Miron Richard J, Bosshardt Dieter D, Gemperli Anja C, Dard Michel, Buser Daniel, Gruber Reinhard, Sculean Anton
Department of Periodontology, School of Dental Medicine, University of Bern, Freiburgstrasse 7, 3010, Bern, Switzerland.
Clin Oral Investig. 2014;18(2):443-51. doi: 10.1007/s00784-013-0977-4. Epub 2013 Apr 26.
Recent studies suggest that a combination of enamel matrix derivative (EMD) with grafting material may improve periodontal wound healing/regeneration. Newly developed calcium phosphate (CaP) ceramics have been demonstrated a viable synthetic replacement option for bone grafting filler materials.
This study aims to test the ability for EMD to adsorb to the surface of CaP particles and to determine the effect of EMD on downstream cellular pathways such as adhesion, proliferation, and differentiation of primary human osteoblasts and periodontal ligament (PDL) cells.
EMD was adsorbed onto CaP particles and analyzed for protein adsorption patterns via scanning electron microscopy and high-resolution immunocytochemistry with an anti-EMD antibody. Cell attachment and cell proliferation were quantified using CellTiter 96 One Solution Cell Assay (MTS). Cell differentiation was analyzed using real-time PCR for genes encoding Runx2, alkaline phosphatase, osteocalcin, and collagen1α1, and mineralization was assessed using alizarin red staining.
Analysis of cell attachment revealed significantly higher number of cells attached to EMD-adsorbed CaP particles when compared to control and blood-adsorbed samples. EMD also significantly increased cell proliferation at 3 and 5 days post-seeding. Moreover, there were significantly higher mRNA levels of osteoblast differentiation markers including collagen1α1, alkaline phosphatase, and osteocalcin in osteoblasts and PDL cells cultured on EMD-adsorbed CaP particles at various time points.
The present study suggests that the addition of EMD to CaP grafting particles may influence periodontal regeneration by stimulating PDL cell and osteoblast attachment, proliferation, and differentiation. Future in vivo and clinical studies are required to confirm these findings.
The combination of EMD and CaP may represent an option for regenerative periodontal therapy in advanced intrabony defects.
近期研究表明，牙釉质基质衍生物（EMD）与移植材料联合使用可能会改善牙周伤口愈合/再生。新开发的磷酸钙（CaP）陶瓷已被证明是骨移植填充材料可行的合成替代选择。
本研究旨在测试EMD吸附到CaP颗粒表面的能力，并确定EMD对下游细胞途径的影响，如原代人成骨细胞和牙周膜（PDL）细胞的黏附、增殖和分化。
将EMD吸附到CaP颗粒上，并通过扫描电子显微镜和使用抗EMD抗体的高分辨率免疫细胞化学分析蛋白质吸附模式。使用CellTiter 96 One Solution Cell Assay（MTS）对细胞附着和细胞增殖进行定量。使用实时PCR分析编码Runx2、碱性磷酸酶、骨钙素和胶原蛋白1α1的基因来分析细胞分化，并使用茜素红染色评估矿化情况。
细胞附着分析显示，与对照和血液吸附样本相比，附着在EMD吸附的CaP颗粒上的细胞数量显著更多。EMD在接种后第3天和第5天也显著增加了细胞增殖。此外，在不同时间点，培养在EMD吸附的CaP颗粒上的成骨细胞和PDL细胞中，包括胶原蛋白1α1、碱性磷酸酶和骨钙素在内的成骨细胞分化标志物的mRNA水平显著更高。
本研究表明，在CaP移植颗粒中添加EMD可能通过刺激PDL细胞和成骨细胞的附着、增殖和分化来影响牙周再生。未来需要进行体内和临床研究来证实这些发现。
EMD和CaP的联合可能是晚期骨内缺损再生性牙周治疗的一种选择。
