RNAseq of gingival fibroblasts exposed to PRF membranes and PRF serum.
Ontology highlight
ABSTRACT:
PROVIDER: PRJNA1115785 | ENA |
REPOSITORIES: ENA
Dataset's files
| Action | DRS | |||
|---|---|---|---|---|
| SRR29150662.fastq.gz | Fastqsanger.gz | |||
| SRR29150663.fastq.gz | Fastqsanger.gz | |||
| SRR29150664.fastq.gz | Fastqsanger.gz | |||
| SRR29150665.fastq.gz | Fastqsanger.gz | |||
| SRR29150666.fastq.gz | Fastqsanger.gz |
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Project description:Platelet-rich fibrin (PRF) is prepared from the coagulated plasma of fractionated blood. When squeezing between two plates, PRF is separated into the solid PRF membranes and a liquid exudate, the PRF serum. The question arises regarding the extent to which the overall PRF activity remains in the membranes and what is lost in the serum. To this aim, we have exposed gingival fibroblasts to lysates prepared from PRF membranes and PRF serum, followed by bulk RNA sequencing. A total of 268 up- and 136 down-regulated genes in gingival fibroblasts exposed to PRF lysates are significantly regulated under the premise of a minimum log2 2.5-fold change and a minus log10 significance level of two, respectively. PRF serum caused 62 up- and 32 down-regulated genes when gingival fibroblasts were exposed to PRF serum, respectively. Among the 61 genes commonly up-regulated by PRF lysate and serum were CXCL1, CXCL5, CXCL6, CXCL8, IL33, and IL6 and PTGS2, STC1. PRF lysate further increased the chemokines CCL2, CCL7, CXCL2, CXCL3, and the IL1R1, IL1RL1, and IL1RN – as well as the paracrine factors IL11, LIF, IGF1, BMP2, BMP6, FGF2, CCN2/CTGF and HAS1, HAS2, HAS3. The 16 up-regulated genes by PRF serum included DKK1. Among the 122 down-regulated genes by PRF lysate were IFIT1, IFIT2, IFIT3, OSR1, OSR2. Among the 32 down-regulated genes by PRF serum were FGF18 and GDF15. Taken together, PRF lysates, compared to PRF serum, cause a more complex response of gingival fibroblasts with a chemokine with an obvious increase in chemokine expression and spectrum of paracrine factors.
2024-05-31 | GSE268248 | GEO
Project description:Platelet-rich fibrin (PRF) is prepared by spontaneous coagulation of fractionated blood. When squeezed between two plates, PRF is separated into solid PRF membranes and a liquid exudate, the PRF serum. The question arises regarding how much the overall activity remains in the PRF membranes and what is discarded into the PRF serum. To this end, we have exposed gingival fibroblasts to lysates prepared from PRF membranes and PRF serum, followed by bulk RNA sequencing. A total of 268 up- and 136 down-regulated genes in gingival fibroblasts exposed to PRF membrane lysates were significantly regulated under the premise of a minimum log2 with 2.5-fold change and a minus log10 significance level of two, respectively. PRF serum only caused 62 up- and 32 down-regulated genes under these conditions. Among the 46 commonly up-regulated genes were CXCL1, CXCL5, CXCL6, CXCL8, IL33, IL6, and PTGS2/COX2, stanniocalcin-1-all linked to an inflammatory response. PRF membrane lysates further increased chemokines CCL2, CCL7, CXCL2, CXCL3, and IL1R1, IL1RL1, and IL1RN, as well as the paracrine factors IL11, LIF, IGF1, BMP2, BMP6, FGF2, and CCN2/CTGF, and all hyaluronan synthases. On the other hand, PRF serum increased DKK1. The genes commonly down-regulated by PRF membrane lysates and PRF serum included interferon-induced protein with tetratricopeptide repeats (IFIT1, IFIT2, IFIT3) and odd-skipped-related transcription factors (OSR1 and OSR2), as well as FGF18 and GDF15, respectively. Taken together, PRF membrane lysates, compared to PRF serum, cause a more complex response in gingival fibroblasts, but each increased chemokine expression in gingival fibroblasts.
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Project description:Considering that PRF is clinically used in combination with dental implants and collagen membranes, we exposed titanium discs and collagen membranes to PRF lysates. We showed here that PRF-derived TGF-β activity adsorbs to titanium implants and collagen membranes indicated by the changes in gene expression and immunoassay analysis. Our study points towards TGF-β as major target of PRF and suggest that TGF-β activity released by PRF adsorbs to titanium surface and collagen membranes.
2020-06-24 | PXD014382 | Pride
Project description:Fibroblasts are widely used cells for regenerative medicine in clinics, such as gingival or facial skin treatment. In fact, fibroblasts are considered as a mixture of various types of cells with "spindle shape" and there is no available clear marker. Gingival and dermal fibroblasts are similar in their morphology and function; however it is considered that the cultured cells retain their original characteristics depending on the origin, which may contribute to the differential therapeutic effects. For example, gingival wounds are known to heal relatively quickly with less scar formation compared with skin, which may imply their higher capability for regeneration as a therapeutic effect. The reason for this phenomenon may be partly due to characteristic differences between gingival and dermal fibroblasts including the expression of migration stimulating factor and matrix formation but these differences remain largely unknown. Recently, the characteristics of dermal fibroblasts have been reported to be different depending on body sites, such as face, trunk and plamoplantar skin. Although the expression of fibronectin and their splicing variants were known to be different between trunk and oral mucosal fibroblasts, there is still no detailed report on the functional differences between gingival and dermal fibroblasts. In this study, we investigated differential gene expression in normal gingival and dermal fibroblasts using DNA microarray to investigate the difference between the vague fibroblast-type cells from different tissue origin to achieve higher therapeutic effect in cell therapy. Gingival and dermal tissues were collected from healthy patients. After successive stages primary culture and RNA extraction and hybridization on Affymetrix microarrays (Genome Focus Array). Dermal and gingival tissues were obtained from healthy volunteers (8 cases for dermal tissue; 6 females, 2 males , average age 48 and 8 cases for buccal gingival tissue; 6 females, 2 males, average age 43) whose informed consent was obtained according to a protocol approved by the ethics committee of Nagoya University Hospital. After enzymatic digestion, tissues were cultured in Dulbecco's modified Eagle's medium (DMEM) containing 10% fetal bovine serum at 37°C in the presence of 5% CO2 for about 4 weeks as reported previously. Total mRNAs were extracted from cells between passages 4-5 by Trizol reagent (Invitrogen, Carlsbad, CA, USA) and were applied to Human Focus Arrays (Affymetrix, Santa Clara, CA, USA) for microarray analysis according to the manufacturer's protocol (http://www.affymetrix.com/support/technical/manuals.affx). The gene expression data were analyzed by Arrayassist (Stratagene, La Jolla, CA, USA). Briefly, 8,500 probes on the array, normalization and scaling (MAS5), flag-positive gene selection, unpaired t-test, and CV selection (<20) resulted in 5,284 genes to analyze. GO (Gene ontology) analysis was performed using the software default settings to find the gene group related to the same category of biological function by searching common key terms that were reported for each gene.
2010-06-08 | E-GEOD-22029 | biostudies-arrayexpress
Project description:Hepatitis E virus (HEV) is an important causative pathogen of acute hepatitis. Because of the absence of an in vitro culture system for HEV, research has been greatly impeded. And interaction between HEV and host cells was mainly studied by tansfection/transinfection system, such as Adeno virus transinfection system. We developed an in vitro culture system for HEV in PLC/PRF/5 cells. With this in vitro culture system, we studied the gene expression profile change by HEV infection. Using a microarray assay, we analysed genes of PLC/PRF/5 cells, whose transcription level could be changed by HEV infection. Five flasks of PLC/PRF/5 cells inoculated with HEV were used as test sample, and five flasks inoculated with serum-free DMEM/199 medium were used as control samples. Both test and control flasks were cultured under the same conditions. Sixty days after inoculation, the test and control flasks was resolved with Trizol and analysed with Affymetrix HG-U133 Plus 2 array.
2014-01-01 | E-GEOD-53731 | biostudies-arrayexpress
Project description:Objective: 2-Hydroxyethyl methacrylate (HEMA), one of the most common components of tooth filling materials, could diffuse throughout dentine, and in gingival and pulp cells, affecting odontoblast vitality. Design: the aim of this study was to assess the differential transcriptome modulation induced by HEMA treatment in human cultured gingival fibroblasts (HGFs) at different exposure time compared to untreated cells. Materials and methods: Gene expression profile was performed by employing a whole gene microarray approach on RNA extracted from cultured HGFs exposed to 3 mmol/l HEMA for 24 or 96 h. Differentially identified transcripts were analyzed by Ingenuity Pathways Analysis software to disclose genes biological functions. Results: Hierarchical clustering analysis of the data set originated from a total of 8 experiments showed the selective presence of two gene clusters, composed by hundreds trandcripts differentially expressed after 24-h and 96-h HEMA treatment. when compared to untreated controls. Functional analysis showed the transcripts involvement mainly in cell survival, proliferation and death, with an unbalance towards cell growth and inflammatory response. Conclusions: Data analysis showed an overall damage induced by HEMA exposure for both HGFs cultures at 24 and 96-h mainly leading to a proliferation impairement. Interestingly 24-h HEMA exposure could induce the cells to trigger repair mechanisms evidencing an early compensatory response and survive while. 96-h incubation might increase apoptosis as a consequence of the chronic damage. Gene expression profile of Human Gingival Fibroblasts (HGFs) exposed to 3mM 2-Hydroxyethyl Methacrylate (HEMA) for 24- and 96-h versus untreated Human Gingival Fibroblasts at 24- and 96-h. A total of 8 experiments was carried out including biological and technical replicates.
2014-12-15 | E-GEOD-61043 | biostudies-arrayexpress
Project description:Fibroblasts are widely used cells for regenerative medicine in clinics, such as gingival or facial skin treatment. In fact, fibroblasts are considered as a mixture of various types of cells with "spindle shape" and there is no available clear marker. Gingival and dermal fibroblasts are similar in their morphology and function; however it is considered that the cultured cells retain their original characteristics depending on the origin, which may contribute to the differential therapeutic effects. For example, gingival wounds are known to heal relatively quickly with less scar formation compared with skin, which may imply their higher capability for regeneration as a therapeutic effect. The reason for this phenomenon may be partly due to characteristic differences between gingival and dermal fibroblasts including the expression of migration stimulating factor and matrix formation but these differences remain largely unknown. Recently, the characteristics of dermal fibroblasts have been reported to be different depending on body sites, such as face, trunk and plamoplantar skin. Although the expression of fibronectin and their splicing variants were known to be different between trunk and oral mucosal fibroblasts, there is still no detailed report on the functional differences between gingival and dermal fibroblasts. In this study, we investigated differential gene expression in normal gingival and dermal fibroblasts using DNA microarray to investigate the difference between the vague fibroblast-type cells from different tissue origin to achieve higher therapeutic effect in cell therapy.
2010-05-28 | GSE22029 | GEO
Project description:Objective: 2-Hydroxyethyl methacrylate (HEMA), one of the most common components of tooth filling materials, could diffuse throughout dentine, and in gingival and pulp cells, affecting odontoblast vitality. Design: the aim of this study was to assess the differential transcriptome modulation induced by HEMA treatment in human cultured gingival fibroblasts (HGFs) at different exposure time compared to untreated cells. Materials and methods: Gene expression profile was performed by employing a whole gene microarray approach on RNA extracted from cultured HGFs exposed to 3 mmol/l HEMA for 24 or 96 h. Differentially identified transcripts were analyzed by Ingenuity Pathways Analysis software to disclose genes biological functions. Results: Hierarchical clustering analysis of the data set originated from a total of 8 experiments showed the selective presence of two gene clusters, composed by hundreds trandcripts differentially expressed after 24-h and 96-h HEMA treatment. when compared to untreated controls. Functional analysis showed the transcripts involvement mainly in cell survival, proliferation and death, with an unbalance towards cell growth and inflammatory response. Conclusions: Data analysis showed an overall damage induced by HEMA exposure for both HGFs cultures at 24 and 96-h mainly leading to a proliferation impairement. Interestingly 24-h HEMA exposure could induce the cells to trigger repair mechanisms evidencing an early compensatory response and survive while. 96-h incubation might increase apoptosis as a consequence of the chronic damage.
2014-12-15 | GSE61043 | GEO
Project description:Treponema denticola is a major pathogen in periodontal disease, frequently isolated from lesions of chronic periodontitis. The ability to adopt its environment is believed to be important for T. denticola in colonizing and proliferating in the gingival crevice. T. denticola use serum as a major nutrient source in the gingival crevices, suggesting that this microorganism utilize serum components to proliferate in gingival crevice. The purpose of this study was to identify T. denticola serum utilization genes. Precultured T. denticola were suspended in tryptone-yeast extract-gelatin-volatile fatty acids medium containing 0, 1 and 10% serum, and incubated anaerobically for 17 h. Total RNA was isolated and T. denticola gene expression was compared by microarray and reverse transcription-polymerase chain reaction. In serum-depleted conditions, the expression of a potential hydroxylamine reductase, several ABC transporters, and phosphoenolpyruvate synthase were increased, while methyl-accepting chemotaxis proteins and a transcriptional regulator were decreased. The results suggest that T. denticola may uptake serum components via ABC transporters. Decreased dmcA expression with decreased serum concentration suggests its involvement in chemotaxis toward serum-rich environments.
2018-07-20 | GSE110856 | GEO
Project description:PLC/PRF/5 cells were harvested for chromatin immunoprecipitation followed by CHIP analysis performed on Aligent platform.
2017-07-10 | GSE100968 | GEO