Thammasat University Research Unit in Dental and Bone Substitute Biomaterials, Faculty of Dentistry, Thammasat University, Pathumthani, Thailand.
Faculty of Dentistry, Thammasat University, Pathumthani, Thailand.
Acta Histochem. 2021 Jan;123(1):151659. doi: 10.1016/j.acthis.2020.151659. Epub 2020 Dec 4.
The main objective of this study was to analyze the changes in osteoblast behaviors by two different methods, a simple digital image analysis method and conventional assays. Osteoblast progenitor cells and osteoblasts were treated with alendronate (ALN; a nitrogen-containing bisphosphonate). Osteoblast behaviors such as the uptake of ALN, cell proliferation, cell differentiation, and mineralized nodule formation were examined. Quantitative assessments were conducted using a publically available ImageJ software along with conventional methods. Furthermore, ImageJ method and conventional assay for mineralized nodule formation were performed simultaneously and were compared in order to demonstrate the reliability of ImageJ analysis. Osteoblast precursors and osteoblasts responded to ALN treatments. The software could identify various colors and allowed for the quantification of staining intensity and area coverage. Both image analysis and conventional assays detected the changes in cell behaviors between treated and untreated samples. For alizarin red S assay, the staining intensity calculated by ImageJ analysis was comparable to the absorbance measured by conventional assay. These findings showed that digital image analysis along with conventional assays could be used for quantitative assessment to evaluate osteoblast alteration by drug treatment. Image analysis method is practical and might be useful for other applications in the field of biology and medical sciences. It could also be employed in a combination with the conventional assays to strengthen the data.
本研究的主要目的是通过两种不同的方法，即简单的数字图像分析方法和常规测定法，来分析成骨细胞行为的变化。用阿伦膦酸盐（ALN；一种含氮双膦酸盐）处理成骨细胞祖细胞和成骨细胞。检测成骨细胞摄取 ALN、细胞增殖、细胞分化和矿化结节形成等行为。使用公共的 ImageJ 软件和常规方法进行定量评估。此外，同时进行 ImageJ 法和矿化结节形成的常规测定，以证明 ImageJ 分析的可靠性。成骨细胞前体和成骨细胞对 ALN 处理有反应。该软件可以识别各种颜色，并允许对染色强度和面积覆盖度进行定量分析。图像分析和常规测定均能检测出药物处理前后细胞行为的变化。对于茜素红 S 测定法，ImageJ 分析计算的染色强度与常规测定法测量的吸光度相当。这些发现表明，数字图像分析与常规测定可用于定量评估药物处理对成骨细胞的改变。图像分析方法实用，可能对生物学和医学科学领域的其他应用有用。它也可以与常规测定法结合使用，以增强数据。
