Developing macroporous bicontinuous materials as scaffolds for tissue engineering.

Calcareous skeletal elements (ossicles) isolated from the seastar, Pisaster giganteus, were characterized and tested as potential biocompatible substrates for cellular attachment. These ossicles have a remarkably robust open-framework architecture with an interconnected network of ca. 10 microm diameter pores. Scanning electron and confocal microscopy was used to characterize the cell-substrate interaction. Cell culturing experiments revealed that the cells firmly attach to the ossicle surface, forming cell aggregates of several layers thick. The anchored cells extended to form 'bridges' between the openings in the bicontinuous framework and the degree of coverage increased as culture time progressed. Osteoblasts grown on the ossicles were found to be viable up to 32 days after initial seeding, as proven by assaying with AlamarBlue and FDA/PI staining indicating the ossicle's potential as an alternative highly effective tissue scaffold. Given the limitation in availability of this natural material, the results presented here should be seen as offering guidelines for future development of synthetic materials with physical and chemical properties strongly conducive to bone repair and restoration.

[1]  A. Meunier,et al.  Tissue-engineered bone regeneration , 2000, Nature Biotechnology.

[2]  V. Goldberg,et al.  The Effect of Implants Loaded with Autologous Mesenchymal Stem Cells on the Healing of Canine Segmental Bone Defects* , 1998, The Journal of bone and joint surgery. American volume.

[3]  M. Heidaran,et al.  Molecular Signaling in Bioengineered Tissue Microenvironments , 2002, Annals of the New York Academy of Sciences.

[4]  L L Hench,et al.  Osteoblast attachment and mineralized nodule formation on rough and smooth 45S5 bioactive glass monoliths. , 2004, Journal of biomedical materials research. Part A.

[5]  N. Forest,et al.  In vitro bone formation on coral granules , 1990, In Vitro Cellular & Developmental Biology.

[6]  H. Akçakaya,et al.  Effects of tricalcium phosphate bone graft materials on primary cultures of osteoblast cells in vitro. , 2004, Clinical oral implants research.

[7]  J B Lian,et al.  Expression of differentiated function by mineralizing cultures of chicken osteoblasts. , 1987, Developmental biology.

[8]  A. Salgado,et al.  Isolation and osteogenic differentiation of bone-marrow progenitor cells for application in tissue engineering. , 2004, Methods in molecular biology.

[9]  E. Shors Coralline bone graft substitutes. , 1999, The Orthopedic clinics of North America.

[10]  D. Wilson,et al.  Comparative study of the osteoinductive properties of bioceramic, coral and processed bone graft substitutes. , 1995, Biomaterials.

[11]  A. Vaccaro The role of the osteoconductive scaffold in synthetic bone graft. , 2002, Orthopedics.

[12]  T. Böhling,et al.  Natural coral as bone-defect-filling material. , 2000, Journal of biomedical materials research.

[13]  R. Ewers,et al.  Histologic findings at augmented bone areas supplied with two different bone substitute materials combined with sinus floor lifting. Report of one case. , 2004, Clinical oral implants research.

[14]  J. Folkman,et al.  Role of cell shape in growth control , 1978, Nature.

[15]  S. Mann,et al.  The potential of biomimesis in bone tissue engineering: lessons from the design and synthesis of invertebrate skeletons. , 2002, Bone.

[16]  H. Ohgushi Coral Derived Porous Framework Having Different Chemical Compositions as a Scaffold for Osteoblastic Differentitation , 1997 .

[17]  T. Böhling,et al.  Bone marrow induced osteogenesis in hydroxyapatite and calcium carbonate implants. , 1996, Biomaterials.

[18]  Dietmar W. Hutmacher,et al.  Evaluation of Ultra-Thin Poly(ε-Caprolactone) Films for Tissue-Engineered Skin , 2001 .

[19]  A I Caplan,et al.  Stem cell technology and bioceramics: from cell to gene engineering. , 1999, Journal of biomedical materials research.

[20]  G. Guillemin,et al.  The use of coral as a bone graft substitute. , 1987, Journal of biomedical materials research.

[21]  E. Brown,et al.  Expression of extracellular calcium-sensing receptor in human osteoblastic MG-63 cell line. , 2001, American journal of physiology. Cell physiology.

[22]  Tianqiu Mao,et al.  Bone graft in the shape of human mandibular condyle reconstruction via seeding marrow-derived osteoblasts into porous coral in a nude mice model. , 2002, Journal of oral and maxillofacial surgery : official journal of the American Association of Oral and Maxillofacial Surgeons.

[23]  C. Laurencin,et al.  Structural and human cellular assessment of a novel microsphere-based tissue engineered scaffold for bone repair. , 2003, Biomaterials.

[24]  P. Stayton,et al.  Biomimetic peptides that engage specific integrin-dependent signaling pathways and bind to calcium phosphate surfaces. , 2003, Journal of biomedical materials research. Part A.

[25]  M. Chapman,et al.  Autogeneic bone marrow and porous biphasic calcium phosphate ceramic for segmental bone defects in the canine ulna. , 1991, Clinical orthopaedics and related research.

[26]  G. Jordan,et al.  High concentrations of dexamethasone suppress the proliferation but not the differentiation or further maturation of human osteoblast precursors in vitro: relevance to glucocorticoid-induced osteoporosis. , 2001, Rheumatology.

[27]  M. Pittenger,et al.  Multilineage potential of adult human mesenchymal stem cells. , 1999, Science.

[28]  R. Yukna,et al.  A 5-year follow-up of 16 patients treated with coralline calcium carbonate (BIOCORAL) bone replacement grafts in infrabony defects. , 1998, Journal of clinical periodontology.

[29]  S. Ahmed,et al.  A new rapid and simple non-radioactive assay to monitor and determine the proliferation of lymphocytes: an alternative to [3H]thymidine incorporation assay. , 1994, Journal of immunological methods.

[30]  G. Muschler,et al.  Bone graft materials. An overview of the basic science. , 2000, Clinical orthopaedics and related research.

[31]  Su‐Li Cheng,et al.  Differentiation of human bone marrow osteogenic stromal cells in vitro: induction of the osteoblast phenotype by dexamethasone. , 1994, Endocrinology.

[32]  G. Schlag,et al.  Biocompatibility of xenogeneic bone, commercially available coral, a bioceramic and tissue sealant for human osteoblasts. , 1994, Biomaterials.

[33]  M Epple,et al.  A thorough physicochemical characterisation of 14 calcium phosphate-based bone substitution materials in comparison to natural bone. , 2004, Biomaterials.