Synergetic effect of chemical and topological signals of gingival regeneration scaffold on the behavior of human gingival fibroblasts.

Achievement of gingiva regeneration poses a substantial challenge for dental aesthetics and periodontal disease repair, but reports of a bioactive and easily available gingival regeneration scaffold are rare. Cell behaviors can be affected by multiple kinds of bioactive signals; thus, it is important to explore the effects of the chemical and topological signals of the scaffold on the behavior of human gingival fibroblasts (HGFs). We hypothesized that the synergetic effect of the chemical and topological scaffold signals were beneficial to gingival regeneration. In this study, HGF behavior on random/aligned poly (e-caprolactone) (PCL) and polydopamine (PDA)-coated PCL electrospun scaffolds was investigated in a common medium and in basic fibroblast growth factor (bFGF) medium. The results showed that the synergistic effect of three signals was better than that of one or two signals. The cell proliferation of the aligned scaffold group was higher than that of the random scaffold, and the PCL/PDA-Aligned+bFGF group showed the highest cell proliferation. Even if two chemical signals were present, the HGFs still maintained an ordered distribution on the aligned scaffold. Cell differentiation and protein secretion analysis indicated that gene and protein expression of focal adhesion kinase and fibronectin were the highest in the PCL/PDA-Aligned+bFGF group. Taken together, the chemical and topographic signals within the electrospun scaffold were considered to display a synergistic effect on HGF behaviors, suggesting the potential usefulness of the PCL/PDA-Aligned+bFGF scaffold for gingiva tissue engineering.

Effect and molecular mechanism of isorhamnetin extracted from Ginkgo biloba on the differentiation of RAW264.7 cells into osteoclasts / 口腔疾病防治

Objective @#To investigate the effect and potential molecular mechanisms of isorhamnetin (ISO) extracted from Ginkgo biloba on the differentiation of osteoclasts.@*Methods@#Osteoclast precursor RAW264.7 cells were induced with RANKL to differentiate into mature osteoclasts. Different concentrations of ISO were added to RAW264.7 cells to determine its effect on osteoclast differentiation. CCK8 was used to evaluate the effect of ISO on cytotoxicity. The impact of ISO on the osteoclast differentiation process was investigated by analyzing tartrate resistance and bone resorption lacuna. Real-time PCR was performed to analyze the levels of differentiation marker genes, including tartrate resistant acid phosphatase (Trap), cathepsin K (Ctsk), and matrix metalloproteinase 9 (MMP-9); differentiation-related transcription factors, including the proto-oncogene protein c-Fos, nuclear factor of activated T-cells cytoplasmic 1(NFATc1); and the levels of downstream NF-κB p65 signaling pathway phosphorylation. Using the above-described method, we verified that ISO exerted an inhibitory effect on osteoclast differentiation and explored related molecular mechanisms. @*Results @#Different concentrations of ISO (1-10 μM) had no cytotoxic effects on RAW264.7 cells, inhibited TRAP activity and decreased the number of bone resorption lacuna during osteoclast differentiation. When applied at a concentration of 10 μM, its inhibitory effect was significant. In addition, ISO significantly reduced the expression levels of Trap, Ctsk, MMP-9, c-Fos, NFATc1 and NF-κB p65 mRNA. @*Conclusion@# ISO extracted from Ginkgo biloba extract exerted an inhibitory effect on osteoclast differentiation, and the mechanism underlying its activity may involve the inhibition of the classical NF-κB pathway.