Effect of Polyhydroxybutyrate/Chitosan/Bioglass nanofiber scaffold on proliferation and differentiation of stem cells from human exfoliated deciduous teeth into odontoblast-like cells

Mater Sci Eng C Mater Biol Appl. 2018 Aug 1:89:128-139. doi: 10.1016/j.msec.2018.03.028. Epub 2018 Mar 30.

Abstract

Scaffolds and their characteristics play a central role in tissue engineering. The purpose of this study was to determine the effects of Polyhydroxybutyrate (PHB)/Chitosan/nano-bioglass (nBG) nanofiber scaffold made using the electrospinning method, on the proliferation and differentiation of stem cells obtained from human exfoliated deciduous teeth into odontoblast-like cells. In this experimental study, the pulps of the molten deciduous teeth were isolated, thereafter, the stem cells from human exfoliated deciduous teeth (SHED) were extracted and then the 3-(4,5-dimethylthiazolyl)-2,5-diphenyltetrazolium bromide (MTT) assay was used to determine the cell viability percentage. The expression of some stem cell genes was studied by flowcytometry. These cells were then subjected to odontoblast by using the bone morphogenetic proteins-2 (BMP2) growth factor in the differentiation medium and for the expression of their specific genes. Primers of collagen type-I, dentin sialophosphoprotein (DSPP) and alkaline phosphatase (ALP) were used and the percentage of differentiation to odontoblast cells in induction scaffolds was investigated using real-time PCR and immunohistochemistry methods. The results revealed a 6-fold increase in the expression of DSPP genes and collagen type-I, and a 2-fold increase in the expression of ALP in scaffold with BMP2 group compared to the scaffold as control group which according to the immunohistochemical test results, showed the extracted SHED to have been differentiated into dentin odontoblast-like cells. As a result, this scaffold can be used as a suitable substrate to apply in dentin tissue engineering.

Keywords: Bone morphogenetic proteins-2 (BMP2); Odontogenic differentiation; Scaffold; Stem cells from human exfoliated deciduous teeth (SHED).

MeSH terms

  • Alkaline Phosphatase / genetics
  • Alkaline Phosphatase / metabolism
  • Biocompatible Materials / chemistry*
  • Biocompatible Materials / pharmacology
  • Bone Morphogenetic Protein 2 / pharmacology
  • Cell Differentiation / drug effects
  • Cell Proliferation / drug effects
  • Cells, Cultured
  • Ceramics / chemistry*
  • Chitosan / chemistry*
  • Extracellular Matrix Proteins / genetics
  • Extracellular Matrix Proteins / metabolism
  • Humans
  • Hydroxybutyrates / chemistry*
  • Nanofibers / chemistry*
  • Odontoblasts / cytology
  • Odontoblasts / metabolism
  • Osteogenesis / drug effects
  • Phosphoproteins / genetics
  • Phosphoproteins / metabolism
  • Polyesters / chemistry*
  • Prohibitins
  • Sialoglycoproteins / genetics
  • Sialoglycoproteins / metabolism
  • Stem Cells / cytology
  • Stem Cells / drug effects
  • Stem Cells / metabolism
  • Tissue Scaffolds / chemistry
  • Tooth, Deciduous / cytology

Substances

  • Biocompatible Materials
  • Bioglass
  • Bone Morphogenetic Protein 2
  • Extracellular Matrix Proteins
  • Hydroxybutyrates
  • PHB protein, human
  • Phosphoproteins
  • Polyesters
  • Prohibitins
  • Sialoglycoproteins
  • dentin sialophosphoprotein
  • poly-beta-hydroxybutyrate
  • Chitosan
  • Alkaline Phosphatase