Bone quality assessment of osteogenic cell cultures by Raman microscopy.
Bone quality assessment of osteogenic cell cultures by Raman microscopy.
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DOI：
10.1002/jrs.5521
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发表时间：
2018-11
期刊：
影响因子：
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通讯作者：
G. Mandair;P. Steenhuis;M. Ignelzi;M. Morris
中科院分区：
文献类型：
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作者：
G. Mandair;P. Steenhuis;M. Ignelzi;M. Morris
使用自体干/祖细胞是修复颅面骨缺损的一种有前途的方法。颅骨被认为是干细胞/祖细胞的可行来源，可以在体外移植形成骨。然而，目前尚不清楚细胞培养中形成的骨是否与天然骨中发现的骨质量相似。在这项研究中，成骨细胞培养物中形成的骨矿物质的质量与出生后小鼠的颅骨进行了比较。鉴于细胞和胶原蛋白光谱之间存在的光谱相似性，还研究了提取细胞活性和骨基质质量信息的可行性。将从胎鼠颅骨分离的干/祖细胞在成骨分化条件下在熔融石英载玻片上培养28天。在特定的时间间隔，取出载玻片并通过拉曼显微镜和矿物染色技术进行分析。我们发现，在第28天的培养中形成的骨类似于出生后1天的小鼠的颅骨，具有可比的矿化、矿物结晶度和胶原交联比。相比之下，与出生后1天的骨相比，在第28天形成的骨含有较低程度的有序胶原纤维。总之，在成骨细胞培养中形成的骨表现出进行性基质成熟和矿化，但不能完全复制在天然骨中发现的高度胶原纤维秩序。
The use of autologous stem/progenitor cells represents a promising approach to the repair of craniofacial bone defects. The calvarium is recognized as a viable source of stem/progenitor cells that can be transplanted in vitro to form bone. However, it is unclear if bone formed in cell culture is similar in quality to that found in native bone. In this study, the quality of bone mineral formed in osteogenic cell cultures were compared against calvarial bone from postnatal mice. Given the spectroscopic resemblance that exists between cell and collagen spectra, the feasibility of extracting information on cell activity and bone matrix quality were also examined. Stem/progenitor cells isolated from fetal mouse calvaria were cultured onto fused-quartz slides under osteogenic differentiation conditions for 28 days. At specific time intervals, slides were removed and analyzed by Raman microscopy and mineral staining techniques. We show that bone formed in culture at Day 28 resembled calvarial bone from 1-day-old postnatal mice with comparable mineralization, mineral crystallinity, and collagen crosslinks ratios. In contrast, bone formed at Day 28 contained a lower degree of ordered collagen fibrils compared with 1-day-old postnatal bone. Taken together, bone formed in osteogenic cell culture exhibited progressive matrix maturation and mineralization but could not fully replicate the high degree of collagen fibril order found in native bone.