Gene Expression Analysis of Chondrogenic Markers in Hair Follicle Dermal Papillae Cells Under the Effect of Laser Photobiomodulation and the Synovial Fluid

Abstract

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.

Near infrared low‐level laser therapy and cell proliferation: The emerging role of redox sensitive signal transduction pathways

“…Such “optical window” accounts for the maximal tissue penetration, due to the lack of massive absorption by tissue chromophores such as hemoglobin and melanin. Moreover, in this range also the light absorption by water is minimal, thus resulting in nearly optical transparency of biological samples . The choice of laser wavelength strongly depends on the biological target of interest: generally, lasers operating in the 600 to 700 nm range are used to treat superficial tissues, while devices operating in the 780 to 950 nm range are used to treat deeper tissues .…”

“…Medical applications of LLLT span over a wide range of clinical settings, such as dentistry, dermatology, rheumatology and physiotherapy, as its use mainly relies on 3 main objectives, namely: wound healing, tissue repair and tissue death prevention, chronic joint disorders (CJD) treatment along with inflammation and edema relief, analgesia and neurological disorders treatment . …”

“…Photostimulation of this enzyme leads to an increase in cellular energy availability, oxygen consumption and signal transduction, along with a transient increase in ROS . The macroscopic result of such stimulation is strongly dependent upon cell type, cell growth phase and associated redox conditions .…”

“…LLLT induced cytochrome c oxidase/NO dissociation results in an enzyme conformation and redox state change that finally modifies the rate of the electron transfer along the respiration chain. As the displaced NO could not easily return to inhibit the enzyme, LLLT mediated effects can thus keep working for some time (hours as well days) after illumination .…”

Aging of lymphoid organs: Can photobiomodulation reverse age‐associated thymic involution via stimulation of extrapineal melatonin synthesis and bone marrow stem cells?

“…These papers demonstrate a variety of effects, including improved blood flow [29], increased mitochondrial activity [30], reduced expression of genes encoding inflammatory mediators [31], altered biochemical functions [26], mobilization, proliferation and differentiation of stem cells [32], neuromodulation [27–30], and a reduction in both acute and chronic pain [33], neuromodulation [34–37], and a reduced incidence of acute and chronic pain [38, 39]. …”