1 / 19

Peter Gal et al.

East-Slovak Institute of Cardiovascular Diseases Pavol Jozef Safarik University. Pharmacological targeting of estrogen receptors-α and -β differently modulates the phenotype of keratinocytes with impact on skin wound healing. Peter Gal et al. Introduction.

howarddouglas
Télécharger la présentation

Peter Gal et al.

An Image/Link below is provided (as is) to download presentation Download Policy: Content on the Website is provided to you AS IS for your information and personal use and may not be sold / licensed / shared on other websites without getting consent from its author. Content is provided to you AS IS for your information and personal use only. Download presentation by click this link. While downloading, if for some reason you are not able to download a presentation, the publisher may have deleted the file from their server. During download, if you can't get a presentation, the file might be deleted by the publisher.

E N D

Presentation Transcript

  1. East-Slovak Institute of Cardiovascular Diseases Pavol Jozef Safarik University Pharmacological targeting of estrogen receptors-α and -β differently modulates the phenotype of keratinocytes with impact on skin wound healing Peter Gal et al.

  2. Introduction increasing average age of (female) population = poor wound healing • USA over 10 billions USD / year - postmenopausalperiod is characterized by: • decrease of estrogen levels • decrease of TGF-β1 levels • increase of MIF levels

  3. Introduction / Aims Estrogen Replacement Therapy (17β estradiol) – various effects on tissue repair → we used specific ER agonists • in vitro:roles of ER-α and -βon fibroblasts, endothelial cells, and keratinocytes (these cells do express ERs) • in vivo part:roles of ER-α and -βin wound healing (excisions = histology + incisions = wound tensile strength measurement)

  4. Methods – in vitro • Human dermal fibroblasts (HDF) / keratinocytes (HaCaT) • isolated from skin residues (DMEM + 10% FBS, ATB, 5%CO2) • ICC: fibronectin+SMA+DAPI / panel of keratins + Ki67 • Human umbilical vein endothelial cells (HUVEC) • isolated from umbilical cords • (M199 + 10% HS + 10% hi NBCS, ECGF, heparin, ATB) • gelatin zymography: MMP-2 a -9 (expresia / aktivita) • PPT / DPN –10nM

  5. Results – in vitro ERs HDFs: HUVECs:

  6. Results – in vitro ERs

  7. Results – in vitro keratinocytes

  8. Results – in vitro fibroblasts SMA Fibronectin a – PPT b – DPN c – control d – TGF-β1

  9. Results – in vitro endothelial cells

  10. Methods – in vivo • female rats • Sprague-Dawley (n=64), OVX – 3 months prior wounding • groups (5 d n=8, 10 d n=8) • NOVX – sham surgery • OVX-C – ovariectomizedvehicle treated • OVX-α – ovariektomized PPT treated (1 mg/kg) • OVX-β – ovariektomized DPN treated (1 mg/kg)

  11. Methods – in vivo histology • left round wound – HE + VG • light microscope (analyzed by QuickPHOTO Micro 2.2) • right wound – FIBR/SMA, CK/VIM, nuclei DAPI • fluorescence mikroscope(analyzed by Lucia 5.1) • 3 channels – RGB:blue – DAPI, green – FITC, red – TRITC

  12. Methods – in vivo TS measurement • both incisions – 2 x 1 cm skin/wound strip • TS=MBS/A, [g/mm2] • wound tensile strength (TS) measuring device

  13. Results – in vivoHE + VG a – OVX-α b – OVX-β c – NOVX-C d – OVX-C

  14. Results – in vivoIHC a – OVX-α b – OVX-β c – NOVX-C d – OVX-C

  15. Results – in vivowound TS

  16. Discussion: incision vs. excision: different treatment ? TGF-β1↑ fb > myofb;mmp13 ↑ wound contraction / scaring ↑ wound TS ↓

  17. Discussion / Conclussion - targeting ER-α(PPT) – excision - targeting ER-β (DPN) – incision + epithelization (open wound) PPT DPN ECM formation + epithelization↑ TGF-β1 fb > myofb;↑ mmp13? / 2 / 9 wound contraction ↑ wound TS ↓ Wound type specific approach !/?

  18. Acknowledgement team post-docs: • Dr. Martin Novotný • Dr. Tomáš Vasilenko • Dr. Martin Slezák • Dr. Ivan Kováč • Dr. Pavol Szabo • Dr. Lenka Varinská colleagues from Prague: • prof. Karel Smetana ml. • Dr. Barbora Dvořánková technician: • Magdaléna Majnušová Ph.D. and M.S. students: Martin Slezak, Martina Polakova, Jan Jakubco, Martina Kostelnikova

  19. Thank you for your attantion!

More Related