title: ADAM28 manipulates proliferation, differentiation, and apoptosis of human dental pulp stem cells.
abstract: BACKGROUND
  The purpose of this study was to investigate the influence of a disintegrin and metalloproteinase 28 (ADAM28) on the proliferation, differentiation, and apoptosis of human dental pulp stem cells (HDPSCs) and possible mechanism.
  
  
  METHODS
  After ADAM28 eukaryotic plasmid and antisense oligodeoxynucleotides (AS-ODNs) were constructed and respectively transfected into HDPSCs by Lipofectamine 2000, the ADAM28 expression levels among diverse groups were estimated by reverse transcription polymerase chain reaction (RT-PCR) and western blotting. Methabenzthiazuron (MTT) and cell cycle assays were used to test the HDPSCs proliferation activity. Annexin V- fluorescein isothiocyanate (FITC)/propidium iodide and alkaline phosphatase analysis were performed respectively to measure apoptosis and the cytodifferentiation level. Immunocytochemistry and western blotting were performed to determine the effects of ADAM28 eukaryotic plasmid on HDPSCs expressing dentin sialophosphoprotein (DSPP), dentin matrix protein 1, and bone sialoprotein.
  
  
  RESULTS
  ADAM28 could be correctly transcribed, translated, and expressed in HDPSCs. The ADAM28 AS-ODN group displayed the highest optical density value, whereas the eukaryotic plasmid group showed the lowest, which suggested that ADAM28 had a negative regulatory effect on the proliferation of HDPSCs. ADAM28 eukaryotic plasmid could significantly inhibit the HDPSC proliferation, promote specific differentiation of HDPSCs, induce apoptosis, and enhance the DSPP expression, whereas ADAM28 AS-ODN produced the opposite effects.
  
  
  CONCLUSIONS
  Our results proved that ADAM28 might actively participate in manipulating the proliferation, differentiation, and apoptosis of HDPSCs.
authors.affiliation: Institute of Stomatology, General Hospital of Chinese People's Liberation Army, Beijing, China. zhaozheng6912@sohu.com
authors.forename: Zheng
authors.initials: Z
authors.lastname: Zhao
authors.forename: Hongchen
authors.initials: H
authors.lastname: Liu
authors.forename: Dongsheng
authors.initials: D
authors.lastname: Wang
mesh_terms: D051722:ADAM Proteins / Q000235:genetics / Q000502:physiology*; D000293:Adolescent; D000328:Adult; D000469:Alkaline Phosphatase / Q000032:analysis; D017304:Annexin A5; D017209:Apoptosis / Q000502:physiology; D002454:Cell Differentiation / Q000502:physiology; D049109:Cell Proliferation; D002478:Cells, Cultured; D004396:Coloring Agents; D003782:Dental Pulp / Q000166:cytology*; D019483:Disintegrins / Q000235:genetics / Q000502:physiology*; D016326:Extracellular Matrix Proteins / Q000032:analysis; D005260:Female; D016650:Fluorescein-5-isothiocyanate / Q000031:analogs & derivatives; D005456:Fluorescent Dyes; D005822:Genetic Vectors / Q000235:genetics; D006801:Humans; D058574:Integrin-Binding Sialoprotein / Q000032:analysis; D008055:Lipids; D008297:Male; D008562:Membrane Glycoproteins / Q000235:genetics / Q000502:physiology*; D020319:Oligodeoxyribonucleotides, Antisense / Q000235:genetics; D010750:Phosphoproteins / Q000032:analysis; D010957:Plasmids / Q000235:genetics; D011419:Propidium; D012795:Sialoglycoproteins / Q000032:analysis; D013234:Stem Cells / Q000502:physiology*; D013778:Tetrazolium Salts; D013844:Thiazoles; D014162:Transfection; D055815:Young Adult; D000099311:Dentin Sialophosphoprotein
chemical_list: D051722:ADAM Proteins; D000469:Alkaline Phosphatase; D017304:Annexin A5; D004396:Coloring Agents; D019483:Disintegrins; D016326:Extracellular Matrix Proteins; D016650:Fluorescein-5-isothiocyanate; D005456:Fluorescent Dyes; D058574:Integrin-Binding Sialoprotein; D008055:Lipids; D008562:Membrane Glycoproteins; D020319:Oligodeoxyribonucleotides, Antisense; D010750:Phosphoproteins; D011419:Propidium; D012795:Sialoglycoproteins; D013778:Tetrazolium Salts; D013844:Thiazoles; D000099311:Dentin Sialophosphoprotein; C513309:DMP1 protein, human; C492760:FITC-annexin A5; C086724:Lipofectamine; C410224:ADAM28 protein, human; C022616:thiazolyl blue