Excellence Center in Regenerative Dentistry, Chulalongkorn University, Bangkok, 10330, Thailand.
Oral Biology Research Center, Faculty of Dentistry, Chulalongkorn University, Bangkok, 10330, Thailand.
Arch Oral Biol. 2018 Apr;88:24-33. doi: 10.1016/j.archoralbio.2018.01.007. Epub 2018 Jan 31.
The aim of the present study was to develop a Jagged1-coated polycaprolactone (PCL) membrane and to evaluate the response of human periodontal ligament cells (hPDL) on this membrane in vitro.
Membranes were prepared from PCL and PCL-incorporated hydroxyapatite (PCL/HA). The membranes' surface roughness, surface wettability, and mechanical properties were examined. An indirect affinity immobilization technique was used to coat the membranes with Jagged1. Membrane cytotoxicity was evaluated using LIVE/DEAD and MTT assays. The morphology of the cells on the membranes was observed using scanning electron microscopy. hPDL alkaline phosphatase (ALP) enzymatic activity and mineral deposition were examined using an ALP assay and Alizarin Red S staining, respectively. Notch target gene mRNA expression was determined using real-time polymerase chain reaction.
The PCL/HA membranes exhibited a significantly reduced surface contact angle, decreased maximum tensile strain, and ultimate tensile stress. However, the surface roughness parameters were significantly increased. The PCL and PCL/HA membranes were not cytotoxic to hPDL in vitro. hPDLs attached and spread on both membrane types. Further, indirect affinity immobilized Jagged1 on the membranes upregulated hPDL Notch target gene expression. After culturing in osteogenic medium, Jagged1-immobilized PCL/HA membranes significantly enhanced hPDL ALP enzymatic activity.
Indirect immobilized Jagged1 PCL/HA membranes could be further developed as an alternative guided tissue regeneration membrane to promote osteogenic differentiation in periodontal defects.
本研究旨在制备一种 Jagged1 涂层的聚己内酯(PCL)膜,并评估其在体外对人牙周膜细胞(hPDL)的反应。
采用聚己内酯和掺入羟基磷灰石的聚己内酯(PCL/HA)制备膜。检测了膜的表面粗糙度、表面润湿性和力学性能。采用间接亲和固定化技术在膜上涂层 Jagged1。通过 LIVE/DEAD 和 MTT 测定法评估膜的细胞毒性。用扫描电子显微镜观察细胞在膜上的形态。通过碱性磷酸酶(ALP)测定和茜素红 S 染色分别检测 hPDL 的碱性磷酸酶(ALP)酶活性和矿化沉积。通过实时聚合酶链反应测定 Notch 靶基因的 mRNA 表达。
PCL/HA 膜的表面接触角显著降低,最大拉伸应变和极限拉伸应力降低。然而,表面粗糙度参数显著增加。PCL 和 PCL/HA 膜在体外对 hPDL 无细胞毒性。hPDL 附着并在两种膜上展开。此外,间接亲和固定化 Jagged1 可上调膜上 hPDL Notch 靶基因的表达。在成骨培养基中培养后,Jagged1 固定化的 PCL/HA 膜显著增强了 hPDL 的 ALP 酶活性。
间接固定化 Jagged1 的 PCL/HA 膜可进一步开发为替代引导组织再生膜,以促进牙周缺损中的成骨分化。