Dynamic perfusion bioreactor system for 3D culture of rat bone marrow mesenchymal stem cells on nanohydroxyapatite/polyamide 66 scaffold in vitro.
The aim of the study was to investigate the biocompatibility and osteogenic effectiveness of the porous nanohydroxyapatite/polyamide 66 (n-HA/PA66) scaffold material that was cultured with the rat bone marrow mesenchymal stem cells (rBMSCs), under the static culture condition and the dynamic perfusion culture condition in vitro, and to investigate whether the 3D perfusion culture condition was better in provoking proliferation of rBMSCs than the 3D static culture condition. The Methyl thiazolyl tetrazolium (MTT) assay, alkaline phosphatase (ALP) activity assay, Osteocalcin (OCN) assay and scanning electron microscope (SEM) were used to observe the proliferation and differentiation of rBMSCs. The samples were respectively harvested at 1st, 3rd, 7th, 14th, and 21st days and effect comparisons were made between the two of the culture conditions. The results showed that values of MTT, ALP, and OCN were increased continuously and revealed a significant difference between the two culture conditions (p < 0.05). On the 14th day, SEM revealed calcified nodules 2-8 μm in diameter in the lamellar structure. Under the static culture condition, the pores were covered with the cells looking like a piece of blanket, but under the perfusion culture condition the cells were observed to have a 3D lamellar structure. In conclusion, the porous n-HA/PA66 scaffold material can be used as a good candidate material for the bone scaffold construction in the tissue engineering because of its excellent 3D structure, which can greatly improve the proliferation and differentiation of rBMSCs and make them proliferate and osteogenesis even better under the perfusion culture condition.
State Key Laboratory of Oral Diseases, Sichuan University, Chengdu, 610041, PR China.
Xu
X
Qian
Fang
F
Yuan
Zhu
Z
Zhimin
Mo
M
Anchun
D000469:Alkaline Phosphatase / Q000378:metabolism; D000818:Animals; D001672:Biocompatible Materials / Q000737:chemistry; D019149:Bioreactors; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D049109:Cell Proliferation; D002478:Cells, Cultured; D017886:Durapatite / Q000737:chemistry; D004867:Equipment Design; D008422:Materials Testing; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000378:metabolism; D008855:Microscopy, Electron, Scanning; D049329:Nanostructures / Q000737:chemistry; D009757:Nylons / Q000737:chemistry; D015675:Osteocalcin / Q000378:metabolism; D010012:Osteogenesis; D010477:Perfusion; D051381:Rats; D023822:Tissue Engineering; D054457:Tissue Scaffolds
D001672:Biocompatible Materials; D009757:Nylons; D015675:Osteocalcin; D017886:Durapatite; D000469:Alkaline Phosphatase
bone graft; dynamic culture; nanohydroxyapatite/polyamide 66 (n-HA/PA66) scaffold; perfusion bioreactor; porosity; tissue engineering