Bi Fei, Tang Huilin, Zhang Zhijun, Lyu Yun, Huo Fangjun, Chen Guoqing, Guo Weihua
Engineering Research Center of Oral Translational Medicine, Ministry of Education, West China Hospital of Stomatology, Sichuan University, Chengdu, China.
National Engineering Laboratory for Oral Regenerative Medicine, West China Hospital of Stomatology, Sichuan University, Chengdu, China.
J Periodontol. 2023 Feb;94(2):263-276. doi: 10.1002/JPER.22-0072. Epub 2022 Nov 6.
Although researchers have been exploring therapeutic strategies of treating serious periodontal tissue loss, including the application of stem cells, tissue regeneration of the periodontal complex involving cementum, periodontium, and alveolar bone has hardly been achieved. Aiming at tackling the problem of severely damaged periodontal complex, it is worth trying to make advantages of Hertwig's epithelial root sheath (HERS) cells to tissue regeneration mimicking the physiological developmental process with their ability of cementum, bone, and periodontium formation.
HERS cells and dental follicle cells (DFCs) were acquired from Sprague Dawley rats' molar germs and identified by immunofluorescence. Alizarin red assay, ALP staining, AKP test, real-time quantitative polymerase chain reaction (RT-qPCR) and Western blot were conducted to confirm the osteogenic potential, epithelial-mesenchymal transition (EMT) character of harvested HERS cells and epithelial-mesenchymal interaction (EMI) with DFCs. An animal model of periodontal defect was constructed to testify the tissue regeneration ability in vivo. Micro-CT and histological examinations were interpreted to unveil the tissue repair outcomes.
HERS cells expressed strong epithelial cell markers CK14 and E-cadherin. The in vitro experiments overall showed the concretely enhanced osteogenic differentiation ability in either HERS group or HERS+DFC group. Meanwhile, the in vivo conduction of rat mandibular periodontal repair experiment showed regenerative effectiveness of periodontal complex structure in both HERS and HERS+DFC group in situ, testified by Micro-CT and histological analysis.
HERS cells show potential for periodontal tissue regeneration which suggests the future possibilities of being considered as one of the cell choices for severely damaged periodontal tissue repair.
尽管研究人员一直在探索治疗严重牙周组织丧失的治疗策略,包括干细胞的应用,但涉及牙骨质、牙周膜和牙槽骨的牙周复合体的组织再生却很难实现。为了解决严重受损的牙周复合体问题,利用赫特维希上皮根鞘(HERS)细胞模拟生理发育过程进行组织再生,发挥其形成牙骨质、骨和牙周膜的能力,值得一试。
从Sprague Dawley大鼠的磨牙胚中获取HERS细胞和牙囊细胞(DFCs),并通过免疫荧光进行鉴定。进行茜素红测定、碱性磷酸酶(ALP)染色、碱性磷酸酶(AKP)测试、实时定量聚合酶链反应(RT-qPCR)和蛋白质免疫印迹法,以确认收获的HERS细胞的成骨潜能、上皮-间质转化(EMT)特性以及与DFCs的上皮-间质相互作用(EMI)。构建牙周缺损动物模型,以验证体内组织再生能力。通过显微CT和组织学检查来揭示组织修复结果。
HERS细胞表达强烈的上皮细胞标志物细胞角蛋白14(CK14)和E-钙黏蛋白。体外实验总体显示,HERS组和HERS+DFC组的成骨分化能力均有具体增强。同时,大鼠下颌牙周修复实验的体内传导显示,HERS组和HERS+DFC组原位牙周复合体结构均有再生效果,显微CT和组织学分析证实了这一点。
HERS细胞显示出牙周组织再生的潜力,这表明其未来有可能被视为严重受损牙周组织修复的细胞选择之一。