Zhai Yue, Wang Yuanyuan, Rao Nanquan, Li Jingzhi, Li Xiaoxia, Fang Tengjiaozi, Zhao Yuming, Ge Lihong
Department of Pediatric Dentistry, Peking University School and Hospital of Stomatology, Beijing, China.
Front Physiol. 2019 Oct 22;10:1304. doi: 10.3389/fphys.2019.01304. eCollection 2019.
Pulpitis in primary teeth, a condition caused by presence of bacteria, is highly prevalent worldwide. The use of biocompatibility materials with anti-inflammatory, anti-bacterial, and regenerative properties is critical for prognosis of this endodontic disease. This study aimed to identify expression of human β defensin 4 (HBD4) in stem cells derived from human exfoliated deciduous teeth (SHED) and characterize the effects of HBD4 on SHED. Quantitative polymerase chain reaction (qPCR) was used to detect HBD4 expression in SHED and the effect of HBD4 on inflammatory factors in lipopolysaccharide (LPS)-stimulated SHED. Affinity measurement was made by the Fortebio Octet System to explore the potential interaction between LPS and HBD4. Western blot analysis was used to explore the effect of HBD4 on mitogen-activated protein kinase (MAPK) pathway. Colony-forming unit methods and scanning electron microscopy were applied to study antimicrobial effect of HBD4 on and . Alkaline phosphatase staining, alizarin red staining, qPCR and western blot were taken to detect effects of HBD4 on osteoblast/odontoblast differentiation of SHED. RT Profiler PCR Array was used to explore the potential signaling pathways involved in the osteogenic/odontogenic differentiation. HBD4 was highly expressed in SHED stimulated by TNF-α and IL-1α. HBD4 could bind to LPS directly and down-regulate IL-1α, IL-1β, IL-6, TNF-α in LPS-stimulated SHED, thus the activation of MAPK pathway decreased. HBD4 was sensitive to and enhanced osteoblast/odontoblast differentiation potential of SHED by modulating Notch pathway. HBD4 was highly expressed in SHED stimulated by proinflammatory cytokines, and possessed anti-inflammatory, anti-bacterial activity. HBD4 promoted osteogenic/odontogenic differentiation of SHED. HBD4 may thus represent a suitable agent for vital pulp therapy in future clinic application.
乳牙牙髓炎是一种由细菌存在引起的疾病，在全球范围内高度流行。使用具有抗炎、抗菌和再生特性的生物相容性材料对于这种牙髓病的预后至关重要。本研究旨在鉴定人β-防御素4（HBD4）在人脱落乳牙干细胞（SHED）中的表达，并表征HBD4对SHED的影响。采用定量聚合酶链反应（qPCR）检测SHED中HBD4的表达以及HBD4对脂多糖（LPS）刺激的SHED中炎症因子的影响。通过Fortebio Octet系统进行亲和力测量，以探索LPS与HBD4之间的潜在相互作用。采用蛋白质免疫印迹分析来探索HBD4对丝裂原活化蛋白激酶（MAPK）途径的影响。应用集落形成单位方法和扫描电子显微镜研究HBD4对[此处原文缺失相关内容]和[此处原文缺失相关内容]的抗菌作用。采用碱性磷酸酶染色、茜素红染色、qPCR和蛋白质免疫印迹检测HBD4对SHED成骨细胞/成牙本质细胞分化的影响。使用RT Profiler PCR Array探索参与成骨/成牙本质分化的潜在信号通路。HBD4在TNF-α和IL-1α刺激的SHED中高表达。HBD4可直接与LPS结合，并下调LPS刺激的SHED中的IL-1α、IL-1β、IL-6、TNF-α，从而降低MAPK途径的激活。HBD4对[此处原文缺失相关内容]敏感，并通过调节Notch途径增强SHED的成骨细胞/成牙本质细胞分化潜能。HBD4在促炎细胞因子刺激的SHED中高表达，具有抗炎、抗菌活性。HBD4促进SHED的成骨/成牙本质分化。因此，HBD4可能是未来临床应用中活髓治疗的合适药物。
