title: Effect of enamel matrix derivative and of proline-rich synthetic peptides on the differentiation of human mesenchymal stem cells toward the osteogenic lineage.
abstract: With the aim of discovering new molecules for induction of bone formation and biomineralization, combination of bioinformatics and simulation methods were used to design the structure of artificial peptides based on proline-rich domains of enamel matrix proteins. In this study, the effect of such peptides on the differentiation toward the osteogenic lineage of human umbilical cord mesenchymal stem cells (hUCMSCs) was evaluated with or without osteogenic supplements (hydrocortisone, β-glycerol phosphate, and ascorbic acid) and compared to the effect of the commercially available enamel matrix derivative (EMD). It was hypothesized that the differentiation toward the osteogenic lineage of hUCMSCs would be promoted by the treatment with the synthetic peptides when combined with differentiation media, or it could even be directed exclusively by the synthetic peptides. Osteoinductivity was assessed by cell proliferation, bone morphogenetic protein-2 secretion, and gene expression of osteogenic markers after 1, 3, and 14 days of treatment. All peptides were safe with the dosages used, showing lower cell toxicity. P2, P4, and P6 reduced cell proliferation with growing media by 10%-15%. Higher expression of early osteoblast markers was found after 3 days of treatment with EMD in combination with osteogenic supplements, while after 14 days of treatment, cells treated by the different synthetic peptides in combination with osteogenic supplements showed higher osteocalcin mRNA levels. We can conclude that osteogenic differentiation of hUCMSCs is promoted by short-term EMD treatment in combination with osteogenic supplements and by long-term treatment by the synthetic peptides in combination with osteogenic supplements, showing similar results for all the peptide variants analyzed in this study.
authors.affiliation: Group of Cell Therapy and Tissue Engineering, Research Institute on Health Sciences-IUNICS, University of the Balearic Islands, Palma de Mallorca, Spain.
authors.forename: Joana Maria
authors.initials: JM
authors.lastname: Ramis
authors.forename: Marina
authors.initials: M
authors.lastname: Rubert
authors.forename: Jiri
authors.initials: J
authors.lastname: Vondrasek
authors.forename: Antoni
authors.initials: A
authors.lastname: Gayà
authors.forename: Staale Petter
authors.initials: SP
authors.lastname: Lyngstadaas
authors.forename: Marta
authors.initials: M
authors.lastname: Monjo
mesh_terms: D000595:Amino Acid Sequence; D015415:Biomarkers / Q000378:metabolism; D055396:Bone Morphogenetic Protein 2 / Q000378:metabolism; D002454:Cell Differentiation / Q000187:drug effects*; D019070:Cell Lineage / Q000187:drug effects*; D049109:Cell Proliferation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D003470:Culture Media; D003746:Dental Enamel Proteins / Q000737:chemistry*; D005786:Gene Expression Regulation / Q000187:drug effects; D006801:Humans; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000201:enzymology; D008958:Models, Molecular; D008969:Molecular Sequence Data; D010012:Osteogenesis / Q000187:drug effects* / Q000235:genetics; D010455:Peptides / Q000737:chemistry / Q000494:pharmacology*; D011392:Proline / Q000737:chemistry*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism
chemical_list: D015415:Biomarkers; D055396:Bone Morphogenetic Protein 2; D003470:Culture Media; D003746:Dental Enamel Proteins; D010455:Peptides; D012333:RNA, Messenger; C109069:enamel matrix proteins; D011392:Proline; D007770:L-Lactate Dehydrogenase