Parameterizing the Transport Pathways for Cell Invasion in Complex Scaffold Architectures

Ashworth, J.; Mehr, Marco; Buxton, Paul; Best, Serena M.; Cameron, Ruth E.

doi:10.1089/ten.tec.2015.0483

Cited by 23 publications

(27 citation statements)

References 19 publications

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“…Micro-CT analysis was also used for numerical analysis of the pore space transport pathways through the structure, by calculation of percolation diameter as described previously [18]. Briefly, the distance accessible in a specified direction, L, to a spherical object of diameter d may be described using the following relationship:…”

Section: Morphological Characterizationmentioning

confidence: 99%

“…This critical diameter describes the characteristic size of the pathways encountered by an invading object in a specified direction [18]. Measurements of L and d were obtained using the Skyscan software CTAn, for object travel in the scaffold z-direction (i.e.…”

Section: Morphological Characterizationmentioning

confidence: 99%

“…These included gelatin [13] and its blend with hydroxyapatite [14], polylactic acid (PLA) [15], polycaprolactone (PCL) [16], and chitosan and alginate [17]. Methods for the characterization of key features of such microporous scaffolds, like the presence and characteristic size of transport pathways and the multi-scale mechanical response, have been developed [18,19]. Nevertheless, the relationships between solid arrangement and scaffold physical properties are still not well understood.…”

Section: Introductionmentioning

confidence: 99%

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Structural determinants of hydration, mechanics and fluid flow in freeze-dried collagen scaffolds

et al. 2016

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Freeze-dried collagen sponges are three-dimensional microporous scaffolds that have been used for a number of exploratory tissue engineering applications. The characterization of the structure-properties relationships of these scaffolds is necessary to understand their biophysical behavior in vivo. In this work, the relationship between morphology and physical properties in the dry and hydrated states was investigated across a range of solid concentrations in the scaffolds. The quantitative results provided can aid the design of scaffolds with a target trade-off between mechanical properties and structural features important for their biological activity.

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Section: Morphological Characterizationmentioning

confidence: 99%

Section: Morphological Characterizationmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Structural determinants of hydration, mechanics and fluid flow in freeze-dried collagen scaffolds

et al. 2016

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“…Micro-computed tomography (MicroCT) provides an obvious tool for the characterization of tissue-engineering scaffolds and development in image analysis, particularly pioneered by Ashworth et al [2][3][4], has allowed for its application as a predictive tool for cell migration. However, the results from any MicroCT analysis should be treated with a degree of caution.…”

Section: Introductionmentioning

confidence: 99%

MicroCT analysis of connectivity in porous structures: optimizing data acquisition and analytical methods in the context of tissue engineering

Nair

Shepherd

Best

et al. 2020

J. R. Soc. Interface.

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Micro-computed X-ray tomography (MicroCT) is one of the most powerful techniques available for the three-dimensional characterization of complex multi-phase or porous microarchitectures. The imaging and analysis of porous networks are of particular interest in tissue engineering due to the ability to predict various large-scale cellular phenomena through the micro-scale characterization of the structure. However, optimizing the parameters for MicroCT data capture and analyses requires a careful balance of feature resolution and computational constraints while ensuring that a structurally representative section is imaged and analysed. In this work, artificial datasets were used to evaluate the validity of current analytical methods by considering the effect of noise and pixel size arising from the data capture, and intrinsic structural anisotropy and heterogeneity. A novel ‘segmented percolation method’ was developed to exclude the effect of anomalous, non-representative features within the datasets, allowing for scale-invariant structural parameters to be obtained consistently and without manual intervention for the first time. Finally, an in-depth assessment of the imaging and analytical procedures are presented by considering percolation events such as micro-particle filtration and cell sieving within the context of tissue engineering. Along with the novel guidelines established for general pixel size selection for MicroCT, we also report our determination of 3 μm as the definitive pixel size for use in analysing connectivity for tissue engineering applications.

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“…

of directing the regeneration of such complex tissues, a detailed understanding of the cell type dependent link between scaffold structure and cell motility is therefore required. [10,11] These transport pathways can be characterized by measurement of percolation diameter, which describes the maximum object diameter that can move through a structure unimpeded. By comparing the invasion behavior of cells from three different sources, in scaffolds of systematically increasing accessibility, we probe the structural requirements for cell movement.

…”

mentioning

confidence: 99%

Towards Cellular Sieving: Exploring the Limits of Scaffold Accessibility for Cell Type Specific Invasion

et al. 2018

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of directing the regeneration of such complex tissues, a detailed understanding of the cell type dependent link between scaffold structure and cell motility is therefore required. In this work, we describe how this cell type dependence may be exploited toward cellular sieving: scaffold structures that permit differing degrees of invasion according to cell type. By comparing the invasion behavior of cells from three different sources, in scaffolds of systematically increasing accessibility, we probe the structural requirements for cell movement. In this way, we demonstrate that highly occlusive structures may be used to allow differential invasion according to cell type, therefore physically separating cells according to their invasive capacity.Freeze-dried collagen scaffolds are promising materials in the tissue engineering field, since their highly tunable structure and properties provide potential for the regeneration of a wide range of tissues. [5][6][7][8][9] In recent work, we have shown that measurement of pore size in these scaffolds is insufficient for predicting accessibility to cell invasion, and that characterizing the transport pathways through the pore space is also necessary. [10,11] These transport pathways can be characterized by measurement of percolation diameter, which describes the maximum object diameter that can move through a structure unimpeded. Percolation diameters below 40 µm have been shown to inhibit the invasion of primary fibroblasts, even in structures of constant pore size and collagen wall orientation. [11] In this study, we present simultaneous analysis of pore size and percolation diameter, meaning that, for the first time, we are able to compare their relative influences on the invasion potential of each cell type. The cells chosen for investigation reflect three distinct tissue engineering applications: human primary fibroblasts, for soft tissue regeneration; [12] the MC3T3 mouse osteoprogenitor cell line, for bone regeneration; [13] and the HT1080 cell line from human fibrosarcoma, reflecting the use of 3D models for studying tumor cell invasion. [14] As well as providing a tool for assaying the invasive capability of different cells in response to defined structures, this approach demonstrates how the informed design of tissue engineering scaffolds could be used to control the directed invasion of multiple cell types. The differing species, phenotype and origin of these cells is also relevant for demonstrating how the application of interest may determine the structural design criteria required for tissue engineering scaffolds, as well as the choice of cell line needed to provide an accurate biological assessment of such materials.Coordinating the behavior of multiple cell types provides a challenge for the tissue engineer, since response to structure is highly cell type dependent. Here, human primary fibroblast invasion is compared with that of the MC3T3 and HT1080 cell lines to demonstrate proof-of-concept that controlled changes in 3D collagen scaffold architecture can m...

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Parameterizing the Transport Pathways for Cell Invasion in Complex Scaffold Architectures

Cited by 23 publications

References 19 publications

Structural determinants of hydration, mechanics and fluid flow in freeze-dried collagen scaffolds

Structural determinants of hydration, mechanics and fluid flow in freeze-dried collagen scaffolds

MicroCT analysis of connectivity in porous structures: optimizing data acquisition and analytical methods in the context of tissue engineering

Towards Cellular Sieving: Exploring the Limits of Scaffold Accessibility for Cell Type Specific Invasion

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