ABSTRACT
BACKGROUND:Low toxicity of Genipin has certain species and cellspecificity. Biocompatibility of Genipin cross-linked type I colagen with human adipose-derived stem cels is essential for construction of tissue-engineered adipose. OBJECTIVE:To investigate the bbiocompatibility of Genipin cross-linked type I colagen with human adipose-derived stem cels. METHODS:Human adipose-derived stem cels were isolated and cultured to the third generation, and the cels were seeded on Genipin cross-linked type I colagen scaffold. MTT assay was used to evaluate the adhesion and proliferation of cels on the scaffold, and the toxic effects of Genipin cross-linked type I colagen on human adipose-derived stem cels. Optical microscopy and scanning electron microscopy were utilized to observe the adhesion and growth process of human adipose-derived stem cels on the scaffold as wel as the morphological changes of cels. RESULTS AND CONCLUSION:Human adipose-derived stem cels could adhere to the scaffold immediately after seeded and increase gradualy on the scaffold, with the average adhesion rate of 86.5%. Optical microscopy and scanning electron microscopy showed that human adipose-derived stem cels adhered wel on the scaffold. The cels increased gradualy over time, and could migrate into the scaffold, and distribute evenly with the passage of time when observed with optical microscopy. The result showed Genipin possesses very low cytotoxicity to the cels, and the outstanding biocompatibility is found between the cels and scaffoldin vitro after cross-linked with Genipin.
ABSTRACT
BACKGROUND:Based on the original advantages of silk fibroin, positive charged water-soluble chitosan modified silk fibroin is modified on surface and could improve celladhesion on the scaffolds. OBJECTIVE:To verify the biocompatibility of chitosan-modified silk fibroin with human adipose-derived stem cells (hADSCs), and feasibility of constructing tissue engineered adipose in vitro. METHODS:The hADSCs at passage 3 were seeded on chitosan-modified silk fibroin at the concentration of 1×107/L, as the experiment group;at the same cellconcentration, hADSCs were seeded in 96-wel plates as the control group. MTT tests were performed to evaluate the adhesion, growth and proliferation of hADSCs on chitosan-modified silk fibroin. Then hADSCs were implanted on the chitosan-modified silk fibroin scaffolds at the concentration of 1×109/L. The hADSCs seeded onto chitosan-modified silk fibroin complexes were respectively cultured with adipogenic differentiation medium and ordinary high-glucose DMEM. The complexes were stained with oil red O, and detected with RT-PCR after cultured 14 days. RESULTS AND CONCLUSION:The hADSCs adhered to and proliferated on the scaffolds. After cultured with adipogenic differentiation medium for 14 days, oil red O staining demonstrated that there were amount of mature adipocytes on the scaffold. The peroxisome proliferator activated receptorγ2 was positively expressed. The chitosan-modified silk fibroin possessed excellent biocompatibility in vitro. The co-cultured hADSCs could be induced to mature adipocytes successful y.