Laser-assisted bioprinting for creating on-demand patterns of human osteoprogenitor cells and nano-hydroxyapatite
Laser-assisted bioprinting for creating on-demand patterns of human osteoprogenitor cells and nano-hydroxyapatite
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DOI：
10.1088/1758-5082/3/2/025001
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发表时间：
2011-06-01
期刊：
影响因子：
9
通讯作者：
Guillemot, Fabien
中科院分区：
文献类型：
--
作者：
Catros, Sylvain;Fricain, Jean-Christophe;Guillemot, Fabien
开发工具来复制和操纵细胞微环境，包括细胞图案的位置和形状，是组织工程的关键。与喷墨打印和压力驱动的机械挤出机并行，激光辅助生物打印(LAB)已成为制造二维和三维组织工程产品的替代技术。本工作的目的是在实验室中确定二维和三维构图和组装纳米羟基磷灰石(NHA)和人骨祖细胞(Hop)的激光打印参数。这项研究中使用的实验室工作站包括一台聚焦在石英带上的红外激光器，该石英带上覆盖着一层薄的钛吸收层和一层生物墨水。扫描系统、石英带和衬底通过专用软件进行试验，允许将不同的生物材料顺序打印成二维和/或三维。采用化学沉淀法合成了纳米羟基磷灰石打印材料(生物墨水)，并用透射电子显微镜、傅里叶变换红外光谱和X射线衍射仪对打印前后的材料进行了表征。HOP生物墨水用3000万个细胞毫升(-1)的悬液在培养液中制备，打印后使用Live/Dead实验和成骨细胞表型标记(碱性磷酸酶和骨钙素)进行鉴定。结果表明，LAB可以打印和组织二维和三维的nha和hop。LAB没有改变NHA的理化性质，也没有改变啤酒花的活力、增殖和表型(长达15天)。本研究表明，LAB是一种适合于二维构图纳米羟基磷灰石和成骨细胞的方法，也适用于三维复合材料的生物制备。
Developing tools to reproduce and manipulate the cell micro-environment, including the location and shape of cell patterns, is essential for tissue engineering. Parallel to inkjet printing and pressure-operated mechanical extruders, laser-assisted bioprinting (LAB) has emerged as an alternative technology to fabricate two-and three-dimensional tissue engineering products. The objective of this work was to determine laser printing parameters for patterning and assembling nano-hydroxyapatite (nHA) and human osteoprogenitors (HOPs) in two and three dimensions with LAB. The LAB workstation used in this study comprised an infrared laser focused on a quartz ribbon that was coated with a thin absorbing layer of titanium and a layer of bioink. The scanning system, quartz ribbon and substrate were piloted by dedicated software, allowing the sequential printing of different biological materials into two and/or three dimensions. nHA printing material (bioink) was synthesized by chemical precipitation and was characterized prior and following printing using transmission electron microscopy, Fourier transformed infrared spectroscopy and x-ray diffraction. HOP bioink was prepared using a 30 million cells ml(-1) suspension in culture medium and cells were characterized after printing using a Live/Dead assay and osteoblastic phenotype markers (alcaline phosphatase and osteocalcin). The results revealed that LAB allows printing and organizing nHA and HOPs in two and three dimensions. LAB did not alter the physico-chemical properties of nHA, nor the viability, proliferation and phenotype of HOPs over time (up to 15 days). This study has demonstrated that LAB is a relevant method for patterning nHA and osteoblastic cells in 2D, and is also adapted to the bio-fabrication of 3D composite materials.