"Introduction. Impairment of extracellular matrix (ECM) remodeling is observed in tumour microenvironment or fibrosis and results in excessive collagen production and/or its decreased degradation by metalloproteinases (MMPs). Due to its pivotal role in tissue architectures and functions, ECM components and the proteins that regulate ECM remodeling are thus promising therapeutic targets for human healthcare. Physical stimuli appear as attractive tools to remodel ECM owing to their local and space-time control effects 1. For that purpose, we assessed the potential of pulsed electric field technology, classically applied to drug and plasmid delivery, to modulate cutaneous cells behavior as well as remodel collagen at tissue scale.
Methodology. Two distinct calibrated electric protocols classically used for gene electrotransfer (GET) (10 square-wave pulses of 5 ms, 1 Hz) and electrochemotherapy (ECT) (8 square-wave pulses of 100 μs, 1 Hz) were applied in our studies without addition of any drugs.
Results. It appeared that dermal fibroblasts proliferation and migration properties were not affected when grown in monolayer, whatever the electric field intensity applied 2. We assessed the ECM remodeling after electroporation was applied onto a self-assembled tissue-engineered human dermal substitute model devoid of exogenous material, by examining genes modulation by transcriptomic and proteins synthesis, as well as MMPs activity. Fourier-Transform Infrared-Attenuated Total Reflectance and Differential Scanning Calorimetry were used as complementary tools to analysed collagen structure. We demonstrated that pulsed electric fields induced 1) a rapid modulation (4h after electrostimulation) of mRNA’s genes composing the matrisome, particularly a down-regulation of pro-collagens and ECM maturation’s enzymes; 2) a transient decrease in pro-collagens production and hydroxyproline tissue content within a week after electrostimulation; 3) a long-lasting ROS-dependent over-activation of MMPs and especially collagenase’s family for at least 48h and 4) a down-regulation of TGF-β1, a key player in pathological fibrosis 3. Our first results also indicate that pro-angiogenic processes occur after pulsed electric field application.
Conclusion. Since electroporation is already used today in clinics (FDA and EMA approved), validated equipment (generator, electrodes, dosimetry) is available and doctors are used to this type of protocol. Our research therefore has a strong applicative potential. Taken together, our results open up realistic and relevant prospects for pulsed electric field technology as a local and effective treatment of skin abnormal ECM.
- Gouarderes, S., Mingotaud, A.-F., Vicendo, P., and Gibot, L. Vascular and extracellular matrix remodeling by physical approaches to improve drug delivery at the tumor site. Expert Opin Drug Deliv. Taylor & Francis, 0, 1, 2020.
- Gouarderes, S., Doumard, L., Vicendo, P., Mingotaud, A.-F., Rols, M.-P., and Gibot, L. Electroporation does not affect human dermal fibroblast proliferation and migration properties directly but indirectly via the secretome. Bioelectrochemistry. 134, 107531, 2020.
- Gouarderes, S., Ober, C., Doumard, L., Dandurand, J., Vicendo, P., Fourquaux, I., et al. Pulsed Electric Fields Induce Extracellular Matrix Remodeling through Matrix Metalloproteinases Activation and Decreased Collagen Production. J Invest Dermatol. 2021."