1Department of Maxillofacial Tissue Regeneration, School of Dentistry and Research Center for Tooth and Periodontal Regeneration (MRC), Kyung Hee University, Seoul, Republic of Korea.
J Biomater Appl. 2014 Mar;28(7):1069-78. doi: 10.1177/0885328213495903. Epub 2013 Jul 9.
The aim of the present study was to fabricate mineralized polycaprolactone nanofibrous scaffold and investigate its ability to elicit odontogenic differentiation of human dental pulp cells, compared to the pure polycaprolactone scaffold. Polycaprolactone nanofibrous scaffold was produced by electrospinning, and the surface was mineralized with apatite. Cellular behaviors on the mineralized polycaprolactone scaffold were assessed in terms of cell adhesion, growth, and odontoblastic differentiation. To evaluate the signal transduction of human dental pulp cells, mRNA expression was analyzed and Western blotting was performed. Mineralized polycaprolactone showed improved cell proliferation, mineralized nodule formation, and expression of odontoblastic marker genes including alkaline phosphatase, osteopontin, osteocalcin, dentin sialophosphoprotein (DSPP), and dentin matrix protein-1, as compared with pure polycaprolactone. Although the cell adhesion on the mineralized polycaprolactone was similar to that of the polycaprolactone, the expression level of proteins including collagen type I and the key adhesion receptor (integrin components α1, α2, and β1) was upregulated in mineralized polycaprolactone compared to polycaprolactone. Especially, cells seeded onto mineralized polycaprolactone scaffolds showed significantly increased levels of phosphorylated focal adhesion kinase, a marker of integrin activation, and downstream pathways, such as phosphor (p)-Akt, p-extracellular signal regulated kinase, p-c Jun N-terminal kinase, nuclear factor-kappa B, c-fos, and c-jun, compared with pure polycaprolactone. The mineralized polycaprolactone scaffold is attractive for dentin tissue engineering by promoting growth and odontogenic differentiation of human dental pulp cells through the integrin-mediated signaling pathway.
本研究旨在制备矿化聚己内酯纳米纤维支架,并与纯聚己内酯支架相比,研究其对人牙髓细胞成牙本质分化的诱导能力。聚己内酯纳米纤维支架通过静电纺丝制备,表面矿化形成磷灰石。通过细胞黏附、生长和牙本质细胞分化来评估矿化聚己内酯支架上的细胞行为。为了评估人牙髓细胞的信号转导,进行了 mRNA 表达分析和 Western 印迹分析。与纯聚己内酯相比,矿化聚己内酯显示出改善的细胞增殖、矿化结节形成和牙本质细胞标志物基因(碱性磷酸酶、骨桥蛋白、骨钙素、牙本质涎磷蛋白(DSPP)和牙本质基质蛋白-1)的表达。尽管矿化聚己内酯上的细胞黏附与聚己内酯相似,但矿化聚己内酯上包括胶原蛋白 I 和关键黏附受体(整合素成分α1、α2 和β1)在内的蛋白质表达水平上调。特别是,与纯聚己内酯相比,接种到矿化聚己内酯支架上的细胞显示出整合素激活标志物磷酸化粘着斑激酶(FAK)以及下游途径(如 p-Akt、p-细胞外信号调节激酶、p-c Jun N-末端激酶、核因子-κB、c-fos 和 c-jun)的水平显著增加。矿化聚己内酯支架通过整合素介导的信号通路促进人牙髓细胞的生长和牙本质分化,是牙本质组织工程的理想选择。