Descrição
In the present work we originally tested the suitability
of corn starch-polycaprolactone (SPCL) scaffolds for
pursuing a cartilage tissue engineering approach. Bovine articular
chondrocytes were seeded on SPCL scaffolds under
dynamic conditions using spinner flasks (total of 4 scaffolds
per spinner flask using cell suspensions of 0.5×106 cells/ml)
and cultured under orbital agitation for a total of 6 weeks.
Poly(glycolic acid) (PGA) non-woven scaffolds and bovine
native articular cartilage were used as standard controls for
the conducted experiments. PGA is a kind of standard in
tissue engineering approaches and it was used as a control
in that sense. The tissue engineered constructs were characterized
at different time periods by scanning electron microscopy
(SEM), hematoxylin-eosin (H&E) and toluidine
blue stainings, immunolocalisation of collagen types I and II,
and dimethylmethylene blue (DMB) assay for glycosaminoglycans
(GAG) quantification assay. SEM results for SPCL
constructs showed that the chondrocytes presented normal
morphological features, with extensive cells presence at the
surface of the support structures, and penetrating the scaffolds
pores. These observations were further corroborated
by H&E staining. Toluidine blue and immunohistochemistry
exhibited extracellular matrix deposition throughout the 3D structure. Glycosaminoglycans, and collagen types I and II
were detected. However, stronger staining for collagen type
II was observed when compared to collagen type I. The PGA
constructs presented similar features toSPCLat the end of the
6 weeks. PGA constructs exhibited higher amounts of matrix
glycosaminoglycans when compared to the SPCL scaffolds.
However, we also observed a lack of tissue in the central
area of the PGA scaffolds. Reasons for these occurrences
may include inefficient cells penetration, necrosis due to high
cell densities, or necrosis related with acidic by-products
degradation. Such situation was not detected in the SPCL
scaffolds, indicating the much better biocompatibility of the
starch based scaffolds.