In vitro assessment of cell penetration into porous hydroxyapatite scaffolds with a central aligned channel.

There is a clinical need for synthetic scaffolds that promote bone regeneration. A common problem encountered when using scaffolds in tissue engineering is the rapid formation of tissue on the outer edge of the scaffold whilst the tissue in the centre becomes necrotic. To address this, the scaffold design should improve nutrient and cell transfer to the scaffold centre. In this study, hydroxyapatite scaffolds with random, open porosity (average pore size of 282+/-11microm, average interconnecting window size of 72+/-4microm) were manufactured using a modified slip-casting methodology with a single aligned channel inserted into the centre. By varying the aligned channel diameter, a series of scaffolds with channel diameters ranging from 170 to 421microm were produced. These scaffolds were seeded with human osteosarcoma (HOS TE85) cells and cultured for 8 days. Analysis of cell penetration into the aligned channels revealed that cell coverage increased with increasing channel diameter; from 22+/-3% in the 170microm diameter channel to 38+/-6% coverage in the 421microm channel. Cell penetration into the middle section of the 421microm diameter channel (average cell area coverage 121x10(3)+/-32x10(3)microm(2)) was significantly greater than that observed within the 170microm channel (average cell area coverage 26x10(3)+/-6x10(3)microm(2)). In addition, the data presented demonstrates that the minimum channel (or pore) diameter required for cell penetration into such scaffolds is approximately 80microm. These results will direct the development of scaffolds with aligned macroarchitecture for tissue engineering bone.

[1]  C. Lohmann,et al.  Migration, Matrix Production and Lamellar Bone Formation of Human Osteoblast-Like Cells in Porous Titanium Implants , 2002, Cells Tissues Organs.

[2]  Fields Rd,et al.  Dual-attribute continuous monitoring of cell proliferation/cytotoxicity. , 1993, American biotechnology laboratory.

[3]  Richard O C Oreffo,et al.  Bone tissue engineering: hope vs hype. , 2002, Biochemical and biophysical research communications.

[4]  K. Burg,et al.  Biomaterial developments for bone tissue engineering. , 2000, Biomaterials.

[5]  J. Bancroft,et al.  Theory and Practice of Histological Techniques , 1990 .

[6]  A. Mikos,et al.  Effect of convection on osteoblastic cell growth and function in biodegradable polymer foam scaffolds. , 2001, Biomaterials.

[7]  Robert E Guldberg,et al.  Microarchitectural and mechanical characterization of oriented porous polymer scaffolds. , 2003, Biomaterials.

[8]  H Oonishi,et al.  Orthopaedic applications of hydroxyapatite. , 1991, Biomaterials.

[9]  Tessa Hadlock,et al.  Manufacture of porous polymer nerve conduits by a novel low-pressure injection molding process. , 2003, Biomaterials.

[10]  Nick Medcalf,et al.  Functional assessment of tissue-engineered meniscal cartilage by magnetic resonance imaging and spectroscopy. , 2003, Tissue engineering.

[11]  G. Moonen,et al.  Poly(D,L-lactide) foams modified by poly(ethylene oxide)-block-poly(D,L-lactide) copolymers and a-FGF: in vitro and in vivo evaluation for spinal cord regeneration. , 2001, Biomaterials.

[12]  M S Chapekar,et al.  Tissue engineering: challenges and opportunities. , 2000, Journal of biomedical materials research.

[13]  Antonios G. Mikos,et al.  Fluid flow increases mineralized matrix deposition in 3D perfusion culture of marrow stromal osteoblasts in a dose-dependent manner , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[14]  A. Grodzinsky,et al.  Fluorometric assay of DNA in cartilage explants using Hoechst 33258. , 1988, Analytical biochemistry.

[15]  C. Perry,et al.  Bone repair techniques, bone graft, and bone graft substitutes. , 1999, Clinical orthopaedics and related research.

[16]  G. Lundborg,et al.  Rat sciatic nerve regeneration through a micromachined silicon chip. , 1997, Biomaterials.

[17]  S. Hollister,et al.  Optimal design and fabrication of scaffolds to mimic tissue properties and satisfy biological constraints. , 2002, Biomaterials.

[18]  L G Griffith,et al.  Survival and function of hepatocytes on a novel three-dimensional synthetic biodegradable polymer scaffold with an intrinsic network of channels. , 1998, Annals of surgery.

[19]  G. Moonen,et al.  Peripheral nerve regeneration using bioresorbable macroporous polylactide scaffolds. , 2000, Journal of biomedical materials research.

[20]  P. Zysset,et al.  Hydroxyapatite cement scaffolds with controlled macroporosity: fabrication protocol and mechanical properties. , 2003, Biomaterials.

[21]  Cato T Laurencin,et al.  Tissue engineered bone: measurement of nutrient transport in three-dimensional matrices. , 2003, Journal of biomedical materials research. Part A.

[22]  Christian Grandfils,et al.  Biodegradable and macroporous polylactide implants for cell transplantation: 1. Preparation of macroporous polylactide supports by solid-liquid phase separation , 1996 .

[23]  M J Yaszemski,et al.  Bone formation by three-dimensional stromal osteoblast culture in biodegradable polymer scaffolds. , 1997, Journal of biomedical materials research.

[24]  T. Einhorn Clinically applied models of bone regeneration in tissue engineering research. , 1999, Clinical orthopaedics and related research.

[25]  M J Yaszemski,et al.  Three-dimensional culture of rat calvarial osteoblasts in porous biodegradable polymers. , 1998, Biomaterials.

[26]  J. Vacanti,et al.  A polymer foam conduit seeded with Schwann cells promotes guided peripheral nerve regeneration. , 2000, Tissue engineering.

[27]  Dietmar W. Hutmacher,et al.  Scaffold design and fabrication technologies for engineering tissues — state of the art and future perspectives , 2001, Journal of biomaterials science. Polymer edition.

[28]  P. Ma,et al.  Microtubular architecture of biodegradable polymer scaffolds. , 2001, Journal of biomedical materials research.

[29]  C. Patrick,et al.  Manufacture of porous biodegradable polymer conduits by an extrusion process for guided tissue regeneration. , 1998, Biomaterials.

[30]  P Ducheyne,et al.  Bioactive ceramics: the effect of surface reactivity on bone formation and bone cell function. , 1999, Biomaterials.

[31]  Scott J Hollister,et al.  Mechanical and in vivo performance of hydroxyapatite implants with controlled architectures. , 2002, Biomaterials.

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