Autor(es):
Oliveira, Joaquim M. 
; Silva, S. S. ; Malafaya, P. B. ; Rodrigues, Márcia T. ; Kotobuki, Noriko ; Hirose, Motohiro ; Gomes, Manuela E. ; Mano, J. F. ; Ohgushi, H. ; Reis, R. L.
Data: 2009
Identificador Persistente: http://hdl.handle.net/1822/20373
Origem: RepositóriUM - Universidade do Minho
Assunto(s): Hydroxyapatite; Bone tissue engineering; Rat bone marrow stromal; Cells; Scaffold; Cell viability
Descrição
In this work, a new methodology is reported
for developing hydroxyapatite (HA) scaffolds using an
organic sacrifice template. The novelty of work consists of
possibility of obtaining porous and highly interconnected
scaffolds mimicking the sacrificial component. Our purpose
consisted of evaluating the physicochemical properties
of the HA scaffolds by means of Fourier transform
infra-red spectroscopy, X-ray diffraction analysis, and
scanning electron microscopy (SEM) attached with an Xray
detector. The HA scaffolds obtained possess a porosity
of !70%, and macropores diameter in the range of
50–600 lm. In contrast, results regarding the microcomputed
tomography analysis have demonstrated both high
pore uniformity and interconnectivity across the scaffolds.
The compressive strength of the HA scaffolds was found
to be 30.2 6 6.0 MPa. Bioactivity of the HA scaffolds was
assessed by immersion into a simulated body fluid solution,
in vitro. SEM observations have showed a deposition
of apatite on the surface of the HA scaffolds, with a ‘‘cauliflower-
like’’ morphology after 1 day, and tend to be
more pronounced with the immersion time. The changes
in calcium and phosphorus concentration were monitored
by inductively-coupled plasma optical emission spectrometry.
Cytotoxicity of the HA scaffolds was preliminarily
investigated by carrying direct observation of mouse
fibroblasts cells (L929 cell-line) death in the inverted
microscope, and then cell viability was determined by
means of carrying out a MTS assay. Complementarily, a
luminescent cell viability assay based on the quantification
of adenosine triphosphate was performed using rat
bone marrow stromal cells (RBMSCs). A LIVE/DEAD
assay and SEM analysis allowed the visualization of the
RBMSCs adhesion and proliferation on the surface of the
HA scaffolds. According to the results obtained from 3D
architecture, mechanical properties, biocompatibility, and
adhesion tests, it is suggested that HA scaffolds has
potential to find applications in bone tissue engineering
scaffolding.
