Three-dimensional culture of rat calvarial osteoblasts in porous biodegradable polymers
Three-dimensional culture of rat calvarial osteoblasts in porous biodegradable polymers
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DOI：
10.1016/s0142-9612(98)00021-0
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发表时间：
1998-08-01
期刊：
影响因子：
14
通讯作者：
Mikos, AG
中科院分区：
文献类型：
--
作者：
Ishaug-Riley, SL;Crane-Kruger, GM;Mikos, AG
新生大鼠颅骨成骨细胞培养在90%的多孔，75：25聚（DL-乳酸-共-乙醇酸）（PLGA）泡沫支架长达56天，以检查细胞接种密度，支架孔径和泡沫厚度对细胞在这个三维环境中的增殖和功能的影响。将成骨细胞以11.1 × 10（5）或22.1 × 10（5）细胞/cm（2）接种到PLGA支架上，所述PLGA支架具有150-300或500-710 μ m的孔径范围，厚度为1.9或3.2 mm。培养1天后，在150-300 μ m孔径的1.9 mm厚支架上，低和高接种密度分别有75.6%和68.6%的接种细胞贴壁和增殖。成骨细胞的数量在整个研究过程中持续增加，并最终在接近56天时趋于平稳，如定量DNA测定所示。低细胞接种密度的成骨细胞/泡沫结构在14天后获得了与高细胞接种密度相当的DNA含量和碱性磷酸酶（ALP 3）活性，56天后获得了矿化结果。在最初用高细胞密度接种的150-300 μ m孔径的成骨细胞/泡沫结构中，56天后达到220 ± 40 μ m的骨组织最大穿透深度。对于500-710 μ m孔径的构建体，在相同条件下的穿透深度为190 +/-40 μ m。支架孔径和厚度对成骨细胞的增殖或功能没有显著影响，如DNA含量、ALP 4活性和矿化组织形成所示。这些数据表明，可比的骨样组织可以在体外工程超过56天的时间内使用不同的大鼠颅骨成骨细胞接种密度到可生物降解的聚合物支架上，孔径范围为150-710 μ m。与之前将类似聚合物支架与骨髓基质细胞接种并培养的研究结果相比，本研究表明，PLGA泡沫是成骨细胞生长和分化功能的合适基质，与细胞来源无关。(C)1998爱思唯尔科技有限公司版权所有
Neonatal rat calvarial osteoblasts were cultured in 90% porous, 75:25 poly(DL-lactic-co-glycolic acid) (PLGA) foam scaffolds for up to 56 days to examine the effects of the cell seeding density, scaffold pore size, and foam thickness on the proliferation and function of the cells in this three-dimensional environment. Osteoblasts were seeded at either 11.1 x 10(5) or 22.1 x 10(5) cells per cm(2) onto PLGA scaffolds having pore sizes in the range of 150-300 or 500-710 mu m with a thickness of either 1.9 or 3.2 mm. After 1 day in culture, 75.6 and 68.6% of the seeded cells attached and proliferated on the 1.9 mm thick scaffolds of 150-300 mu m pore size for the low and high seeding densities, respectively. The number of osteoblasts continued to increase throughout the study and eventually leveled off near 56 days, as indicated by a quantitative DNA assay. Osteoblast/foam constructs with a low cell seeding density achieved comparable DNA content and alkaline phosphatase (ALPase) activity after 14 days, and mineralization results after 56 days to those with a high cell seeding density. A maximum penetration depth of osseous tissue of 220 +/- 40 mu m was reached after 56 days in the osteoblast/foam constructs of 150-300 mu m pore size initially seeded with a high cell density. For constructs of 500-710 mu m pore size, the penetration depth was 190 +/- 40 mu m under the same conditions. Scaffold pore size and thickness did not significantly affect the proliferation or function of osteoblasts as demonstrated by DNA content, ALPase activity, and mineralized tissue formation. These data show that comparable bone-like tissues can be engineered in vitro over a 56 day period using different rat calvarial osteoblast seeding densities onto biodegradable polymer scaffolds with pore sizes in the range of 150-710 mu m. When compared with the results of a previous study where similar polymer scaffolds were seeded and cultured with marrow stromal cells, this study demonstrates that PLGA foams are suitable substrates for osteoblast growth and differentiated function independent of cell source. (C) 1998 Elsevier Science Ltd. All rights reserved