title: Rapid fibrin plug formation within cutaneous ablative fractional CO2 laser lesions.
abstract: BACKGROUND
  Ablative fractional laser procedures have been shown to facilitate topical drug delivery into the skin. Past studies have mainly used ex vivo models to demonstrate enhanced drug delivery and in vivo studies have investigated laser created channels over a time course of days and weeks rather than within the first few minutes and hours after exposures. We have noticed rapid in vivo fibrin plug formation within ablative fractional laser lesions impairing passage through the laser created channels.
  
  
  METHODS
  In vivo laser exposures were performed in a porcine model. A fractional CO2 laser (AcuPulse™ system, AcuScan 120™ handpiece, Lumenis, Inc., Yokneam, Israel) was programmed in quasi-continuous wave (QCW) mode, at 40W, 50 mJ per pulse, 5% coverage, nominal 120 µm spot size, 8 × 8 mm square pattern, 169 MTZs per scan. Six millimeters punch biopsies were procured at 0, 2, 5, 10, 15, 30, 60, 90 minutes after completion of each scan, then fixed in 10% formalin. 12 repeats were performed of each time point. Skin samples were processed for serial vertically cut paraffin sections (5 μm collected every 25 μm) then H&E and special immunohistochemistry staining for fibrin and platelet. Dimensions of Microscopic Treatment Zones (MTZs) and extent of fibrin plug were assessed and quantified histologically. Ex vivo laser exposures of the identical laser parameter were performed on porcine and human skin at different storage conditions.
  
  
  RESULTS
  Histology procured at various predetermined time intervals after in vivo fractional CO2 laser exposures revealed a rapidly forming fibrin plug initiating at the bottom of the MTZ lesions. At longer time intervals, the fibrin plug was extending towards the superficial sections. Within the first 5 minutes, more than 25% length of the entire laser-ablated channel was filled with a fibrin plug. With increased time intervals, the cavity was progressively filled with a fibrin plug. At 90 minutes, more than 90% length of the entire laser-ablated channel was occluded. Ex vivo exposures failed to produce any significant fibrin plug formation.
  
  
  CONCLUSIONS
  The current study has demonstrated rapid fibrin plug formation after ablative fractional laser procedures. It was shown that the passage through laser created pathways is critically time dependent for in vivo exposures. In contrast, ex vivo exposures do not exhibit such time dependent passage capacity. In particular, drug, substance, and cell delivery studies for ablative fractional laser treatments should take early fibrin plug formation into consideration and further investigate the impact on transdermal delivery.
authors.affiliation: Cutaneous Biology Research Center, Department of Dermatology, Massachusetts General Hospital Research Institute, Charlestown, Massachusetts, 02129.|Harvard Medical School, Boston, Massachusetts, 02115.
authors.forename: Garuna
authors.initials: G
authors.lastname: Kositratna
authors.affiliation: Cutaneous Biology Research Center, Department of Dermatology, Massachusetts General Hospital Research Institute, Charlestown, Massachusetts, 02129.|Harvard Medical School, Boston, Massachusetts, 02115.
authors.forename: Michael
authors.initials: M
authors.lastname: Evers
authors.affiliation: Cutaneous Biology Research Center, Department of Dermatology, Massachusetts General Hospital Research Institute, Charlestown, Massachusetts, 02129.|Harvard Medical School, Boston, Massachusetts, 02115.
authors.forename: Amir
authors.initials: A
authors.lastname: Sajjadi
authors.affiliation: Cutaneous Biology Research Center, Department of Dermatology, Massachusetts General Hospital Research Institute, Charlestown, Massachusetts, 02129.|Harvard Medical School, Boston, Massachusetts, 02115.
authors.forename: Dieter
authors.initials: D
authors.lastname: Manstein
mesh_terms: D000279:Administration, Cutaneous; D000818:Animals; D015415:Biomarkers / Q000378:metabolism; D001706:Biopsy; D016503:Drug Delivery Systems / Q000295:instrumentation / Q000379:methods*; D005260:Female; D005337:Fibrin / Q000378:metabolism*; D006801:Humans; D054020:Lasers, Gas; D012867:Skin / Q000378:metabolism / Q000473:pathology*; D013552:Swine; D013997:Time Factors
chemical_list: D015415:Biomarkers; D005337:Fibrin
keywords: ablative fractional lasers; cell delivery; drug delivery; fibrin plug formation; gateways; laser-assisted; substance delivery; time course; time dependence; transcutaneous; transdermal; transport