Medical Hydrology and Balneology: Environmental Aspects

Vetriolo's (Trentino, Italy) thermal water
modulate the negative effect of interleukin-1ß in
human osteoarthrtic chondrocytes
Fioravanti A (1) , Tenti S (1) , Lamboglia A (1) , De Nobili E (2) , Pascarelli N (1) ,
Moretti E (3) , Manica P (2) , Collodel G (3)
(1) Department of Clinical Medicine and Immunological Sciences, Rheumatology Unit,
University of Siena, Italy
(2) Terme di Levico e Vetriolo, Trentino, Italy
(3) Department of Biomedical Sciences, Applied Biology Section, University of Siena, Italy
fioravanti7@virgilio.it
Introduction and Objectives
Spa therapy is one of the most commonly used non-pharmacological approaches
for osteoarthrite (OA). The water “Debole di Vetriolo” (VW) is known for its
therapeutic properties in the treatment of OA. This is a highly mineralized water,
strongly acidic, sulfate, rich in iron, it is also rich in other minerals. The aim of our
study was to investigate the possible in vitro effects of the VW in human OA chondrocytes
cultivated in the presence or in the absence of Interleukin-1 beta (IL-1β).
Materials and Methods
Human OA chondrocytes were detached from the culture flasks by a mild trypsin
treatment and then seeded at 4x104 cells into wells containing 2.0 ml of either
DMEM medium, whose chemical constituents had been dissolved in Deionised
Water (DW) (controls in DW-DMEM), or in one four different in VW, in which
DW had been totally (100%) or in part (25% or 50%) substituted with VW. All
samples were analyzed before and after treatment with IL-1β at a concentration of 5
ng/ml Cells viability was evaluated by MTT assay, metalloproteinase-3 (MMP-3)
by a solid phase Enzyme Amplified Sensitivity Immunoassay (EASIA) and nitrite
concentrations by the Griess method. Immunocytochemistry was also performed to
localize the inducible nitric oxide synthase (iNOS). The incidence of apoptosis and
necrosis were evaluated by Annexin-V-FITC and Propidium Iodide assay and a
morphological analysis was performed by transmission electron microscopy (TEM).
Results
In chondrocyte cultures reconstituted with DW-DMEM or with VW, low levels
of MMP-3 and NO were detected at basal conditions, while in the presence of IL-1β
the levels of MMP-3 and NO were significantly increased (p