title: [Experimental study on the in vitro osteogenic differentiation of dental pulp stem cells encapsulated in Pluronic F-127 hydrogel].
abstract: OBJECTIVE
  To evaluate the biocompatibility and viability of nonionic triblock copolymer Pluronic F-127 as a cell scaffold for osteogenic differentiation of dental pulp stem cells(DPSC).
  
  
  METHODS
  DPSC were obtained via enzymatic digestion method and purified bylimited dilution method. The freeze dried hydrogel of 20% Pluronic F-127 was prepared and itsstructurewas observed usingscanning electron microscopy(SEM). After the encapsulation of cells of passage 3 in Pluronic F-127, the effects of hydrogel on the proliferations of DPSC were assessed with methyl thiazolyl terazolium(MTT) after one day and 3, 5, 7 days of incubations, respectively. On day 14, osteogenic abilities of DPSC encapsulated in the hydrogel were estimated by means of alizarin red S, immunocytochemical staining and real-time quantitative PCR(RT-qPCR).
  
  
  RESULTS
  DPSC were isolated and cultured successfully in the present study. SEM observations showed that porous structures which might be suitable for cell culture. A570 values of MTT were then normalized. A570 values of the cells in 2D cultures were 0.30±0.06, 0.30±0.17, 0.35±0.04 and 0.25±0.06 and A570 values of DPSC in 3D cultures were 0.36±0.06, 0.54±0.18, 0.70±0.10 and 0.32±0.10 on day 1, 3, 5 and 7, respectively. A570 value peaks were found on day 5 in both groups. The proliferation of 3D cultured DPSC was higher than that of 2D cultured cells(P<0.05). After 14 days of osteogenic induction, there were no calcium nodules observed in the control group and the numbers of calcium nodulesin the 2D and 3D groups had no significant difference(P>0.05). Inmmunocytochemical staining demonstrated strong expression of osteoblast marker Runt-related transcription factor 2(RUNX2), type Ⅰ collagen(Col-Ⅰ) and relatively low expression of osteocalcin(OCN). Moreover, RT-qPCR showed no differences between the relative expression of ALP, RUNX-2, OCN in the 2D and 3D groups (P>0.05), but a higher relative expression of Col-Ⅰ was observed in the 3D group(P=0.023).
  
  
  CONCLUSIONS
  Pluronic F-127 is a promising cell scaffold or cell carrier for the osteobalst differentiation of dental pulp stem cells.
authors.affiliation: Department of Stomatology, People's Hospital of Xinjiang Uygur Autonomous Region, Urumqi 830001, China.
authors.forename: Abudureheman
authors.initials: A
authors.lastname: Paerhati
authors.affiliation: Department of Stomatology, People's Hospital of Xinjiang Uygur Autonomous Region, Urumqi 830001, China.
authors.forename: Huojia
authors.initials: H
authors.lastname: Muhetaer
authors.affiliation: Department of Stomatology, People's Hospital of Xinjiang Uygur Autonomous Region, Urumqi 830001, China.
authors.forename: Wufuer
authors.initials: W
authors.lastname: Duolikun
authors.affiliation: Department of Stomatology, People's Hospital of Xinjiang Uygur Autonomous Region, Urumqi 830001, China.
authors.forename: Halike
authors.initials: H
authors.lastname: Maimaitiyiming
authors.affiliation: Department of Stomatology, People's Hospital of Xinjiang Uygur Autonomous Region, Urumqi 830001, China.
authors.forename: X W
authors.initials: XW
authors.lastname: Liu
mesh_terms: D018929:Cell Culture Techniques; D002454:Cell Differentiation; D002478:Cells, Cultured; D024042:Collagen Type I / Q000378:metabolism; D050659:Core Binding Factor Alpha 1 Subunit / Q000378:metabolism; D003782:Dental Pulp / Q000166:cytology*; D006801:Humans; D020136:Hydrogel, Polyethylene Glycol Dimethacrylate; D010006:Osteoblasts / Q000378:metabolism; D015675:Osteocalcin / Q000378:metabolism; D010012:Osteogenesis; D020442:Poloxamer; D013234:Stem Cells / Q000166:cytology*; D054457:Tissue Scaffolds
chemical_list: D024042:Collagen Type I; D050659:Core Binding Factor Alpha 1 Subunit; C493709:RUNX2 protein, human; D015675:Osteocalcin; D020442:Poloxamer; D020136:Hydrogel, Polyethylene Glycol Dimethacrylate