背景:最近的研究表明,在再生牙髓治疗(RET)中用于消毒根管的肛门内抗微生物剂可能对根尖乳头(SCAP)的干细胞具有细胞毒性,导致治疗结果不一致。然而,尚未研究在亚致死浓度下肛门内抗微生物剂对SCAP牙源性分化能力的影响.这项研究的目的是使用临床相关浓度范围(0.1-0.8mg/mL)确定肛门内抗微生物剂对SCAP活力和牙源性分化能力的影响。
方法:从71例患者中收集未成熟的人类第三磨牙,并收获顶端乳头以形成单细胞悬浮液。肛门内抗微生物剂包括双抗生素糊剂(DAP)的细胞毒性作用,三重或改良三重抗生素糊剂(TAP或MTAP),在将细胞以0.1至0.8mg/mL暴露于处理组7天后,使用AlamarBlue和Live/Dead测定评估STRO-1+SCAP上的氢氧化钙(Ca(OH)2)。通过牙本质基质蛋白1和牙本质唾液酸磷蛋白表达的免疫细胞化学染色评估STRO-1SCAP的牙源性分化潜力。
结果:所有浓度的TAP都显着降低了STRO-1SCAP的活力和牙源性分化(P<.001),而DAP浓度无显著细胞毒性。Ca(OH)2和MTAP浓度低于0.4mg/mL和0.2mg/mL,分别,没有显着降低生存能力。DAP,MTAP,Ca(OH)2对STRO-1+SCAP成牙分化能力无明显影响。
结论:肛门内抗微生物剂对STRO-1+SCAP的体外影响不同,提示对当前根管消毒方案的修改可能会提高RET的成功率。
BACKGROUND: 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).
METHODS: 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)2) 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.
RESULTS: All concentrations of TAP significantly reduced STRO-1+ SCAP viability and odontogenic differentiation (P < .001), whereas no DAP concentrations were significantly cytotoxic. Ca(OH)2 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)2 did not significantly impact the
odontogenic differentiation capacity of STRO-1+ SCAP.
CONCLUSIONS: 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.