The data processing regarding AFM nanoindentation experiments on biological samples relies on the basic contact mechanics models like the Hertz model and the Oliver & Pharr analysis. Despite the fact that the two aforementioned techniques are assumed to provide equivalent results since they are based on the same underlying theory of contact mechanics, significant differences regarding the Young’s modulus calculation even on the same tested sample have been presented in the literature. The differences can be even greater than 30% depending on the used model. In addition, when the Oliver & Pharr analysis is used, a systematic greater Young’s modulus value is always calculated compared to the Hertzian analysis. In this paper, the two techniques are briefly described and two possible reasons that accurately explain the observed differences in the calculated value of the Young’s modulus are presented.

Atomic force microscopy (AFM) is an easy-to-use, powerful, high-resolution microscope that allows the user to image any surface and under any aqueous condition. AFM has been used in the investigation of the structural and mechanical properties of a wide range of biological matters including biomolecules, biomaterials, cells, and tissues. It provides the capacity to acquire high-resolution images of biosamples at the nanoscale and allows at readily carrying out mechanical characterization. The capacity of AFM to image and interact with surfaces, under physiologically relevant conditions, is of great importance for realistic and accurate medical and pharmaceutical applications. The aim of this paper is to review recent trends of the use of AFM on biological materials related to health and sickness. First, we present AFM components and its different imaging modes and we continue with combined imaging and coupled AFM systems. Then, we discuss the use of AFM to nanocharacterize collagen, the major fibrous protein of the human body, which has been correlated with many pathological conditions. In the next section, AFM nanolevel surface characterization as a tool to detect possible pathological conditions such as osteoarthritis and cancer is presented. Finally, we demonstrate the use of AFM for studying other pathological conditions, such as Alzheimer’s disease and human immunodeficiency virus (HIV), through the investigation of amyloid fibrils and viruses, respectively. Consequently, AFM stands out as the ideal research instrument for exploring the detection of pathological conditions even at very early stages, making it very attractive in the area of bio- and nanomedicine.

AFM nanoindentation is a modern technique which is used for the determination of the mechanical properties of materials and biomaterials. Several models of applied mechanics can be used for data processing regarding the nanoindentation procedure such as the Hertz model which is used in the case of spherical indenters. The purpose of this review is to provide a complete presentation of the Hertz model theory, which is the most commonly used model in the fields of Biophysics and Biomaterials as well as related applications. In addition, the theory which correlates the Hertz model and the general nanoindentation equation for the Young’s modulus determination is provided.

Atomic Force Microscopy nanoindentation method is a powerful technique that can be used for the nano-mechanical characterization of bio-samples. Significant scientific efforts have been performed during the last two decades to accurately determine the Young’s modulus of collagen fibrils at the nanoscale, as it has been proven that mechanical alterations of collagen are related to various pathological conditions. Different contact mechanics models have been proposed for processing the force–indentation data based on assumptions regarding the shape of the indenter and collagen fibrils and on the elastic or elastic–plastic contact assumption. However, the results reported in the literature do not always agree; for example, the Young’s modulus values for dry collagen fibrils expand from 0.9 to 11.5 GPa. The most significant parameters for the broad range of values are related to the heterogeneous structure of the fibrils, the water content within the fibrils, the data processing errors, and the uncertainties in the calibration of the probe. An extensive discussion regarding the models arising from contact mechanics and the results provided in the literature is presented, while new approaches with respect to future research are proposed.