Q-VAT: Quantitative Vascular Analysis Tool

Callewaert, Bram; Gsell, Willy; Himmelreich, Uwe; Jones, Elizabeth A. V.

doi:10.3389/fcvm.2023.1147462

Cited by 4 publications

(2 citation statements)

References 32 publications

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“…To assess vascular density, the Quantitative Vascular Analysis Tool (Q-VAT) 29 was utilized. The samples were placed inside the chicken egg for CAM assay, and the images were acquired using a digital microscope.…”

Section: Methodsmentioning

confidence: 99%

“…This assay provides insights into the biological activity and neovascularization potential of each construct, through quantitative evaluation of vascular density around engineered grafts After 4 days, we observed an extensive vessel formation in the periphery of both MSOD-B and MSOD-BΔV1 eECMs (Figure 2B). Quantification of vascular densities 29 suggested a reduced vessel formation in MSOD-BΔV1 samples but without reaching significance (Figure 2C and Supplementary Figure 2A). Thus, the CAM assay indicated that both MSOD-B and MSOD-BΔV1 cartilages retain angiogenic potentials.…”

Section: Vegf Knockout Cartilage Tissues Retain Bone Remodeling Capac...mentioning

confidence: 99%

See 1 more Smart Citation

Compositional editing of extracellular matrices by CRISPR/Cas9 engineering of human mesenchymal stem cell lines

Prithiviraj,

Garcia,

Linderfalk

et al. 2023

Preprint

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Tissue engineering strategies predominantly rely on the production of living substitutes, whereby implanted cells actively participate in the regenerative process. Beyond cost and delayed graft availability, the patient-specific performance of engineered tissues poses serious concerns on their clinical translation ability. A more exciting paradigm consist in exploiting cell-laid, engineered extracellular matrices (eECM), which can be used as off-the-shelf materials. Here, the regenerative capacity solely relies on the preservation of the eECM structure and embedded signals to instruct an endogenous repair. We recently described the possibility to exploit custom human stem cell lines for eECM manufacturing. In addition to the conferred standardization, the availability of such cell lines opened avenues for the design of tailored eECMs by applying dedicated genetic tools. In this study, we aim at demonstrating the possibility of exploiting CRISPR/Cas9 as a high precision system for editing the composition and function of eECMs. To this end, human mesenchymal stromal/stem cell (hMSC) lines were modified to knockout VEGF and RUNX2 and assessed for their capacity to generate osteoinductive cartilage matrices. We report the successful editing of hMSCs, subsequently leading to targeted VEGF and RUNX2-knockout cartilage eECMs. Despite the absence of VEGF, eECMs retained full capacity to instruct ectopic endochondral ossification. Conversely, RUNX2-edited eECMs exhibited impaired hypertrophy which resulted in delayed bone formation in vivo. In summary, our approach can be harnessed to identify the necessary eECM factors driving endogenous repair. Our work paves the road towards the compositional eECMs editing and their exploitation in broad regenerative contexts.

show abstract

Section: Methodsmentioning

confidence: 99%

Section: Vegf Knockout Cartilage Tissues Retain Bone Remodeling Capac...mentioning

confidence: 99%

Compositional editing of extracellular matrices by CRISPR/Cas9 engineering of human mesenchymal stem cell lines

Prithiviraj,

Garcia,

Linderfalk

et al. 2023

Preprint

View full text Add to dashboard Cite

show abstract

Intravoxel incoherent motion as a surrogate marker of perfused vascular density in rat brain

Callewaert,

Gsell,

Lox

et al. 2024

NMR in Biomedicine

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Intravoxel incoherent motion (IVIM) MRI has emerged as a valuable technique for the assessment of tissue characteristics and perfusion. However, there is limited knowledge about the relationship between IVIM‐derived measures and changes at the level of the vascular network. In this study, we investigated the potential use of IVIM MRI as a noninvasive tool for measuring changes in cerebral vascular density. Variations in quantitative immunohistochemical measurements of the vascular density across different regions in the rat brain (cortex, corpus callosum, hippocampus, thalamus, and hypothalamus) were related to the pseudo‐diffusion coefficient D* and the flowing blood fraction f in healthy Wistar rats. We assessed whether region‐wise differences in the vascular density are reflected by variations in the IVIM measurements and found a significant positive relationship with the pseudo‐diffusion coefficient (p < 0.05, β = 0.24). The effect of cerebrovascular alterations, such as blood–brain barrier (BBB) disruption on the perfusion‐related IVIM parameters, is not well understood. Therefore, we investigated the effect of BBB disruption on the IVIM measures in a rat model of metabolic and vascular comorbidities (ZSF1 obese rat) and assessed whether this affects the relationship between the cerebral vascular density and the noninvasive IVIM measurements. We observed increased vascular permeability without detecting any differences in diffusivity, suggesting that BBB leakage is present before changes in the tissue integrity. We observed no significant difference in the relationship between cerebral vascular density and the IVIM measurements in our model of comorbidities compared with healthy normotensive rats.

show abstract

Compositional editing of extracellular matrices by CRISPR/Cas9 engineering of human mesenchymal stem cell lines

Prithiviraj,

Garcia,

Linderfalk

et al. 2024

Preprint

View full text Add to dashboard Cite

Tissue engineering strategies predominantly rely on the production of living substitutes, whereby implanted cells actively participate in the regenerative process. Beyond cost and delayed graft availability, the patient-specific performance of engineered tissues poses serious concerns on their clinical translation ability. A more exciting paradigm consist in exploiting cell-laid, engineered extracellular matrices (eECM), which can be used as off-the-shelf materials. Here, the regenerative capacity solely relies on the preservation of the eECM structure and embedded signals to instruct an endogenous repair. We recently described the possibility to exploit custom human stem cell lines for eECM manufacturing. In addition to the conferred standardization, the availability of such cell lines opened avenues for the design of tailored eECMs by applying dedicated genetic tools. In this study, we demonstrated the exploitation of CRISPR/Cas9 as a high precision system for editing the composition and function of eECMs. Human mesenchymal stromal/stem cell (hMSC) lines were modified to knockout VEGF and RUNX2 and assessed for their capacity to generate osteoinductive cartilage matrices. We report the successful editing of hMSCs, subsequently leading to targeted VEGF and RUNX2-knockout cartilage eECMs. Despite the absence of VEGF, eECMs retained full capacity to instruct ectopic endochondral ossification. Conversely, RUNX2-edited eECMs exhibited impaired hypertrophy, reduced ectopic ossification and superior cartilage repair in a rat osteochondral defect. In summary, our approach can be harnessed to identify the necessary eECM factors driving endogenous repair. Our work paves the road towards the compositional eECMs editing and their exploitation in broad regenerative contexts.

show abstract

Q-VAT: Quantitative Vascular Analysis Tool

Cited by 4 publications

References 32 publications

Compositional editing of extracellular matrices by CRISPR/Cas9 engineering of human mesenchymal stem cell lines

Compositional editing of extracellular matrices by CRISPR/Cas9 engineering of human mesenchymal stem cell lines

Intravoxel incoherent motion as a surrogate marker of perfused vascular density in rat brain

Compositional editing of extracellular matrices by CRISPR/Cas9 engineering of human mesenchymal stem cell lines

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