[Culture of chondrocytes using collagen-hydroxyapatite composite scaffolds in vitro].
OBJECTIVE
To evaluate the feasibility and the value of the layered cylindric collagen-hydroxyapatite composite as a scaffold for the cartilage tissue engineering after an observation of how it absorbs the chondrocytes and affects the cell behaviors.


METHODS
The chondrocytes were isolated and multiplied in vitro, and then the chondrocytes were seeded onto the porous collagen/hydroxyapatite composite scaffold and were cultured in a three-dimensional environment for 3 weeks. The effects of the composite scaffold on the cell adhesivity, proliferation, morphological changes, and synthesis of the extracellular matrix were observed by the phase-contrast microscopy, histology, scanning electron microscopy, and immunohistochemistry.


RESULTS
The pore diameter of the upper layer of the collagen-hydroxyapatite composite scaffold was about 147 microm, and the porosity was 89%; the pore diameter of the bottom layer was about 85 microm and the porosity was 85%. The layered cylindric collagen-hydroxyapatite composite scaffold had good hydrophilia. The chondrocytes that adhered to the surface of the scaffold, proliferated and migrated into the scaffold after 24 hours. The chondrocytes attached to the wall of the microholes of the scaffold maintained a rounded morphology and could secrete the extracellular matrix on the porous scaffold.


CONCLUSIONS
The layered cylindric collagen-hydroxyapatite composite scaffold has a good cellular compatibility, and it is stronger in the mechanical property than the pure collagen. It will be an ideal scaffold for the cartilage tissue engineering.
Lu
Huading
H
Department of Orthopaedics, 3rd Affiliated hospital, Sun Yat-Sen University, Guangzhou Guangdong 510630, P R China. skyforest@medmail.com.cn
Cai
Daozhang
D
Fen
Zhiying
Z
Wu
Gang
G
Zeng
Chun
C
Shi
Dehai
D
Li
Xiaofeng
X
D000818:Animals; D018929:Cell Culture Techniques; D019902:Chondrocytes / Q000166:cytology*; D003094:Collagen / Q000494:pharmacology; D017886:Durapatite / Q000494:pharmacology; D011817:Rabbits; D023822:Tissue Engineering / Q000379:methods*
D003094:Collagen; D017886:Durapatite