Faculty of Dentistry, Department of Periodontology, Selcuk University, Konya, Turkey.
Department of Biotechnology, Middle East Technical University, Ankara, Turkey.
J Periodontol. 2024 May;95(5):456-468. doi: 10.1002/JPER.23-0246. Epub 2023 Oct 3.
Stem cell-based approaches in regenerative periodontal therapy have been used in different experimental models. In this study, the effect of local application of gingival mesenchymal stem cells (GMSC) in fibroin/chitosan oligosaccharide lactate hydrogel (F/COS) on periodontal regeneration was evaluated using experimental periodontitis model in rats.
Mesenchymal stem cells were isolated from the gingiva of rats and characterized. Viability tests and confocal imaging of GMSC in hydrogels were performed. Healthy control without periodontitis (Health; H; n=10), control with periodontitis but no application (Periodontitis; P; n=10), only hydrogel application (F/COS; n=10), and GMSC+F/COS (n=10) four groups were formed for in vivo studies. Experimental periodontitis was created with silk sutures around the maxillary second molars. GMSC labeled with green fluorescent protein (GFP) (250,000 cells/50 μL) in F/COS were applied to the defect. Animals were sacrificed at 2nd and 8th weeks and maxillae of the animals were evaluated by micro-computed tomography (micro-CT) and histologically. The presence of GFP-labeled GMSC was confirmed at the end of 8 weeks.
Micro-CT analysis showed statistically significant new bone formation in the F/COS+GMSC treated group compared with the P group at the end of 8 weeks (p < 0.05). New bone formation was also observed in the F/COS group, but the statistical analysis revealed that this difference was not significant when compared with the P group (p > 0.05). Long junctional epithelium formation was less in the F/COS+GMSC group compared with the P group. Periodontal ligament and connective tissue were well-organized in F/COS+GMSC group.
The results showed that local GMSC application in hydrogel contributed to the formation of new periodontal ligament and alveolar bone in rats with experimental periodontitis. Since gingiva is easly accessible tissue, it is promising for autologous cell-based treatments in clinical applications.
基于干细胞的再生牙周治疗方法已在不同的实验模型中得到应用。在这项研究中,使用大鼠实验性牙周炎模型评估了局部应用牙龈间充质干细胞(GMSC)在丝素/壳聚糖低聚糖乳酸水凝胶(F/COS)中对牙周再生的影响。
从大鼠牙龈中分离间充质干细胞并进行鉴定。进行了 GMSC 在水凝胶中的活力测试和共聚焦成像。未发生牙周炎的健康对照组(Health;H;n=10)、未应用但发生牙周炎的对照组(Periodontitis;P;n=10)、仅应用水凝胶组(F/COS;n=10)和 GMSC+F/COS 组(n=10)形成了用于体内研究的四组。用丝线环绕上颌第二磨牙制作实验性牙周炎。将绿色荧光蛋白(GFP)标记的 GMSC(250,000 个细胞/50 μL)加入 F/COS 中应用于缺损处。在第 2 周和第 8 周处死动物,并通过微计算机断层扫描(micro-CT)和组织学评估动物的上颌骨。在第 8 周结束时证实了 GFP 标记的 GMSC 的存在。
微 CT 分析显示,在第 8 周结束时,与 P 组相比,F/COS+GMSC 治疗组的新骨形成有统计学显著差异(p<0.05)。F/COS 组也观察到新骨形成,但统计学分析显示与 P 组相比,这一差异无统计学意义(p>0.05)。与 P 组相比,F/COS+GMSC 组的长结合上皮形成较少。F/COS+GMSC 组的牙周韧带和结缔组织组织排列良好。
结果表明,局部应用 GMSC 于水凝胶中有助于形成大鼠实验性牙周炎中新的牙周韧带和牙槽骨。由于牙龈是易于接近的组织,因此它有望在临床应用中用于自体细胞为基础的治疗。