Department of Oral and Maxillofacial Surgery, College of Stomatology, Ninth People’s Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai 200011, PR China.
Eur Cell Mater. 2011 Jul 15;22:68-82; discussion 83. doi: 10.22203/ecm.v022a06.
The purpose of this study was to investigate the effects of akermanite as compared to β-TCP on attachment, proliferation, and osteogenic differentiation of human periodontal ligament cells (hPDLCs). Scanning electron microscopy (SEM) and actin filament labeling were used to reveal attachment and growth of hPDLCs seeded on β-TCP and akermanite ceramic. Cell proliferation was tested by lactic acid production and MTT analysis, while osteogenic differentiation was assayed by alkaline phosphatase (ALP) expression and real-time polymerase chain reaction (PCR) analysis on markers of osteopontin (OPN), dentin matrix acidic phosphoprotein-1 (DMP-1), and osteocalcin (OCN), and further detected by enzyme-linked immunosorbent analysis (ELISA) analysis for OCN expression. Besides, the ions released from akermanite and their effect on hPDLCs was also measured by inductively coupled plasma atomic emission spectroscopy (ICP-AES), MTT analysis, ALP expression and real-time PCR analysis. hPDLCs attached well on both ceramics, but showed better spreading on akermanite. hPDLCs proliferated more rapidly on akermanite than β-TCP. Importantly, osteogenic differentiation of hPDLCs was enhanced on akermanite compared to β-TCP. Besides, Ca, Mg and Si ions were released from akermanite, while only Ca ions were released from β-TCP. Moreover, more pronounced proliferation and higher osteogenic gene expression for hPDLCs cultured with akermanite extract were detected as compared to cells cultured on akermanite. Therefore, akermanite ceramic showed an enhanced effect on proliferation and osteogenic differentiation of hPDLCs, which might be attributed to the release of ions containing Ca, Mg and Si from the material. It is suggested that akermanite ceramics may serve as a potential material for periodontal bone regeneration.
本研究旨在探究钙镁硅灰石(akermanite)与 β-TCP 相比对人牙周膜细胞(hPDLCs)黏附、增殖和成骨分化的影响。通过扫描电子显微镜(SEM)和肌动蛋白丝标记揭示了 hPDLCs 在 β-TCP 和钙镁硅灰石陶瓷上的黏附与生长。通过乳酸产量和 MTT 分析测试细胞增殖,通过碱性磷酸酶(ALP)表达和实时聚合酶链反应(PCR)分析对骨桥蛋白(OPN)、牙本质基质酸性磷酸蛋白-1(DMP-1)和骨钙素(OCN)的标志物进行成骨分化分析,并用酶联免疫吸附分析(ELISA)检测 OCN 表达。此外,还通过电感耦合等离子体原子发射光谱(ICP-AES)、MTT 分析、ALP 表达和实时 PCR 分析测量了钙镁硅灰石释放的离子及其对 hPDLCs 的影响。hPDLCs 在两种陶瓷上黏附良好,但在钙镁硅灰石上表现出更好的铺展。hPDLCs 在钙镁硅灰石上的增殖速度明显快于β-TCP。重要的是,与 β-TCP 相比,hPDLCs 在钙镁硅灰石上的成骨分化增强。此外,钙镁硅灰石释放 Ca、Mg 和 Si 离子,而β-TCP 仅释放 Ca 离子。此外,与培养在钙镁硅灰石上的细胞相比,培养在钙镁硅灰石浸提液中的 hPDLCs 增殖更明显,成骨基因表达更高。因此,钙镁硅灰石陶瓷对 hPDLCs 的增殖和成骨分化有增强作用,这可能归因于材料中释放的含 Ca、Mg 和 Si 的离子。提示钙镁硅灰石陶瓷可能成为牙周骨再生的潜在材料。