Amir Lisa R, Soeroso Yuniarti, Fatma Dewi, Sunarto Hari, Sulijaya Benso, Idrus Erik, Rahdewati Herlis, Tjokrovonco Angelia M, Izumi Kenji, Abbas Basril, Latief Fourier D E
Department of Oral Biology, Faculty of Dentistry, Universitas Indonesia, Jakarta, Indonesia.
Department of Periodontology, Faculty of Dentistry, Universitas Indonesia, Jakarta, Indonesia.
Eur J Dent. 2020 Mar;14(2):306-314. doi: 10.1055/s-0040-1709955. Epub 2020 May 12.
The aim of this study was to examine the potential of periodontal ligament (PDL) cells sheet and arginine-glycyl-aspartic acid (RGD)-modified chitosan scaffold for periodontal tissue regeneration in horizontal periodontal defect model.
PDL cell cytotoxicity was tested with 3-[4,5- dimethylthiazol-2yl]-2,5-diphenyl-2H-tetrazolium bromide assay. Cell migration toward the chitosan-based materials was analyzed with trans-well migration assay. Horizontal periodontal defect model was created in four maxillary and mandibular lateral incisors of . Following periodontal therapy, the sites were transplanted with various regenerative materials: (1) chitosan, (2) RGD-modified chitosan, (3) PDL cell sheet with chitosan, (4) PDL cell sheet with RGD-modified chitosan. The periodontal tissue regeneration was evaluated clinically and radiographically. Gingival crevicular fluids were collected each week to evaluate cementum protein-1 (CEMP-1) expression with enzyme-linked immunosorbent assay, while the biopsies were retrieved after 4 weeks for histological and microcomputed tomography evaluation.
Data was statistically analyzed using GraphPad Prism 6 for MacOS X. Normality was tested using the Shapiro-Wilk normality test. The Kruskal-Wallis test was used to compare the groups. Significance was accepted when < 0.05.
Clinical examination revealed more epithelial attachment was formed in the group with PDL cell sheet with RGD-modified chitosan. Similarly, digital subtraction radiography analysis showed higher gray scale, an indication of higher alveolar bone density surrounded the transplanted area, as well as higher CEMP-1 protein expression in this group. The incorporation of RGD peptide to chitosan scaffold in the group with or without PDL cells sheet reduced the distance of cement-enamel junction to the alveolar bone crest; hence, more periodontal tissue formed.
Horizontal periodontal defect model could be successfully created in model. Combination of PDL cell sheet and RGD-modified chitosan resulted in the higher potential for periodontal tissue regeneration. The results of this study highlight the PDL cell sheet and RGD-modified chitosan as a promising approach for future clinical use in periodontal regeneration.
本研究旨在探讨牙周膜（PDL）细胞片和精氨酸 - 甘氨酸 - 天冬氨酸（RGD）修饰的壳聚糖支架在水平型牙周缺损模型中促进牙周组织再生的潜力。
采用3 - [4,5 - 二甲基噻唑 - 2 - 基] - 2,5 - 二苯基 - 2H - 四氮唑溴盐法检测PDL细胞的细胞毒性。通过Transwell迁移实验分析细胞向壳聚糖基材料的迁移情况。在 的四颗上颌和下颌侧切牙上建立水平型牙周缺损模型。牙周治疗后，将不同的再生材料移植到这些部位：（1）壳聚糖，（2）RGD修饰的壳聚糖，（3）含壳聚糖的PDL细胞片，（4）含RGD修饰壳聚糖的PDL细胞片。从临床和影像学方面评估牙周组织的再生情况。每周收集龈沟液，用酶联免疫吸附测定法评估牙骨质蛋白 - 1（CEMP - 1）的表达，4周后取组织活检进行组织学和显微计算机断层扫描评估。
使用适用于MacOS X的GraphPad Prism 6对数据进行统计分析。使用Shapiro - Wilk正态性检验来检验数据的正态性。采用Kruskal - Wallis检验对各组进行比较。当P < 0.05时认为具有统计学意义。
临床检查显示，含RGD修饰壳聚糖的PDL细胞片组形成了更多的上皮附着。同样，数字减影放射摄影分析显示该组灰度更高，表明移植区域周围牙槽骨密度更高，且CEMP - 1蛋白表达也更高。在有或无PDL细胞片的组中，将RGD肽掺入壳聚糖支架均减小了牙骨质 - 釉质界到牙槽嵴顶的距离；因此，形成了更多的牙周组织。
在 模型中可成功建立水平型牙周缺损模型。PDL细胞片和RGD修饰的壳聚糖联合使用具有更高的牙周组织再生潜力。本研究结果突出了PDL细胞片和RGD修饰的壳聚糖作为未来牙周再生临床应用的一种有前景的方法。
