Preparation of a three-dimentional acellular cartilage matrix scaffold and its characteristics
- VernacularTitle:脱细胞软骨基质三维支架的制备及特性研究
- Author:
Long YU
;
Yunyu HU
;
Long BI
- Publication Type:Journal Article
- Keywords:
tissue engineered cartilage,decellularization,scaffold,genipin
- From:
Orthopedic Journal of China
2006;0(09):-
- CountryChina
- Language:Chinese
-
Abstract:
[Objective]To prepare a cartilage acellular matrix scaffold and to explore its feasibility in cartilage tissue engineering. [Methods]Microparticles about 100 ?m~154 ?m were prepared after calf cartilage physically shattered and experienced gradient centrifugation,and then treated by a modified Courtman's four-step method which was improved to produce acellular cartilage matrix.After this treatment the microparticles were made into 3% suspension which was placed into moulds.With the freeze-drying method,3-D cartilage acellular matrix (CACM) was prepared.The scaffolds were cross-linked by a neotype crosslinking agent genepin for 48h,and then placed into glycine solution server times for removing redundant genepin.The freeze-drying method was used to prepare CACM.The scaffolds were investigated by gross observation,histological staining (haematoxylin-eosin,toluidine blue) ,scanning electron microscope (SEM) observation and porosity measurement,water absorption rate and degradation rate analysis.After being cultivated for ten days,bone marrow stranal cells (BMSCs) of rabbit were seeded into the scaffold.MTT test and SEM were done to assess the growth and proliferation of BMSCs.[Results]Gross observation showed the scaffolds had a loosely porous and dark blue appearance after being cross-linked by genepin.The histological staining (haematoxylin-eosin,toluidine blue staining) showed that there were no chondrocyte fragments in the scaffold.The CACM scaffold had 90% porosity,(1314?337) % water absorption rate,and (13.69?7.3)% or (25.99?8.9) % degradation rate at 2 or 4 weeks.MTT test showed that BMSCs grew well in the 3-D CACM scaffolds of logarithmic trend,supporting that the scaffolds had no cytotoxic effect on BMSCs.SEM micrographs indicated that the scaffolds were porous and the cells covered the scaffolds firmly with cell processes.[Conclusion]The improved Courtman's four-step method makes a more thoroughly acellular scaffold.The 3-D CACM scaffold retains most of extracelluar matrix.After being cross-linked by genepin,the 3-D CACM scaffold has good biocompatibility and degradation rate of the scaffolds is decreased,which makes it a suitable carrier for cartilage tissue engineering.
