Zhu Minwen, Miao Bo, Zhu Jianhua, Wang Haiyan, Zhou Zengtong
Department of Oral Mucosal Diseases, Shanghai Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200011, P.R. China.
Department of Stomatology, Aviation General Hospital of China Medical University, Beijing 100012, P.R. China.
Mol Med Rep. 2017 Sep;16(3):2455-2460. doi: 10.3892/mmr.2017.6913. Epub 2017 Jul 4.
Periodontitis is an oral chronic inflammatory disease induced by microorganisms that can destroy tooth‑supporting structures. Human β‑defensin‑3 (HBD‑3) is a type of endogenous antimicrobial peptide that inhibits a broad spectrum of microorganisms. The objectives of the present study were to transfect human periodontal ligament cells (HPDLCs) and human bone marrow stromal cells (HBMSCs) with lentivirus containing the HBD‑3 gene, determine the transfection efficiency, and investigate the antimicrobial activity of the experimental cells against periodontal pathogens. Fluorescence microscopy was used to calculated the transfection efficiency. Western blot analysis and ELISA were conducted to confirm the expression of HBD‑3 at the protein level. The effect of the HBD‑3 gene on the antimicrobial activity of the cells were demonstrated by antimicrobial tests. The results of the present study demonstrated that the transfected HPDLCs and HBMSCs stably expressed HBD‑3. In addition, periodontal pathogens and caries‑causing bacteria were susceptible to the antimicrobial activity of the cells. Both HPDLCs and HBMSCs hold potential for use as seeding cells in cell‑ and gene‑based therapies for periodontal disease. The lentiviral vector containing HBD‑3 resulted in broad‑spectrum antimicrobial activity against a variety of oral organisms, and could potentially be applied in the treatment of oral infectious diseases, including periodontitis.
牙周炎是一种由微生物引起的口腔慢性炎症性疾病,可破坏牙齿支持结构。人β-防御素-3(HBD-3)是一种内源性抗菌肽,可抑制多种微生物。本研究的目的是用携带HBD-3基因的慢病毒转染人牙周膜细胞(HPDLCs)和人骨髓基质细胞(HBMSCs),确定转染效率,并研究实验细胞对牙周病原体的抗菌活性。用荧光显微镜计算转染效率。进行蛋白质印迹分析和酶联免疫吸附测定以确认HBD-3在蛋白质水平的表达。通过抗菌试验证明HBD-3基因对细胞抗菌活性的影响。本研究结果表明,转染的HPDLCs和HBMSCs稳定表达HBD-3。此外,牙周病原体和致龋菌对细胞的抗菌活性敏感。HPDLCs和HBMSCs都有潜力用作牙周疾病细胞和基因治疗的种子细胞。携带HBD-3的慢病毒载体对多种口腔生物具有广谱抗菌活性,并有可能应用于包括牙周炎在内的口腔感染性疾病的治疗。