IADR Abstract Archives
Biological Effect of CEMP1 Peptide by Human Periodontal Ligament Cells
Objectives: Cementum is a specialized connective mineralized tissue. Cementum seems to be critical to achieve the regeneration of the periodontal structures lost due periodontal disease. Human periodontal ligament cells (hPLC), contain heterogeneous cell subpopulations and mesenchymal stem cells that might be involved in the periodontium homeostasis and regeneration of the periodontal tissues. However, cementum proteins such as Cementum protein 1 (CEMP1) has been shown to play a key role on human periodontal ligament cells and their commitment to cementoblast/osteoblast lineages, through promoting the proliferation and differentiation of hPLC cells by ERK1/2 kinase activation. Nowadays peptides are widely used to induce cellular responses. The identification of a CEMP1 mimic peptide, applied on mineralization inductive processes will allow us to consider new strategies for promote cementogenesis.
OBJECTIVE.-To determine the effect of a CEMP1 peptide on the differentiation of hPLC cells in vitro through ERK1/2 and p38 MAP kinases activation.
Methods: The hPLC cells were cultured in DMEM medium. The positive and negative controls were determined and the experimental group was stimulated with different concentrations of CEMP1 peptide. MTT assay was performed and the effective dose was determined. Differentiation tests were performed; they included alizarin red staining, qRT-PCR real time and western blotting for BSP, CAP, CEMP1, AMEL and AMBN, and activity of alkaline phosphatase, every assay was measured on 3, 7 and 14 days of culture (triplicate assays). Time course was also performed at short and long times and western blotting to determine the phosphorylation of ERK 1/2 and p38 kinases.
Results: CEMP1 peptide promotes proliferation of hPLC and induces differentiation at transcriptional and translational level toward a mineralizing phenotype; also increases the ALP activity and calcium nodule formation through phosphorylation of MAPK kinases.
Conclusions: CEMP1 peptide shows to be biomimetic for the functions before described for the whole CEMP1 recombinant protein.
OBJECTIVE.-To determine the effect of a CEMP1 peptide on the differentiation of hPLC cells in vitro through ERK1/2 and p38 MAP kinases activation.
Methods: The hPLC cells were cultured in DMEM medium. The positive and negative controls were determined and the experimental group was stimulated with different concentrations of CEMP1 peptide. MTT assay was performed and the effective dose was determined. Differentiation tests were performed; they included alizarin red staining, qRT-PCR real time and western blotting for BSP, CAP, CEMP1, AMEL and AMBN, and activity of alkaline phosphatase, every assay was measured on 3, 7 and 14 days of culture (triplicate assays). Time course was also performed at short and long times and western blotting to determine the phosphorylation of ERK 1/2 and p38 kinases.
Results: CEMP1 peptide promotes proliferation of hPLC and induces differentiation at transcriptional and translational level toward a mineralizing phenotype; also increases the ALP activity and calcium nodule formation through phosphorylation of MAPK kinases.
Conclusions: CEMP1 peptide shows to be biomimetic for the functions before described for the whole CEMP1 recombinant protein.