title: Cementum protein 1 (CEMP1) induces differentiation by human periodontal ligament cells under three-dimensional culture conditions.
abstract: PDL (periodontal ligament) is a source of multi-potent stem cells in humans and their differentiation potential to a cementoblast and osteoblast phenotypes has been shown. Tissue construction from PDL-derived cells could be considered as a valuable technique for periodontal regenerative medicine. On these basis, we determined the role of CEMP1 (cementum protein 1) as a factor to induce differentiation of human PDL cells in a 3D (three-dimensional) fashion. Human PDL cells were grown in an RCCS (rotary cell culture system) D-410 RWV (rotating wall vessel) bioreactor, and maintained in either experimental (CEMP1 2.5 μg/ml) or control media during 4 weeks. Cell proliferation in the presence of CEMP1 was determined. The tissue-like structure formed was analysed histologically, stained with Alizarin Red and Alcian Blue. ALP (alkaline phosphatase)-specific activity, immunostaining, RT-PCR (reverse transcription-PCR) and Western blotting were performed to determine the expression of BSP (bone sialoprotein), enamel [AMBN (ameloblastin) and AMEL (amelogenin)], cementum [CAP (cementum attachment protein) and CEMP1] and cartilage-related proteins (Sox9, aggrecan, types II and X collagens). Our results show that hrCEMP1 (human recombinant CEMP1) promoted cell proliferation by human PDL cells in 3D cultures and induced the formation of a tissue-like structure resembling bone and/or cementum and material similar to cartilage. The addition of hrCEMP1 to the 3D human PDL cell cultures increased ALP-specific activity by 2.0-fold and induced the expression of markers for the osteogenic, cementogenic and chondrogenic phenotypes at the mRNA and protein levels. Our data show that human PDL cells in 3D cultures with the addition of CEMP1 has the potential to be used for the bioengineering reconstruction of periodontal tissues and cartilage since our results suggest that CEMP1 stimulates human PDL cells to differentiate towards different phenotypes.
authors.affiliation: Laboratorio de Biologa Periodontal y Tejidos Mineralizados, Facultad de Odontología, Universidad Nacional Autónoma de México, Mexico City, Mexico.
authors.forename: Lía
authors.initials: L
authors.lastname: Hoz
authors.forename: Enrique
authors.initials: E
authors.lastname: Romo
authors.forename: Margarita
authors.initials: M
authors.lastname: Zeichner-David
authors.forename: Mariano
authors.initials: M
authors.lastname: Sanz
authors.forename: Javier
authors.initials: J
authors.lastname: Nuñez
authors.forename: Luis
authors.initials: L
authors.lastname: Gaitán
authors.forename: Gabriela
authors.initials: G
authors.lastname: Mercado
authors.forename: Higinio
authors.initials: H
authors.lastname: Arzate
mesh_terms: D053674:Aggrecans / Q000378:metabolism; D000469:Alkaline Phosphatase / Q000235:genetics / Q000378:metabolism; D053523:Amelogenin / Q000378:metabolism; D018929:Cell Culture Techniques; D002454:Cell Differentiation / Q000187:drug effects*; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D024043:Collagen Type II / Q000378:metabolism; D024121:Collagen Type X / Q000378:metabolism; D003746:Dental Enamel Proteins / Q000378:metabolism; D006801:Humans; D058574:Integrin-Binding Sialoprotein / Q000378:metabolism; D010513:Periodontal Ligament / Q000166:cytology*; D017027:Protein Tyrosine Phosphatases / Q000378:metabolism; D011506:Proteins / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology*; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology; D055758:SOX9 Transcription Factor / Q000378:metabolism; D013234:Stem Cells / Q000166:cytology / Q000378:metabolism; D023822:Tissue Engineering
chemical_list: C490889:AMBN protein, human; D053674:Aggrecans; D053523:Amelogenin; C510886:CEMP1 protein, human; D024043:Collagen Type II; D024121:Collagen Type X; D003746:Dental Enamel Proteins; D058574:Integrin-Binding Sialoprotein; D011506:Proteins; D011994:Recombinant Proteins; D055758:SOX9 Transcription Factor; C527932:SOX9 protein, human; D000469:Alkaline Phosphatase; C404489:HACD1 protein, human; D017027:Protein Tyrosine Phosphatases