ABSTRACT
Numerous studies have indicated dental pulp stem cells (DPSCs) potency to differentiate into several types of cell lineages. Oligodendrocyte lineage transcription factor 2 (OLIG2) plays an important role in the oligodendrogenic pathway. In this study, a tetracycline (Tet)-inducible system expressing OLIG2 gene was transfected into human DPSCs to direct their differentiation toward oligodendrocyte progenitor cells (OPCs). Following induction, the expression of stage-specific markers was studied by Reverse Transcription quantitative Polymerase Chain Reaction (RT-qPCR), immunocytochemistry and western blotting. In the following, the cells were transplanted into the mouse model of local sciatic demyelination damage by lysolecithin. Recovery of lysolecithin-induced lesions in sciatic nerve was studied by treadmill exercise, von Frey filament test and hind paw withdrawal in response to a thermal stimulus. Improvement of behavioral symptoms was efficiently observed from the second week to the sixth week post-transplantation. Our findings showed that exogenous expression of the OLIG2 gene by a Tet-regulated system could be used as an efficient way to induce the differentiation of DPSCs into functional oligodendrocytes. Meanwhile, the DPSC-derived OPCs have relevant therapeutic potential in the animal model of sciatic nerve injury and therefore might represent a valuable tool for stem cell-based therapy in inflammatory and degenerative diseases of the peripheral and central nervous systems (CNSs).
Subject(s)
Basic Helix-Loop-Helix Transcription Factors/metabolism , Gene Expression/drug effects , Nerve Regeneration/physiology , Nerve Tissue Proteins/metabolism , Protein Synthesis Inhibitors/pharmacology , Stem Cell Transplantation/methods , Stem Cells/drug effects , Tetracycline/pharmacology , Animals , Basic Helix-Loop-Helix Transcription Factors/genetics , Cell Differentiation , Cells, Cultured , Dental Pulp/cytology , Disease Models, Animal , Humans , Lysophosphatidylcholines/toxicity , Male , Mice , Mice, Inbred BALB C , Motor Activity/physiology , Nerve Tissue Proteins/genetics , Oligodendrocyte Transcription Factor 2 , Oligodendroglia/physiology , Sciatic Neuropathy/chemically induced , Sciatic Neuropathy/physiopathology , Sciatic Neuropathy/surgery , Stem Cells/physiology , Time FactorsABSTRACT
An Iranian field isolate (IR01) of Infectious bursal disease virus (IBDV) was characterized by sequence analysis of its VP2 gene and protein. Comparison of the obtained sequences with those of IBDV isolates from other countries revealed that IR01 was similar to very virulent IBDV (vvIBDV) strains with the identities at nucleotide and amino acid levels reaching 98.198.9% and 99.199.3%, respectively. On the other hand, it was less similar to non-vvIBDV strains; with nucleotide and amino acid identities of 95.295.7% and 96.097.3%, respectively. Out of nine unique nucleotide differences found between IR01 and some other serotype 1 strains only two resulted in amino acid substitutions (Ile296Val and Thr359Lys). In phylogenetic analysis, IR01 was closely related to Asian and European vvIBDV strains. Based on these results, IR01 obviously belongs to vvIBDV strains.
