Objective To investigate the osteogenic differentiation potential of human β-defensin 3 (hβD-3) transfected adipose stem cells (ADSCs) and its effect on anti-periodontitis pathogenic in vitro.
Methods The human abdominal adipose tissue was extracted in vitro, and the immunophenotype (CD29/CD44) of human adipose stem cells (hADSCs) was identified by flow cytometry after separation. The hADSCs were cultured in vitro, and the cells were divided into control group, no-load group and experimental group. The control group was untreated. The empty group was transfected with a lentiviral vector containing no hβD-3 gene. The experimental group used a plasmid to construct a lentiviral vector containing the hβD-3 gene and was transfected hADSCs. Both the no-load group and the experimental group were added with osteogenic differentiation inducer, and the control group was not treated. The expressions of osteogenic related genesalkaline phosphatase (ALP), type Ⅰ collagen (Col-Ⅰ), osteopontin (OPN), osteocalcin (OCN) of the three groups were detected by real-time fluorescent quantitative polymerase chain reaction after 7 days and 14 days of induction to evaluate the differentiation ability of hADSCs. The diameter of antibacterial zone was measured, and antibiotic positive controls (minocycline group and ciprofloxacin group) were set, and the antibacterial activity of the transfected hADSCs against the periodontal pathogens such as Staphylococcus and Porphyromonas was detected.
Results At 7 and 14 days of induction, the relative expressions of ALP, Col-Ⅰ, OPN and OCN mRNA in the no-load group and the experimental group were significantly higher than those in the control group (P < 0.05); the relative expressions of ALP, Col-Ⅰ, OPN and OCN mRNA in the no-load group and the experimental group at 14 days induction were significantly higher than those of 7 days after induction(P < 0.05). In the Staphylococcal antibacterial experiment, the inhibition zone of the low-density group was significantly smaller than that of the minocycline group and ciprofloxacin group, the zone of inhibition in the medium-density group was significantly greater than that in the ciprofloxacin group, the zone of inhibition in the high-density group was significantly greater than that of the minocycline group and ciprofloxacin group (P < 0.05). In the bacteriostasis experiment of Porphyromonas, the inhibition zone of the low-density group was significantly smaller than that of the minocycline group and the ciprofloxacin group, the inhibition zone of medium-density group group was significantly higher than that of minocycline group, and was significantly higher in the high-density group than that of the minocycline group and the ciprofloxacin group (P < 0.05).
Conclusion hβD-3 transfection can promote the osteogenic differentiation of hADSCs, and has good antibacterial activity against periodontal pathogens such as Staphylococcus and Porphyromonas.
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