Department of Dental Clinical Sciences, Faculty of Dentistry, Dalhousie University, Halifax, Nova Scotia, Canada.
Department of Dental Clinical Sciences, Faculty of Dentistry, Dalhousie University, Halifax, Nova Scotia, Canada.
J Endod. 2022 Jul;48(7):880-886. doi: 10.1016/j.joen.2022.04.003. Epub 2022 Apr 9.
Recent studies have indicated that intracanal antimicrobials used to disinfect the root canal in regenerative endodontic therapies (RETs) may be cytotoxic to stem cells from the apical papilla (SCAP), leading to inconsistent treatment outcomes. However, the effects of intracanal antimicrobial agents on the odontogenic differentiation capacity of SCAP at sub-lethal concentrations have not been investigated. The aim of this study was to determine the effects of intracanal antimicrobials on SCAP viability and odontogenic differentiation capacity using a clinically relevant concentration range (0.1-0.8 mg/mL).
Immature human third molars were collected from 71 patients and the apical papillae were harvested to form single-cell suspensions. The cytotoxic effects of intracanal antimicrobials including double antibiotic paste (DAP), triple or modified-triple antibiotic paste (TAP or MTAP), and calcium hydroxide (Ca(OH)) on STRO-1 SCAP were assessed using AlamarBlue and Live/Dead assays after exposing cells to treatment groups for 7 days at 0.1 to 0.8 mg/mL. The odontogenic differentiation potential of STRO-1 SCAP was evaluated by immunocytochemistry staining of dentin matrix protein-1 and dentin sialophosphoprotein expression.
All concentrations of TAP significantly reduced STRO-1 SCAP viability and odontogenic differentiation (P < .001), whereas no DAP concentrations were significantly cytotoxic. Ca(OH) and MTAP concentrations below 0.4 mg/mL and 0.2 mg/mL, respectively, did not significantly reduce viability. The DAP, MTAP, and Ca(OH) did not significantly impact the odontogenic differentiation capacity of STRO-1 SCAP.
The varying effects of intracanal antimicrobials on STRO-1 SCAP in vitro suggest amendments to the current root canal disinfection protocol may improve the success of RETs.
最近的研究表明，再生牙髓治疗（RETs）中用于根管消毒的根管内抗菌剂可能对根尖乳头干细胞（SCAP）具有细胞毒性，导致治疗结果不一致。然而，亚致死浓度的根管内抗菌剂对 SCAP 牙源性分化能力的影响尚未得到研究。本研究旨在使用临床相关浓度范围（0.1-0.8mg/ml）确定根管内抗菌剂对 SCAP 活力和牙源性分化能力的影响。
从 71 名患者中采集未成熟的第三磨牙，收获根尖乳头形成单细胞悬液。用 AlamarBlue 和 Live/Dead 测定法在 0.1 至 0.8mg/ml 的治疗组中暴露细胞 7 天后，评估根管内抗菌剂包括双抗生素糊剂（DAP）、三或改良三抗生素糊剂（TAP 或 MTAP）和氢氧化钙（Ca（OH））对 STRO-1 SCAP 的细胞毒性作用。通过牙本质基质蛋白-1和牙本质涎磷蛋白表达的免疫细胞化学染色评估 STRO-1 SCAP 的牙源性分化潜能。
所有浓度的 TAP 均显著降低 STRO-1 SCAP 的活力和牙源性分化（P<0.001），而无 DAP 浓度具有显著细胞毒性。Ca（OH）和 MTAP 浓度分别低于 0.4mg/ml 和 0.2mg/ml 时，活力无明显降低。DAP、MTAP 和 Ca（OH）对 STRO-1 SCAP 的牙源性分化能力无显著影响。
根管内抗菌剂对 STRO-1 SCAP 的体外作用不同，表明对当前根管消毒方案进行修改可能会提高 RETs 的成功率。
