title: Pulsed acoustic cellular treatment induces expression of proangiogenic factors and chemokines in muscle flaps.
abstract: BACKGROUND
  Pulsed Acoustic Cellular Expression (PACE) treatment is a novel technology with potential to improve tissue perfusion, but the mechanism of this action is unknown. We assessed in vivo the effect of PACE therapy on muscle microcirculatory hemodynamics, neovascularization, and proangiogenic and proinflammatory gene expression.
  
  
  METHODS
  Cremaster muscles were prepared for standard intravital microscopy in 42 Lewis rats divided into five groups: (1) control (n = 10); acute PACE treatment 15 minutes before surgery with (2) 200 impulses (n = 8) and (3) 500 impulses (n = 8); and PACE treatment 24 hours before surgery with (4) 200 impulses (n = 8) and (5) 500 impulses (n = 8).Microcirculatory hemodynamics of red blood cell velocity and capillary perfusion were recorded for 4 hours. Gene expression levels of proinflammatory (inductible nitric oxide synthase [iNOS]) and proangiogenic factors (endothelial nitric oxide synthase [eNOS], vascular endothelial growth factor [VEGF], chemokine (C-X-C motif) ligand 5 [CXCL5], chemokine (C-C motif) ligand 2 [CCL2], and chemokine (C-C motif) receptor 2 [CCR2] were measured using Taqman real-time Polymerase Chain Reaction (PCR). Immunohistochemistry assessed expression of proangiogenic factors: VEGF, von Willebrand factor (vWF), and vessel density by CD31.
  
  
  RESULTS
  PACE treatment resulted in an increase of arteriolar diameters in acute groups 2 and 3 (p < 0.05). In group 5, vessel densities assessed by CD31, VEGF, and vWF expression increased significantly 24 hours after PACE treatment compared with control (p < 0.05). PACE application downregulated proinflammatory iNOS gene expression and upregulated proangiogenic genes expression of eNOS, VEGF, CXCL5, and CCL2.
  
  
  CONCLUSIONS
  Application of PACE treatment, applied as short time acting preconditioning and conditioning treatment, resulted in upregulation of proangiogenic chemokines gene expression in the muscle and showed upregulation of expression of proangiogenic factors such as VEGF and vWF on the vessel endothelium.
authors.affiliation: Department of Plastic Surgery, Cleveland Clinic, Cleveland, Ohio, USA.
authors.forename: Lukasz
authors.initials: L
authors.lastname: Krokowicz
authors.forename: Joanna
authors.initials: J
authors.lastname: Cwykiel
authors.forename: Aleksandra
authors.initials: A
authors.lastname: Klimczak
authors.forename: Mariusz
authors.initials: M
authors.lastname: Mielniczuk
authors.forename: Maria
authors.initials: M
authors.lastname: Siemionow
mesh_terms: D000818:Animals; D018932:Chemokine CCL2 / Q000235:genetics / Q000378:metabolism; D054365:Chemokine CXCL5 / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D006439:Hemodynamics; D019220:High-Energy Shock Waves; D006801:Humans; D007124:Immunoenzyme Techniques; D008297:Male; D008833:Microcirculation; D018482:Muscle, Skeletal / Q000378:metabolism*; D018919:Neovascularization, Physiologic; D052247:Nitric Oxide Synthase Type II / Q000235:genetics / Q000378:metabolism; D052250:Nitric Oxide Synthase Type III / Q000235:genetics / Q000378:metabolism; D019408:Platelet Endothelial Cell Adhesion Molecule-1 / Q000235:genetics / Q000378:metabolism; D051381:Rats; D011917:Rats, Inbred Lew; D054390:Receptors, CCR2 / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D018709:Statistics, Nonparametric; D013524:Surgical Flaps; D042442:Vascular Endothelial Growth Factors / Q000235:genetics / Q000378:metabolism; D014841:von Willebrand Factor / Q000235:genetics / Q000378:metabolism
chemical_list: D018932:Chemokine CCL2; D054365:Chemokine CXCL5; D019408:Platelet Endothelial Cell Adhesion Molecule-1; D054390:Receptors, CCR2; D042442:Vascular Endothelial Growth Factors; D014841:von Willebrand Factor; D052247:Nitric Oxide Synthase Type II; D052250:Nitric Oxide Synthase Type III