title: Extracorporeal shock wave-mediated changes in proliferation, differentiation, and gene expression of human osteoblasts.
abstract: BACKGROUND
  The goal of this study was to determine whether cell proliferation, differentiation, and gene expression of primary human osteoblasts (hOB) are influenced by shock wave application (SWA).
  
  
  METHODS
  Osteoblast cultures were isolated from cancellous bone fragments and treated with 500 impulses of energy flux densities of 0.06 mJ/mm, 0.18 mJ/mm, 0.36 mJ/mm, and 0.50 mJ/mm. Twenty-four hours and 96 hours after SWA cell proliferation, alkaline phosphatase activity, and mineralization were analyzed. The global gene expression profiling was determined 96 hours after SWA employing Affymetrix HG-U133A microarrays.
  
  
  RESULTS
  After 24 hours, hOB showed a dose-dependent increase in cell proliferation from 68.7% (at 0.06 mJ/mm, p = 0.002) up to 81.6% (at 0.5 mJ/mm, p = 0.001), which also persisted after 96 hours. Numbers of alkaline phosphatase-positive hOB increased after SWA treatment with peak levels of response between 0.18 mJ/mm and 0.5 mJ/mm after 24 hours. Mineralization was significantly higher in all groups compared with controls. Microarray analyses revealed SWA-induced differential expression of 94 genes involved in physiologic processes, cell homeostasis, and bone formation. Most intriguing was the up-regulation of multiple genes involved in skeletal development and osteoblast differentiation (e.g., PTHrP, prostaglandin E2-receptor EP3, BMP-2 inducible kinase, chordin, cartilage oligomeric matrix protein, matrillin).
  
  
  CONCLUSIONS
  We showed that shock waves have direct dose-dependent stimulatory effects on proliferation and differentiation of osteoblasts from normal human cancellous bone. We demonstrated that several genes critical for osteoblast differentiation and function are regulated after SWA. Overall, data presented herein will aid further understanding of the osteogenic effect of shock waves and, in addition, will enhance current knowledge of the SWA-mediated gene expression.
authors.affiliation: Department of Trauma Surgery, Johannes Gutenberg University School of Medicine, Mainz, Germany. hofmann.trauma-surgery@gmx.net
authors.forename: Alexander
authors.initials: A
authors.lastname: Hofmann
authors.forename: Ulrike
authors.initials: U
authors.lastname: Ritz
authors.forename: Martin Henri
authors.initials: MH
authors.lastname: Hessmann
authors.forename: Mauro
authors.initials: M
authors.lastname: Alini
authors.forename: Pol Maria
authors.initials: PM
authors.lastname: Rommens
authors.forename: Jan-Dirk
authors.initials: JD
authors.lastname: Rompe
mesh_terms: D000469:Alkaline Phosphatase / Q000378:metabolism; D015519:Bone Density / Q000235:genetics / Q000502:physiology; D002454:Cell Differentiation / Q000235:genetics*; D002455:Cell Division / Q000235:genetics*; D015870:Gene Expression; D020869:Gene Expression Profiling; D019220:High-Energy Shock Waves; D006801:Humans; D066298:In Vitro Techniques; D020411:Oligonucleotide Array Sequence Analysis; D010006:Osteoblasts / Q000166:cytology*; D020133:Reverse Transcriptase Polymerase Chain Reaction
chemical_list: D000469:Alkaline Phosphatase