title: Pullulan: a new cytoadhesive for cell-mediated cartilage repair.
abstract: BACKGROUND
  Local delivery of mesenchymal stem cells (MSCs) to the acutely injured or osteoarthritic joint retards cartilage destruction. However, in the absence of assistive materials the efficiency of engraftment of MSCs to either intact or fibrillated cartilage is low and localization is further reduced by natural movement of the joint surfaces. It is hypothesised that enhanced engraftment of the delivered MSCs at the cartilage surface will increase their reparative effect and that the application of a bioadhesive to the degraded cartilage surface will provide improved cell retention. Pullulan is a structurally flexible, non-immunogenic exopolysaccharide with wet-stick adhesive properties and has previously been used for drug delivery via the wet surfaces of the buccal cavity. In this study, the adhesive character of pullulan was exploited to enhance MSC retention on the damaged cartilage surface.
  
  
  METHODS
  MSCs labeled with PKH26 were applied to pullulan-coated osteoarthritic cartilage explants to measure cell retention. Cytocompatability was assessed by measuring the effects of prolonged exposure to the bioadhesive on MSC viability and proliferation. The surface phenotype of the cells was assessed by flow cytometry and their multipotent nature by measuring osteogenic, adipogenic and chrondrogenic differentiation. Experiments were also carried out to determine expression of the C-type lectin Dectin-2 receptor.
  
  
  RESULTS
  MSCs maintained a stable phenotype following exposure to pullulan in terms of metabolic activity, proliferation, differentiation and surface antigen expression. An increase in osteogenic activity and Dectin-2 receptor expression was seen in MSCs treated with pullulan. Markedly enhanced retention of MSCs was observed in explant culture of osteoarthritic cartilage.
  
  
  CONCLUSIONS
  Pullulan is a biocompatible and effective cytoadhesive material for tissue engraftment of MSCs. Prolonged exposure to pullulan has no negative impact on the phenotype, viability and differentiation potential of the cells. Pullulan dramatically improves the retention of MSCs at the fibrillated surface of osteoarthritic articular cartilage. Pullulan causes an upregulation in expression of the Dectin-2 C-type lectin transmembrane complex.
authors.affiliation: Regenerative Medicine Institute, National University of Ireland Galway, Biosciences, Dangan, Galway, Ireland. sarah.bulman@smith-nephew.com.|Smith & Nephew, York Science Park, Heslington, York, YO10 5DF, UK. sarah.bulman@smith-nephew.com.
authors.forename: Sarah E
authors.initials: SE
authors.lastname: Bulman
authors.affiliation: Regenerative Medicine Institute, National University of Ireland Galway, Biosciences, Dangan, Galway, Ireland. cynthia.coleman@nuigalway.ie.
authors.forename: Cynthia M
authors.initials: CM
authors.lastname: Coleman
authors.affiliation: Regenerative Medicine Institute, National University of Ireland Galway, Biosciences, Dangan, Galway, Ireland. mary.murphy@nuigalway.ie.
authors.forename: J Mary
authors.initials: JM
authors.lastname: Murphy
authors.affiliation: School of Mechanical and Manufacturing Engineering, Loughborough University, Leicestershire, LE11 3TU, UK. n.medcalf@lboro.ac.uk.
authors.forename: Nicholas
authors.initials: N
authors.lastname: Medcalf
authors.affiliation: Regenerative Medicine Institute, National University of Ireland Galway, Biosciences, Dangan, Galway, Ireland. aideen.ryan@nuigalway.ie.
authors.forename: Aideen E
authors.initials: AE
authors.lastname: Ryan
authors.affiliation: Regenerative Medicine Institute, National University of Ireland Galway, Biosciences, Dangan, Galway, Ireland. frank.barry@nuigalway.ie.
authors.forename: Frank
authors.initials: F
authors.lastname: Barry
mesh_terms: D050156:Adipogenesis / Q000502:physiology; D000293:Adolescent; D000328:Adult; D001672:Biocompatible Materials / Q000494:pharmacology; D002356:Cartilage / Q000166:cytology*; D002448:Cell Adhesion / Q000187:drug effects; D002478:Cells, Cultured; D019902:Chondrocytes / Q000378:metabolism; D020219:Chondrogenesis / Q000502:physiology*; D005936:Glucans / Q000494:pharmacology*; D006801:Humans; D037181:Lectins, C-Type / Q000096:biosynthesis; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000378:metabolism*; D010003:Osteoarthritis / Q000378:metabolism; D014014:Tissue Adhesives / Q000494:pharmacology*; D014945:Wound Healing / Q000187:drug effects; D055815:Young Adult
chemical_list: D001672:Biocompatible Materials; C000593630:CLEC6A protein, human; D005936:Glucans; D037181:Lectins, C-Type; D014014:Tissue Adhesives; C009109:pullulan