3D analysis and grading of calcifications from ex vivo human meniscus

Hellberg, Iida; Karjalainen, Ville-Pauli; Finnilä, Mikko; Jonsson, Erland; Turkiewicz, Aleksandra; Önnerfjord, Patrik; Hughes, V.; Tjörnstrand, Jon; Englund, Martin; Saarakkala, Simo

doi:10.1016/j.joca.2022.10.016

Cited by 6 publications

(6 citation statements)

References 43 publications

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“…33,51 Moreover, we have seen similar calcifications in our previous study where we showed the feasibility of quantitative studies of the calcified meniscus in 3D with µCT and found that calcification volume in the meniscus increases with increasing histopathological degeneration. 24 Our findings show that all three samples inhabiting calcifications also had the highest ratio of the vascular fraction of the anterior horn in adults, suggesting correlation a between meniscus angiogenesis and ossification in the medial anterior horn. While it is possible to do quantitative analysis of the meniscal calcifications using µCT imaging, characterizing the crystal type requires another modality, for example, Raman spectroscopy or x-ray diffraction.…”

Section: Cpd-based Sample Drying Protocol Together With High-resoluti...supporting

confidence: 51%

“…Few adult menisci were found inhabiting calcified structures in the µCT images. We refined our previously used calcification analysis method where we had identified calcifications from meniscus together with µCT and histology 24 . In this study, we identified similar particles and identified them as calcifications based on their high density compared to surrounding soft tissue, their location inside the meniscus, and their crystal‐shaped morphology and size.…”

Section: Methodsmentioning

confidence: 99%

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Age and anatomical region‐related differences in vascularization of the porcine meniscus using microcomputed tomography imaging

Karjalainen,

Herrera Millar,

Modina

et al. 2024

Journal Orthopaedic Research

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Meniscal lesions in vascularized regions are known to regenerate while lack of vascular supply leads to poor healing. Here, we developed and validated a novel methodology for three‐dimensional structural analysis of meniscal vascular structures with high‐resolution microcomputed tomography (µCT). We collected porcine medial menisci from 10 neonatal (not‐developed meniscus, n‐) and 10 adults (fully developed meniscus, a‐). The menisci were cut into anatomical regions (anterior horn (n‐AH and a‐AH), central body (n‐CB and a‐CB), and posterior horn (n‐PH and a‐PH). Specimens were cut in half, fixed, and one specimen underwent critical point drying and µCT imaging, while other specimen underwent immunohistochemistry and vascularity biomarker CD31 staining for validation of µCT. Parameters describing vascular structures were calculated from µCT. The vascular network in neonatal spread throughout meniscus, while in adult was limited to a few vessels in outer region, mostly on femoral side. n‐AH, n‐CB, and n‐PH had 20, 17, and 11 times greater vascular volume fraction than adult, respectively. Moreover, thickness of blood vessels, in three regions, was six times higher in adults than in neonatal. a‐PH appeared to have higher vascular fraction, longer and thicker blood vessels than both a‐AH and a‐CB. Overall, neonatal regions had a higher number of blood vessels, more branching, and higher tortuosity compared to adult regions. For the first time, critical point drying‐based µCT imaging allowed detailed three‐dimensional visualization and quantitative analysis of vascularized meniscal structures. We showed more vascularity in neonatal menisci, while adult menisci had fewer and thicker vascularity especially limited to the femoral surface.

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Section: Cpd-based Sample Drying Protocol Together With High-resoluti...supporting

confidence: 51%

Section: Methodsmentioning

confidence: 99%

Age and anatomical region‐related differences in vascularization of the porcine meniscus using microcomputed tomography imaging

Karjalainen,

Herrera Millar,

Modina

et al. 2024

Journal Orthopaedic Research

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“…In early OA, increased expression of VEGF and angiogenesis promotes vascular content leading to ossification in the articular cartilage 29,48 . Moreover, in our previous study, we found that calcifications in the meniscus increase with increasing histopathological degeneration 49 . Our findings show that all three samples inhabiting calcifications had also the highest ratio of the vascular fraction of the anterior horn in adults, suggesting correlation a between meniscus angiogenesis and ossification in the medial anterior horn.…”

Section: Discussionmentioning

confidence: 71%

“…Our findings show that all three samples inhabiting calcifications had also the highest ratio of the vascular fraction of the anterior horn in adults, suggesting correlation a between meniscus angiogenesis and ossification in the medial anterior horn. We have previously shown that with µCT, you can quantitatively study the calcified tissue in the meniscus in 3D 49 . Thus, the presented method of µCT imaging enables imaging of hard tissues like calcification together with soft tissues like vascularization and could be used to study the effects of ossification and angiogenesis in the meniscus in the future.…”

Section: Discussionmentioning

confidence: 99%

Age and anatomical region related differences in vascularization of the porcine meniscus using micro-computed tomography imaging

Karjalainen,

Herrera M,

Modina

et al. 2023

Preprint

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Meniscal lesions in vascularized regions are known to regenerate while lack of vascular supply leads to poor healing. Here we developed and validated novel methodology for three-dimensional structural analysis of meniscal vascular structures with high-resolution micro-computed tomography (µCT).We collected porcine medial menisci from 10 neonatal (not-developed meniscus, n-) and 10 adults (fully developed meniscus, a-). The menisci were cut into anatomical regions (anterior horn (n-AH & a-AH), central body (n-CB & a-CB), and posterior horn (n-PH & a-PH).Specimens were cut in half, fixed, and one specimen underwent critical point drying and µCT imaging, while other specimen underwent immunohistochemistry and vascularity biomarker CD31 staining for validation of µCT. Parameters describing vascular structures were calculated from µCT.The vascular network in neonatal spread throughout meniscus, while in adult was limited to a few vessels in outer region, mostly on femoral side. a-AH, a-CB, and a-PH had three, five, and seven times greater vascular volume than neonate, respectively. Moreover, thickness of blood vessels, in three regions, was six times higher in adult than in newborn. Finally, a-PH appeared to have thicker blood vessels than both a-AH and a-CB.For the first time, critical point drying-based µCT imaging allowed detailed three-dimensional visualization and quantitative analysis of vascularized meniscal structures. We showed more vascularity in neonatal menisci, while adult menisci had fewer and thicker vascularity especially limited to the femoral surface which is involved in load transmission response, thus suggesting how nutritional support in this area of the outer zone is more necessary.

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“…The analysis of CT datasets permits to define several parameters and geometrical variables, which are relevant for describing the structure and properties of a sample 24 , 25 . In studies focused on meniscal analysis, micro-CT technology has been applied to a variety of investigations: geometry and dimensions 26 – 28 , calcification detection 29 – 31 , fibrous microstructure 11 , 32 – 36 , fluid flow regimes 37 , and contrast agent diffusion kinematics 38 . Despite this extensive application of micro-CT to various aspects of meniscal research, none of these studies have yet utilized this promising technique specifically to analyze vascularization within the meniscus.…”

Section: Introductionmentioning

confidence: 99%

Revealing the complexity of meniscus microvasculature through 3D visualization and analysis

Orellana,

Grassi,

Hlushchuk

et al. 2024

Sci Rep

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Three-dimensional information is essential for a proper understanding of the healing potential of the menisci and their overall role in the knee joint. However, to date, the study of meniscal vascularity has relied primarily on two-dimensional imaging techniques. Here we present a method to elucidate the intricate 3D meniscal vascular network, revealing its spatial arrangement, connectivity and density. A polymerizing contrast agent was injected into the femoral artery of human cadaver legs, and the meniscal microvasculature was examined using micro-computed tomography at different levels of detail and resolution. The 3D vascular network was quantitatively assessed in a zone-base analysis using parameters such as diameter, length, tortuosity, and branching patterns. The results of this study revealed distinct vascular patterns within the meniscus, with the highest vascular volume found in the outer perimeniscal zone. Variations in vascular parameters were found between the different circumferential and radial meniscal zones. Moreover, through state-of-the-art 3D visualization using micro-CT, this study highlighted the importance of spatial resolution in accurately characterizing the vascular network. These findings, both from this study and from future research using this technique, improve our understanding of microvascular distribution, which may lead to improved therapeutic strategies.

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3D analysis and grading of calcifications from ex vivo human meniscus

Cited by 6 publications

References 43 publications

Age and anatomical region‐related differences in vascularization of the porcine meniscus using microcomputed tomography imaging

Age and anatomical region‐related differences in vascularization of the porcine meniscus using microcomputed tomography imaging

Age and anatomical region related differences in vascularization of the porcine meniscus using micro-computed tomography imaging

Revealing the complexity of meniscus microvasculature through 3D visualization and analysis

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