Effect of lentivirus‐mediated BMP2 from autologous tooth on the proliferative and osteogenic capacity of human periodontal ligament cells

• Published in: Journal of periodontal research 2022-08, Vol.57 (4), p.869-879
• Main Authors: Qin, Ruoshan, Cui, Ziwei, Zhou, Hongli, Guo, Ru, Yao, Xuanxuan, Wang, Tao, Qin, Xiaodong, He, Xiangyi
• Format: Article
• Language: English
• Subjects: Alkaline phosphatase; Bone growth; Bone healing; Bone morphogenetic protein 2; Bone sialoprotein; Bones; Cholecystokinin; Collagen (type I); Cytology; Gene expression; Growth factors; human periodontal ligament cells; lentivirus; Ligaments; Maxillofacial; Osteocalcin; Osteogenesis; osteogenic differetiation; Osteopontin; Periodontal ligament; Periodontitis; Phosphatase; Regeneration; Tissue engineering
• ISSN: 0022-3484; 1600-0765
• DOI: 10.1111/jre.13025

Background and objective Periodontitis is a chronic progressive inflammation that invades periodontal supporting tissues, in which periodontal tissue regeneration engineering offers new hope for prevention and treatment, including seed cells, scaffolds, and growth factors. In recent years, scholars have shown that autologous teeth can be used as new bone tissue repair materials for periodontal regeneration and bone tissue repair. The aim of this study was to establish a human periodontal ligament cell line that expresses the human bone morphogenetic protein 2 gene (BMP2) in a stable manner using lentiviral mediation in order to explore the effect of BMP2 from autologous tooth on the proliferative and osteogenic capacity of human periodontal ligament cells (hPDLCs). Materials and methods Human periodontal ligament cells were cultured, subcultured, and identified, and then homologous recombinant lentivirus plasmid plv‐BMP2 was constructed and transfected into the third passage (P3) hPDLCs. After that, the effect of BMP2 on its proliferation was detected by CCK‐8, at the same time, the osteogenic induction of hPDLCs was carried out at 7, 14, and 21 days, and then the effect of BMP2 on its osteogenic ability was detected by alizarin red staining, alkaline phosphatase activity determination, and the mRNA expression levels of osteogenic‐related genes using real‐time fluorescence quantitative PCR, including alkaline phosphatase, runt‐related transcription factor 2, bone sialoprotein, osteocalcin, osteopontin, and collagen I. Finally, spss26.0 software was used for statistical processing. Results The results showed that cells transfected with the homologous recombinant lentiviral plasmid pLV‐BMP2 had a similar morphology to normal hPDLCs, showing a typical radial arrangement; the cell proliferative capacity of the pLV‐BMP2 group as measured by CCK‐8 was enhanced compared with the control group and the pLV‐puro group (p