Employing Nanostructured Scaffolds to Investigate the Mechanical Properties of Adult Mammalian Retinae Under Tension

Numerous eye diseases are linked to biomechanical dysfunction of the retina. However, the underlying forces are almost impossible to quantify experimentally. Here, we show how biomechanical properties of adult neuronal tissues such as porcine retinae can be investigated under tension in a home-built tissue stretcher composed of nanostructured TiO2 scaffolds coupled to a self-designed force sensor. The employed TiO2 nanotube scaffolds allow for organotypic long-term preservation of adult tissues ex vivo and support strong tissue adhesion without the application of glues, a prerequisite for tissue investigations under tension. In combination with finite element calculations we found that the deformation behavior is highly dependent on the displacement rate which results in Young’s moduli of (760–1270) Pa. Image analysis revealed that the elastic regime is characterized by a reversible shear deformation of retinal layers. For larger deformations, tissue destruction and sliding of retinal layers occurred with an equilibration between slip and stick at the interface of ruptured layers, resulting in a constant force during stretching. Since our study demonstrates how porcine eyes collected from slaughterhouses can be employed for ex vivo experiments, our study also offers new perspectives to investigate tissue biomechanics without excessive animal experiments. © 2020 by the authors. Licensee MDPI, Basel, Switzerland

Novel approaches to study the biomechanics of intact central nervous tissue

In nature, cells are not randomly clustered to form tissues. The tissue is a more complicated system with functions that go beyond what any single cell type could accomplish. While studying single-cell mechanics and dynamics is relevant from an investigative point of view, this approach loses, or fail to gather information about the tissue. The tissue investigated in this study is the neurosensory retina which seeing as extension of the brain is a very convenient model for the central nervous system due to its accessibility. The retina is constantly subjected to different mechanical stresses from development to adulthood. Although the majority of the phenomena where mechanical stresses are involved are well-studied, the mechanics behind them is not well understood. However, knowledge about the ability of the retina to adjust to mechanical stresses is essential, for example, for improving retinal surgery. Establishing a method to mechanically probe the retina is a challenge due to the extremely delicate nature of this multilayered neural tissue and to the short-time survival ex vivo. The organotypic tissue culture is a powerful tool because it allows to maintain with high accuracy the complex multicellular anatomy and the microenvironment of the original tissue. One of the limitations of the organotypic culture techniques has been until recently due to the ability to use only post-natal/juvenile tissues for long-term culture. The importance of using adult tissue is incontestable when the investigation focuses on age-related pathologies such as vitreous shrinkage or macula degeneration. In this work, TiO2 nanotube arrays are presented as the innovative substrate for long-term organotypic culture of adult neural tissue. The retinal whole-mount of adult guinea pig and the brain slices of adult mouse were cultures for 14 days without showing any sign of edema or swelling. Furthermore, in order to study the behavior of the retinal tissue under shear stress new set-ups were designed. For the first time, the behavior of the retinal layers were observed showing that the retina does not act as an homogeneous material in response to an applied stress. The methods developed here can be used for future quantitative studies, to provide an exact knowledge of retinal biomechanics which will help retinal surgeons to optimize their methods

Novel approaches to study the biomechanics of intact central nervous tissue

In nature, cells are not randomly clustered to form tissues. The tissue is a more complicated system with functions that go beyond what any single cell type could accomplish. While studying single-cell mechanics and dynamics is relevant from an investigative point of view, this approach loses, or fail to gather information about the tissue. The tissue investigated in this study is the neurosensory retina which seeing as extension of the brain is a very convenient model for the central nervous system due to its accessibility. The retina is constantly subjected to different mechanical stresses from development to adulthood. Although the majority of the phenomena where mechanical stresses are involved are well-studied, the mechanics behind them is not well understood. However, knowledge about the ability of the retina to adjust to mechanical stresses is essential, for example, for improving retinal surgery. Establishing a method to mechanically probe the retina is a challenge due to the extremely delicate nature of this multilayered neural tissue and to the short-time survival ex vivo. The organotypic tissue culture is a powerful tool because it allows to maintain with high accuracy the complex multicellular anatomy and the microenvironment of the original tissue. One of the limitations of the organotypic culture techniques has been until recently due to the ability to use only post-natal/juvenile tissues for long-term culture. The importance of using adult tissue is incontestable when the investigation focuses on age-related pathologies such as vitreous shrinkage or macula degeneration. In this work, TiO2 nanotube arrays are presented as the innovative substrate for long-term organotypic culture of adult neural tissue. The retinal whole-mount of adult guinea pig and the brain slices of adult mouse were cultures for 14 days without showing any sign of edema or swelling. Furthermore, in order to study the behavior of the retinal tissue under shear stress new set-ups were designed. For the first time, the behavior of the retinal layers were observed showing that the retina does not act as an homogeneous material in response to an applied stress. The methods developed here can be used for future quantitative studies, to provide an exact knowledge of retinal biomechanics which will help retinal surgeons to optimize their methods

Annexin A2 in Inflammation and Host Defense

Annexin A2 (AnxA2) is a multifunctional calcium2+ (Ca2+) and phospholipid-binding protein that is expressed in a wide spectrum of cells, including those participating in the inflammatory response. In acute inflammation, the interaction of AnxA2 with actin and adherens junction VE-cadherins underlies its role in regulating vascular integrity. In addition, its contribution to endosomal membrane repair impacts several aspects of inflammatory regulation, including lysosome repair, which regulates inflammasome activation, and autophagosome biogenesis, which is essential for macroautophagy. On the other hand, AnxA2 may be co-opted to promote adhesion, entry, and propagation of bacteria or viruses into host cells. In the later stages of acute inflammation, AnxA2 contributes to the initiation of angiogenesis, which promotes tissue repair, but, when dysregulated, may also accompany chronic inflammation. AnxA2 is overexpressed in malignancies, such as breast cancer and glioblastoma, and likely contributes to cancer progression in the context of an inflammatory microenvironment. We conclude that annexin AnxA2 normally fulfills a spectrum of anti-inflammatory functions in the setting of both acute and chronic inflammation but may contribute to disease states in settings of disordered homeostasis

Employing Nanostructured Scaffolds to Investigate the Mechanical Properties of Adult Mammalian Retinae Under Tension

Numerous eye diseases are linked to biomechanical dysfunction of the retina. However, the underlying forces are almost impossible to quantify experimentally. Here, we show how biomechanical properties of adult neuronal tissues such as porcine retinae can be investigated under tension in a home-built tissue stretcher composed of nanostructured TiO2 scaffolds coupled to a self-designed force sensor. The employed TiO2 nanotube scaffolds allow for organotypic long-term preservation of adult tissues ex vivo and support strong tissue adhesion without the application of glues, a prerequisite for tissue investigations under tension. In combination with finite element calculations we found that the deformation behavior is highly dependent on the displacement rate which results in Young’s moduli of (760–1270) Pa. Image analysis revealed that the elastic regime is characterized by a reversible shear deformation of retinal layers. For larger deformations, tissue destruction and sliding of retinal layers occurred with an equilibration between slip and stick at the interface of ruptured layers, resulting in a constant force during stretching. Since our study demonstrates how porcine eyes collected from slaughterhouses can be employed for ex vivo experiments, our study also offers new perspectives to investigate tissue biomechanics without excessive animal experiments

Employing Nanostructured Scaffolds to Investigate the Mechanical Properties of Adult Mammalian Retinae Under Tension

Numerous eye diseases are linked to biomechanical dysfunction of the retina. However, the underlying forces are almost impossible to quantify experimentally. Here, we show how biomechanical properties of adult neuronal tissues such as porcine retinae can be investigated under tension in a home-built tissue stretcher composed of nanostructured TiO2 scaffolds coupled to a self-designed force sensor. The employed TiO2 nanotube scaffolds allow for organotypic long-term preservation of adult tissues ex vivo and support strong tissue adhesion without the application of glues, a prerequisite for tissue investigations under tension. In combination with finite element calculations we found that the deformation behavior is highly dependent on the displacement rate which results in Young’s moduli of (760–1270) Pa. Image analysis revealed that the elastic regime is characterized by a reversible shear deformation of retinal layers. For larger deformations, tissue destruction and sliding of retinal layers occurred with an equilibration between slip and stick at the interface of ruptured layers, resulting in a constant force during stretching. Since our study demonstrates how porcine eyes collected from slaughterhouses can be employed for ex vivo experiments, our study also offers new perspectives to investigate tissue biomechanics without excessive animal experiments

Employing Nanostructured Scaffolds to Investigate the Mechanical Properties of Adult Mammalian Retinae Under Tension

Numerous eye diseases are linked to biomechanical dysfunction of the retina. However, the underlying forces are almost impossible to quantify experimentally. Here, we show how biomechanical properties of adult neuronal tissues such as porcine retinae can be investigated under tension in a home-built tissue stretcher composed of nanostructured TiO2 scaffolds coupled to a self-designed force sensor. The employed TiO2 nanotube scaffolds allow for organotypic long-term preservation of adult tissues ex vivo and support strong tissue adhesion without the application of glues, a prerequisite for tissue investigations under tension. In combination with finite element calculations we found that the deformation behavior is highly dependent on the displacement rate which results in Young’s moduli of (760–1270) Pa. Image analysis revealed that the elastic regime is characterized by a reversible shear deformation of retinal layers. For larger deformations, tissue destruction and sliding of retinal layers occurred with an equilibration between slip and stick at the interface of ruptured layers, resulting in a constant force during stretching. Since our study demonstrates how porcine eyes collected from slaughterhouses can be employed for ex vivo experiments, our study also offers new perspectives to investigate tissue biomechanics without excessive animal experiments

Employing Nanostructured Scaffolds to Investigate the Mechanical Properties of Adult Mammalian Retinae Under Tension

Numerous eye diseases are linked to biomechanical dysfunction of the retina. However, the underlying forces are almost impossible to quantify experimentally. Here, we show how biomechanical properties of adult neuronal tissues such as porcine retinae can be investigated under tension in a home-built tissue stretcher composed of nanostructured TiO2 scaffolds coupled to a self-designed force sensor. The employed TiO2 nanotube scaffolds allow for organotypic long-term preservation of adult tissues ex vivo and support strong tissue adhesion without the application of glues, a prerequisite for tissue investigations under tension. In combination with finite element calculations we found that the deformation behavior is highly dependent on the displacement rate which results in Young’s moduli of (760–1270) Pa. Image analysis revealed that the elastic regime is characterized by a reversible shear deformation of retinal layers. For larger deformations, tissue destruction and sliding of retinal layers occurred with an equilibration between slip and stick at the interface of ruptured layers, resulting in a constant force during stretching. Since our study demonstrates how porcine eyes collected from slaughterhouses can be employed for ex vivo experiments, our study also offers new perspectives to investigate tissue biomechanics without excessive animal experiments