title: Microcirculatory effect of topical vapocoolants.
abstract: BACKGROUND
  Vapocoolant sprays are commonly used to minimize pain following minor interventions such as venipuncture, shave biopsy or needle insertion. Although these sprays have been widely used in clinical practice, little is known about their effect on microcirculation or cutaneous blood flow.
  
  
  OBJECTIVE
  To evaluate the real-time effect of a topical vapocoolant using a well-established, rat cremaster muscle microcirculatory model, allowing direct measurement of changes in vessel diameter, capillary density and leukocyte behaviour.
  
  
  METHODS
  Fifty rats were divided into a control and four experimental groups: group 1: 4 s spray with vapocoolant at 18 cm distance; group 2: 10 s spray at 18 cm distance; group 3: 4 s spray at 8 cm distance; and group 4: 10 s spray at 8 cm distance. Vessel diameters, capillary density and leukocyte behaviour were monitored for 1 h thereafter. Muscle was harvested for immunohistochemistry analysis of proangiogenic markers (vascular endothelial growth factor and von Willebrand factor), leukocyte behaviour markers (E-selectin, vascular cell adhesion molecule, intercellular adhesion molecule), pimonidazole-hypoxia staining and ApopTag (Millipore, USA) staining for apoptosis. Gene expression for inflammatory markers (interleukin [IL]-1β, IL-2, IL-4, IL-6, IL-10, tumour necrosis factor-alpha and interferon-gamma) was evaluated using polymerase chain reaction and myeloperoxidase assay for inflammation was performed.
  
  
  RESULTS
  The use of refrigerant spray decreased vessel diameter and capillary density initially, although none of these decreases were statistically significant. Polymerase chain reaction showed no significant changes. The myeloperoxidase assay showed statistically significant increase in myeloperoxidase activity in groups 2, 3 and 4. Immunohistochemistry was negative for angiogenic and proinflammatory markers.
  
  
  CONCLUSIONS
  The lack of statistically significant changes in vessel diameter and inflammatory markers corroborated the safety on microcirculation.
authors.affiliation: Department of Plastic Surgery, Loma Linda University, Loma Linda, California;
authors.forename: Izabela
authors.initials: I
authors.lastname: Galdyn
authors.affiliation: Department of Plastic and Reconstructive Surgery, The Johns Hopkins Hospital, The Johns Hopkins University School of Medicine, Baltimore, Maryland;
authors.forename: Edward
authors.initials: E
authors.lastname: Swanson
authors.affiliation: Department of Plastic and Reconstructive Surgery, The Johns Hopkins Hospital, The Johns Hopkins University School of Medicine, Baltimore, Maryland;
authors.forename: Chad
authors.initials: C
authors.lastname: Gordon
authors.affiliation: Department of Plastic Surgery, Cleveland Clinic, Cleveland, Ohio;
authors.forename: Grzegorz
authors.initials: G
authors.lastname: Kwiecien
authors.affiliation: Quantitative Health Sciences, Cleveland Clinic, Cleveland, Ohio;
authors.forename: James
authors.initials: J
authors.lastname: Bena
authors.affiliation: Department of Orthopaedic Surgery, University of Illinois at Chicago, Chicago, Illinois, USA.
authors.forename: Maria
authors.initials: M
authors.lastname: Siemionow
authors.affiliation: Department of Plastic Surgery, Cleveland Clinic, Cleveland, Ohio;
authors.forename: James
authors.initials: J
authors.lastname: Zins
keywords: Cremaster muscle; Microcirculation; Refrigerant; Vapocoolant