title: Multiple Inoculations of Bone Marrow Stromal Cells into Beta-Tricalcium Phosphate/Chitosan Scaffolds Enhances the Formation and Reconstruction of New Bone.
abstract: OBJECTIVE
  To compare the effects of different cell inoculation strategies on cellular attachment and viability as well as on osteogenic differentiation capacity and subsequent bone graft functionality after implantation of cellularized scaffolds in a mandibular defect rabbit model.
  
  
  METHODS
  Bone marrow stromal cells (BMSCs) were inoculated into the scaffolds of beta-tricalcium phosphate/chitosan (β-TCP/CS) either once or multiple times and their efficiency assessed based on cell viability, initial seeding efficiency, alkaline phosphatase activity in cell lysates, mineralization activity in cryosections of cell-seeded scaffolds by von Kossa staining, and osteogenic differentiation on reverse transcriptase polymerase chain reaction.
  
  
  RESULTS
  Compared with one-time inoculation of BMSCs, multiple inoculations led to higher cell viability, increased proliferation, more homogeneous distribution in the scaffold, better in vitro osteogenic differentiation, and improved in vivo bone repair in a rabbit model of mandibular defect.
  
  
  CONCLUSIONS
  Multiple inoculations have many advantages over the one-time inoculation method and should be further researched for bone tissue engineering.
authors.forename: Gu
authors.initials: G
authors.lastname: Cheng
authors.forename: Zhi
authors.initials: Z
authors.lastname: Li
authors.forename: Xin
authors.initials: X
authors.lastname: Xing
authors.forename: Dian-Qi
authors.initials: DQ
authors.lastname: Li
authors.forename: Zu-Bing
authors.initials: ZB
authors.lastname: Li
mesh_terms: D000469:Alkaline Phosphatase / Q000032:analysis; D000818:Animals; D001861:Bone Regeneration / Q000502:physiology*; D002113:Calcification, Physiologic / Q000502:physiology; D002130:Calcium Phosphates / Q000737:chemistry*; D002448:Cell Adhesion / Q000502:physiology; D002452:Cell Count; D018929:Cell Culture Techniques; D002454:Cell Differentiation / Q000502:physiology; D049109:Cell Proliferation; D002470:Cell Survival / Q000502:physiology; D048271:Chitosan / Q000737:chemistry*; D004247:DNA / Q000032:analysis; D004195:Disease Models, Animal; D008297:Male; D008336:Mandibular Diseases / Q000601:surgery; D045164:Mesenchymal Stem Cell Transplantation / Q000379:methods; D059630:Mesenchymal Stem Cells / Q000502:physiology*; D018613:Microscopy, Confocal; D015675:Osteocalcin / Q000032:analysis; D010012:Osteogenesis / Q000502:physiology*; D015676:Osteonectin / Q000032:analysis; D053244:Osteoprotegerin / Q000032:analysis; D011817:Rabbits; D054457:Tissue Scaffolds / Q000737:chemistry*
chemical_list: D002130:Calcium Phosphates; D015676:Osteonectin; D053244:Osteoprotegerin; C485817:beta-tricalcium phosphate; D015675:Osteocalcin; D004247:DNA; D048271:Chitosan; D000469:Alkaline Phosphatase