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Updated: Dec 12, 2025
Assessing Therapeutic Angiogenesis in a Murine Model of Hindlimb Ischemia
Published on: June 8, 2019
Systematic Interrogation of Angiogenesis in the Ischemic Mouse Hind Limb: Vulnerabilities and Quality Assurance
Jason J Lee1,2,3, John-Michael Arpino1,3, Hao Yin1
1Robarts Research Institute (J.J.L., J.-M.A., H.Y., Z.N., A.S., J.C., C.O., J.G.P.), Western University, London, Ontario, Canada.
Objective:
There has been little success in translating preclinical studies of mouse hind limb ischemia into benefit for patients with peripheral artery disease. Using systematic strategies, we sought to define the injury and angiogenesis landscapes in mice subjected to hind limb ischemia and ascertain whether published studies to date have used an analysis strategy concordant with these data. Approach and Results: Maps of ischemic injury were generated from 22 different hind limb muscles and 33 muscle territories in 12-week-old C57BL/6 mice, based on loss or centralization of myofiber nuclei. Angiogenesis was similarly mapped based on CD (cluster of differentiation) 31-positive capillary content. Only 10 of 33 muscle territories displayed consistent muscle injury, with the distal anterior hind limb muscles most reliably injured. Angiogenesis was patchy and exclusively associated with zones of regenerated muscle (central nuclei). Angiogenesis was not observed in normal appearing muscle, necrotic muscle, or injury border zones. Systematic review of mouse hind limb angiogenesis studies identified 5147 unique publications, of which 509 met eligibility criteria for analysis. Only 7% of these analyzed manuscripts evaluated angiogenesis in distal anterior hind limb muscles and only 15% consistently examined for angiogenesis in zones of muscle regeneration.
Conclusions:
In 12-week C57BL/6 mice, angiogenesis postfemoral artery excision proceeds exclusively in zones of muscle regeneration. Only a minority of studies to date have analyzed angiogenesis in regions of demonstrably regenerating muscle or in high-likelihood territories. Quality assurance standards, informed by the atlas and mapping data herein, could augment data reliability and potentially help translate mouse hind limb ischemia studies to patient care.
Insights
Mouse hind limb ischemia studies often fail to translate to patient benefit. Angiogenesis, or new blood vessel growth, primarily occurs in regenerating muscle zones, a factor rarely analyzed in published research.
Area of Science:
- Cardiovascular Research
- Regenerative Medicine
- Preclinical Models
Background:
- Translating preclinical mouse hind limb ischemia studies to peripheral artery disease patient benefit remains a challenge.
- Understanding the precise injury and angiogenesis patterns in mouse models is crucial for improving study relevance.
Purpose of the Study:
- To map ischemic injury and angiogenesis in mouse hind limbs.
- To assess if existing studies analyze angiogenesis concordantly with injury and regeneration patterns.
Main Methods:
- Generated injury maps based on myofiber nuclei loss/centralization in 22 hind limb muscles.
- Mapped angiogenesis using CD31-positive capillary content.
- Conducted a systematic review of 509 mouse hind limb angiogenesis studies.
Main Results:
- Consistent muscle injury was observed in only 10 of 33 muscle territories, predominantly distal anterior hind limb muscles.
- Angiogenesis was exclusively associated with regenerating muscle zones (central nuclei), not necrotic or normal muscle.
- Only 7% of reviewed studies analyzed angiogenesis in distal anterior hind limb muscles, and 15% examined regenerating zones.
Conclusions:
- Angiogenesis in mouse hind limb ischemia models occurs exclusively in regenerating muscle zones.
- Current preclinical studies often fail to analyze angiogenesis in relevant muscle regeneration areas or high-likelihood injury territories.
- Implementing quality assurance standards based on detailed mapping can improve data reliability and translational potential.
