Abstract：
BACKGROUND: The dental pulp\'s environment is essential for the regulation of mesenchymal stem cells\' homeostasis and thus, it is of great importance to evaluate the materials used in regenerative procedures.
OBJECTIVE: To assess in vitro (i) the effect of chitosan nanoparticles, 0.2% chitosan irrigation solution, Dual Rinse®, 17% EDTA, 10% citric acid and 2.5% NaOCl on DSCS viability; (ii) the effect of different concentrations of TGF-β1 on DCSC proliferation; and (iii) whether treatment with TGF-β1 following exposure to the different irrigation solutions could compensate for their negative effects.
METHODS: (i) DSCS were treated with three dilutions (1:10, 1:100 and 1:1000) of the six irrigation solutions prepared in DMEM for 10 and 60 min to assess the effect on viability. (ii) The effect of different concentrations (0, 1, 5 and 10 ng/mL) of TGF-β1 on DCSC proliferation was assessed at 1, 3 and 7 days. (iii) The proliferative effect of TGF-β1 following 10-min exposure to 1:10 dilution of each irrigation solution was also tested. We used MTT assay to assess viability and proliferation. We performed statistical analysis using Prism software.
RESULTS: (i) The different endodontic irrigation solutions tested showed a significant effect on cell viability (p ≤ .0001). Significant interactions between the endodontic irrigation solutions and their dilutions were also found for all parameters (p ≤ .0001). Chitosan nanoparticles and 0.2% chitosan irrigation solution were the least cytotoxic to DSCS whilst 2.5% NaOCl was the most cytotoxic followed by 17% EDTA. (ii) TGF-β1 at concentrations of 1 and 5 ng/mL resulted in significantly higher proliferation compared to the control group. (iii) Exposure to 17% EDTA or 2.5% NaOCl for 10 min was sufficient to make DSCS cells refractory to the proliferative effects of TGF-β1. DSCS groups treated with TGF-β1 following exposure to chitosan nanoparticles, 0.2% chitosan irrigation solution, Dual Rinse® and 10% CA demonstrated significantly higher proliferation compared to non-TGF-β1-treated groups (p ≤ .0001, p ≤ .0001, p ≤ .0001 and p = .01 respectively).
CONCLUSIONS: The current study offers data that can be implemented to improve the outcome of regenerative endodontic procedures by using less toxic irrigation solutions and adding TGF-β1 to the treatment protocol.
OBJECTIVE: To assess in vitro (i) the effect of chitosan nanoparticles, 0.2% chitosan irrigation solution, Dual Rinse®, 17% EDTA, 10% citric acid and 2.5% NaOCl on DSCS viability; (ii) the effect of different concentrations of TGF-β1 on DCSC proliferation; and (iii) whether treatment with TGF-β1 following exposure to the different irrigation solutions could compensate for their negative effects.
METHODS: (i) DSCS were treated with three dilutions (1:10, 1:100 and 1:1000) of the six irrigation solutions prepared in DMEM for 10 and 60 min to assess the effect on viability. (ii) The effect of different concentrations (0, 1, 5 and 10 ng/mL) of TGF-β1 on DCSC proliferation was assessed at 1, 3 and 7 days. (iii) The proliferative effect of TGF-β1 following 10-min exposure to 1:10 dilution of each irrigation solution was also tested. We used MTT assay to assess viability and proliferation. We performed statistical analysis using Prism software.
RESULTS: (i) The different endodontic irrigation solutions tested showed a significant effect on cell viability (p ≤ .0001). Significant interactions between the endodontic irrigation solutions and their dilutions were also found for all parameters (p ≤ .0001). Chitosan nanoparticles and 0.2% chitosan irrigation solution were the least cytotoxic to DSCS whilst 2.5% NaOCl was the most cytotoxic followed by 17% EDTA. (ii) TGF-β1 at concentrations of 1 and 5 ng/mL resulted in significantly higher proliferation compared to the control group. (iii) Exposure to 17% EDTA or 2.5% NaOCl for 10 min was sufficient to make DSCS cells refractory to the proliferative effects of TGF-β1. DSCS groups treated with TGF-β1 following exposure to chitosan nanoparticles, 0.2% chitosan irrigation solution, Dual Rinse® and 10% CA demonstrated significantly higher proliferation compared to non-TGF-β1-treated groups (p ≤ .0001, p ≤ .0001, p ≤ .0001 and p = .01 respectively).
CONCLUSIONS: The current study offers data that can be implemented to improve the outcome of regenerative endodontic procedures by using less toxic irrigation solutions and adding TGF-β1 to the treatment protocol.
摘要：
背景：牙髓的环境对于调节间充质干细胞的体内平衡至关重要，因此，评估再生程序中使用的材料非常重要。
目的：为了评估体外（i）壳聚糖纳米粒的作用，0.2%壳聚糖灌洗液，双冲洗®,17%EDTA,10%柠檬酸和2.5%NaOCl对DSCS活力的影响;(ii)不同浓度的TGF-β1对DCSC增殖的影响;和(iii)在暴露于不同冲洗溶液之后用TGF-β1处理是否可以补偿其负面影响。
方法:(i)将DSCS用在DMEM中制备的六种冲洗溶液的三种稀释液(1:10、1:100和1:1000)处理10和60分钟以评估对生存力的影响。（ii）在第1、3和7天评估了不同浓度（0、1、5和10ng/mL）的TGF-β1对DCSC增殖的影响。(iii)还测试了在暴露于每种冲洗溶液的1:10稀释10分钟后TGF-β1的增殖作用。我们使用MTT测定来评估活力和增殖。我们使用Prism软件进行统计分析。
结果：（i）测试的不同牙髓冲洗溶液对细胞活力具有显着影响（p≤.0001）。对于所有参数（p≤0.0001），还发现了牙髓冲洗溶液及其稀释液之间的显着相互作用。壳聚糖纳米颗粒和0.2％壳聚糖冲洗溶液对DSCS的细胞毒性最小，而2.5％NaOCl的细胞毒性最大，其次是17％EDTA。(ii)与对照组相比,浓度为1和5ng/mL的TGF-β1导致显著更高的增殖。(iii)暴露于17%EDTA或2.5%NaOCl10分钟足以使DSCS细胞对TGF-β1的增殖作用难以反应。暴露于壳聚糖纳米颗粒后，用TGF-β1处理的DSCS组，0.2%壳聚糖灌洗液，与非TGF-β1治疗组相比，双冲洗®和10％CA显示出明显更高的增殖（分别为p≤0.0001，p≤0.0001，p≤0.0001和p=0.01）。
结论：本研究提供的数据可以通过使用毒性较小的冲洗溶液和在治疗方案中添加TGF-β1来改善再生牙髓手术的结果。
目的：为了评估体外（i）壳聚糖纳米粒的作用，0.2%壳聚糖灌洗液，双冲洗®,17%EDTA,10%柠檬酸和2.5%NaOCl对DSCS活力的影响;(ii)不同浓度的TGF-β1对DCSC增殖的影响;和(iii)在暴露于不同冲洗溶液之后用TGF-β1处理是否可以补偿其负面影响。
方法:(i)将DSCS用在DMEM中制备的六种冲洗溶液的三种稀释液(1:10、1:100和1:1000)处理10和60分钟以评估对生存力的影响。（ii）在第1、3和7天评估了不同浓度（0、1、5和10ng/mL）的TGF-β1对DCSC增殖的影响。(iii)还测试了在暴露于每种冲洗溶液的1:10稀释10分钟后TGF-β1的增殖作用。我们使用MTT测定来评估活力和增殖。我们使用Prism软件进行统计分析。
结果：（i）测试的不同牙髓冲洗溶液对细胞活力具有显着影响（p≤.0001）。对于所有参数（p≤0.0001），还发现了牙髓冲洗溶液及其稀释液之间的显着相互作用。壳聚糖纳米颗粒和0.2％壳聚糖冲洗溶液对DSCS的细胞毒性最小，而2.5％NaOCl的细胞毒性最大，其次是17％EDTA。(ii)与对照组相比,浓度为1和5ng/mL的TGF-β1导致显著更高的增殖。(iii)暴露于17%EDTA或2.5%NaOCl10分钟足以使DSCS细胞对TGF-β1的增殖作用难以反应。暴露于壳聚糖纳米颗粒后，用TGF-β1处理的DSCS组，0.2%壳聚糖灌洗液，与非TGF-β1治疗组相比，双冲洗®和10％CA显示出明显更高的增殖（分别为p≤0.0001，p≤0.0001，p≤0.0001和p=0.01）。
结论：本研究提供的数据可以通过使用毒性较小的冲洗溶液和在治疗方案中添加TGF-β1来改善再生牙髓手术的结果。
