title: A new co-ultramicronized composite including palmitoylethanolamide and luteolin to prevent neuroinflammation in spinal cord injury.
abstract: BACKGROUND
  It has recently been demonstrated that palmitoylethanolamide (PEA), an endogenous lipid amide belonging to the N-acylethanolamine family, exerts neuroprotection in central nervous system (CNS) pathologies. In recent studies, we have demonstrated that treatment with PEA significantly reduced inflammatory secondary events associated with spinal cord injury (SCI). Since oxidative stress is considered to play an important role in neuroinflammatory disorders, in the present work we studied a new composite, a formulation including PEA and the antioxidant compound luteolin (Lut), subjected to an ultramicronization process, co-ultraPEALut. We investigated the effect of co-ultraPEALut (in the respective fixed doses of 10:1 in mass) in both an ex vivo organotypic spinal cord culture model and an in vivo model of SCI.
  
  
  METHODS
  For the organotypic cultures, spinal cords were prepared from mice at postnatal day 6 and were cut into transverse slices of 400 μm thickness to generate the lumbar organotypic slice cultures. After 7 days of culturing, the slices were mechanically injured onto the center of the slice and the co-ultraPEALut was applied at different concentrations (0.00009, 0.0009 and 0.009 g/l) 1 hour before damage. For in vivo studies, SCI was induced in mice through spinal cord compression by the application of vascular clips (force of 24 g) to the dura via a four-level T5 to T8 laminectomy, and co-ultraPEALut (1 mg/kg ip) was administered at 1 and 6 hours after SCI. At 24 hours after SCI, mice were sacrificed and the spinal cords were collected for further evaluation. Additional animals were treated similarly and sacrificed 10 days after SCI.
  
  
  RESULTS
  Pretreatment with co-ultraPEALut significantly reduced cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS) expression in a concentration-dependent manner, restored neuronal nitric oxide synthase (nNOS) expression at all three tested concentrations, and protected cells by cell death (MTT assay) in spinal cord organotypic cultures. Moreover, we demonstrated in vivo that co-ultraPEALut 1 mg/kg reduced the severity of trauma induced by compression and improved the motor activity evaluated at 10 days post-injury.
  
  
  CONCLUSIONS
  The present study demonstrates that the protective effect of PEA on SCI-associated neuroinflammation could be improved by co-ultramicronization with Lut possibly due to its antioxidant properties.
authors.affiliation: Department of Biological and Environmental Sciences, University of Messina, Viale Ferdinando Stagno D'Alcontres, 98166 Messina, Italy.
authors.forename: Irene
authors.initials: I
authors.lastname: Paterniti
authors.forename: Daniela
authors.initials: D
authors.lastname: Impellizzeri
authors.forename: Rosanna
authors.initials: R
authors.lastname: Di Paola
authors.forename: Michele
authors.initials: M
authors.lastname: Navarra
authors.forename: Salvatore
authors.initials: S
authors.lastname: Cuzzocrea
authors.forename: Emanuela
authors.initials: E
authors.lastname: Esposito
mesh_terms: D000577:Amides; D000818:Animals; D015153:Blotting, Western; D002470:Cell Survival / Q000187:drug effects; D004396:Coloring Agents; D051546:Cyclooxygenase 2 / Q000096:biosynthesis; D004338:Drug Combinations; D063388:Endocannabinoids / Q000008:administration & dosage / Q000627:therapeutic use*; D004983:Ethanolamines / Q000008:administration & dosage / Q000627:therapeutic use*; D007249:Inflammation / Q000209:etiology* / Q000473:pathology / Q000517:prevention & control*; D047311:Luteolin / Q000008:administration & dosage / Q000627:therapeutic use*; D051379:Mice; D053758:Nanoparticles; D052248:Nitric Oxide Synthase Type I / Q000096:biosynthesis; D052247:Nitric Oxide Synthase Type II / Q000096:biosynthesis; D009573:Nitrites / Q000378:metabolism; D009924:Organ Culture Techniques; D010169:Palmitic Acids / Q000008:administration & dosage / Q000627:therapeutic use*; D019999:Pharmaceutical Solutions; D013119:Spinal Cord Injuries / Q000150:complications* / Q000473:pathology*; D013778:Tetrazolium Salts; D013844:Thiazoles
chemical_list: D000577:Amides; D004396:Coloring Agents; D004338:Drug Combinations; D063388:Endocannabinoids; D004983:Ethanolamines; D009573:Nitrites; D010169:Palmitic Acids; D019999:Pharmaceutical Solutions; D013778:Tetrazolium Salts; D013844:Thiazoles; C005958:palmidrol; D052248:Nitric Oxide Synthase Type I; D052247:Nitric Oxide Synthase Type II; D051546:Cyclooxygenase 2; C022616:thiazolyl blue; D047311:Luteolin