Fabrication of tubular tissue constructs by centrifugal casting of cells suspended in an in situ crosslinkable hyaluronan‐gelatin hydrogel

Achieving the optimal cell density and desired cell distribution in scaffolds is a major goal of cell seeding technologies in tissue engineering. In order to reach this goal, a novel centrifugal casting technology was developed using in situ crosslinkable hyaluronan-based (HA) synthetic extracellular matrix (sECM). Living cells were suspended in a viscous solution of thiol-modified HA and thiol-modified gelatin, a polyethyleneglycol diacrylate crosslinker was added, and a hydrogel was formed during rotation. The tubular tissue constructs consisting of a densely packed cell layer were fabricated with the rotation device operating at 2000 rpm for 10 min. The majority of cells suspended in the HA mixture before rotation were located inside the layer after centrifugal casting. Cells survived the effect of the centrifugal forces experienced under the rotational regime employed. The volume cell density (65.6%) approached the maximal possible volume density based on theoretical sphere packing models. Thus, centrifugal casting allows the fabrication of tubular constructs with the desired redistribution, composition and thickness of cell layers that makes the maximum efficient use of available cells. Centrifugal casting in this sECM would enable rapid fabrication of tissue-engineered vascular grafts, as well as other tubular and planar tissue-engineered constructs.

[1]  B. Gupta,et al.  Biomechanics of human common carotid artery and design of novel hybrid textile compliant vascular grafts. , 1997, Journal of biomedical materials research.

[2]  Glenn D Prestwich,et al.  Attachment and spreading of fibroblasts on an RGD peptide-modified injectable hyaluronan hydrogel. , 2004, Journal of biomedical materials research. Part A.

[3]  Glenn D Prestwich,et al.  Disulfide cross-linked hyaluronan hydrogels. , 2002, Biomacromolecules.

[4]  Vladimir Mironov,et al.  Organ printing: computer-aided jet-based 3D tissue engineering. , 2003, Trends in biotechnology.

[5]  R R Markwald,et al.  Mixed cultures of avian blastoderm cells and the quail mesoderm cell line QCE-6 provide evidence for the pluripotentiality of early mesoderm. , 1997, Developmental biology.

[6]  G Olivetti,et al.  Morphometric Study of Early Postnatal Development in the Left and Right Ventricular Myocardium of the Rat: II. Tissue Composition, Capillary Growth, and Sarcoplasmic Alterations , 1980, Circulation research.

[7]  Fabio Palumbo,et al.  Disulfide-crosslinked hyaluronan-gelatin hydrogel films: a covalent mimic of the extracellular matrix for in vitro cell growth. , 2003, Biomaterials.

[8]  J. Vacanti,et al.  Tissue engineering : Frontiers in biotechnology , 1993 .

[9]  N Ohshima,et al.  Novel cell immobilization method utilizing centrifugal force to achieve high-density hepatocyte culture in porous scaffold. , 2001, Journal of biomedical materials research.

[10]  R M Nerem,et al.  Vascular tissue engineering. , 2001, Annual review of biomedical engineering.

[11]  W McIntosh,et al.  Transdermal photopolymerization for minimally invasive implantation. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[12]  T. Okano,et al.  Cell sheet engineering for myocardial tissue reconstruction. , 2003, Biomaterials.

[13]  G. Prestwich,et al.  Biocompatibility and stability of disulfide-crosslinked hyaluronan films. , 2005, Biomaterials.

[14]  E Bell,et al.  A blood vessel model constructed from collagen and cultured vascular cells. , 1986, Science.

[15]  A. Atala,et al.  A novel use of centrifugal force for cell seeding into porous scaffolds. , 2004, Biomaterials.

[16]  R Cancedda,et al.  Computer-based technique for cell aggregation analysis and cell aggregation in in vitro chondrogenesis. , 1997, Cytometry.

[17]  M. Shoichet,et al.  Creating porous tubes by centrifugal forces for soft tissue application. , 2001, Biomaterials.

[18]  Thomas M Truskett,et al.  Is random close packing of spheres well defined? , 2000, Physical review letters.

[19]  Glenn D Prestwich,et al.  In situ crosslinkable hyaluronan hydrogels for tissue engineering. , 2004, Biomaterials.

[20]  Smadar Cohen,et al.  Optimization of cardiac cell seeding and distribution in 3D porous alginate scaffolds. , 2002, Biotechnology and bioengineering.

[21]  R Langer,et al.  Functional arteries grown in vitro. , 1999, Science.

[22]  F A Auger,et al.  A completely biological tissue‐engineered human blood vessel , 1998, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.