Xenogeneic acellular collagen matrices represent a safe alternative to autologous soft tissue transplants in periodontology and implant dentistry. Here, we aimed to investigate the adsorption and release of growth factors from four porcine-derived collagen matrices using enzyme-linked immunosorbent assay. Non-crosslinked collagen matrix (NCM), crosslinked collagen matrix (CCM), dried acellular dermal matrix (DADM), and hydrated acellular dermal matrix (HADM) adsorbed each of the following growth factors, TGF-β1, FGF-2, PDGF-BB, GDF-5 and BMP-2, with an efficiency close to 100%. Growth factor release for a 13-day period was in the range of 10–50% of the adsorbed protein, except for the BMP-2 release that was in the range of 5–7%. Generally, protein release occurred in two phases. Phase I was arbitrary defined by the highest release from the matrices, usually within 24 h. Phase II, spanning the period immediately after the peak release until day 13, corresponded to the delayed release of the growth factors from the deeper layers of the matrices. HADM showed significantly (P < 0.001) higher TGF-β1, FGF-2, and PDGF-BB release in phase II, compared to the rest of the matrices. NCM exhibited significantly (P < 0.001) higher FGF-2 release in phase II, compared to CCM and DADM as well as a characteristic second peak in PDGF-BB release towards the middle of the tested period. In contrast to NCM and HADM, CCM and DADM showed a gradual and significantly higher release of GDF-5 in the second phase. Several burst releases of BMP-2 were characteristic for all matrices. The efficient adsorption and sustained protein release in the first 13 days, and the kinetics seen for HADM, with a burst release within hours and high amount of released growth factor within a secondary phase, may be beneficial for the long-term tissue regeneration following reconstructive periodontal surgery.
ObjectiveIn human gingival fibroblasts (HGFs), TLR4 recognises Pathogen-associated molecular patterns (PAMPs), such as LPS, and subsequently activates downstream signals that lead to the production of pro-inflammatory cytokines. The aim of this study was to explore the mechanisms of LPS-induced miRNA-146 regulation of TLR4 signals in HGFs.Materials and methodsHGFs were treated with Porphyromonas gingivalis (P.g) LPS, the cells were harvested, and kinase phosphorylation levels were detected by western blot. Selective pharmacological inhibitors and agonists were used to block or activate the relevant kinases, miRNA-146a/b expression levels were detected by real-time PCR, and IL-1, IL-6, and TNF-α production were measured by enzyme-linked immunosorbent assays (ELISA). A luciferase reporter plasmid containing miRNA-146a/b promoter was tested in terms of p50/p65 regulation.ResultsAfter P.g LPS treatment, NF-κB and Erk1/2 were strongly activated in HGFs. miRNA-146a/b, IL-1, IL-6 and TNF-α levels were down-regulated when NF-κB inhibitor was used. p50/p65 strongly activated miRNA-146a/b promoter as measured with the luciferase assay.ConclusionIn TLR4 signalling in HGFs, both miRNA-146a and miRNA-146b are downstream targets of NF-κB, but not of AP-1 signalling. miRNA-146a/b expression was specifically dependent on NF-κB but not Erk1/2 or JNK signalling.
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