Early morphology observation of human periodontal fibroblast cell line in the presence of human salivary exosomes

@#Human salivary exosomes have been identified as a highly informative nanovesicle with clinical-relevant information for variation of diagnostic purposes. As a continued effort from previous studies on human salivary exosomes effect at gene expression level, this study is carried out to observe the morphology of human periodontal fibroblast (HPdLF) treated with exosomes cells under the same period of changes in genotypic level occurred. In vitro, HPdLF cells were cultured for 24 hours with 10 µg/ml of human salivary exosomes. The morphology of HPdLF cells was examined under inverted light microscopy and scanning electron microscopy (SEM) for both control samples and samples treated with human salivary exosomes, while the cell count was performed via trypan blue staining. There was no significant difference in the morphology under the inverted light microscopy and the cell number of HPdLF cells for both treated and untreated cells with exosomes. However, for SEM, the treated HPdLF with salivary exosomes showed slight observable changes on the filopodia, lamellipodia, cytoplasmic vesicles and the cytoskeleton of the cells. Even within a short period (24 hours) of culturing time for cells with human salivary exosomes, the samples showed minimal changes which positively suggested a simultaneous event of exchanging materials from human salivary exosomes to cells had occurred, hence, potentially proving that human salivary exosomes can enhance cell proliferation

Inhibition of Akt/PKB signaling pathway promotes hypoxia-induced apoptosis of human periodontal ligament fibroblasts / 实用口腔医学杂志

Objective: To investigate the effects of the Akt/PKB signaling pathway on hypoxia-induced apoptosis. Methods: The cell proliferation, apoptosis, Akt/PKB signaling pathway and HIF-1α expression in periodontal ligament fibroblasts(hPDLFs) under normoxic and hypoxic conditions were evaluated by MTT assay, flowoytometry, qRT-PCR, Western blot and Lipofectamin 2000 transfection respectively. Results: The cell proliferation and the Akt/PKB pathway in hPDLFs were inhibited by hypoxia. Hypoxia promoted apoptosis and increased the levels of HIF-1α of hPDLFs. Akt/PKB signaling pathway inhibition inhibited cell proliferation and promoted hypoxia-induced apoptosis of hPDLFs. Conclusion: The inhibition of Akt/PKB signaling pathway may promote hypoxia-induced apoptosis of hPDLFs under hyoxia condtition.

The effect of Er:YAG laser radiation on the proliferation of human periodontal ligament fibroblasts / 实用口腔医学杂志

Objective:To observe the effect of Er:YAG laser on the proliferation of human periodontal ligament fibroblasts(hP-DLFs).Methods:Human periodontal ligament fibroblasts were cultured in vitro and identified by immunohistochemistry.The cells of 5th passage were divided into 5 groups.The cells in group A without treatment were used as the controls,in group B,C,D and E were treated with Er:YAG laser of 10 Hz at 50 mJ,100 mJ,150 mJ and E-200 mJ for 1 s respectively.The proliferation of the cells was examined on day 1,3,5,7,9 by CCK-8.Results:The proliferation of hPDLFs in group B,C,D and E increased more than that in group A(P <0.05)5 d after Er:YAG laser radiation.Conclusion:Low intensity of Er:YAG laser radiation can promote the prolifera-tion of human periodontal ligament fibroblasts.

Periodontal tissue engineering by hPDLF seeding on scaffold / 대한치주과학회지

Human periodontal ligament fibroblasts (hPDLF) are very important for curing the periodontal tissue because they can be differentiated into various cells. A tissue engineering approach using a cell-scaffold is essential for comprehending today's periodontal tissue regeneration procedure. This study examined the possibility of using an acellular dermal matrix as a scaffold for human periodontal ligament fibroblast (hPDLF). The hPDLF was isolated from the middle third of the root of periodontally healthy teeth extracted for orthodontic reasons. The cells were cultured in a medium containing Dulbecco's modified Eagle medium supplemented with 10% fetal bovine serum at 37degrees C in humidified air with 5% CO2. The acellular dermal matrix(ADM) was provided by the US tissue banks(USA). Second passage cells were used in this study. The hPDLF cells were cultured with the acellular dermal matrix for 2 days, and the dermal matrix cultured by the hPDLF was transferred to a new petri dish and used as the experimental group. The control group was cultured without the acellular dermal matrix. The control and experimental cells were cultured for six weeks. The hPDLF cultured on the acellular dermal matrix was observed by Transmission Electron microscopy (TEM). Electron micrography shows that the hPDLF was proliferated on the acellular dermal matrix. This study suggests that the acellular dermal matrix can be used as a scaffold for hPDLF.

Profiling of genes in healthy hGF, aging hGF, healthy hPDLF and inflammatory hPDLF by DNA microarray / 대한치주과학회지

No abstract available.

Effects of different tension stress on the shape and F-actin of human peridontal ligament fibroblast in vitro / 第三军医大学学报

Objective To investigate the shape and F-actin of human periodontal ligament fibroblasts(HPDLF) under different tension stress in vitro so as to learn the stress-biological effects of HPDLF. Methods HPDLF were cultivated for 6 passages and observed morphologically and identified by immunocytochemistry to be positive anti-vimentin and negative anti-keratin. HPDLF were divided into four groups: control (without tension stress), static tension stress (5 kPa), dynamic tension stress group 1 (5 to 0 to 5 kPa at the frequency of 3/min) and dynamic tension stress group 2 (5 to 2.5 to 5 kPa at the frequency of 3/min). The cells of each group were observed at the different time points of 16, 24, 32 h. The projection areas and shapes of cells as well as the structure of F-actin were examined by laser scanning confocal microscope and immunity fluorescence technique. The relationship among tension stress, time, shape and the structure of F-actin of HPDLF was analysed. Results In the dynamic tension stress group 1 and 2, the shape and the arrangement of F-actin of some HPDLF underwent regular changes at 16, 24 h, but the changes appeared to be obvious at 32 h. In the static tension stress group, it was found the increase of interspace of HPDLF, but no obvious changes in the structure of F-actin. Conclusion Different tension stress patterns had different effects on the shape and F-actin of HPDLF. Especially in the dynamic tension stress group 2, it showed direct ratio between the cell projection areas, the average fluorescence density and different loading time.