title: Procaine Inhibits Osteo/Odontogenesis through Wnt/β-Catenin Inactivation.
abstract: BACKGROUND
  Periodontitis is a complex pathology characterized by the loss of alveolar bone. The causes and the mechanisms that promote this bone resorption still remain unknown. The knowledge of the critical regulators involved in the alteration of alveolar bone homeostasis is of great importance for developing molecular therapies. Procaine is an anesthetic drug with demethylant properties, mainly used by dentists in oral surgeries. The inhibitor role of Wnt signaling of procaine was described in vitro in colon cancer cells.
  
  
  METHODS
  In this work we evaluated the role of procaine (1 uM) in osteo/odontogenesis of rat bone marrow mesenchymal stem cells. Similarly, the mechanisms whereby procaine achieves these effects were also studied.
  
  
  RESULTS
  Procaine administration led to a drastic decrease of calcium content, alkaline phosphatase activity, alizarin red staining and an increase in the expression of Matrix Gla Protein. With respect to osteo/odontogenic markers, procaine decreased early and mature osteo/odontogenic markers. In parallel, procaine inhibited canonical Wnt/β-catenin pathway, observing a loss of nuclear β-catenin, a decrease in Lrp5 and Frizzled 3, a significant increase of sclerostin and Gsk3β and an increase of phosphorylated β-catenin. The combination of osteo/odontogenic stimuli and Lithium Chloride decreased mRNA expression of Gsk3β, recovered by Procaine. Furthermore it was proved that Procaine alone dose dependently increases the expression of Gsk3β and β-catenin phosphorylation. These effects of procaine were also observed on mature osteoblast. Interestingly, at this concentration of procaine no demethylant effects were observed.
  
  
  CONCLUSIONS
  Our results demonstrated that procaine administration drastically reduced the mineralization and osteo/odontogenesis of bone marrow mesenchymal stem cells inhibiting Wnt/β-catenin pathway through the increase of Gsk3β expression and β-catenin phosphorylation.
authors.affiliation: Instituto Maimónides para la Investigación Biomédica de Córdoba (IMIBIC)/Hospital Universitario Reina Sofía/Universidad de Córdoba, Serv Nefrologia, Córdoba, Spain.
authors.forename: Carmen
authors.initials: C
authors.lastname: Herencia
authors.affiliation: Instituto Maimónides para la Investigación Biomédica de Córdoba (IMIBIC)/Hospital Universitario Reina Sofía/Universidad de Córdoba, Serv Nefrologia, Córdoba, Spain.
authors.forename: Juan Miguel
authors.initials: JM
authors.lastname: Diaz-Tocados
authors.affiliation: Instituto Maimónides para la Investigación Biomédica de Córdoba (IMIBIC)/Hospital Universitario Reina Sofía/Universidad de Córdoba, Serv Nefrologia, Córdoba, Spain.
authors.forename: Lidia
authors.initials: L
authors.lastname: Jurado
authors.affiliation: Instituto Maimónides para la Investigación Biomédica de Córdoba (IMIBIC)/Hospital Universitario Reina Sofía/Universidad de Córdoba, Serv Nefrologia, Córdoba, Spain.
authors.forename: Addy
authors.initials: A
authors.lastname: Montes de Oca
authors.affiliation: Nephrology Lab, Fundación Jiménez Díaz, REDinREN, Madrid, Spain.
authors.forename: Maria Encarnación
authors.initials: ME
authors.lastname: Rodríguez-Ortiz
authors.affiliation: Instituto Maimónides para la Investigación Biomédica de Córdoba (IMIBIC)/Hospital Universitario Reina Sofía/Universidad de Córdoba, Serv Nefrologia, Córdoba, Spain.
authors.forename: Carmen
authors.initials: C
authors.lastname: Martín-Alonso
authors.affiliation: Instituto Maimónides para la Investigación Biomédica de Córdoba (IMIBIC)/Hospital Universitario Reina Sofía/Universidad de Córdoba, Serv Nefrologia, Córdoba, Spain.
authors.forename: Julio M
authors.initials: JM
authors.lastname: Martínez-Moreno
authors.affiliation: Instituto Maimónides para la Investigación Biomédica de Córdoba (IMIBIC)/Hospital Universitario Reina Sofía/Universidad de Córdoba, Serv Nefrologia, Córdoba, Spain.
authors.forename: Noemi
authors.initials: N
authors.lastname: Vergara
authors.affiliation: Instituto Maimónides para la Investigación Biomédica de Córdoba (IMIBIC)/Hospital Universitario Reina Sofía/Universidad de Córdoba, Serv Nefrologia, Córdoba, Spain.
authors.forename: Mariano
authors.initials: M
authors.lastname: Rodríguez
authors.affiliation: Lipids and Atherosclerosis Unit, (CIBEROBN), Hosp Univ Reina Sofia, IMIBIC, REDinREN, Córdoba, Spain.
authors.forename: Yolanda
authors.initials: Y
authors.lastname: Almadén
authors.affiliation: Instituto Maimónides para la Investigación Biomédica de Córdoba (IMIBIC)/Hospital Universitario Reina Sofía/Universidad de Córdoba, Serv Nefrologia, Córdoba, Spain.
authors.forename: Juan R
authors.initials: JR
authors.lastname: Muñoz-Castañeda
mesh_terms: D000818:Animals; D002118:Calcium / Q000378:metabolism; D002454:Cell Differentiation / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D019175:DNA Methylation / Q000187:drug effects; D008297:Male; D059630:Mesenchymal Stem Cells / Q000187:drug effects / Q000378:metabolism; D053652:Nuclear Transfer Techniques; D009805:Odontogenesis / Q000187:drug effects; D010006:Osteoblasts / Q000187:drug effects; D010012:Osteogenesis / Q000187:drug effects; D011343:Procaine / Q000494:pharmacology*; D051381:Rats; D017208:Rats, Wistar; D020133:Reverse Transcriptase Polymerase Chain Reaction; D051153:Wnt Proteins / Q000378:metabolism*; D051176:beta Catenin / Q000378:metabolism*
chemical_list: D051153:Wnt Proteins; D051176:beta Catenin; D011343:Procaine; D002118:Calcium