Cell-Cell Communication Between Fibroblast and 3T3-L1 Cells Under Co-culturing in Oxidative Stress Condition Induced by H2O2

Abstract

The present study was carried out to understand the interaction between fibroblast and 3T3-L1 preadipocyte cells under HO-induced oxidative stress condition. HO (40 μM) was added in co-culture and monoculture of fibroblast and 3T3-L1 cell. The cells in the lower well were harvested for analysis and the process was carried out for both cells. The cell growth, oxidative stress markers, and antioxidant enzymes were analyzed. Additionally, the mRNA expressions of caspase-3 and caspase-7 were selected for analysis of apoptotic pathways and TNF-α and NF-κB were analyzed for inflammatory pathways. The adipogenic marker such as adiponectin and PPAR-γ and collagen synthesis markers such as LOX and BMP-1 were analyzed in the co-culture of fibroblast and 3T3-L1 cells. Cell viability and antioxidant enzymes were significantly increased in the co-culture compared to the monoculture under stress condition. The apoptotic, inflammatory, adipogenic, and collagen-synthesized markers were significantly altered in HO-induced co-culture of fibroblast and 3T3-L1 cells when compared with the monoculture of HO-induced fibroblast and 3T3-L1 cells. In addition, the confocal microscopical investigation indicated that the co-culture of HO-induced 3T3-L1 and fibroblast cells increases collagen type I and type III expression. From our results, we suggested that co-culture of fat cell (3T3-L1) and fibroblast cells may influence/regulate each other and made the cells able to withstand against oxidative stress and aging. It is conceivable that the same mechanism might have been occurring from cell to cell while animals are stressed by various environmental conditions.

Fabrication of nanofiber coated with l-arginine via electrospinning technique: a novel nanomatrix to counter oxidative stress under crosstalk of co-cultured fibroblasts and satellite cells

“…Previous study showing that co-cultured satellite cells with fibroblasts could increase cell growth when compared to monoculture of satellite cells (Subramaniyan et al 2016). Similarly, PU-dextran nanofibrous scaffolds have increased cell growth of fibroblasts compared to PU scaffolds (Unnithan et al 2012).…”

“…Moreover, less cell viability was found in monoculture of satellite cells, indicating buildup of oxidative damage due to their inadequate ROS scavenging nature compared to co-culture (Rather et al 2018). Fibroblasts are the most common cells for collagen synthesis and SOD production during oxidative stress (Subramaniyan et al 2016). The same mechanism might have occurred in the treatment with L-arginine during co-culture results attenuate oxidative stress through the suppression of ROS production.…”

“…Such secreted molecules including SOD, adiponectin, and PPAR-c can downregulate apoptosis and inflammation through attenuation of caspases, TNF-a, and NF-jB. Adiponectin might be able to penetrate from adipocytes to other neighboring cells to improve cell growth under oxidative stress condition (Subramaniyan et al 2016). Co-culture of liver cells can elevate or reduce gene transcription compared to monoculture (Holen et al 2014).…”

“…Cytotoxicity and cell viability of nanofiber were measured by 2% trypan blue staining using published method of Subramaniyan et al (2016). The number of viable cells was estimated in each group by counting in a Neubauer chamber.…”

“…However, when ROS cellular overproduction overwhelms the intrinsic antioxidant capacity, oxidative stress will occur, causing damage to biomolecules of normal cells and tissues due to activation of apoptotic proteins and cellular necrosis (Atilano et al 2005). ROS encompass a variety of free radicals, including the superoxide anion (O 2À ), hydroxyl radical (OH ), singlet oxygen ( 1 O 2 ), and hydrogen peroxide (H 2 O 2 ) (Subramaniyan et al 2016). H 2 O 2 is a molecule can readily penetrate cell membranes and generate the most reactive form of oxygen and OH radical, could inducing oxidative stress in in vitro models (Schmidley 1990).…”