Evaluation of bioreactor-cultivated bone by magnetic resonance microscopy and FTIR micro spectroscopy
Evaluation of bioreactor-cultivated bone by magnetic resonance microscopy and FTIR micro spectroscopy
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DOI：
10.1016/j.bone.2006.10.020
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发表时间：
2007-04-01
期刊：
影响因子：
4.1
通讯作者：
Potter, Kimberlee
中科院分区：
文献类型：
--
作者：
Chesnick, Ingrid E.;Avallone, Francis A.;Potter, Kimberlee
我们提出了一个基于中空纤维生物反应器（HFBR）的三维矿化模型，该生物反应器接种了从雏鸡颅骨中分离出的原代成骨细胞。使用非侵入性磁共振显微镜 (MRM)，在 9 周的时间内监测单个纤维周围矿化组织的生长和发育。完整组织的水质子 MRM 特性的空间图（分辨率为 78 μm）用于确定发育过程中组织成分的变化。分别通过质子密度（PD）和磁化传递比（MTR）图以高特异性检测组织中矿物质和胶原蛋白含量的独特变化。在生长期结束时，通过组织学验证了骨样组织的存在，并通过选区电子衍射和电子探针X射线微量分析验证了低结晶磷灰石的形成。 FTIR 显微光谱证实了所形成的骨样组织的异质性。 FTIR 衍生的磷酸盐图证实，PD 值最低的位置含有最多的矿物质，而 FTIR 衍生的胶原蛋白图证实 NITR 图上的明亮像素对应于高胶原蛋白含量的区域。总之，通过 FTIR 显微光谱绘制的组织成分空间图证实了非侵入性 MRM 测量的结果，并支持 MRM 在监测体外骨形成过程中的作用。 (c) 2006 Elsevier Inc. 保留所有权利。
We present a three-dimensional mineralizing model based on a hollow fiber bioreactor (HFBR) inoculated with primary osteoblasts isolated from embryonic chick calvaria. Using non-invasive magnetic resonance microscopy (MRM), the growth and development of the mineralized tissue around the individual fibers were monitored over a period of 9 weeks. Spatial maps of the water proton MRM properties of the intact tissue, with 78 mu m resolution, were used to determine changes in tissue composition with development. Unique changes in the mineral and collagen content of the tissue were detected with high specificity by proton density (PD) and magnetization transfer ratio (MTR) maps, respectively. At the end of the growth period, the presence of a bone-like tissue was verified by histology and the formation of poorly crystalline apatite was verified by selected area electron diffraction and electron probe X-ray microanalysis. FTIR microspectroscopy confirmed the heterogeneous nature of the bone-like tissue formed. FTIR-derived phosphate maps confirmed that those locations with the lowest PD values contained the most mineral, and FTIR-derived collagen maps confirmed that bright pixels on NITR maps corresponded to regions of high collagen content. In conclusion, the spatial mapping of tissue constituents by FTIR micro spectroscopy corroborated the findings of non-invasive MRM measurements and supported the role of MRM in monitoring the bone formation process in vitro. (c) 2006 Elsevier Inc. All rights reserved.