Method for quantitative analysis of glycosaminoglycan distribution in cultured natural and engineered cartilage
Method for quantitative analysis of glycosaminoglycan distribution in cultured natural and engineered cartilage
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DOI：
10.1114/1.205
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发表时间：
1999-09-01
影响因子：
3.8
通讯作者：
Vunjak-Novakovic, G
中科院分区：
文献类型：
--
作者：
Martin, I;Obradovic, B;Vunjak-Novakovic, G
软骨组织工程可以为组织发育的受控研究提供有价值的工具。例如，在不同培养条件下工程化的软骨组织切片中糖胺聚糖（GAG）的空间分布分析可用于将环境因素的影响与再生组织的结构相关联。在本文中，我们描述了一种基于计算机的技术，用于定量分析番红-O染色的组织切片，使用低放大倍数的光学显微镜图像。我们确定了一个参数来量化切片中红色的强度，这反过来又与相应组织样品中GAG的生化测定湿重分数成比例，并将GAG的空间分布描述为深度的函数。部分边缘。然后使用折线回归模型确定具有较低GAG分数的外部区域的厚度和GAG含量的空间变化率。将该方法应用于在三种不同容器中培养6周的天然和工程软骨样品中GAG分布的定量：静态烧瓶、混合烧瓶和旋转生物反应器。(C)1999年生物医学工程学会。[S0090-6964（99）00505-6]。
Cartilage tissue engineering can provide a valuable tool for controlled studies of tissue development. As an example, analysis of the spatial distribution of glycosaminoglycans (GAG) in sections of cartilaginous tissues engineered under different culture conditions could be used to correlate the effects of environmental factors with the structure of the regenerated tissue. In this paper we describe a computer-based technique for quantitative analysis of safranin-O stained histological sections, using low magnification light microscopy images. We identified a parameter to quantify the intensity of red color in the sections, which in turn was proportional to the biochemically determined wet weight fraction of GAG in corresponding tissue samples, and to describe the spatial distribution of GAG as a function of depth from. the section edge. A broken line regression model was then used to determine the thickness of an external region, with lower GAG fractions, and the spatial rate of change in GAG content. The method was applied to the quantitatation of GAG distribution in samples of natural and engineered cartilage, cultured for 6 weeks in three different vessels: static flasks, mixed flasks, and rotating bioreactors. (C) 1999 Biomedical Engineering Society. [S0090-6964(99)00505-6].