Design and Fabrication of Anatomical Bioreactor Systems Containing Alginate Scaffolds for Cartilage Tissue Engineering

The aim of the present study was to develop a tissue-engineering approach through alginate gel molding to mimic cartilage tissue in a three-dimensional culture system. The perfusion biomimetic bioreactor was designed to mimic natural joint. The shear stresses exerting on the bioreactor chamber were calculated by Computational Fluid Dynamic (CFD). Several alginate/bovine chondrocyte constructs were prepared, and were cultured in the bioreactor. Histochemical and immunohistochemical staining methods for the presence of glycosaminoglycan(GAG), overall matrix production and type II collagen protein were performed, respectively. The dynamic mechanical device applied a linear mechanical displacement of 2 mm to 10 mm. The CFD modeling indicated peak velocity and maximum wall shear stress were 1.706×10−3 m/s and 0.02407 dyne/cm 2, respectively. Histochemical and immunohistochemical analysis revealed evidence of cartilage-like tissue with lacunas similar to those of natural cartilage and the production of sulfated GAG of matrix by the chondrons, metachromatic territorial matrix-surrounded cells and accumulation of type II collagen around the cells. The present study indicated that when chondrocytes were seeded in alginate hydrogel and cultured in biomimetic cell culture system, cells survived well and secreted newly synthesized matrix led to improvement of chondrogenesis.

[1]  X. Sherry Liu,et al.  Engineering anatomically shaped human bone grafts , 2009, Proceedings of the National Academy of Sciences.

[2]  T. Okano,et al.  Thermo-responsive culture dishes allow the intact harvest of multilayered keratinocyte sheets without dispase by reducing temperature. , 2001, Tissue engineering.

[3]  Y. Tabata,et al.  Perfusion culture enhances osteogenic differentiation of rat mesenchymal stem cells in collagen sponge reinforced with poly(glycolic Acid) fiber. , 2005, Tissue engineering.

[4]  Charles A Vacanti,et al.  Tissue Engineering of Autologous Cartilage for Craniofacial Reconstruction by Injection Molding , 2003, Plastic and reconstructive surgery.

[5]  Hisatoshi Kobayashi,et al.  Ectopic bone formation in collagen sponge self-assembled peptide-amphiphile nanofibers hybrid scaffold in a perfusion culture bioreactor. , 2006, Biomaterials.

[6]  G. Ateshian,et al.  Anatomically shaped osteochondral constructs for articular cartilage repair. , 2003, Journal of biomechanics.

[7]  W. Bugbee,et al.  Shaped, Stratified, Scaffold-free Grafts for Articular Cartilage Defects , 2008, Clinical orthopaedics and related research.

[8]  F. Lim,et al.  Microencapsulated islets as bioartificial endocrine pancreas. , 1980, Science.

[9]  Masayuki Yamato,et al.  Functional bioengineered corneal epithelial sheet grafts from corneal stem cells expanded ex vivo on a temperature-responsive cell culture surface , 2004, Transplantation.

[10]  G. Ateshian,et al.  A layered agarose approach to fabricate depth‐dependent inhomogeneity in chondrocyte‐seeded constructs , 2005, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[11]  H. Park,et al.  Novel type of alginate gel-based adsorbents for heavy metal removal , 2004 .

[12]  R. T. Tran,et al.  Scaffold Sheet Design Strategy for Soft Tissue Engineering , 2010, Nature materials.

[13]  B. Obradovic,et al.  Evaluation of alginate hydrogels under in vivo–like bioreactor conditions for cartilage tissue engineering , 2010, Journal of materials science. Materials in medicine.

[14]  P. Chang Encapsulation for Somatic Gene Therapy , 1999, Annals of the New York Academy of Sciences.

[15]  Jochen Ringe,et al.  Stem cells for regenerative medicine: advances in the engineering of tissues and organs , 2002, Naturwissenschaften.

[16]  Gordana Vunjak-Novakovic,et al.  CHAPTER 13 – TISSUE ENGINEERING BIOREACTORS , 2000 .

[17]  Scott Hollister,et al.  Tissue formation and vascularization in anatomically shaped human joint condyle ectopically in vivo. , 2009, Tissue engineering. Part A.

[18]  Jason P. Gleghorn,et al.  Integration of layered chondrocyte-seeded alginate hydrogel scaffolds. , 2007, Biomaterials.

[19]  D J Mooney,et al.  Injection molding of chondrocyte/alginate constructs in the shape of facial implants. , 2001, Journal of biomedical materials research.

[20]  Joseph P Vacanti,et al.  Reconstruction of mandibular defects with autologous tissue-engineered bone. , 2004, Journal of oral and maxillofacial surgery : official journal of the American Association of Oral and Maxillofacial Surgeons.

[21]  Koichi Masuda,et al.  A novel two‐step method for the formation of tissue‐engineered cartilage by mature bovine chondrocytes: The alginate‐recovered‐chondrocyte (ARC) method , 2003, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[22]  Anthony Ratcliffe,et al.  Bioreactors and Bioprocessing for Tissue Engineering , 2002, Annals of the New York Academy of Sciences.

[23]  Jerry C. Hu,et al.  Articular Cartilage Tissue Engineering , 2009 .

[24]  D. Mooney Tissue Engineering of Autologous Cartilage for Craniofacial Reconstruction by Injection Molding; Sophia C. N. Chang, M.D., Ph.D., Geoffrey Tobias, M.D., Amit K. Roy, Ph.D., Charles A. Vacanti, M.D., and Lawrence J. Bonassar, Ph.D. , 2003 .

[25]  Jason P. Gleghorn,et al.  Adhesive properties of laminated alginate gels for tissue engineering of layered structures. , 2008, Journal of biomedical materials research. Part A.

[26]  Laurent David,et al.  Multi-membrane hydrogels , 2008, Nature.

[27]  M. Sefton,et al.  Tissue engineering. , 1998, Journal of cutaneous medicine and surgery.

[28]  Robert Langer,et al.  Principles of tissue engineering , 2014 .

[29]  C. Vacanti,et al.  Tissue-engineered composites of bone and cartilage for mandible condylar reconstruction. , 2001, Journal of oral and maxillofacial surgery : official journal of the American Association of Oral and Maxillofacial Surgeons.

[30]  Vassilios Sikavitsas,et al.  Tissue Engineering Bioreactors , 2006 .

[31]  K Masuda,et al.  Tissue engineering of stratified articular cartilage from chondrocyte subpopulations. , 2003, Osteoarthritis and cartilage.

[32]  Yasuhiko Tabata,et al.  Enhanced ectopic bone formation using a combination of plasmid DNA impregnation into 3-D scaffold and bioreactor perfusion culture. , 2006, Biomaterials.

[33]  O. Smidsrod,et al.  Molecular basis for some physical properties of alginates in the gel state , 1974 .

[34]  M. Gümüşderelioğlu,et al.  Computational fluid dynamics modeling of momentum transport in rotating wall perfused bioreactor for cartilage tissue engineering. , 2010, Journal of biotechnology.

[35]  J. Mao,et al.  Tissue-engineered Neogenesis of Human-shaped Mandibular Condyle from Rat Mesenchymal Stem Cells , 2003, Journal of dental research.

[36]  J. Vacanti,et al.  De Novo Cartilage Generation Using Calcium Alginate‐Chondrocyte Constructs , 1996, Plastic and reconstructive surgery.

[37]  G. Skjåk-Bræk,et al.  Alginate as immobilization matrix for cells. , 1990, Trends in biotechnology.

[38]  A. Pinzano,et al.  Designing a three-dimensional alginate hydrogel by spraying method for cartilage tissue engineering , 2010 .

[39]  Kathryn E. Smith,et al.  Influence of hydrostatic and distortional stress on chondroinduction. , 2008, Biorheology.

[40]  Jørgen Møller,et al.  A Study of the Constitution of Alginic Acid by Partial Acid Hydrolysis. , 1966 .