Sociedad Americana de Hirudoterapia

Investigating the therapeutic potential of medical leech and leech saliva extract in flap survival: an in vivo study using rats

Ünal K, Emre Erol M, Dayanır D, Deniz E, Ayhan H, Fındıkçıoğlu K (2025) · Journal of Complementary & Integrative Medicine · n=0

RCT evidence detailTrial reference
GRADE Very LowInsufficient evidence

Study Profile

Design
controlled animal experiment with three groups of female Wistar albino rats subjected to a dorsal random skin flap model (Gazi University Faculty of Medicine, Ankara, Turkey)
Sample size (n)
0
Intervention
Three groups: control (flap only), medicinal leech therapy (MLT) group, and leech saliva extract (LSE) injection group; histological, immunohistochemical (VEGF), and ELISA-based biochemical analyses performed at postoperative day 7
Comparator
Sham/control group (flap only without leech intervention)
Primary endpoint
Flap necrosis area, VEGF-positive cell percentage, neovascularization, epithelial regeneration, granulation tissue thickness, and inflammatory cell infiltration on postoperative day 7
Primary result
Flap necrosis area was significantly lower in both MLT and LSE groups versus control (p<0.05); VEGF-positive cells, neovascularization, epithelial regeneration, and granulation tissue thickness all significantly higher in MLT and LSE groups versus control (p<0.05); inflammatory cells substantially lower in LSE group versus control (p<0.05); first study to demonstrate the effect of medicinal leech extract injection in a flap model
Follow-up duration
7 postoperative days

Key Findings

  • First in vivo rat study to investigate medicinal leech saliva extract injection in a dorsal random flap model
  • Both medicinal leech therapy (MLT) and leech saliva extract (LSE) injection significantly reduced flap necrosis area
  • VEGF-positive cells, neovascularization, epithelial regeneration, and granulation tissue thickness were all enhanced in both leech groups
  • Leech saliva extract injection significantly reduced inflammatory cell infiltration
  • Provides molecular and histological mechanistic support for the clinical use of leech therapy in flap salvage

Limitations

  • Animal model (rat dorsal random flap) - direct human translation requires further study
  • Short follow-up (7 postoperative days) - long-term outcomes untested
  • Small sample size per arm (not specified per group in abstract)
  • Saliva extract concentration and standardization details unclear
  • Mechanism characterization limited to selected markers (VEGF, histology)

Clinical Implications

Ünal 2025 is a Turkish preclinical animal study that documents both whole-leech therapy and leech saliva extract injection produce significant histological and biochemical improvements in a dorsal random flap rat model. For ASH editorial purposes, the trial supports the mechanistic basis of leech therapy in flap salvage and adds preclinical support to the K040187-cleared indication. For US clinicians, the trial does not change practice but does provide modern animal-model evidence that the venous-decongestion mechanism is biologically operative. The saliva extract finding also supports future research on purified leech-derived biologics as alternatives to whole-leech device therapy.

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