Effect of transforming growth factor-β2 on biological regulation of multilayer primary chondrocyte culture

Abstract

Cytokines are extremely potent biomolecules that regulate cellular functions and play multiple roles in initiation and inhibition of disease. These highly specialised macromolecules are actively involved in control of cellular proliferation, apoptosis, cell migration and adhesion. This work, investigates the effect of transforming growth factor-beta2 (TGF-β2) on the biological regulation of chondrocyte and the repair of a created model wound on a multilayer culture system. Also the effect of this cytokine on cell length, proliferation, and cell adhesion has been investigated. Chondrocytes isolated from knee joint of rats and cultured at 4 layers. Each layer consisted of 2 × 105 cells/ml with and without TGF-β2. The expression of mRNA and protein levels of TGF-β receptors and Smad1, 3, 4, and 7 have been analysed by RT-PCR and western blot analysis. The effect of different supplementations in chondrocyte cell proliferation, cell length, adhesion, and wound repair was statistically analysed by One-way ANOVA test. Our results showed that the TGFβ2 regulates mRNA levels of its own receptors, and of Smad3 and Smad7. Also the TGF-β2 caused an increase in chondrocyte cell length, but decreased its proliferation rate and the wound healing process. TGF-β2 also decreased cell adhesion ability to the surface of the culture flask. Since, TGF-β2 increased the cell size, but showed negative effect on cell proliferation and adhesion of CHC, the effect of manipulated TGF-β2 with other growth factors and/or proteins needs to be investigated to finalize the utilization of this growth factor and design of scaffolding in treatment of different types of arthritis.

Lumican mediates HTB94 chondrosarcoma cell growth via an IGF‑IR/Erk1/2 axis

“…TGF-β2 has previously been shown to negatively mediate the growth of physiological chondrocytes ( 28 , 29 ) and human chondrosarcoma cells ( 30 ). In the present study, treatment of the HTB94 cells with TGF-β2 resulted in a potent decrease in growth (P≤0.001; Fig.…”

The role of TGF-β2 in cartilage development and diseases

“…Therefore, these findings indicated that the role of TGF-β2 in promoting cartilage defect repair was very important. On the contrary, Khaghani et al 96 indicated that TGF-β2 repressed the wound healing process of chondrocytes by scratch assay. This meant that TGF-β2 might have a negative regulatory effect on chondrocyte migration.…”

“…Interestingly, in recent studies, TGF-β2 had a negative effect on cell proliferation. For instance, Khaghani et al 96 showed that TGF-β2 exerted an inhibitory effect on chondrocyte cell proliferation and might have synergistic effects on joint chondrocytes with other growth factors or proteins, such as basic fibroblast growth factor (bFGF). 97 Moreover, cyclin B2 (the onset of proliferation) and proliferating cell nuclear antigen (PCNA) are regarded as proliferation markers.…”

Growth Factors, Reactive Oxygen Species, and Metformin—Promoters of the Wound Healing Process in Burns?

“…The TGF-β superfamily includes three isoforms, TGF-β1, -β2, and -β3, coded by genes with different locations in various chromosomes [86]. TGF-β are soluble extracellular proteins, which regulate the development in both vertebrates and invertebrates [87]. TGF-α belongs to the EGF family; it is a 50-amino-acid polypeptide, which in humans has 10 members, isolated from a conditioned medium of virally transformed cells and tumor cells [88].…”

“…TGF-α belongs to the EGF family; it is a 50-amino-acid polypeptide, which in humans has 10 members, isolated from a conditioned medium of virally transformed cells and tumor cells [88]. TGF-β members regulate cell functions such as migration, apoptosis, proliferation, and differentiation [87]. TGF-β is secreted in a latent form by most cells and is activated via TGF-β receptor signaling [89].…”