Li Weiguang, Tian Weidong, Wu Yafei, Guo Shujuan
Department of Periodontics, West China Hospital of Stomatology, Sichuan University, Chengdu, P.R. China.
State Key Laboratory of Oral Diseases, West China Hospital of Stomatology, Sichuan University, Chengdu, P.R. China.
Tissue Eng Part A. 2023 Dec;29(23-24):620-632. doi: 10.1089/ten.TEA.2023.0112. Epub 2023 Sep 8.
Periodontium is the rally of soft and hard tissues, which will be devastated continuously by the compromise of periodontitis. Current periodontal therapeutic methods cannot effectively reconstruct periodontal ligament (PDL), which is oriented at an angle with tooth root and combined hard tissues to form cementum-PDL-alveolar bone complex. Hence, it is urgent to find new techniques for PDL reconstruction to achieve functional regeneration of periodontium. Herein, we developed a novel method to manipulate the distribution and growth of periodontal ligament stem cells (PDLSCs) by utilizing highly paralleled static magnetic field (SMF) and magnetic nanoparticles (MNPs). PDLSCs were incubated with MNPs to label with them. Meanwhile, CCK8 and live/dead cell staining assay were used to detect the impact of SMF and MNPs on cell viability. The directional migration and growth of PDLSCs were visualized under microscope. Furthermore, real-time quantitative PCR and western blot were utilized to calculate the expression level of PDL-related genes. The results showed that PDLSCs could rapidly take up MNPs without compromising cell proliferation and viability, consequently endowed with the ability to respond via magnetic force. The cell migration analysis indicated that PDLSCs could move along the magnetic induction line, testifying that SMF exerted forces on PDLSCs that labeled with MNPs. It was demonstrated that collective application of SMF and MNPs not only induced PDLSCs organized and grew directionally, but also initiated elongation of cells and nucleus. Furthermore, the morphological alteration of the nucleus could also effectively enhance the gene and protein expression of Collagen Ⅰα2, Collagen Ⅲ, and Periostin, suggesting the capability of PDLSCs to differentiate into PDL. In conclusion, this study exhibits a new approach for directional reconstruction of PDL to obtain physiological and functional regeneration of periodontium. The Clinical Trial Registration number: WCHSIRB-D-2022-458.
牙周组织是软硬组织的集合,会因牙周炎的损害而持续遭到破坏。目前的牙周治疗方法无法有效重建牙周韧带(PDL),牙周韧带与牙根呈一定角度排列,并与硬组织结合形成牙骨质 - 牙周韧带 - 牙槽骨复合体。因此,迫切需要找到新的牙周韧带重建技术,以实现牙周组织的功能再生。在此,我们开发了一种新方法,通过利用高度平行的静磁场(SMF)和磁性纳米颗粒(MNPs)来操控牙周韧带干细胞(PDLSCs)的分布和生长。将PDLSCs与MNPs孵育以对其进行标记。同时,使用CCK8和活/死细胞染色试验来检测SMF和MNPs对细胞活力的影响。在显微镜下观察PDLSCs的定向迁移和生长。此外,利用实时定量PCR和蛋白质印迹法来计算牙周韧带相关基因的表达水平。结果表明,PDLSCs能够快速摄取MNPs,而不影响细胞增殖和活力,从而赋予其通过磁力做出反应的能力。细胞迁移分析表明,PDLSCs可以沿着磁感应线移动,证明SMF对标记有MNPs的PDLSCs施加了力。结果表明,SMF和MNPs的联合应用不仅诱导PDLSCs定向组织和生长,还引发细胞和细胞核的伸长。此外,细胞核的形态改变还能有效增强Ⅰα2型胶原、Ⅲ型胶原和骨膜蛋白的基因和蛋白表达,表明PDLSCs具有分化为牙周韧带的能力。总之,本研究展示了一种用于牙周韧带定向重建以实现牙周组织生理和功能再生的新方法。临床试验注册号:WCHSIRB-D-2022-458。