Atomic Force Microscopy Micro-Indentation Methods for Determining the Elastic Modulus of Murine Articular Cartilage.
Atomic Force Microscopy Micro-Indentation Methods for Determining the Elastic Modulus of Murine Articular Cartilage.
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DOI：
10.3390/s23041835
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发表时间：
2023-02-07
期刊：
影响因子：
--
通讯作者：
Westendorf JJ
中科院分区：
文献类型：
--
作者：
Arnold KM;Sicard D;Tschumperlin DJ;Westendorf JJ
生物组织的力学特性影响其功能，并可以在大体组织学或生理变化被检测到之前预测退行性疾病。对于像关节软骨这样的结构组织来说尤其如此，关节软骨具有主要的机械功能，在损伤后和骨关节炎的早期阶段会下降。虽然原子力显微镜（AFM）已被用于测试关节软骨的弹性模量之前，没有一致或一致性的方法报道。对于小鼠关节软骨，不同的方法主要有两个方面：实验参数的选择和样品的制备。影响AFM结果的实验参数包括压痕力和悬臂刚度；这些取决于尖端，样品和使用的仪器。本项目的目的是优化这些实验参数，用AFM微压痕测量小鼠关节软骨弹性模量。我们首先研究了实验参数对对照材料聚二甲基硅氧烷凝胶（PDMS）的影响，该材料具有与关节软骨相同数量级的弹性模量。在此对照材料上缩小实验参数，然后在野生型C57BL/6J小鼠关节软骨样本上确定实验参数，该样本采用一种新的技术制备，可以在不脱钙的情况下对关节软骨和长骨的骨骺段进行冷冻切片。该技术有助于AFM测量在小鼠关节软骨基质上的精确定位，并且消除了将软骨与底层骨组织分离的需要，这在小鼠骨骼中由于其小尺寸而具有挑战性。总之，新的样品制备方法和优化的实验参数为测量小鼠关节软骨弹性模量的微尺度变化提供了可靠的标准操作程序。
The mechanical properties of biological tissues influence their function and can predict degenerative conditions before gross histological or physiological changes are detectable. This is especially true for structural tissues such as articular cartilage, which has a primarily mechanical function that declines after injury and in the early stages of osteoarthritis. While atomic force microscopy (AFM) has been used to test the elastic modulus of articular cartilage before, there is no agreement or consistency in methodologies reported. For murine articular cartilage, methods differ in two major ways: experimental parameter selection and sample preparation. Experimental parameters that affect AFM results include indentation force and cantilever stiffness; these are dependent on the tip, sample, and instrument used. The aim of this project was to optimize these experimental parameters to measure murine articular cartilage elastic modulus by AFM micro-indentation. We first investigated the effects of experimental parameters on a control material, polydimethylsiloxane gel (PDMS), which has an elastic modulus on the same order of magnitude as articular cartilage. Experimental parameters were narrowed on this control material, and then finalized on wildtype C57BL/6J murine articular cartilage samples that were prepared with a novel technique that allows for cryosectioning of epiphyseal segments of articular cartilage and long bones without decalcification. This technique facilitates precise localization of AFM measurements on the murine articular cartilage matrix and eliminates the need to separate cartilage from underlying bone tissues, which can be challenging in murine bones because of their small size. Together, the new sample preparation method and optimized experimental parameters provide a reliable standard operating procedure to measure microscale variations in the elastic modulus of murine articular cartilage.
影响因子：
3.7
作者：
Golubeva YG;Smith RM;Sternberg LR
通讯作者：
Sternberg LR