title: Leukocyte-platelet-rich plasma (L-PRP) impairs the osteoconductive capacity of the autograft associated to changes in the immunolocalization of TGF-β1 and its co-expression with Wnt10b and CD34 cells.
abstract: BACKGROUND
  This study evaluated the osteoconductive effect of an autograft, in the presence or absence of the L-PRP, using histomorphometric analysis of the bone formed, and we compared the results in the presence of TGF-β1, Wnt10b and CD34 detected by immunohistochemistry.
  
  
  METHODS
  Two bone defects were produced in the calvaria of 20 rabbits. The defects were treated with autograft and autograft combined with L-PRP. The animals were euthanized at 15 and 40 days post-surgery. Data were analyzed by Student-Newman-Keuls (p ≤ 0.05) test for histomorphometric and immunohistochemical interpretation.
  
  
  RESULTS
  The results revealed that the presence of bone matrix was significantly less in the defects treated with L-PRP. These results coincided with changes of the immunolocalization of the TGF-β1. In the L-PRP-free groups the TGF-β1 was restricted to bone matrix while the CD34 was scarce and the Wnt10b occurred in peritrabecular cells. In contrast, in defects that received L-PRP the presence of TGF-β1 occurred in cells, which occupied whole area of defect. These TGF-β1+ cells also were co-expressed to Wnt10b and CD34.
  
  
  CONCLUSIONS
  These results suggest that L-PRP induces a cross-reaction between TGF-β1 and Wnt10b, which stimulates the self-renewal and maintenance of CD34+ stem cells immunophenotype, impairing the osteoconductivity properties of the autograft.
authors.affiliation: Positivo University, Rua Pedro Viriato Parigot de Souza #5300, Curitiba, Paraná 81280-330, Brazil. Electronic address: afgiovanini@gmail.com.
authors.forename: Allan Fernando
authors.initials: AF
authors.lastname: Giovanini
authors.forename: Tatiana Miranda
authors.initials: TM
authors.lastname: Deliberador
authors.forename: José Eduardo
authors.initials: JE
authors.lastname: Tannuri Nemeth
authors.forename: Viviane Rozeira
authors.initials: VR
authors.lastname: Crivellaro
authors.forename: Giovanna Schirmer
authors.initials: GS
authors.lastname: Portela
authors.forename: Marco Antonio
authors.initials: MA
authors.lastname: de Oliveira Filho
authors.forename: Melissa Rodrigues
authors.initials: MR
authors.lastname: de Araujo
authors.forename: João Cesar
authors.initials: JC
authors.lastname: Zielak
authors.forename: Carla Castiglia
authors.initials: CC
authors.lastname: Gonzaga
mesh_terms: D000818:Animals; D018952:Antigens, CD34 / Q000032:analysis*; D064592:Autografts / Q000502:physiology / Q000637:transplantation*; D001847:Bone Diseases / Q000601:surgery; D001853:Bone Marrow / Q000473:pathology; D001857:Bone Matrix / Q000473:pathology; D016025:Bone Transplantation / Q000379:methods*; D005260:Female; D007091:Image Processing, Computer-Assisted / Q000379:methods; D007150:Immunohistochemistry; D007962:Leukocytes / Q000276:immunology / Q000502:physiology*; D059630:Mesenchymal Stem Cells / Q000502:physiology; D010012:Osteogenesis / Q000502:physiology; D053657:Platelet-Rich Plasma / Q000502:physiology*; D011817:Rabbits; D013997:Time Factors; D053773:Transforming Growth Factor beta1 / Q000032:analysis*; D051153:Wnt Proteins / Q000032:analysis*
chemical_list: D018952:Antigens, CD34; D053773:Transforming Growth Factor beta1; D051153:Wnt Proteins
keywords: Bone regeneration; CD34; Platelet-rich plasma; TGF-β1; Wnt10b