Group of Cell Therapy and Tissue Engineering, Research Institute on Health Sciences-IUNICS, University of the Balearic Islands, Palma de Mallorca, Spain.
Tissue Eng Part A. 2012 Jun;18(11-12):1253-63. doi: 10.1089/ten.tea.2011.0404. Epub 2012 Mar 19.
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.
为了发现新的诱导骨形成和生物矿化的分子，本研究结合生物信息学和模拟方法，基于釉基质蛋白富含脯氨酸的结构域来设计人工肽的结构。在本研究中，评估了这些肽对人脐带间充质干细胞（hUCMSCs）向成骨谱系分化的影响，同时评估了有无成骨补充剂（氢化可的松、β-甘油磷酸和抗坏血酸）的影响，并与市售的釉基质衍生物（EMD）进行了比较。研究假设，当与分化培养基联合使用时，这些合成肽将促进 hUCMSCs 向成骨谱系分化，或者甚至可以完全由这些合成肽来指导。通过细胞增殖、骨形态发生蛋白-2 分泌和成骨标志物基因表达的测定，在 1、3 和 14 天的治疗后评估成骨诱导能力。所有肽都在使用的剂量下是安全的，表现出较低的细胞毒性。P2、P4 和 P6 在基础培养基中使细胞增殖减少 10%-15%。在用成骨补充剂联合 EMD 处理 3 天后，早期成骨标志物的表达更高，而在用成骨补充剂联合不同合成肽处理 14 天后，细胞中的骨钙素 mRNA 水平更高。我们可以得出结论，hUCMSCs 的成骨分化被短期的 EMD 处理联合成骨补充剂所促进，并且被长期的合成肽处理联合成骨补充剂所促进，在本研究中分析的所有肽变体都显示出相似的结果。
