NANOSTRUCTURED BIOMATERIALS: NANOHYDROXYAPATITE/COLLAGEN

SCAFFOLDS SUPPLEMENTED WITH SPARC AS FACILITATORS IN BONE

ENGINEERING
A. Y. Pataquiva-Mateus1,2,3 H.-C. Wu1,4 C. Lucchesi 1 M. P. Ferraz 5 F. J. Monteiro 2,3 M.
Spector 1,6
1 Tissue Engineering, VA Boston Healthcare System, Boston, Massachusetts
2 Laboratrio de Biomateriais, INEB Instituto de Engenharia Biomdica, Porto, Portugal
3 Departamento de Engenharia Metalrgica e Materiais, Universidade do Porto, Porto, Portugal
4 Institute of Biomedical Engineering, National Taiwan University, Taipei, Taiwan
5 Universidade Fernando Pessoa, Porto, Portugal
6 Orthopaedic Research, Brigham and Womens Hospital, Harvard Medical School, Boston,
Massachusetts

Key Words: collagen, mineralization, stem cells, biomaterials availability, bone.

Abstract. Bone exhibits natural hydroxyapatite crystals with needle-like or rod-like

shapes, well arranged within the polymeric matrix of collagen type I. These natural
nanoparticles formed in physiological environment have a more dynamic response by
bone cells when compared to synthetic material with larger particle size. Added to this,
the nonstructural extracellular matrix-associated protein, SPARC (Secreted Protein
Acidic and Rich in Cysteine) is known to play a role in the mineralization of collagen in
bone formation. Thus, the objective of this study was to determine the degree to which
SPARC supplementation of type I collagen scaffolds in vitro enhanced the uptake of
calcium and phosphorus when the scaffolds were soaked in a calcium phosphate
solution and a suspension of hydroxyapatite nanoparticles. For this, hydroxyapatite
nanoparticles were prepared using a chemical precipitation synthesis based on
H3PO4, Ca(OH)2 and a surfactant, SDS (sodium dodecylsulphate), as starting
reagents; and later added to collagen type 1 sponges. The calcium and phosphorus
contents of the scaffolds were evaluated by inductively coupled plasma analysis, and
the elastic modulus of the scaffolds determined by unconfined compression testing.
Scaffolds were seeded with goat bone marrow-derived mesenchymal stem cells and
the cell-seeded constructs grown in osteogenic medium. Supplementation of the
scaffolds with as little as 0.008 % by wt. of collagen resulted in a increase in the
binding of hydroxyapatite nanoparticles to the scaffold, but had no effect on
incorporation of calcium or phosphorus from a calcium phosphate solution. The
incorporation of hydroxyapatite nanoparticles into the scaffolds did not result in an
increase in modulus. Supplementation of the scaffolds with SPARC and the increase in
the binding of hydroxyapatite nanoparticles did not affect the proliferation of
mesenchymal stem cells.
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