The Role of TGF-ß3 in Radiation Response.

Transforming growth factor-beta 3 (TGF-ß3) is a ubiquitously expressed multifunctional cytokine involved in a range of physiological and pathological conditions, including embryogenesis, cell cycle regulation, immunoregulation, and fibrogenesis. The cytotoxic effects of ionizing radiation are employed in cancer radiotherapy, but its actions also influence cellular signaling pathways, including that of TGF-ß3. Furthermore, the cell cycle regulating and anti-fibrotic effects of TGF-ß3 have identified it as a potential mitigator of radiation- and chemotherapy-induced toxicity in healthy tissue. This review discusses the radiobiology of TGF-ß3, its induction in tissue by ionizing radiation, and its potential radioprotective and anti-fibrotic effects.

Cytokine Levels in Saliva Are Associated with Salivary Gland Fibrosis and Hyposalivation in Mice after Fractionated Radiotherapy of the Head and Neck.

Cytokines are mediators of inflammation that could lead to fibrosis. The aim was to monitor cytokine levels in saliva and serum after locally fractionated radiotherapy of the head and neck in mice and investigate associations with salivary gland fibrosis and hyposalivation. C57BL/6 mice were randomized to sham or X-ray irradiation of 66 Gy in 10 fractions over 5 days. Blood and saliva were collected on days -7, 5, 35, 80, and 105 following cytokine analysis. The harvested submandibular salivary gland was assessed for the presence of fibrosis. Decision tree regression analysis was used to investigate whether cytokine levels could predict late endpoints in terms of hyposalivation or fibrosis. Significant formation of fibrosis in gland tissue and reduced saliva production was found after irradiation. The pro-inflammatory cytokines IL-1α, TNF, TIMP1, G-CSF, KC, and MIP-1α showed increased levels in saliva in irradiated mice and a strong correlation with late endpoints. The decision tree analysis largely separated controls from irradiated animals, with IL-1α being the strongest predictor. Pro-inflammatory cytokines in saliva, but not in serum, were associated with late endpoints. This indicates that cytokine expression in saliva is a good biomarker for local salivary gland damage with IL-1α as the strongest single predictor.

Low-Dose-Rate Radiation-Induced Secretion of TGF-ß3 Together with an Activator in Small Extracellular Vesicles Modifies Low-Dose Hyper-Radiosensitivity through ALK1 Binding.

Hyper-radiosensitivity (HRS) is the increased sensitivity to low doses of ionizing radiation observed in most cell lines. We previously demonstrated that HRS is permanently abolished in cells irradiated at a low dose rate (LDR), in a mechanism dependent on transforming growth factor ß3 (TGF-ß3). In this study, we aimed to elucidate the activation and receptor binding of TGF-ß3 in this mechanism. T-47D cells were pretreated with inhibitors of potential receptors and activators of TGF-ß3, along with addition of small extracellular vesicles (sEVs) from LDR primed cells, before their radiosensitivity was assessed by the clonogenic assay. The protein content of sEVs from LDR primed cells was analyzed with mass spectrometry. Our results show that sEVs contain TGF-ß3 regardless of priming status, but only sEVs from LDR primed cells remove HRS in reporter cells. Inhibition of the matrix metalloproteinase (MMP) family prevents removal of HRS, suggesting an MMP-dependent activation of TGF-ß3 in the LDR primed cells. We demonstrate a functional interaction between TGF-ß3 and activin receptor like kinase 1 (ALK1) by showing that TGF-ß3 removes HRS through ALK1 binding, independent of ALK5 and TGF-ßRII. These results are an important contribution to a more comprehensive understanding of the mechanism behind TGF-ß3 mediated removal of HRS.

Effects of ionizing irradiation and interface backscatter on human mesenchymal stem cells cultured on titanium surfaces.

Radiotherapy to the head and neck region negatively influences the osseointegration and survival of dental implants. The effects of cobalt 60 (60 Co) ionizing radiation and the impact of backscatter rays were investigated on human mesenchymal stem cells cultured on titanium surfaces. Bone marrow-derived human mesenchymal stem cells were seeded on titanium (Ti), fluoride-modified titanium (TiF), and tissue culture plastic. Cells were exposed to ionizing γ-radiation in single doses of 2, 6, or 10 Gy using a 60 Co source. Density and distribution of cells were evaluated using confocal laser-scanning microscopy, 21 d post-irradiation. Lactate dehydrogenase concentration and the levels of total protein and cytokines/chemokines were measured in the cell-culture medium on days 1, 3, 7, 14, and 21 post-irradiation. Unirradiated cells were used as the control. Irradiation had no effect on cell viability, collagen and actin expression, or cell distribution, but induced an initial increase in the secretion of interleukin (IL)-6, IL-8, monocyte chemotactic protein 1 (MCP-1), and vascular endothelial growth factor (VEGF), followed by a decrease in secretion after 3 or 7 d. Irradiation resulted in secretion of a lower amount of all analytes examined compared with controls on day 21, irrespective of radiation dose and growth surface. Backscattering from titanium did not influence the cell response significantly, suggesting a clinical potential for achieving successful osseointegration of dental implants placed before radiotherapy.