Zantedeschia aethiopica is an evergreen perennial plant cultivated worldwide and commonly used for ornamental and medicinal purposes including the treatment of bacterial infections. However, the current understanding of molecular and physiological mechanisms in this plant is limited, in comparison to other non-model plants. In order to improve understanding of the biology of this botanical species, RNA-Seq technology was used for transcriptome assembly and characterization. Following Z. aethiopica spathe tissue RNA extraction, high-throughput RNA sequencing was performed with the aim of obtaining both abundant and rare transcript data. Functional profiling based on KEGG Orthology (KO) analysis highlighted contigs that were involved predominantly in genetic information (37%) and metabolism (34%) processes. Predicted proteins involved in the plant circadian system, hormone signal transduction, secondary metabolism and basal immunity are described here. In silico screening of the transcriptome data set for antimicrobial peptide (AMP) –encoding sequences was also carried out and three lipid transfer proteins (LTP) were identified as potential AMPs involved in plant defense. Spathe predicted protein maps were drawn, and suggested that major plant efforts are expended in guaranteeing the maintenance of cell homeostasis, characterized by high investment in carbohydrate, amino acid and energy metabolism as well as in genetic information.
Aim
To evaluate in vitro whether MTA Repair HP can induce repair processes at a distance, including its effects on biofilm, cell viability, migration, production of TGF‐β, phosphate and ALP, evaluated through MTA diluted extracts.
Methodology
Initially, antibacterial tests were performed with the bacterium Streptococcus mutans (ATCC 25175) in the presence of MTA extracts (dilutions of 1:1, 1:2 and 1:4). Growth inhibition assay by microdilution in broth, antibiofilm plate assay of young biofilm and antibiofilm assay in confocal microscopy of mature biofilm were carried out. Then, pulp cells were stimulated in the presence of several MTA dilutions, and cell viability (MTT assay), proliferation and migration capacity (scratch assay) were evaluated. To evaluate the capacity of 1:1, 1:2 and 1:4 dilutions of MTA Repair HP to promote the production of important agents of odontogenic differentiation and mineralization, ALP activity, TGF‐β secretion and phosphate quantification were measured. Statistical differences were verified using one‐way and two‐way anova and Tukey's post‐tests.
Results
The test dilutions of MTA Repair HP did not inhibit planktonic S. mutans growth but were able to reduce young and mature S. mutans biofilm (p < 0.001). In addition, none of the MTA Repair HP dilutions was cytotoxic for pulp cells. The 1:2 and 1:4 dilutions of MTA Repair HP induced migration and proliferation of pulp cells (p < 0.05). ALP activity and TGF‐β secretion were independent of the tested dilution (p < 0.001). Diluted 1:4 MTA Repair HP produced less phosphate than the more concentrated 1:1 and 1:2 MTA dilutions (p < 0.001).
Conclusions
Undiluted MTA Repair HP reduced S. mutans biofilm, when compared to 1:2 and 1:4 MTA dilutions. Furthermore, none of the tested dilutions was cytotoxic to pulp cells. MTA Repair HP promoted cell migration and proliferation at a distance, assessed through the dilution of the MTA. Even from a distance, MTA Repair HP has the ability to participate in some events related to repair, such as migration, proliferation and TGF production.
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