Hertwig's epithelial root sheath cells show potential for periodontal complex regeneration
Hertwig's epithelial root sheath cells show potential for periodontal complex regeneration
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DOI：
10.1002/jper.22-0072
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发表时间：
2022-11-06
影响因子：
4.3
通讯作者：
Guo, Weihua
中科院分区：
文献类型：
--
作者：
Bi, Fei;Tang, Huilin;Guo, Weihua
背景虽然研究人员一直在探索治疗严重牙周组织缺损的治疗策略，包括应用干细胞，但涉及牙骨质、牙周组织和牙槽骨的牙周复合体的组织再生几乎没有实现。针对牙周复合体严重受损的问题，利用Hertwig's epithelial root sheath（HERS）细胞的牙骨质、骨和牙周组织形成能力，模拟生理发育过程进行组织再生是值得尝试的。方法从Sprague道利大鼠磨牙牙胚中分离培养HERS细胞和牙囊细胞，免疫荧光法鉴定。采用茜素红法、碱性磷酸酶（ALP）染色、碱性磷酸酶（AKP）检测、实时定量聚合酶链反应（RT-qPCR）和Western blot等方法检测HERS细胞的成骨能力、上皮-间充质转化（EMT）特征以及与DFC的上皮-间充质相互作用（EMI）。建立牙周缺损动物模型，验证其体内组织再生能力。显微CT和组织学检查进行解释，以揭示组织修复的结果。结果HERS细胞表达上皮细胞标志物CK 14和E-cadherin。体外实验结果表明，无论是HERS组还是HERS+DFC组，成骨分化能力都有明显的增强。同时，大鼠下颌骨牙周组织原位修复实验中，通过Micro-CT和组织学分析，证实了HERS组和HERS+DFC组牙周复合体结构原位再生的有效性。结论HERS细胞具有良好的牙周组织再生能力，有望成为牙周组织修复的理想细胞之一。
Background 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. Methods 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. Results 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. Conclusions 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.