STRO-1 selected rat dental pulp stem cells transfected with adenoviral-mediated human bone morphogenetic protein 2 gene show enhanced odontogenic differentiation.
Dental pulp stem cells harbor great potential for tissue-engineering purposes. However, previous studies have shown variable results, and some have reported only limited osteogenic and odontogenic potential.Because bone morphogenetic proteins (BMPs) are well-established agents to induce bone and dentin formation,in this study STRO-1-selected rat dental pulp-derived stem cells were transfected with the adenoviral mediated human BMP-2 gene. Subsequently, the cells were evaluated for their odontogenic differentiation ability in medium not containing dexamethasone or other stimuli. Cultures were investigated using light microscopy and scanning electron microscopy (SEM) and evaluated for cell proliferation, alkaline phosphatase(ALP) activity, and calcium content. Real-time polymerase chain reaction (PCR) was performed for gene expression of Alp, osteocalcin, collagen type I, bone sialoprotein, dentin sialophosphoprotein, and dentin matrix acidic phosphoprotein 1. Finally, an oligo-microarray was used to profile the expression of odontogenesis-related genes. Results of ALP activity, calcium content, and real-time PCR showed that only BMP2-transfected cells had the ability to differentiate into the odontoblast phenotype and to produce a calcified extracellular matrix. SEM and oligo-microarray confirmed these results. In contrast, the non-transfected cells represented a less differentiated cell phenotype. Based on our results, we concluded that the adenovirus can transfect STRO-1 selected cells with high efficacy. After BMP2 gene transfection, these cells had the ability to differentiate into odontoblast phenotype, even without the addition of odontogenic supplements to the medium.
Yang
Xuechao
X
Department of Periodontology and Biomaterials, Radboud University Nijmegen Medical Centre, Nijmegen, The Netherlands.
van der Kraan
Peter M
PM
van den Dolder
Juliette
J
Walboomers
X Frank
XF
Bian
Zhuan
Z
Fan
Mingwen
M
Jansen
John A
JA
D000256:Adenoviridae / Q000235:genetics; D000469:Alkaline Phosphatase / Q000032:analysis; D000818:Animals; D055396:Bone Morphogenetic Protein 2; D019485:Bone Morphogenetic Proteins / Q000235:genetics*; D002113:Calcification, Physiologic / Q000502:physiology; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D049109:Cell Proliferation; D002478:Cells, Cultured; D024042:Collagen Type I / Q000378:metabolism; D003470:Culture Media; D004247:DNA / Q000032:analysis; D003782:Dental Pulp / Q000166:cytology* / Q000378:metabolism / Q000502:physiology* / Q000648:ultrastructure; D005109:Extracellular Matrix / Q000378:metabolism; D016326:Extracellular Matrix Proteins / Q000378:metabolism; D005456:Fluorescent Dyes / Q000378:metabolism; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D006801:Humans; D058574:Integrin-Binding Sialoprotein; D009930:Organic Chemicals / Q000378:metabolism; D015675:Osteocalcin / Q000378:metabolism; D010750:Phosphoproteins / Q000378:metabolism; D011498:Protein Precursors / Q000378:metabolism; D051381:Rats; D012795:Sialoglycoproteins / Q000378:metabolism; D013234:Stem Cells / Q000166:cytology* / Q000378:metabolism / Q000502:physiology* / Q000648:ultrastructure; D013997:Time Factors; D014162:Transfection; D016212:Transforming Growth Factor beta / Q000235:genetics*; D000099311:Dentin Sialophosphoprotein