Three-dimensional dynamic culture of pre-osteoblasts seeded in HA-CS/Col/nHAP composite scaffolds and treated with α-ZAL  

作　　者：Lu Liu Yong Guo Xuezhong Chen Ruixin Li Zhihong Li Liang Wang Zongming Wan Jianyu Li Qingxin Hao Hao Li Xizheng Zhang 

机构地区：[1]Institute of Medical Equipment,Academy of Military Medical Sciences,Tianjin 300161,China [2]Medical College,Chinese People's Armed Police Forces,Tianjin 300162,China [3]Shenzhen Pingle Orthopaedic Hospital,Shenzhen 518000,China

出　　处：《Acta Biochimica et Biophysica Sinica》2012年第8期669-677,共9页生物化学与生物物理学报（英文版）

基　　金：Funding This work was supported by a grant from the National Nature Science Foundation of China （No. 10832012）.

摘　　要：Pre-osteoblast MC3T3-E1 cells were cultured in hyaluronic acid-modified chitosan/collagen/nano-hydroxyapatite （HA-CS/Coi/nHAP） composite scaffolds and treated with phytoestrogen α-zearalanol （α-ZAL） to improve bone tissue formation for bone tissue engineering. Perfusion and dynamic strain were applied to three-dimensional （3D） cultured cells, which simulates mechanical microenvironment in bone tissue and solves mass transfer issues. The morphology of cell-scaffold constructs in vitro was then examined and markers of osteogenesis were assessed by immunohistochemistry staining and western blotting. The results showed that cells expanded their pseudopodia in an irregular manner and dispersed along the walls in 3D-dynamic culture. Osteogenic phenotype was increased or maintained by enhanced collagen I （COLI） levels, decreased osteopontin expression and having little effect on osteocalcin expression during the 12 days of in vitro culture. In response to α-ZAL, the cell-scaffold constructs showed inhibited cellular proliferation, enhanced the alkaline phosphatase （ALP） activity and increased ratio of osteoprotegerin to receptor activator of nuclear factor kappa B （NF-KB） ligand （RANKL）. Application of perfusion and dynamic strain to cells-scaffold constructs treated with α-ZAL represents a promising approach in the studies of osteogenesis stimulation of bone tissue engineering.Pre-osteoblast MC3T3-E1 cells were cultured in hyaluronic acid-modified chitosan/collagen/nano-hydroxyapatite （HA-CS/Coi/nHAP） composite scaffolds and treated with phytoestrogen α-zearalanol （α-ZAL） to improve bone tissue formation for bone tissue engineering. Perfusion and dynamic strain were applied to three-dimensional （3D） cultured cells, which simulates mechanical microenvironment in bone tissue and solves mass transfer issues. The morphology of cell-scaffold constructs in vitro was then examined and markers of osteogenesis were assessed by immunohistochemistry staining and western blotting. The results showed that cells expanded their pseudopodia in an irregular manner and dispersed along the walls in 3D-dynamic culture. Osteogenic phenotype was increased or maintained by enhanced collagen I （COLI） levels, decreased osteopontin expression and having little effect on osteocalcin expression during the 12 days of in vitro culture. In response to α-ZAL, the cell-scaffold constructs showed inhibited cellular proliferation, enhanced the alkaline phosphatase （ALP） activity and increased ratio of osteoprotegerin to receptor activator of nuclear factor kappa B （NF-KB） ligand （RANKL）. Application of perfusion and dynamic strain to cells-scaffold constructs treated with α-ZAL represents a promising approach in the studies of osteogenesis stimulation of bone tissue engineering.

关 键 词：OSTEOBLAST composite scaffold α-zearalano BIOREACTOR OSTEOGENESIS 

分 类 号：Q813.11[生物学—生物工程] S432.1[农业科学—植物病理学]