Department of Restorative Dentistry, Division of Endodontics, Piracicaba Dental School, State University of Campinas, Piracicaba, São Paulo, Brazil.
Department of Morphology, Division of Histology and Embryology, Piracicaba Dental School, State University of Campinas, Piracicaba, São Paulo, Brazil.
J Endod. 2018 Mar;44(3):438-445. doi: 10.1016/j.joen.2017.10.006. Epub 2017 Dec 21.
This study aimed to evaluate the time required for bone marrow-derived cells (BMDCs) from transgenic green fluorescent protein (GFP)+ donor mice (GFP+ mice) to migrate into the dental pulp of wild-type GFP- recipient mice (GFP- mice) by using bone marrow transplantation (BMT) as an in vivo model for tracking BMDCs from GFP+ mice (GFP+ BMDCs).
GFP+ BMDCs were injected into irradiated GFP- mice. Maxillary arches, tibiae, and femora from GFP- mice were isolated and processed at 24 hours, 48 hours, 4, 7, and 14 days, and 7 weeks after BMT. Confocal laser microscopy analyses were performed to assess the presence of GFP+ BMDCs in the dental pulp, and flow cytometry of BM was performed to confirm the efficiency of engraftment of GFP+ BMDCs.
Confocal laser microscopy analyses evidenced the presence of GFP+ BMDCs in the dental pulp of GFP- mice from 14 days to 7 weeks after BMT. There was no presence of GFP+ BMDCs at 24 hours, 48 hours, 4 days, and 7 days. Flow cytometry of the BM of GFP- mice demonstrated a constant increase in the presence of GFP+ BMDCs at 24 hours, 48 hours, and 4 days after BMT, which stabilized from 7 days to 7 weeks.
The study demonstrated the presence of GFP+ BMDCs in the dental pulp from 14 days to 7 weeks after BMT and the feasibility of using GFP+ animals and BMT as an in vivo model for tracking GFP+ BMDCs.
本研究旨在通过骨髓移植（BMT）作为追踪 GFP+ 供体鼠（GFP+ 鼠）骨髓源性细胞（BMDCs）的体内模型，评估 GFP+ 鼠 BMDC 迁移到野生型 GFP- 受体鼠（GFP- 鼠）牙髓所需的时间。
将 GFP+ BMDC 注射到照射后的 GFP- 鼠中。在 BMT 后 24 小时、48 小时、4 天、7 天和 14 天以及 7 周时，分离并处理 GFP- 鼠的上颌弓、胫骨和股骨。进行共聚焦激光显微镜分析以评估 GFP+ BMDC 是否存在于牙髓中，并进行 BM 的流式细胞术以确认 GFP+ BMDC 的植入效率。
共聚焦激光显微镜分析表明，在 BMT 后 14 天至 7 周，GFP- 鼠的牙髓中存在 GFP+ BMDCs。在 24 小时、48 小时、4 天和 7 天均未发现 GFP+ BMDCs。GFP- 鼠 BM 的流式细胞术表明，在 BMT 后 24 小时、48 小时和 4 天，GFP+ BMDCs 的存在持续增加，从 7 天到 7 周稳定。
本研究表明，在 BMT 后 14 天至 7 周，GFP+ BMDC 存在于牙髓中，并且使用 GFP+ 动物和 BMT 作为追踪 GFP+ BMDC 的体内模型是可行的。
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