Université de Reims Champagne Ardenne, BIOS EA 4691, Reims, France; Université de Reims Champagne Ardenne, UFR d'Odontologie, Reims, France.
Université de Reims Champagne Ardenne, LRN EA 4682, Reims, France.
Nanomedicine. 2020 Oct;29:102256. doi: 10.1016/j.nano.2020.102256. Epub 2020 Jun 29.
In bone tissue engineering, stem cells are known to form inhomogeneous bone-like nodules on a micrometric scale. Herein, micro- and nano-infrared (IR) micro-spectroscopies were used to decipher the chemical composition of the bone-like nodule. Histological and immunohistochemical analyses revealed a cohesive tissue with bone-markers positive cells surrounded by dense mineralized type-I collagen. Micro-IR gathered complementary information indicating a non-mature collagen at the top and periphery and a mature collagen within the nodule. Atomic force microscopy combined to IR (AFM-IR) analyses showed distinct spectra of "cell" and "collagen" rich areas. In contrast to the "cell" area, spectra of "collagen" area revealed the presence of carbohydrate moieties of collagen and/or the presence of glycoproteins. However, it was not possible to determine the collagen maturity, due to strong bands overlapping and/or possible protein orientation effects. Such findings could help developing protocols to allow a reliable characterization of in vitro generated complex bone tissues.
在骨组织工程中，已知干细胞在微米尺度上形成不均匀的类骨结节。在此，微纳红外（IR）微光谱用于解析类骨结节的化学成分。组织学和免疫组织化学分析显示，一种有凝聚力的组织，阳性骨标志物细胞被密集矿化的 I 型胶原包围。微 IR 收集了互补信息，表明在顶部和外围是非成熟的胶原，而在结节内是成熟的胶原。原子力显微镜与红外（AFM-IR）分析相结合，显示了“细胞”和“胶原”丰富区域的不同光谱。与“细胞”区域相比，“胶原”区域的光谱显示了胶原的碳水化合物部分的存在和/或糖蛋白的存在。然而，由于强带重叠和/或可能的蛋白质取向效应，无法确定胶原的成熟度。这些发现可以帮助开发协议，以允许对体外生成的复杂骨组织进行可靠的表征。
