Phase-contrast enhanced synchrotron micro-tomography of human meniscus tissue

Einarsson, Emma; Pierantoni, M.; Novák, Vladimír; Svensson, Jonas; Isaksson, Hanna; Englund, Martin

doi:10.1016/j.joca.2022.06.003

Cited by 11 publications

(7 citation statements)

References 35 publications

(43 reference statements)

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“…For image analysis, 5 sub‐volumes of the full meniscus depth × 500 × 500 µm 3 were cropped from each tomographic reconstruction for collagen fiber orientation analysis. The orientation was computed using the structure gradient tensor method, [ 64 , 65 ] as previously applied to static imaging of human meniscus tissue [ 35 ] and rat Achilles tendons. [ 44 ] Briefly, images were initially smoothed using a Gaussian filter (standard deviation of 1).…”

Section: Methodsmentioning

confidence: 99%

“…For soft tissues, such differences can be several orders of magnitude higher than in conventional absorption. [ 27 ] This technique has previously been used to characterize the microstructure of rodent, [ 28 , 29 ] bovine, [ 30 ] and human cartilage, [ 18 , 31 , 32 , 33 , 34 ] as well as human meniscus, [ 35 ] allowing to differentiate cellular and collagenous fiber structures, and showing some potential to distinguish some signs of OA‐related degeneration. However, these studies have focused on static measurements, without considering the tissues’ mechanical and microstructural responses under loading conditions.…”

Section: Introductionmentioning

confidence: 99%

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In Situ Loading and Time‐Resolved Synchrotron‐Based Phase Contrast Tomography for the Mechanical Investigation of Connective Knee Tissues: A Proof‐of‐Concept Study

Dejea,

Pierantoni,

Orozco

et al. 2024

Advanced Science

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Articular cartilage and meniscus transfer and distribute mechanical loads in the knee joint. Degeneration of these connective tissues occurs during the progression of knee osteoarthritis, which affects their composition, microstructure, and mechanical properties. A deeper understanding of disease progression can be obtained by studying them simultaneously. Time‐resolved synchrotron‐based X‐ray phase‐contrast tomography (SR‐PhC‐µCT) allows to capture the tissue dynamics. This proof‐of‐concept study presents a rheometer setup for simultaneous in situ unconfined compression and SR‐PhC‐µCT of connective knee tissues. The microstructural response of bovine cartilage (n = 16) and meniscus (n = 4) samples under axial continuously increased strain, or two steps of 15% strain (stress–relaxation) is studied. The chondrocyte distribution in cartilage and the collagen fiber orientation in the meniscus are assessed. Variations in chondrocyte density reveal an increase in the top 40% of the sample during loading, compared to the lower half. Meniscus collagen fibers reorient perpendicular to the loading direction during compression and partially redisperse during relaxation. Radiation damage, image repeatability, and image quality assessments show little to no effects on the results. In conclusion, this approach is highly promising for future studies of human knee tissues to understand their microstructure, mechanical response, and progression in degenerative diseases.

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Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

In Situ Loading and Time‐Resolved Synchrotron‐Based Phase Contrast Tomography for the Mechanical Investigation of Connective Knee Tissues: A Proof‐of‐Concept Study

Dejea,

Pierantoni,

Orozco

et al. 2024

Advanced Science

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show abstract

“…PhC-micr was able to visualize the human meniscus with good contrast and resolution and the ages were able to show the individual collagen fibers and the crimping pattern of th bers. The study also found that the quantitative parameters related to fiber orientation From the same research group, the following year, Einarsson et al [47] investigated the feasibility of synchrotron radiation-based PhC-microCT for the imaging of the human meniscus. Quantitative parameters pertaining to both fiber orientation and crimping were systematically assessed as prospective indicators of tissue degeneration.…”

Section: Knee Joint and Surrounding Tendonsmentioning

confidence: 96%

“…These findings suggest that fresh frozen samples are the prefe sample preparation method for imaging tendon collagen fibers as it preserves the n structure of the fibers and minimizes the introduction of artifacts. From the same research group, the following year, Einarsson et al [47] investig the feasibility of synchrotron radiation-based PhC-microCT for the imaging of the hu meniscus. Quantitative parameters pertaining to both fiber orientation and crimping w systematically assessed as prospective indicators of tissue degeneration.…”

Section: Knee Joint and Surrounding Tendonsmentioning

confidence: 99%

High-Resolution Phase-Contrast Tomography on Human Collagenous Tissues: A Comprehensive Review

Furlani,

Riberti,

Gatto

et al. 2023

Tomography

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Phase-contrast X-ray imaging is becoming increasingly considered since its first applications, which occurred almost 30 years ago. Particular emphasis was placed on studies that use this technique to investigate soft tissues, which cannot otherwise be investigated at a high resolution and in a three-dimensional manner, using conventional absorption-based settings. Indeed, its consistency and discrimination power in low absorbing samples, unified to being a not destructive analysis, are pushing interests on its utilization from researchers of different specializations, from botany, through zoology, to human physio-pathology research. In this regard, a challenging method for 3D imaging and quantitative analysis of collagenous tissues has spread in recent years: it is based on the unique characteristics of synchrotron radiation phase-contrast microTomography (PhC-microCT). In this review, the focus has been placed on the research based on the exploitation of synchrotron PhC-microCT for the investigation of collagenous tissue physio-pathologies from solely human samples. Collagen tissues’ elasto-mechanic role bonds it to the morphology of the site it is extracted from, which could weaken the results coming from animal experimentations. Encouraging outcomes proved this technique to be suitable to access and quantify human collagenous tissues and persuaded different researchers to approach it. A brief mention was also dedicated to the results obtained on collagenous tissues using new and promising high-resolution phase-contrast tomographic laboratory-based setups, which will certainly represent the real step forward in the diffusion of this relatively young imaging technique.

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“…Several studies have used SR PhC-µCT to investigate the microstructure of unstained human organs (Walsh et al, 2021, Einarsson et al, 2022, Lee et al, 2022). Some have acquired images from human brain samples with sub-cellular detail and successfully segmented neurons to study their three-dimensional spatial distributions (Hieber et al, 2006, Frost et al, 2023).…”

Section: Introductionmentioning

confidence: 99%

Protocol for 3D Virtual Histology of Unstained Human Brain Tissue using Synchrotron Radiation Phase-Contrast Microtomography

Lee,

Donato,

Mack

et al. 2023

Preprint

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X-ray phase-contrast micro computed tomography using synchrotron radiation (SR PhC-µCT) offers unique 3D imaging capabilities for visualizing microstructure of the human brain. Its applicability for unstained soft tissue is an area of active research. Acquiring images from a tissue block without needing to section it into thin slices, as required in routine histology, allows for investigating the microstructure in its natural 3D space. This paper presents a detailed step-by-step guideline for imaging unstained human brain tissue at resolutions of a few micrometers with SR PhC-µCT implemented at SYRMEP, the hard X-ray imaging beamline of Elettra, the Italian synchrotron facility. We present examples of how blood vessels and neurons appear in the images acquired with isotropic 5 µm and 1 µm voxel sizes. Furthermore, the proposed protocol can be used to investigate important biological substrates such as neuromelanin or corpora amylacea. Their spatial distribution can be studied using specifically tailored segmentation tools that are validated by classical histology methods. In conclusion, SR PhC-µCT using the proposed protocols, including data acquisition and image processing, offers viable means of obtaining information about the anatomy of the human brain at the cellular level in 3D.

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Phase-contrast enhanced synchrotron micro-tomography of human meniscus tissue

Cited by 11 publications

References 35 publications

In Situ Loading and Time‐Resolved Synchrotron‐Based Phase Contrast Tomography for the Mechanical Investigation of Connective Knee Tissues: A Proof‐of‐Concept Study

In Situ Loading and Time‐Resolved Synchrotron‐Based Phase Contrast Tomography for the Mechanical Investigation of Connective Knee Tissues: A Proof‐of‐Concept Study

High-Resolution Phase-Contrast Tomography on Human Collagenous Tissues: A Comprehensive Review

Protocol for 3D Virtual Histology of Unstained Human Brain Tissue using Synchrotron Radiation Phase-Contrast Microtomography

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