Soe Zar Chi, Nan Daneeya Na, Wahyudi Rahman, Vongsutilers Vorasit, Kamolratanakul Paksinee, Everts Vincent, Osathanon Thanaphum, Limjeerajarus Chalida Nakalekha, Limjeerajarus Nuttapol
Faculty of Dentistry, Graduate Program in Oral Biology, Chulalongkorn University, Bangkok, Thailand.
Center of Excellence in Regenerative Dentistry, Faculty of Dentistry, Chulalongkorn University, Bangkok, Thailand.
J Endod. 2025 Jul;51(7):931-938. doi: 10.1016/j.joen.2025.04.004. Epub 2025 Apr 15.
Asiaticoside (AS) demonstrated potential in wound healing and anti-inflammation. However, its therapeutic applications are limited due to poor solubility and low bioavailability, which make it difficult to use as dental pulp capping agent. The nanosponges (Ns) are nanosized carriers capable of carrying small drug molecules. We proposed a method to encapsulate AS in Ns (asiaticoside-loaded nanosponges) and incorporate it into hydrogel (asiaticoside-loaded nanosponges hydrogel [AS/Ns-gel]).
Using different concentrations of polymer and carbopol four different fractions of asiaticoside-loaded nanosponges (N1-N4) and AS/Ns-gel (G1-G4) were prepared. The optimal fraction was determined by characterizing physiochemical properties and in vitro release kinetics. An in vitro model of inflammatory human dental pulp cells (hDPCs) was induced using a cytokine cocktail and/or lipopolysaccharide prior to application of AS/Ns-gel. Messenger ribonucleic acid (mRNA) and protein expression of inflammatory cytokines were measured. To assess the wound healing potential of AS/Ns-gel, an in vitro scratch test was performed.
N1/G3 AS/Ns-gel exhibited the most optimized and uniform particle size distribution, with good solubility, sustained AS release, and effective encapsulation. In an in vitro study of hDPCs pretreated with lipopolysaccharide or cytokine cocktail, the AS/Ns-gel downregulated interleukin 6 and interleukin 8 mRNA expression while upregulating interleukin 10 expression. Reverse transcription quantitative polymerase chain reaction and western blot analysis revealed a time-dependent increase in transforming growth factor β1, collagen type 1 and matrix metalloproteinase 9 mRNA/protein levels. Additionally, AS/Ns-gel accelerated hDPCs migration.
We successfully developed an AS/Ns-gel that reduced the expression of inflammatory cytokines in an inflamed pulp model in vitro. AS-loaded hydrogels have sustained release properties, promote cell proliferation and cell migration, thus suggesting its potential to be used in regenerative endodontic therapy.
积雪草苷（AS）在伤口愈合和抗炎方面显示出潜力。然而，由于其溶解性差和生物利用度低，其治疗应用受到限制，这使得它难以用作牙髓盖髓剂。纳米海绵（Ns）是能够携带小药物分子的纳米级载体。我们提出了一种将AS包裹在Ns中（载积雪草苷纳米海绵）并将其掺入水凝胶中的方法（载积雪草苷纳米海绵水凝胶[AS/Ns-凝胶]）。
使用不同浓度的聚合物和卡波姆制备了四种不同级分的载积雪草苷纳米海绵（N1-N4）和AS/Ns-凝胶（G1-G4）。通过表征理化性质和体外释放动力学来确定最佳级分。在应用AS/Ns-凝胶之前，使用细胞因子混合物和/或脂多糖诱导炎症性人牙髓细胞（hDPCs）的体外模型。测量炎症细胞因子的信使核糖核酸（mRNA）和蛋白质表达。为了评估AS/Ns-凝胶的伤口愈合潜力，进行了体外划痕试验。
N1/G3 AS/Ns-凝胶表现出最优化且均匀的粒径分布，具有良好的溶解性、AS的持续释放和有效的包封。在对用脂多糖或细胞因子混合物预处理的hDPCs的体外研究中，AS/Ns-凝胶下调白细胞介素6和白细胞介素8的mRNA表达，同时上调白细胞介素10的表达。逆转录定量聚合酶链反应和蛋白质印迹分析显示转化生长因子β1、I型胶原和基质金属蛋白酶9的mRNA/蛋白质水平呈时间依赖性增加。此外，AS/Ns-凝胶加速了hDPCs的迁移。
我们成功开发了一种AS/Ns-凝胶，其在体外炎症牙髓模型中降低了炎症细胞因子的表达。载AS水凝胶具有持续释放特性，促进细胞增殖和细胞迁移，因此表明其在再生牙髓治疗中的应用潜力。
