Introduction: Regarding the limited ability of the damaged cartilage cells to self-renew, which is due to their specific tissue structure, subtle damages can usually cause diseases such as osteoarthritis. In this work, using laser photobiomodulation and an interesting source of growth factors cocktail called the synovial fluid, we analyzed the chondrogenic marker genes in treated hair follicle dermal papilla cells as an accessible source of cells with relatively high differentiation potential. Methods: Dermal papilla cells were isolated from rat whisker hair follicle (Rattus norvegicus) and established cell cultures were treated with a laser (gallium aluminum arsenide diode Laser (λ=780 nm, 30 mW) at 5 J/cm2 ), the synovial fluid, and a combination of both. After 1, 4, 7, and 14 days, the morphological changes were evaluated and the expression levels of four chondrocyte marker genes (Col2a1, Sox-9, Col10a1, and Runx-2) were assessed by the quantitative real-time polymerase chain reaction. Results: It was monitored that treating cells with laser irradiation can accelerate the rate of proliferation of cells. The morphology of the cells treated with the synovial fluid altered considerably as in the fourth day they surprisingly looked like cultured articular chondrocytes. The gene expression analysis showed that all genes were up-regulated until the day 14 following the treatments although not equally in all the cell groups. Moreover, the cell groups treated with both irradiation and the synovial fluid had a significantly augmented expression in gene markers. Conclusion: Based on the gene expression levels and the morphological changes, we concluded that the synovial fluid can have the potential to make the dermal papilla cells to most likely mimic the chondrogenic and/or osteogenic differentiation, although this process seems to be augmented by the irradiation of the low-level laser.
Combining biosensors with nanoscience provides great advantages such as being label-free and real time, highly sensitive, and small in size, as well as providing a low limit of detection and integration to other systems. That is why plasmonics finds various applications in drug detection, food safety, agriculture, photothermal therapy, etc. In this paper, we have fabricated a two-dimensional plasmonic grating biosensor using a soft lithography technique, which has eliminated some disadvantages of conventional plasmonic structures like expensive fabrication cost, inflexibility, and lack of mass production. On the other hand, we benefited from infrared neural stimulation for regulating membrane depolarization, which was based on photothermal mechanism and provided a contact-free and high spatial/temporal resolution. Eventually, the membrane depolarization of two different cell types of Hep G2 and mesenchymal stem cells cultured on two-dimensional plasmonic structure has been investigated under infrared neural stimulation. After preparing the soft plasmonic crystal, its reflection spectra and respective ellipsometry parameters were analyzed before and after cell culture with/without stimulation (near-infrared immune region ∼1450 nm). By comparing the obtained ellipsometry results for HEP G2 and mesenchymal stem cells, it is observed that the behavior of two cell types with respect to IR stimulation was the same as well as providing us the possibility of distinguishing the level of membrane depolarization under various stimulating frequencies. The strength of this integrated system for membrane depolarization detection has been shown experimentally, which can open new avenues toward neuroplasmonic application in the future.
Background Cancer-associated fibroblasts (CAFs), one of the major components of the tumor stroma, contribute to an immunosuppressive tumor microenvironment (TME) through the induction and functional polarization of protumoral macrophages. We have herein investigated the contribution of CAFs to monocyte recruitment and macrophage polarization. We also sought to identify a possible paracrine mechanism by which CAF-educated monocytes affect breast cancer (BC) cell progression. Methods Monocytes were educated by primary CAFs and normal fibroblast (NF); the phenotypic alterations of CAF- or NF-educated monocytes were measured by flow cytometry. Exosomes isolated from the cultured conditioned media of the educated monocytes were characterized. An in vivo experiment using a subcutaneous transplantation tumor model in athymic nude mice was conducted to uncover the effect of exosomes derived from CAF- or NF-educated monocytes on breast tumor growth. Gain- and loss-of-function experiments were performed to explore the role of miR-181a in BC progression with the involvement of the AKT signaling pathway. Western blotting, enzyme-linked immunosorbent assay, RT-qPCR, flow cytometry staining, migration assay, immunohistochemical staining, and bioinformatics analysis were performed to reveal the underlying mechanisms. Results We illustrated that primary CAFs recruited monocytes and established pro-tumoral M2 macrophages. CAF may also differentiate human monocyte THP-1 cells into anti-inflammatory M2 macrophages. Besides, we revealed that CAFs increased reactive oxygen species (ROS) generation in THP-1 monocytes, as differentiating into M2 macrophages requires a level of ROS for proper polarization. Importantly, T-cell proliferation was suppressed by CAF-educated monocytes and their exosomes, resulting in an immunosuppressive TME. Interestingly, CAF-activated, polarized monocytes lost their tumoricidal abilities, and their derived exosomes promoted BC cell proliferation and migration. In turn, CAF-educated monocyte exosomes exhibited a significant promoting effect on BC tumorigenicity in vivo. Of clinical significance, we observed that up-regulation of circulating miR-181a in BC was positively correlated with tumor aggressiveness and found a high level of this miRNA in CAF-educated monocytes and their exosomes. We further clarified that the pro-oncogenic effect of CAF-educated monocytes may depend in part on the exosomal transfer of miR-181a through modulating the PTEN/Akt signaling axis in BC cells. Conclusions Our findings established a connection between tumor stromal communication and tumor progression and demonstrated an inductive function for CAF-educated monocytes in BC cell progression. We also proposed a supporting model in which exosomal transfer of miR-181a from CAF-educated monocytes activates AKT signaling by regulating PTEN in BC cells.
Multidrug resistance is one of the major obstacles in the treatment of cancers. This undesirable feature increases the mortality rate of cancers, including breast cancer. Circular RNA (CircRNA)/microRNA (miRNA)/messenger RNA (mRNA) is one of the important axes with major roles in the promotion and resistance of breast cancer. This heterogeneous pathway includes mRNA of oncogenes and tumor suppressors, which are controlled by miRNAs and CircRNAs. Unfortunately, this network could be easily deregulated, resulting in drug resistance and tumor development. Therefore, understanding these dysregulations may thus help to identify effective therapeutic targets. On this basis, we try to review the latest findings in the field, which could help us to better comprehend this significant axis in breast cancer.
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