Shockwave therapy combined with hiPSC-MSC exosomes enhances perfusion recovery in murine hindlimb ischaemia
M Ng, M Belau, R Storz, S Y Liao, H F TseAbstract
Background
Peripheral arterial disease (PAD) is characterised by limb ischaemia and impaired perfusion, with limited regenerative options. Exosomes from mesenchymal stem cells have pro-angiogenic potential, while extracorporeal shockwave therapy promotes neovascularisation. Whether combining biophysical stimulation with exosome-based therapy augments vascular recovery is unknown.
Purpose
To evaluate the effects of shockwave therapy and human iPSC-derived mesenchymal stem cell (hiPSC-MSC) exosomes, alone and in combination, on perfusion recovery in a murine hindlimb ischaemia model.
Methods
Hindlimb ischaemia was induced by femoral artery ligation in ICR mice, which were randomised into four groups (n=6/group): control, hiPSC-MSC exosomes, shockwave, and combination therapy. Exosomes (35 μg in saline) were injected intramuscularly immediately after ischaemia induction. Shockwave therapy (0.20 mJ/mm², 5 Hz) was applied in four sessions: immediately post-operatively and at weeks 1, 2, and 3. Blood perfusion was assessed by laser Doppler imaging immediately post-operatively and at 1, 2, 3, and 4 weeks. At 4 weeks, hindlimb tissues were collected for Masson’s trichrome and immunohistochemical staining. One-way ANOVA with post hoc testing was used for statistical analysis.
Results
At week 4, both exosome and shockwave monotherapies significantly improved perfusion versus control (314.74 perfusion units [PU] vs 177.03 PU and 343.48 PU vs 177.03 PU, respectively; both p<0.05). Combination therapy achieved the highest perfusion (441.36 PU), significantly greater than exosomes alone (441.36 PU vs 314.74 PU; p<0.05). Histology demonstrated reduced fibrosis and increased neovascularisation in the combination group compared with all other groups.
Conclusion
Shockwave therapy improves perfusion recovery in murine hindlimb ischaemia and provides an additive benefit when combined with hiPSC-MSC–derived exosomes. This combinatorial approach integrating biophysical stimulation with exosome-based therapy may represent a promising regenerative strategy for ischaemic vascular disease.