title: Effect of sustained gene delivery of platelet-derived growth factor or its antagonist (PDGF-1308) on tissue-engineered cementum.
abstract: BACKGROUND
  Cementum, a mineralized tissue lining the tooth root surface, is destroyed during the inflammatory process of periodontitis. Restoration of functional cementum is considered a criterion for successful regeneration of periodontal tissues, including formation of periodontal ligament, cementum, and alveolar bone. Short-term administration of platelet-derived growth factor (PDGF) has been shown to partially regenerate periodontal structures. Nonetheless, the role of PDGF in cementogenesis is not well understood. The aim of the present study was to determine the effect of sustained PDGF gene transfer on cementum formation in an ex vivo ectopic biomineralization model.
  
  
  METHODS
  Osteocalcin (OC) promoter-driven SV40 transgenic mice were used to obtain immortalized cementoblasts (OCCM). The OCCM cells were transduced with adenoviruses (Ad) encoding either PDGF-A, an antagonist of PDGF signaling (PDGF-1308), a control virus (green fluorescent protein, GFP), or no treatment (NT). The transduced cells were incorporated into polymer scaffolds and implanted subcutaneously into severe combined immunodeficient (SCID) mice. The implants were harvested at 3 and 6 weeks for histomorphometric analysis of the newly formed mineralized tissues. Northern blot analysis was performed to determine the expression levels of mineral-associated genes including bone sialoprotein (BSP), OC, and osteopontin (OPN) in the cell-implant specimens at 3 and 6 weeks.
  
  
  RESULTS
  The results indicated mineralization was significantly reduced in both the Ad/PDGF-A and Ad/PDGF-1308 treated specimens when compared to the NT or Ad/GFP groups at 3 and 6 weeks (P<0.01). In addition, the size of the implants treated with Ad/PDGF-A and Ad/PDGF-1308 was significantly reduced compared to implants from Ad/GFP and NT groups at 3 weeks (P<0.05). At 6 weeks, the size of implants and mineral formation increased in NT, Ad/GFP, and Ad/PDGF-A groups, while the Ad/PDGF-1308 treated implants continued to decrease in size and mineral formation (P<0.01). Northern blot analysis revealed that in the Ad/PDGF-A treated implants OPN was increased, whereas OC gene expression was downregulated at 3 weeks. In the Ad/PDGF-1308 treated implants, BSP, OC, and OPN were all downregulated at 3 weeks. At 3 weeks, the Ad/PDGF-A treated implants contained significantly higher multinucleated giant cell (MNGC) density compared to NT, Ad/GFP, and Ad/PDGF-1308 specimens. The MNGC density in NT, Ad/GFP, and Ad/PDGF-A treated groups reduced over time, while the Ad/PDGF-1308 transduced implants continued to exhibit significantly higher MNGC density compared with the other treatment groups at 6 weeks.
  
  
  CONCLUSIONS
  The results showed that continuous exposure to PDGF-A had an inhibitory effect on cementogenesis, possibly via the upregulation of OPN and subsequent enhancement of MNGCs at 3 weeks. On the other hand, Ad/PDGF-1308 inhibited mineralization of tissue-engineered cementum possibly due to the observed downregulation of BSP and OC and a persistence of stimulation of MNGCs. These findings suggest that continuous exogenous delivery of PDGF-A may delay mineral formation induced by cementoblasts, while PDGF is clearly required for mineral neogenesis.
authors.affiliation: Center for Craniofacial Regeneration, Department of Periodontics/Prevention/Geriatrics, School of Dentistry, University of Michigan, Ann Arbor, MI 48109-1078, USA.
authors.forename: Orasa
authors.initials: O
authors.lastname: Anusaksathien
authors.forename: Qiming
authors.initials: Q
authors.lastname: Jin
authors.forename: Ming
authors.initials: M
authors.lastname: Zhao
authors.forename: Martha J
authors.initials: MJ
authors.lastname: Somerman
authors.forename: William V
authors.initials: WV
authors.lastname: Giannobile
mesh_terms: D000818:Animals; D002452:Cell Count; D002460:Cell Line; D025021:Cementogenesis / Q000187:drug effects* / Q000235:genetics; D003739:Dental Cementum / Q000187:drug effects* / Q000378:metabolism; D015536:Down-Regulation; D018014:Gene Transfer Techniques; D015726:Giant Cells / Q000187:drug effects; D049452:Green Fluorescent Proteins; D007202:Indicators and Reagents; D058574:Integrin-Binding Sialoprotein; D019344:Lactic Acid; D008164:Luminescent Proteins; D051379:Mice; D016513:Mice, SCID; D008822:Mice, Transgenic; D015675:Osteocalcin / Q000235:genetics; D053495:Osteopontin; D010982:Platelet-Derived Growth Factor / Q000037:antagonists & inhibitors / Q000235:genetics / Q000494:pharmacology*; D011100:Polyglycolic Acid; D000077182:Polylactic Acid-Polyglycolic Acid Copolymer; D011108:Polymers; D012795:Sialoglycoproteins / Q000235:genetics; D013539:Simian virus 40 / Q000235:genetics; D023822:Tissue Engineering
chemical_list: C546872:Ibsp protein, mouse; D007202:Indicators and Reagents; D058574:Integrin-Binding Sialoprotein; D008164:Luminescent Proteins; D010982:Platelet-Derived Growth Factor; D011108:Polymers; D012795:Sialoglycoproteins; C506896:Spp1 protein, mouse; C075278:platelet-derived growth factor A; D015675:Osteocalcin; D053495:Osteopontin; D049452:Green Fluorescent Proteins; D000077182:Polylactic Acid-Polyglycolic Acid Copolymer; D011100:Polyglycolic Acid; D019344:Lactic Acid