Effects of initial cell density and hydrodynamic culture on osteogenic activity of tissue-engineered bone grafts.
Effects of initial cell density and hydrodynamic culture on osteogenic activity of tissue-engineered bone grafts.
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初始细胞密度和水动力培养对组织工程骨移植物成骨活性的影响
DOI：
10.1371/journal.pone.0053697
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发表时间：
2013
期刊：
影响因子：
3.7
通讯作者：
Xu JZ
中科院分区：
文献类型：
--
作者：
Luo F;Hou TY;Zhang ZH;Xie Z;Wu XH;Xu JZ
本研究旨在探讨细胞初始密度和体外培养方法对组织工程骨构建及成骨活性的影响。将人骨髓间充质干细胞（hMSCs）接种到由脱钙骨基质（DBM）制备的立方体支架上，通过三种方法-静态，流体动力学和纤维蛋白水凝胶辅助接种。通过静态烧瓶培养或流体动力学培养体外培养所得的细胞-支架构建体。分析构建体的初始细胞密度和随后的体外增殖和碱性磷酸酶活性。构建体也被皮下植入裸鼠中以检查它们的体内成骨活性。水凝胶辅助播种的播种效率最高，其次是流体动力学和传统的静态播种。在体外培养过程中，流体动力学培养产生更高的高原细胞密度，碱性磷酸酶（ALP）的活动，和细胞外基质的生产比静态培养。在裸鼠皮下植入后，通过水凝胶辅助接种和流体动力学培养相结合制备的植入物产生比其他方法制备的植入物更高的湿重和骨密度。结果表明，水凝胶辅助接种可以显著增加支架中的初始种子细胞密度。随后的流体动力学培养可以促进种子细胞的增殖和成骨分化。相应地，通过这两种方法的组合产生的骨移植物在所采用的三种方法中实现了最高的成骨活性。
This study aimed to study the effects of initial cell density and in vitro culture method on the construction of tissue-engineered bone grafts and osteogenic activities. Human mesenchymal stem cells (hMSCs) were seeded onto cubic scaffolds prepared from demineralized bone matrix (DBM) by three methods - static, hydrodynamic, or fibrin hydrogel-assisted seeding. The resulting cell-scaffold constructs were cultured in vitro by static flask culture or hydrodynamic culture. The initial cell density and the subsequent in vitro proliferation and alkaline phosphate activities of the constructs were analyzed. The constructs were also subcutaneously implanted in nude mice to examine their in vivo osteogenic activities. Hydrogel-assisted seeding gave the highest seeding efficiency, followed by hydrodynamic and conventional static seeding. During in vitro culture, hydrodynamic culture produced higher plateau cell densities, alkaline phosphatase (ALP) activities, and extracellular matrix production than static culture. After subcutaneous implantation in nude mice, the implants prepared by the combination of hydrogel-assisted seeding and hydrodynamic culture produced higher wet weight and bone mineral density than implants prepared by other methods. The results suggest that the hydrogel-assisted seeding can substantially increase the initial seed cell density in scaffolds. Subsequent hydrodynamic culture can promote the proliferation and osteoblastic differentiation of the seeded cells. Correspondingly, bone grafts produced by the combination of these two methods achieved the highest osteogenic activity among the three methods employed.
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影响因子：
14
作者：
Aubin H;Nichol JW;Hutson CB;Bae H;Sieminski AL;Cropek DM;Akhyari P;Khademhosseini A
通讯作者：
Khademhosseini A
影响因子：
3.7
作者：
Borriello A;Caldarelli I;Basile MA;Bencivenga D;Tramontano A;Perrotta S;Della Ragione F;Oliva A
通讯作者：
Oliva A
影响因子：
9.7
作者：
Jiang, Tao;Nukavarapu, Syam P.;Laurencin, Cato T.
通讯作者：
Laurencin, Cato T.
影响因子：
4.1
作者：
Liu, Yang;Liu, Tianqing;Cui, Zhanfeng
通讯作者：
Cui, Zhanfeng
影响因子：
3.8
作者：
Ouyang, Anli;Yang, Shang-Tian
通讯作者：
Yang, Shang-Tian