title: Evaluation of oral keratinocyte progenitor and T-lymphocite cells response during early healing after augmentation of keratinized gingiva with a 3D collagen matrix - a pilot study.
abstract: BACKGROUND
  The aim of the present study is to analyze the behavior of selected populations of oral keratinocytes and T-lymphocytes, responsible for re-constructing and maintaining the oral epithelial tissue architecture, following augmentation of the keratinized oral mucosa using a 3D-collagen matrix.
  
  
  METHODS
  Different groups of oral keratinocytes were isolated from biopsies harvested from 3 patients before the surgical procedure, as well as 7 and 14 days after the augmentation procedure. T-lymphocytes were isolated from peripheral blood at same timepoints. Keratinocytes were characterized for stem and differentiation markers, such as p63, cytokeratin 10 and 14, and in vitro parameters, such as cell viability, cell size and colony-forming efficiency. T-lymphocytes were analyzed for viability and the expression of various cluster of differentiation markers. The methods included magnetic separation of cell populations, immunofluorescence, flow cytometry, and histology of oral biopsies.
  
  
  RESULTS
  Both at 7 and 14 days, the majority of cells that repopulate the matrix were actively proliferating/progenitor oral keratinocytes with the phenotype integrin alfa6beta4 + CD71+. These cells display in vitro characteristics similar to the progenitor cells analyzed before the matrix placement. T-lymphocytes expressed CD8 and CD69 markers, while CD25 was absent.
  
  
  CONCLUSIONS
  The study shows that two weeks after the collagen membrane placement, the healing process appeared to be histologically complete, with no abnormal immune response induced by the matrix, however, with a higher than usual content of active proliferating cells, the majority of keratinocytes being characterized as transit amplifying cells.
authors.affiliation: Department of Periodontology, Victor Babes University of Medicine and Pharmacy, Bv. Revolutiei 1989, Nr. 9, 300041, Timisoara, Romania.
authors.forename: Darian
authors.initials: D
authors.lastname: Rusu
authors.affiliation: Department of Biochemistry, Faculty of Dental Medicine, University of Medicine and Pharmacy, Carol Davila, Blvd-ul Eroii Sanitari, No 8, Bucharest, Romania. bcalenic@yahoo.co.uk.|Victor Babes National Institute of Pathology, Biochemistry-Proteomics Department, Blv. Splaiul Independenţei nr. 99 - 101, Bucharest, Romania. bcalenic@yahoo.co.uk.
authors.forename: Bogdan
authors.initials: B
authors.lastname: Calenic
authors.affiliation: Department of Biochemistry, Faculty of Dental Medicine, University of Medicine and Pharmacy, Carol Davila, Blvd-ul Eroii Sanitari, No 8, Bucharest, Romania.
authors.forename: Maria
authors.initials: M
authors.lastname: Greabu
authors.affiliation: Department of Periodontology, Victor Babes University of Medicine and Pharmacy, Bv. Revolutiei 1989, Nr. 9, 300041, Timisoara, Romania.
authors.forename: Alexander
authors.initials: A
authors.lastname: Kralev
authors.affiliation: Department of Odontotherapy and Endodontics, Victor Babes University of Medicine and Pharmacy, Bv. Revolutiei 1989, 9, 300041, Timisoara, Romania.
authors.forename: Marius
authors.initials: M
authors.lastname: Boariu
authors.affiliation: Department of Functional Sciences-Immunology, Victor Babes University of Medicine and Pharmacy, Pta Eftimie Murgu nr.2, 300041, Timisoara, Romania.
authors.forename: Florina
authors.initials: F
authors.lastname: Bojin
authors.affiliation: Department of Functional Sciences-Immunology, Victor Babes University of Medicine and Pharmacy, Pta Eftimie Murgu nr.2, 300041, Timisoara, Romania.
authors.forename: Simona
authors.initials: S
authors.lastname: Anghel
authors.affiliation: Department of Functional Sciences-Immunology, Victor Babes University of Medicine and Pharmacy, Pta Eftimie Murgu nr.2, 300041, Timisoara, Romania.
authors.forename: Virgil
authors.initials: V
authors.lastname: Paunescu
authors.affiliation: Dental Clinic Dr.Stratul, Str.Emanoil Gojdu, nr.5, 300176, Timisoara, Romania.
authors.forename: Octavia
authors.initials: O
authors.lastname: Vela
authors.affiliation: Department of Dentistry, Faculty of Medicine and Pharmacy, University of Oradea, Pta 1 Decembrie nr.10, Oradea, Romania.
authors.forename: Horia
authors.initials: H
authors.lastname: Calniceanu
authors.affiliation: Department of Periodontology, Victor Babes University of Medicine and Pharmacy, Bv. Revolutiei 1989, Nr. 9, 300041, Timisoara, Romania.
authors.forename: Stefan-Ioan
authors.initials: SI
authors.lastname: Stratul
mesh_terms: D002478:Cells, Cultured; D003094:Collagen / Q000378:metabolism*; D005881:Gingiva; D006801:Humans; D015603:Keratinocytes; D010865:Pilot Projects
chemical_list: D003094:Collagen
keywords: Collagen matrix; Keratinocytes; Oral mucosa; T-lymphocytes