High energy shock waves enhance the cytotoxic effect of doxorubicin and methotrexate to human osteosarcoma cell lines

Abstract

Abstract. Despite recent advances, the prognosis of relapsed osteosarcoma patients remains very poor. Application of high energy shock waves may change the tumour cell growth and increase the cytotoxic effect of in vivo and in vitro chemotherapeutic agents. We studied the effect of their association with doxorubicin or methotrexate on three in vitro osteosarcoma cell lines. The effect of these combinations on SJSA-1, MG-63 and HOS human osteosarcoma cell lines were evaluated through incubation with doxorubicin or methotrexate and high energy shock wave treatment with 1000 shots at 0.22 mJ/mm 2 and an evaluation of the cell number, cell proliferation and apoptosis at days 1, 3 and 6 from the start of treatment. The combination of high energy shock waves and doxorubicin induced a statistically significant advantage in terms of a decrease in cell number on the SJSA-1 and HOS cell lines, a decrease of cell proliferation on all three cell lines and an increase of apoptosis only on the SJSA-1 cell line. The combination of high energy shock waves with methotrexate induced a decrease of the cell number only in the SJSA-1 and in the HOS cell lines, of the cell proliferation in the SJSA-1 and in the MG-63 cell lines, and an increase of apoptosis only on the SJSA-1 cell line. In conclusion, these experiments show an interesting effect of high energy shock waves on osteosarcoma cell lines, which could lead to future studies of the in vivo effects of high energy shock waves in the murine model as well. IntroductionOsteosarcoma (OS) is the most frequent bone tumour and is most common between the ages of 15 and 25. A high number of patients have a high-grade tumour, which prognosis has dramatically improved from 10-15% to 65-70% in the last 25 years, thanks to the use of adjuvant and neoadjuvant chemotherapy (1,2). Nevertheless, despite continuing scientific research into the treatment, the prognosis of the patients with OS in metastatic relapse, especially to bones (3), is still very poor; even with a recently concluded Italian and Scandinavian phase II protocol including high-dose chemotherapy the 3-year overall survival and the 3-year disease free-survival rates were only 20% and 12% respectively (4).The search of new treatments for improving the survival of the patients without increasing the collateral effects of the chemotherapy has led us to study new ways to increase the chemo-sensitivity of the OS cells.High energy shock waves (HESW) are routinely used in clinical practise in orthopaedic and in urology to treat a number of different diseases, such as urinary stones, non-unions of the bone fractures, tennis elbow, calcifying tendonitis, pseudoarthrosis and necrosis of the femoral head (5,6).In the past, many studies of HESW were conducted on different human and animal tumour cell lines, describing an alteration of the cell growth (7-11). The in vitro effects of shock waves are due to a suppression of cell proliferation correlated with an apoptotic cell death process; moreover, HESW treatment has been shown to cause a t...

Doxorubicin induces ZAKα overexpression with a subsequent enhancement of apoptosis and attenuation of survivability in human osteosarcoma cells

“…Furthermore, DOX treatment can decrease the number of human OS cells. However, the apoptotic effect of this treatment is not yet known, and the treatment can increase relative RNA expression of ZAK . To determine the effect of ZAKα and the DOX treatment on human OS cells, we treated human OS cells with DOX.…”

“…However, the role of ZAKα overexpression in human OS cells is not yet known. Recently, it was shown that DOX could provide a statistically significant advantage in terms of a decrease in cell number and cell proliferation but not in an increase in apoptosis in human OS cell lines . Based on previous data, we have formed a hypothesis that the DOX treatment can induce ZAK overexpression with a subsequent decrease in cell proliferation and increase in apoptosis in human OS cells.…”

Combining doxorubicin-nanobubbles and shockwaves for anaplastic thyroid cancer treatment: preclinical study in a xenograft mouse model

“…ESWs can be focused with high precision in depth, and they determine permeabilization of plasma membranes (Lauer et al 1997, Kodama et al 2002, Frairia et al 2003, Catalano et al 2007. As a consequence of ESWinduced cell permeability, ESWs increase the cytotoxic effects of different anti-cancer drugs (Frairia et al 2003, Palmero et al 2006, Catalano et al 2007, Canaparo et al 2008. All these features make ESWs an ideal tool to be used in combination with drug-loaded NBs.…”

Shock wave-induced ATP release from osteosarcoma U2OS cells promotes cellular uptake and cytotoxicity of methotrexate

“…Recent advances suggest that shock wave treatment may sensitize osteosarcoma cells to chemotherapeutic drugs [ 8 ]. Shock waves are elicited by a transient pressure disturbance and characterized by high positive pressure.…”