title: Synergistic effects of chitosan scaffold and TGFβ1 on the proliferation and osteogenic differentiation of dental pulp stem cells derived from human exfoliated deciduous teeth.
abstract: OBJECTIVE
  Multipotent stem cells derived from human exfoliated deciduous teeth (SHED) represent a promising cell source for tissue regeneration. In the present study we decided to test the inductive effect of chitosan and transforming growth factor-β1 (TGFβ1) as a scaffold/factor combination on SHED proliferation and osteogenic differentiation.
  
  
  METHODS
  Cell proliferation was quantitatively assessed by PrestoBlue, live/dead assay was performed and cell attachment to chitosan scaffold was examined by scanning electron microscopy (SEM). For osteogenic differentiation analysis, alkaline phosphatase activity was quantified, cells were stained with Alizarin Red, and the lineage specific genes/proteins ALP, COL I, BSP, and OCN were analysed by real-time PCR and Western blot.
  
  
  RESULTS
  SHED remained viable and attached well to the chitosan structure. Moreover, TGFβ1 significantly enhanced the proliferative activity of SHED on the chitosan scaffold. Our data further revealed that chitosan and TGFβ1 enhanced the osteogenic differentiation of SHED, as evidenced by high ALP activity, strong mineral deposition, and the up-regulation of ALP, COL I, BSP, and OCN gene/protein expression.
  
  
  CONCLUSIONS
  Together, data from our study indicate that the combination of chitosan scaffolds and TGFβ1 enhanced proliferation and osteogenic differentiation of SHED. These findings suggest that the combined application of chitosan scaffold and TGFβ1 in conjunction with SHED might be beneficial for in vivo bone regeneration.
authors.affiliation: Conservative Department, School of Dental Sciences, Universiti Sains Malaysia, 16150 Kubang Kerian, Kelantan, Malaysia. Electronic address: ghorafi44@yahoo.com.
authors.forename: Manal
authors.initials: M
authors.lastname: Farea
authors.affiliation: Conservative Department, School of Dental Sciences, Universiti Sains Malaysia, 16150 Kubang Kerian, Kelantan, Malaysia. Electronic address: adamkck@usm.my.
authors.forename: Adam
authors.initials: A
authors.lastname: Husein
authors.affiliation: Reconstructive Sciences Unit, School of Medical Sciences, Universiti Sains Malaysia, 16150 Kubang Kerian, Kelantan, Malaysia. Electronic address: ashalim@usm.my.
authors.forename: Ahmad Sukari
authors.initials: AS
authors.lastname: Halim
authors.affiliation: Oral Biology Unit, School of Dental Sciences, Universiti Sains Malaysia, 16150 Kubang Kerian, Kelantan, Malaysia.
authors.forename: Nurul Asma
authors.initials: NA
authors.lastname: Abdullah
authors.affiliation: Oral Biology Unit, School of Dental Sciences, Universiti Sains Malaysia, 16150 Kubang Kerian, Kelantan, Malaysia.
authors.forename: Khairani Idah
authors.initials: KI
authors.lastname: Mokhtar
authors.affiliation: Skin Research and Innovation Centre, Research and Development Division, WIPRO Manufacturing Services Sdn Bhd, Persiaran Subang Permai, Taman Perindustrian Subang, 47610 Subang Jaya, Selangor, Malaysia.
authors.forename: Chin Keong
authors.initials: CK
authors.lastname: Lim
authors.affiliation: Conservative Department, School of Dental Sciences, Universiti Sains Malaysia, 16150 Kubang Kerian, Kelantan, Malaysia.
authors.forename: Zurairah
authors.initials: Z
authors.lastname: Berahim
authors.affiliation: Conservative Department, School of Dental Sciences, Universiti Sains Malaysia, 16150 Kubang Kerian, Kelantan, Malaysia.
authors.forename: Kasmawati
authors.initials: K
authors.lastname: Mokhtar
mesh_terms: D000469:Alkaline Phosphatase / Q000378:metabolism; D015153:Blotting, Western; D002448:Cell Adhesion; D002454:Cell Differentiation / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D048271:Chitosan / Q000494:pharmacology*; D003782:Dental Pulp / Q000166:cytology*; D006801:Humans; D008855:Microscopy, Electron, Scanning; D010012:Osteogenesis / Q000187:drug effects*; D012313:RNA / Q000032:analysis; D060888:Real-Time Polymerase Chain Reaction; D013234:Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D023822:Tissue Engineering; D054457:Tissue Scaffolds; D014094:Tooth, Deciduous / Q000166:cytology*; D053773:Transforming Growth Factor beta1 / Q000494:pharmacology*
chemical_list: D053773:Transforming Growth Factor beta1; D012313:RNA; D048271:Chitosan; D000469:Alkaline Phosphatase
keywords: Chitosan; Osteogenic differentiation; Proliferation; SHED; TGFβ(1)