ABSTRACT
This study aimed to investigate the immunomodulatory effect of two different forms of phosphate-based glass microspheres (solid and porous), on human macrophages. Human THP-1 monocytes were converted to M0 macrophages after being treated with 100 ng/mL phorbol 12-myristate 13-acetate for 48 h. The differentiated cells were analysed for the CD14 marker using flow cytometry. The adhesion, spreading, and viability of M0 macrophages grown directly or indirectly (extracts) at varying concentrations of solid and porous glass microspheres (GMs) were analysed via phase contrast microscopy, confocal microscopy, and XTT assay. The expression of IL-8, IL-1ß, IL-6, IL-10, TNF-α, and IL-12p70 cytokines was investigated using flow cytometry. The conversion to M0 macrophages was confirmed by their adherent nature, increased granularity, and CD14 expression. The results showed that both solid and porous GMs or extracts favored the attachment, spreading, and proliferation of macrophages in a comparable manner to cells grown in a normal tissue culture medium. Only the higher concentration of porous GMs (10 mg/mL) changed the morphology of M0 macrophages and increased the expression of IL-1ß and IL-8 pro-inflammatory cytokines; this could be related to the fast degradation nature of porous GMs. Of the six cytokines analysed, M0 macrophages grown directly or indirectly with GMs only expressed IL-1ß, IL-10, and IL-8. Accordingly, solid microspheres may have advantages as regenerative agents due to their controlled degradation.
ABSTRACT
This study aimed to develop polycaprolactone (PCL) electrospun membranes coated with mineral trioxide aggregate/hydroxyapatite (MTA/HA) as a potential material for dental pulp capping. Initially, the PCL membrane was prepared by an electrospinning process, which was further surface coated with MTA (labeled as PCLMTA) and HA (labeled as PCLHA). The physico-chemical characterization of the fabricated membranes was carried out using field emission scanning electron microscopy (FE-SEM)/Energy dispersive X-ray (EDX), X-ray diffraction (XRD), Raman spectroscopy, and contact angle analysis. The biocompatibility of the human dental pulp stem cells (hDPSCs) on the fabricated membranes was checked by XTT assay, and the hDPSCs adhesion and spreading were assessed by FE-SEM and confocal microscopy. The wound healing ability of hDPSCs in response to different electrospun membrane extracts was examined by scratch assay. The surface morphology analysis of the membranes by FE-SEM demonstrated a uniform nanofibrous texture with an average fiber diameter of 594 ± 124 nm for PCL, 517 ± 159 nm for PCLHA, and 490 ± 162 nm for PCLMTA. The elemental analysis of the PCLHA membrane indicated the presence of calcium and phosphorous elements related to HA, whereas the PCLMTA membrane showed the presence of calcium and silicate, related to MTA. The presence of MTA and HA in the PCL membranes was also confirmed by Raman spectroscopy. The water contact analysis demonstrated the hydrophobic nature of the membranes. The results indicated that PCL, PCLHA, and PCLMTA membranes were biocompatible, while PCLMTA exhibited better cell adhesion, spreading, and migration.
ABSTRACT
Ficus latex is rich in polyphenolic compounds and hence considered as an antioxidant and anti-proliferative. Many studies are available on Ficus carica (common fig) whereas Ficus salicifolia is less studied. F. salicifolia grows in a harsh dry environment, therefore its latex was selected in the current study along with the F. carica for their comparative anti-cancer potential and the involved molecular mechanism. Triple-negative breast cancer (TNBC) derived MDA-MB-231 cells were used in the study. MTT and morphological studies indicated that the latex of both plants has anti-proliferative effects. To know their anti-metastatic effects, a wound-healing assay was performed. Both species were able to maintain the wound size compared to the untreated cells indicating their anti-metastatic effects. Using a DNA damage assay kit, we found that both fig species have genotoxic and cytotoxic effects in MDA-MB-231 cells compared to the untreated control. To know the potential molecular mechanism involved, we used a human kinase array kit. We found that ERK2, CREB, and AKT2 were downregulated after treatment the MDA-Mb-231 cells with the latex of F. carica. We assumed that F. salicifolia will also affect the same pathways, however after confirmation through real-time (RT)-PCR, downregulations of the above mentioned pathways were confirmed in cells treated with F. carica latex, however, in cells treated with F. salicifolia the selected genes were upregulated at the transcriptional level. We conclude that latex of both species of ficus have anti-cancer effects in MDA-MB-231 cells, however differ in their level of toxicity and the mechanism of action at the molecular level.
