Department of Operative Dentistry and Endodontics, Faculty of Dentistry, Mahidol University, 6 Yothi Road, Ratchathewi, Bangkok, Thailand.
Dental Biomaterial Analysis and Research Center, Faculty of Dentistry, Mahidol University, Bangkok, Thailand.
Clin Oral Investig. 2023 May;27(5):1973-1980. doi: 10.1007/s00784-023-04919-1. Epub 2023 Feb 15.
To evaluate the effect of EDTA and saline as the final irrigation in regenerative endodontic procedures (REPS) on the attachment, proliferation, migration, and differentiation of stem cells from the apical papilla (SCAPs).
Dentin specimens from 140 human third molars were irrigated with various protocols-group 1: normal sterile saline (NSS), group 2: EDTA, group 3: EDTA then 5 mL NSS, or group 4: EDTA then 20 mL NSS. The specimens were used in cell assays. For cell proliferation, SCAPs were seeded on dentin, and the cell viability on days 1, 3, and 7 was determined using an MTT assay. At day 3, the attached cells' morphology was observed using SEM, and cell migration was investigated using a transwell migration assay. The ALP activity and odonto/osteogenic differentiation gene expression were evaluated at days 7, 14, and 21 using an ALP activity assay and RT-qPCR.
On days 3 and 7, group 4 demonstrated more viable cells than group 1 (p < 0.01). The amount of migrated cells in groups 2, 3, and 4 was greater compared with group 1 (p < 0.05). Moreover, SCAP differentiation was similar between groups.
Irrigating dentin with EDTA alone or with EDTA then NSS promoted SCAP migration. However, a final irrigation with 20 mL NSS after EDTA promoted SCAP proliferation without affecting their differentiation.
When using a blood clot as a scaffold, a final flushing with 20 mL NSS after EDTA could be beneficial for clinical REP protocols.
评估 EDTA 和生理盐水作为再生牙髓治疗（REPS）最终冲洗液对根尖乳头干细胞（SCAPs）黏附、增殖、迁移和分化的影响。
从 140 颗人类第三磨牙的牙本质标本中用不同方案冲洗：第 1 组：正常无菌生理盐水（NSS），第 2 组：EDTA，第 3 组：EDTA 后用 5 mL NSS，或第 4 组：EDTA 后用 20 mL NSS。这些标本用于细胞实验。对于细胞增殖，将 SCAP 接种在牙本质上，使用 MTT 测定法在第 1、3 和 7 天测定细胞活力。在第 3 天，使用 SEM 观察附着细胞的形态，使用 Transwell 迁移实验研究细胞迁移。使用 ALP 活性测定法和 RT-qPCR 在第 7、14 和 21 天评估 ALP 活性和牙/骨向分化基因表达。
第 3 天和第 7 天，第 4 组的活细胞数多于第 1 组（p<0.01）。与第 1 组相比，第 2 组、第 3 组和第 4 组的迁移细胞数量更多（p<0.05）。此外，各组间 SCAP 分化情况相似。
EDTA 单独冲洗或 EDTA 后冲洗 NSS 均可促进 SCAP 迁移。然而，EDTA 后用 20 mL NSS 冲洗最终可促进 SCAP 增殖而不影响其分化。
在使用血凝块作为支架时，EDTA 后用 20 mL NSS 冲洗最终可能有益于临床 REP 方案。
