Autor(es):
Ribeiro, V. P. 
; Martins, A. R. ; Marques, A. P. ; Bonifácio, G. ; Oliveira, A. L. ; Sousa, R. A. ; Reis, R. L.
Data: 2012
Identificador Persistente: http://hdl.handle.net/1822/24083
Origem: RepositóriUM - Universidade do Minho
Assunto(s): 3D biotextiles; Bone tissue engineering; Human adipose-derived stem cells; Silk
Descrição
Human Adipose-derived Stem Cells (hASCs) became an emerging possibility
for tissue replacement therapies, such as bone tissue regeneration.
Due to their osteogenic differentiation potential, easy isolation,
expansion and in vitro proliferation, they have become a highly potential
source of seed cells to be seeded in bone tissue engineering (TE)
constructs and have demonstrated promising prospects in bone regeneration
[1, 2]. To date several strategies have been proposed with more
or less success to prepare porous three-dimensional biodegradable scaffolds
for bone TE. Among them, textile technologies are particularly
interesting since they can allow for producing finely tuned, fibre-based
complex structures, offering superior control over the design (ex: size,
shape, porosity, fibre alignment), manufacturing and reproducibility.
The aim of this work is to evaluate the potential of recently developed
silk-based biotextile structures [3] to promote hASCs adhesion, proliferation
and osteoblastic differentiation. Natural silk yarns were processed
into different 3D structures using standard knitting or warpknitting
technologies to increase the scaffold’s tridimensionality. In the
latter case two knitted silk layers are assembled and spaced by a monofilament
of polyethylene terephthalate (PET). These constructs were
characterized in terms of their morphology by Microcomputed Tomography
(l-CT) and scanning electron microscopy (SEM). The mechanical
properties were investigated through compressive tests and
dynamic mechanical analysis (DMA). All constructs disclose a biocompatible
behavior, assessed using a mouse fibroblastic cell line (L929;
ECACC, UK). hASCs were seeded onto the scaffolds and cultured for
14, 21 and 28 days in osteogenic medium. All textile constructs were
analysed in terms of cell adhesion, proliferation and differentiation
potential influence through the biological assays, alkaline phosphatase
(ALP), DNA and Ca2+ quantification and histological, confocal, SEM
and Real-Time PCR analysis. The obtained constructs present very
reproducible intra-architectural scaffold geometry with high surface
area and exhibiting a wide range of porosities. By the above mentioned
assays it was possible validate the developed constructs as suitable for
hASCs adhesion, proliferation and differentiation into an osteoblastic
lineage. The positive influence of the developed 2D/3D textile structures
on the osteoblastic differentiation potential of hACSs is an important
outcome that validates future bone tissue enginnering approaches
using these fibre-based architectures.
