Elshahat Sara, Elgendy Abeer Abdelhakim, Elsewify Tarek
Endodontic Department, Faculty of Dentistry, Ain Shams University, Cairo, Egypt.
Restorative Dental Sciences Department, College of Dentistry, Gulf Medical University, Ajman, UAE.
Eur J Dent. 2024 May;18(2):665-671. doi: 10.1055/s-0043-1777044. Epub 2024 Mar 5.
The aim of this study was to evaluate the osteogenic differentiation ability and proliferation of apical papilla stem cells (SCAPs) using chitosan-coated nanohydroxyapatite and bioactive glass nanoparticles.
Hydroxyapatite, chitosan-coated nanohydroxyapatite, and bioactive glass 45S5 nanoparticles were prepared and characterized using a transmission electron microscope and X-ray diffraction. SCAPs were harvested from freshly extracted impacted wisdom teeth, cultured, and characterized using flow cytometric analysis. Tested nanomaterials were mixed and samples were classified into five equal groups as follows: negative control group: SCAP with Dulbecco's modified eagle's medium, positive control group: SCAP with inductive media, first experimental group: nanohydroxyapatite with SCAP, second experimental group: chitosan-coated nanohydroxyapatite with SCAP, third experimental group: bioactive glass nanoparticles with SCAP. Osteoblastic differentiation was assessed using an alkaline phosphatase (ALP) assay. Receptor activator of nuclear factor kappa beta ligand (RANKL) expression was evaluated using specific polyclonal antibodies by fluorescence microscope. The proliferation of SCAP was assessed using cell count and viability of trypan blue in addition to an 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay.
Isolated SCAP showed a nonhematopoietic origin. Chitosan-coated nanohydroxyapatite showed the highest ALP concentration followed by nanobioactive glass, nanohydroxyapatite, and negative control. Chitosan-coated nanohydroxyapatite showed the highest H score followed by nanobioactive glass, nanohydroxyapatite, and negative control in RANKL expression. Chitosan-coated nanohydroxyapatite showed the highest viable cell count.
SCAP isolation is achievable from extracted fully impacted immature third molars. All tested biomaterials have the ability to induce osteogenic differentiation and proliferation of SCAP. Composite nanoparticle materials show better osteogenic differentiation and proliferation of SCAP than single nanoparticles.
本研究旨在评估使用壳聚糖包覆的纳米羟基磷灰石和生物活性玻璃纳米颗粒对根尖乳头干细胞（SCAPs）的成骨分化能力和增殖情况。
制备羟基磷灰石、壳聚糖包覆的纳米羟基磷灰石和生物活性玻璃45S5纳米颗粒，并使用透射电子显微镜和X射线衍射进行表征。从新鲜拔除的阻生智齿中获取SCAPs，进行培养，并使用流式细胞术分析进行表征。将测试的纳米材料混合，样本分为五个相等的组，如下所示：阴性对照组：SCAP与杜氏改良伊格尔培养基；阳性对照组：SCAP与诱导培养基；第一实验组：纳米羟基磷灰石与SCAP；第二实验组：壳聚糖包覆的纳米羟基磷灰石与SCAP；第三实验组：生物活性玻璃纳米颗粒与SCAP。使用碱性磷酸酶（ALP）测定法评估成骨细胞分化。通过荧光显微镜使用特异性多克隆抗体评估核因子κB受体激活剂配体（RANKL）的表达。除了3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐（MTT）测定法外，还使用细胞计数和台盼蓝活力评估SCAP的增殖。
分离出的SCAP显示非造血来源。壳聚糖包覆的纳米羟基磷灰石显示出最高的ALP浓度，其次是纳米生物活性玻璃、纳米羟基磷灰石和阴性对照。在RANKL表达方面，壳聚糖包覆的纳米羟基磷灰石显示出最高的H评分，其次是纳米生物活性玻璃、纳米羟基磷灰石和阴性对照。壳聚糖包覆的纳米羟基磷灰石显示出最高的活细胞计数。
从拔除的完全阻生的未成熟第三磨牙中可实现SCAP的分离。所有测试的生物材料都有能力诱导SCAP的成骨分化和增殖。复合纳米颗粒材料比单一纳米颗粒显示出更好的SCAP成骨分化和增殖能力。
