinct differences were noted in tissue morphology, including more intense staining for proteoglycans by safranin-O and alignment of cells in the direction of media flow. Culture of chondrocyte seeded matrices thus offers the possibility of rapid in vitro expansion of donor cartilage for the repair of structural defects, tracheal injury, and vascularized tissue damage.
Links

DOI: 10.1021/bp000082v

PubMed: 11027186

WoS: 000089797600032
History

Accepted: 2000-07-18

Cited Works (5)

Year	Entry
1988	Gray ML, Pizzanelli AM, Grodzinsky AJ, Lee RC. Mechanical and physicochemical determinants of the chondrocyte biosynthetic response. J Orthop Res. November 1988;6(6):777-792.
1993	Freed LE, Marquis JC, Nohria A, Emmanual J, Mikos AG, Langer R. Neocartilage formation in vitro and in vivo using cells cultured on synthetic biodegradable polymers. J Biomed Mater Res. January 1993;27(1):11-23.
1995	Kim Y-J, Bonassar LJ, Grodzinsky AJ. The role of cartilage streaming potential, fluid flow and pressure in the stimulation of chondrocyte biosynthesis during dynamic compression. J Biomech. September 1995;28(9):1055-1066.
1988	Kim Y-J, Sah RLY, Doong J-YH, Grodzinsky AJ. Fluorometric assay of DNA in cartilage explants using Hoechst 33258. Anal Ciochem. October 1988;174(1):168-176.
1961	Woessner JF Jr. The determination of hydroxyproline in tissue and protein samples containing small proportions of this imino acid. Arch Biochem Biophys. May 1961;93(2):440-447.

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2003	Guilak F, Setton LA. Functional tissue engineering and the role of biomechanical signaling in articular cartilage repair. In: Guilak F, Butler DL, Goldstein SA, Mooney DJ, eds. Functional Tissue Engineering. New York, NY: Inc., Springer-Verlag; 2003:277-290.
2002	Mauck RL, Seyhan SL, Ateshian GA, Hung CT. Influence of seeding density and dynamic deformational loading on the developing structure/function relationships of chondrocyte-seeded agarose hydrogels. Ann Biomed Eng. September 2002;30(8):1046-1056.
2004	Hung CT, Mauck RL, Wang CC-B, Lima EG, Ateshian GA. A paradigm for functional tissue engineering of articular cartilage via applied physiologic deformational loading. Ann Biomed Eng. January 2004;32(1):35-49.
2016	Huang BJ, Hu JC, Athanasiou KA. Cell-based tissue engineering strategies used in the clinical repair of articular cartilage. Biomaterials. August 2016;98:1-22.
2001	Guilak F, Butler DL, Goldstein SA. Functional tissue engineering: the role of biomechanics in articular cartilage repair. Clin Orthop Relat Res. October 2001;391(suppl):S295-S305.
2003	Hung CT, Lima EG, Mauck RL, Taki E, LeRoux MA, Lu HH, Stark RG, Guo XE, Ateshian GA. Anatomically shaped osteochondral constructs for articular cartilage repair. J Biomech. December 2003;36(12):1853-1864.
2004	Kisiday JD, Jin M, DiMicco MA, Kurz B, Grodzinsky AJ. Effects of dynamic compressive loading on chondrocyte biosynthesis in self-assembling peptide scaffolds. J Biomech. May 2004;37(5):595-604.
2004	Leddy HA, Awad HA, Guilak F. Molecular diffusion in tissue-engineered cartilage constructs: effects of scaffold material, time, and culture conditions. J Biomed Mater Res. August 15, 2004;B70(2):397-406.
2003	Mauck RL, Wang CC-B, Oswald ES, Ateshian GA, Hung CT. The role of cell seeding density and nutrient supply for articular cartilage tissue engineering with deformational loading. Osteoarthritis Cartilage. December 2003;11(12):879-890.
2014	Bhumiratana S, Eton RE, Oungoulian SR, Wan LQ, Ateshian GA, Vunjak-Novakovic G. Large, stratified, and mechanically functional human cartilage grown in vitro by mesenchymal condensation. Proc Natl Acad Sci USA. May 13, 2014;111(19):6940-6945.
2003	Darling EM, Athanasiou KA. Articular cartilage bioreactors and bioprocesses. Tissue Eng. February 2003;9(1):9-26.
2003	Mauck RL, Nicoll SB, Seyhan SL, Ateshian GA, Hung CT. Synergistic action of growth factors and dynamic loading for articular cartilage tissue engineering. Tissue Eng. August 2003;9(4):597-611.
2006	Gemmiti CV, Guldberg RE. Fluid flow increases type II collagen deposition and tensile mechanical properties in bioreactor-grown tissue-engineered cartilage. Tissue Eng. March 2006;12(3):469-479.
2015	Whitney GA. Characterization of the Frictional-Shear Damage Properties of Scaffold-Free Engineered Cartilage and Reduction of Damage Susceptibility by Upregulation of Collagen Content [PhD thesis]. Cleveland, OH: Case Western Reserve University; January 2015.
2003	Mauck RL. Functional Tissue Engineering of Articular Cartilage: The Effect of Physical Forces on the in Vitro Growth of Engineered Constructs [PhD thesis]. Columbia University; 2003.
2006	Chahine NO. Multi-Scale Measurements of the Mechanical and Transport Properties of Native and Engineered Articular Cartilage [PhD thesis]. Columbia University; 2006.
2007	Jiang J. The Role of Cellular Interactions in Osteochondral Graft Design and the Maintenance of a Stable Osteochondral Interface [PhD thesis]. Columbia University; 2007.
2007	Wan Q. Fundamental Theoretical and Experimental Studies for Applications Toward Functional Tissue Engineering of Articular Cartilage [PhD thesis]. Columbia University; 2007.
2008	Lima EG. The Tissue-Engineering of Functional Osteochondral Constructs for Cartilage Repair [PhD thesis]. Columbia University; 2008.
2014	Tan AR. Towards Clinical Use of Engineered Tissues for Cartilage Repair [PhD thesis]. Columbia University; 2014.
2017	Roach BL. Modulation of the in Vitro Mechanical and Chemical Environment for the Optimization of Tissue-Engineered Articular Cartilage [PhD thesis]. Columbia University; 2017.
2006	Roy R. Non-Enzymatic Glycation for the Processing of Type I Collagen Gels for Cartilage Tissue Engineering [PhD thesis]. Ithaca, NY: Cornell University; January 2006.
2010	Babalola OM. Effects of Physical Stimuli on Extracellular Matrix Assembly by Chondrocytes and Mesenchymal Stem Cells: Experimental and Finite Element Analyses [PhD thesis]. Ithaca, NY: Cornell University; May 2010.
2010	Degala S. Fluid Flow Regulates Articular Chondrocyte Mechanotransduction Pathways in 3D Culture [PhD thesis]. Ithaca, NY: Cornell University; May 2010.
2014	Puetzer JL. Development of Engineered Menisci With Native-Like Organization Using High Density Collagen Gels and Mechanical Stimulation [PhD thesis]. Ithaca, NY: Cornell University; August 2014.
2005	Case ND. Oscillatory Compressive Loading Effects on Mesenchymal Progenitor Cells Undergoing Chondrogenic Differentiation in Hydrogel Suspension [PhD thesis]. Atlanta, GA: Georgia Institute of Technology; May 2005.
2006	Gemmiti CV. Effect of Fluid Flow on Tissue-Engineered Cartilage in a Novel Bioreactor [PhD thesis]. Atlanta, GA: Georgia Institute of Technology; December 2006.
2008	Wang L. Tissue Engineering the TMJ Condyle Using Human Umbilical Cord Mesenchymal Stromal Cells [PhD thesis]. Lawrence, KS: University of Kansas; 2008.
2011	Yuan T-Y. Innovative Methods to Determine Material Properties of Cartilaginous Tissues and Application for Tissue Engineering [PhD thesis]. University of Miami; August 2011.
2003	Kisiday JD. In Vitro Culture of a Chondrocyte-Seeded Peptide Hydrogel and the Effects of Dynamic Compression [PhD thesis]. Cambridge, MA: Massachusetts Institute of Technology; June 2003.
2012	Brenner JM. Development and Validation of Large-Sized Engineered Cartilage Constructs in Full –thickness Chondral Defects in a Rabbit Model [Master's thesis]. Queen's University; January 2012.
2012	Khan AA. Effects of Additional Sodium Bicarbonate on Extra/intra Cellular Factors in a Continuous Flow Bioreactor for the Production of Tissue Engineered Articular Cartilage [PhD thesis]. Queen's University; October 2012.
2004	Darling EM. Phenotype of Passaged, Zonal Articular Chondrocytes [PhD thesis]. Houston, TX: Rice University; July 2004.
2005	Scott CC. Interferometry of Chondrocytes and Impact of Articular Cartilage [PhD thesis]. Houston, TX: Rice University; August 2005.
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2007	Koay EJ. Chondrocyte Biomechanics and Chondrogenesis [PhD thesis]. Houston, TX: Rice University; December 2007.
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2006	Nugent GE. Biomechanical Regulation of Articular Cartilage Metabolism of Proteoglycan 4 and Articular Surface Integrity [PhD thesis]. San Diego (UCSD), University of California; 2006.
2009	Hsieh-Bonassera ND. Cartilage Tissue Engineering With Human Chondrocytes from Osteoarthritic Knees and a Semi-Permeable Membrane [PhD thesis]. San Diego (UCSD), University of California; 2009.
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2014	Pourmohammadali H. Mechanical and Hydromechanical Stimulation of Chondrocytes for Articular Cartilage Tissue Engineering [PhD thesis]. University of Waterloo; 2014.