Department of Dental Medicine, Karolinska Institutet, Huddinge, Sweden.
J Dent Res. 2012 Aug;91(8):789-94. doi: 10.1177/0022034512452430. Epub 2012 Jun 26.
The success of tissue-engineering therapies is dependent on the ability of scaffolds to guide differentiation of progenitor cells. Here we present a new approach using a biomimetic construct composed of hydroxyapatite modified with an in vitro-derived extracellular matrix (HA-ECM) and seeded with periodontal ligament progenitor cells (PDLCs). The study aimed to investigate the effect of HA-ECM on osteogenic differentiation of PDLCs and in vivo evaluation of the PDLC-seeded HA-ECM constructs using a rat calvarial critical-sized defect model. After flow-cytometric phenotyping of PDLCs for typical mesenchymal stem cell markers, the PDLCs were cultured on HA-ECM or HA alone in osteogenic media and assessed by MTT, alkaline phosphatase (ALP) assays, and real-time qPCR at different time intervals after seeding. New bone formation induced by PDLC-seeded constructs was assessed by histomorphometric analysis at 12 weeks post-operatively. The PDLCs seeded on HA-ECM showed significantly higher ALP activity and up-regulation of bone-related genes. The treatment with PDLC-seeded HA-ECM significantly improved calvarial bone repair, with the highest amount of newly formed bone elicited by cell-seeded constructs cultured for 14 days. Our results highlight the PDLC-seeded HA-ECM constructs as a promising tool for craniofacial bone regeneration.
组织工程疗法的成功取决于支架引导祖细胞分化的能力。在这里，我们提出了一种新的方法，使用由体外衍生的细胞外基质（HA-ECM）修饰的仿生构建体和牙周韧带祖细胞（PDLCs）进行种子处理。本研究旨在研究 HA-ECM 对 PDLC 成骨分化的影响，并使用大鼠颅骨临界尺寸缺损模型对 PDLC 种子 HA-ECM 构建体进行体内评估。在对 PDLC 进行典型间充质干细胞标志物的流式细胞表型分析后，将 PDLC 接种在 HA-ECM 或 HA 上，在成骨培养基中培养，并在接种后不同时间点通过 MTT、碱性磷酸酶（ALP）测定和实时 qPCR 进行评估。术后 12 周通过组织形态计量学分析评估由 PDLC 种子构建体诱导的新骨形成。在 HA-ECM 上接种的 PDLC 表现出明显更高的 ALP 活性和骨相关基因的上调。PDLC 种子 HA-ECM 的治疗显著改善了颅骨骨修复，用培养 14 天的细胞种子构建体引起的新骨形成量最高。我们的结果强调了 PDLC 种子 HA-ECM 构建体作为颅面骨再生的有前途的工具。
