Division of Endodontics, Department of Restorative Dentistry, Piracicaba Dental School, University of Campinas, São Paulo, Brazil.
Int Endod J. 2012 Oct;45(10):907-14. doi: 10.1111/j.1365-2591.2012.02047.x. Epub 2012 May 3.
To examine the feasibility of using the pOBCol3.6GFPtpz [3.6-green fluorescent protein (GFP)] transgenic mice as an in vivo model for studying the biological sequence of events during pulp healing and reparative dentinogenesis.
Pulp exposures were created in the first maxillary molar of 12-16-week-old 3.6-GFP transgenic mice with CD1 and C57/Bl6 genetic background. Direct pulp capping on exposed teeth was performed using mineral trioxide aggregate followed by restoration with a light-cured adhesive system (AS) and composite resin. In control teeth, the AS was placed in direct contact with the pulp. Animals were euthanized at various time points after pulp exposure and capping. The maxillary arch was isolated, fixed and processed for histological and epifluorescence analysis to examine reparative dentinogenesis.
Analysis of teeth immediately after pulp exposure revealed absence of odontoblasts expressing 3.6-GFP at the injury site. Evidence of reparative dentinogenesis was apparent at 4 weeks in 3.6-GFP mice in CD1 background and at 8 weeks in 3.6-GFP mice with C57/Bl6 background. The reparative dentine with both groups contained newly formed atubular-mineralized tissue resembling a dentine bridge and/or osteodentine that was lined by cells expressing 3.6-GFP as well as 3.6-GFP expressing cells embedded within the atubular matrix.
This study was conducted in a few animals and did not allow statistical analysis. The results revealed that the 3.6-GFP transgenic animals provide a unique model for direct analysis of cellular and molecular mechanisms of pulp repair and tertiary dentinogenesis in vivo. The study also shows the effects of the capping material and the genetic background of the mice in the sequence and timing of reparative dentinogenesis.
研究 pOBCol3.6GFPtpz[3.6-绿色荧光蛋白(GFP)]转基因小鼠是否可作为研究牙髓愈合和修复性牙本质形成过程中生物序列事件的体内模型。
用 CD1 和 C57/Bl6 遗传背景的 12-16 周龄 3.6-GFP 转基因小鼠建立第一上颌磨牙牙髓暴露模型。用矿化三氧化物凝聚体(mineral trioxide aggregate,MTA)直接覆盖暴露的牙齿,然后用光固化粘接系统(adhesive system,AS)和复合树脂进行修复。在对照牙中,AS 直接与牙髓接触。牙髓暴露和覆盖后,动物在不同时间点处死。分离上颌弓,固定,进行组织学和荧光分析,以研究修复性牙本质形成。
牙髓暴露后立即分析牙齿,发现损伤部位无表达 3.6-GFP 的成牙本质细胞。在 CD1 背景的 3.6-GFP 小鼠中,在 4 周时,在 C57/Bl6 背景的 3.6-GFP 小鼠中,在 8 周时,均可观察到修复性牙本质形成的证据。两组的修复性牙本质均含有新形成的无小管-矿化组织,类似于牙本质桥和/或骨样牙本质,由表达 3.6-GFP 的细胞以及嵌入无小管基质内的表达 3.6-GFP 的细胞所包绕。
本研究在少数动物中进行,未进行统计学分析。结果表明,3.6-GFP 转基因动物为体内直接分析牙髓修复和第三期牙本质形成的细胞和分子机制提供了独特的模型。该研究还表明,盖髓材料和小鼠的遗传背景对修复性牙本质形成的顺序和时间有影响。