SproutAngio: an open-source bioimage informatics tool for quantitative analysis of sprouting angiogenesis and lumen space

Beter, Mustafa; Abdollahzadeh, Ali; Pulkkinen, H. H.; Huang, H-Y.; Orsenigo, Fabrizio; Magnusson, Peetra U.; Ylä‐Herttuala, Seppo; Tohka, Jussi; Laakkonen, Johanna P.

doi:10.1038/s41598-023-33090-6

Cited by 3 publications

(3 citation statements)

References 51 publications

(70 reference statements)

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“…Particularly popular platforms for angiogenetic analysis are Sprout Analysis plugin to ImageJ, 102 Matlab-based AngioQuant for in vitro assays, 103 or AngioIQ, 104 which uses a graphic interface, as well as SproutAngio tool written in Python. 105 Carpentier et al developed Angiogenesis Analyzer 108 and recently applied it to fibrin-based assays. 41 There is also a commercially available IKOSA AI tool, 106 which exploits machine-learning algorithms.…”

Section: Discussionmentioning

confidence: 99%

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Long-term day-by-day tracking of microvascular networks sprouting in fibrin gels: From detailed morphological analyses to general growth rules

Rojek,

Wrzos,

Żukowski

et al. 2024

APL Bioengineering

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Understanding and controlling of the evolution of sprouting vascular networks remains one of the basic challenges in tissue engineering. Previous studies on the vascularization dynamics have typically focused only on the phase of intense growth and often lacked spatial control over the initial cell arrangement. Here, we perform long-term day-by-day analysis of tens of isolated microvasculatures sprouting from endothelial cell-coated spherical beads embedded in an external fibrin gel. We systematically study the topological evolution of the sprouting networks over their whole lifespan, i.e., for at least 14 days. We develop a custom image analysis toolkit and quantify (i) the overall length and area of the sprouts, (ii) the distributions of segment lengths and branching angles, and (iii) the average number of branch generations—a measure of network complexity. We show that higher concentrations of vascular endothelial growth factor (VEGF) lead to earlier sprouting and more branched networks, yet without significantly affecting the speed of growth of individual sprouts. We find that the mean branching angle is weakly dependent on VEGF and typically in the range of 60°–75°, suggesting that, by comparison with the available diffusion-limited growth models, the bifurcating tips tend to follow local VEGF gradients. At high VEGF concentrations, we observe exponential distributions of segment lengths, which signify purely stochastic branching. Our results—due to their high statistical relevance—may serve as a benchmark for predictive models, while our new image analysis toolkit, offering unique features and high speed of operation, could be exploited in future angiogenic drug tests.

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Section: Discussionmentioning

confidence: 99%

“…We have performed a basic comparison between our software and the three chosen software: Sprout Analyzer, 102 Sprout Angio, 105 and IKOSA AI. 106 The details of the comparison are given in the supplementary material, and here, we provide a short summary.…”

Section: Discussionmentioning

confidence: 99%

Long-term day-by-day tracking of microvascular networks sprouting in fibrin gels: From detailed morphological analyses to general growth rules

Rojek,

Wrzos,

Żukowski

et al. 2024

APL Bioengineering

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“…In addition, quantification of relative erythrocyte area (i.e. segmented area containing erythrocytes/total plug area) was performed on H&E-stained sections using an automated intensity-based SproutAngio quantification tool ( https://github.com/mbbio/SproutAngio , Beter et al, 2023 ) from whole plug images scanned with an Olympus VS200 slide scanner using a 20×/0.5 Plan Fluor objective.…”

Section: Methodsmentioning

confidence: 99%

ErbB signaling is a potential therapeutic target for vascular lesions with fibrous component

Jauhiainen

Ilmonen

Vuola

et al. 2023

eLife

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Background: Sporadic venous malformation (VM) and angiomatosis of soft tissue (AST) are benign, congenital vascular anomalies affecting venous vasculature. Depending on the size and location of the lesion, symptoms vary from motility disturbances to pain and disfigurement. Due to high recurrence of the lesions more effective therapies are needed. Methods: As targeting stromal cells has been an emerging concept in anti-angiogenic therapies, here, by using VM/AST patient samples, RNA-sequencing, cell culture techniques and a xenograft mouse model, we investigated the crosstalk of endothelial cells (EC) and fibroblasts and its effect on vascular lesion growth. Results: We report, for the first time, expression and secretion of transforming growth factor A (TGFA) in ECs or intervascular stromal cells in AST and VM lesions. TGFA induced secretion of VEGF-A paracrinally, and regulated EC proliferation. Oncogenic PIK3CA variant in p.H1047R, a common somatic mutation found in these lesions, increased TGFA expression, enrichment of hallmark hypoxia, and in a mouse xenograft model, lesion size and vascularization. Treatment with afatinib, a pan-ErbB tyrosine-kinase inhibitor, decreased vascularization and lesion size in mouse xenograft model with ECs expressing oncogenic PIK3CA p.H1047R variant and fibroblasts. Conclusions: Based on the data, we suggest that targeting of both intervascular stromal cells and ECs is a potential treatment strategy for vascular lesions having a fibrous component. Funding: Academy of Finland, Ella and Georg Ehnrooth foundation, the ERC grants, Sigrid Jusélius Foundation, Finnish Foundation for Cardiovascular Research, Jane and Aatos Erkko Foundation, and Department of Musculosceletal and Plastic Surgery, Helsinki University Hospital.

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A Comprehensive Look at In Vitro Angiogenesis Image Analysis Software

Pereira,

Pinto,

Arteaga

et al. 2023

IJMS

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One of the complex challenges faced presently by tissue engineering (TE) is the development of vascularized constructs that accurately mimic the extracellular matrix (ECM) of native tissue in which they are inserted to promote vessel growth and, consequently, wound healing and tissue regeneration. TE technique is characterized by several stages, starting from the choice of cell culture and the more appropriate scaffold material that can adequately support and supply them with the necessary biological cues for microvessel development. The next step is to analyze the attained microvasculature, which is reliant on the available labeling and microscopy techniques to visualize the network, as well as metrics employed to characterize it. These are usually attained with the use of software, which has been cited in several works, although no clear standard procedure has been observed to promote the reproduction of the cell response analysis. The present review analyzes not only the various steps previously described in terms of the current standards for evaluation, but also surveys some of the available metrics and software used to quantify networks, along with the detection of analysis limitations and future improvements that could lead to considerable progress for angiogenesis evaluation and application in TE research.

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SproutAngio: an open-source bioimage informatics tool for quantitative analysis of sprouting angiogenesis and lumen space

Cited by 3 publications

References 51 publications

Long-term day-by-day tracking of microvascular networks sprouting in fibrin gels: From detailed morphological analyses to general growth rules

Long-term day-by-day tracking of microvascular networks sprouting in fibrin gels: From detailed morphological analyses to general growth rules

ErbB signaling is a potential therapeutic target for vascular lesions with fibrous component

A Comprehensive Look at In Vitro Angiogenesis Image Analysis Software

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