Introduction: Chlorhexidine (CHX) has been used for some time in clinical practice as a local antiseptic agent with excellent efficacy. The combination of CHX with APRF (Advanced-platelet rich fibrin) membrane has the potential to stimulate tissue regeneration and to provide a bactericidal effect. We hypothesize that this may reduce the rate of infections development and protect cell viability. Aim: The aim of this study was two-fold—to create a stable APRF membrane treated with different concentrations of CHX (0.01% and 0.02%) and to monitor its effect on the viability of PDL cells in vitro. This benefits the introduction of a new protocol for APRF membrane production -CHX-PRF and enriches the available evidence on the effect of this antiseptic agent on PDL (Periodontal ligament) cells. Materials and methods: APRF membranes were prepared by the addition of two concentrations (0.01% and 0.02%) of CHX. Membranes without the antiseptic were also prepared and used as control samples. PDL cells were cultivated on the membranes for 72 h. Cell number and vitality were examined by fluorescent cell viability assays. Results: Our results demonstrated that a concentration of 0.01% CHX allowed the production of a stable APRF membrane. This concentration slightly reduced the viability of PDL cells to 96.7%, but significantly decreased the average number of cells attached to the membrane—149 ± 16.5 cells/field compared to controls −336 ± 26.9 cells/field. APRF-CHX 0.02% membranes were unstable, indicating a dose-dependent cytotoxic effect of CHX. Conclusions: The introduced novel protocol leads to the production of a new type of APRF membrane—CHX-PRF. The incorporation of an antiseptic into the APRF membrane can improve its bactericidal activity and might serve as an important step for the prevention of postoperative infections.
Germination of seeds of root parasites like broomrapes (Orobanchaceae) is tightly regulated by chemical products exuded from the roots of the host plant, known as germination stimulants (GSs). Changes in the levels of synthesis and emission of GS can allow the development of practical measures for control of the crops-harming parasitic species. However, the genes encoding enzymes responsible for GS biosynthesis are still unknown. We performed a large-scale screening of 62,000 Arabidopsis activation-tag mutants for alteration in susceptibility to Phelipanche ramosa and to identify lines with altered GS production among them. After five successive screenings we identified 36 lines with altered susceptibility to P. ramosa. Seven of them displayed altered levels of GS production. By using a combination of Southern blot and thermal asymmetric interlaced polymerase chain reaction (TAIL-PCR), we pinpointed the location of activation-tag constructs in these lines. A combination of differential display and quantitative real-time PCR (qRT-PCR) allowed us to identify several affected genes. Two of them are directly involved in isoprenoid biosynthetic pathway in chloroplasts, and we believe that their activation led to increased levels of GS production. We believe that these genes are responsible for increased GS production in five of the Arabidopsis lines resistant to P. ramosa.
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