A sulfate-arsenical-ferruginous

Overview

This source page is a mechanical bulk-ingest record for a PDF in the research-pulls corpus. It preserves source-level identity, routeable product/analyte scope, and exact extracted numeric lines for later human or fresh-context audit. It does not derive HMTc thresholds, percentiles, or brand-by-brand comparisons.

Key numbers

The worker extracted the full PDF text with layout preservation twice and compared extraction hashes before commit. The following lines are copied from numeric/table-bearing regions of the PDF and retain the source units and wording where legible:

Collodel G, Moretti E, Carta S and Independent Researcher, Siena, Italy, 2 Health Resort of Levico and Vetriolo, Levico Terme, Trento,
bone sclerosis, and synovial membrane inﬂammation (22). Senese (Siena, Italy) (Supplementary Table S1). OA grades deﬁned
Multiple factors are involved in its complex pathogenesis, such as by Mankin degree, ranged from 3 to 7 (34).
in OA has been reported, particularly at its late stage (24). A role contained 10% fetal bovine serum (FBS) (Euroclone, Milan, Italy),
for noncoding RNAs, such as microRNA (miRNA), has also been 2% penicillin/streptomycin (P/S) (Sigma-Aldrich, Milan, Italy). OA
starting density of 1 × 105 cells/well, until 85% conﬂuence, in Cells were then harvested, collected into cytometry tubes, and
DMEM, 10% FBS, and 2% P/S. LW was provided by Levico Terme centrifuged. Then the pellet was dissolved in saline solution before
reported in Supplementary Table S2. expressed as median ﬂuorescence.
was added after 2 h of preincubation with 50% or 25% of LW,
chondrocytes were ﬁxed in 4% paraformaldehyde (ThermoFisher
as the mean ± standard deviation (SD) of triplicate values The incubation of the cells with 50% or 25% of LW alone, for
using 2CT values for each sample. All tests were provided treatment of the cells with LW at 50% (P < 0.05) (Figures 1a–c,
by the SAS System (SAS Institute Inc., Cary, NC, USA) 2a–c). Furthermore, 25% of LW resulted in being able to
Chondrocytes were incubated for 24 h with Levico water (LW), at 50% or 25%, in the presence or not of interleukin (IL)-1β (10 ng/ml). (a) Evaluation of
PCR. The data analysis was calculated as fold change to control culture (CTRL = equal to 100 or 1). Data were represented as mean ± standard
Chondrocytes were incubated for 48 h with Levico water (LW), at 50% or 25%, in the presence or not of interleukin (IL)-1β (10 ng/ml). (a) Evaluation of
PCR. The data analysis was calculated as fold change to control culture (CTRL = equal to 100 or 1). Data were represented as mean ± standard
that 50% or 25% of LW, tested alone at 24 h and 48 h, signiﬁcantly expression were shown in chondrocyte cultures stimulated with IL-
reduced the production of mitochondrial superoxide anion in 1β (Figure 3 and Figure 4). The pre-incubation of the cells with 50%
the analyzed antioxidant enzymes were found (Figures 1d–g, 2d–g). P < 0.01). 25% of LW was able to signiﬁcantly counteract IL-1β
LW at 50% or 25% tested alone, did not alter the transcriptional
As shown in Figure 3 and in Figure 4, 50% or 25% of LW, tested signiﬁcantly reduced by the pre-incubation of chondrocytes with
alone, didn’t modify the transcriptional levels of IL-1β, IL-6, and 50% of LW, for the expression of miR-34a, miR-140, and miR-181a,
quantitative real time PCR. The data analysis was calculated as fold change to control culture (CTRL = equal to 1). Data were represented as mean ±
(Figure 5, Figure 6). In contrast, the 50% LW concentration signiﬁcantly changed
this trend (Table 1, Figure 7E). Speciﬁcally, pre-treatment with
3.4 Immunoﬂuorescence analysis and gene signal (Table 1, Figure 7F).
To elucidate the potential mechanism by which LW might that the incubation of OA chondrocytes with 50% or 25% of LW,
of NF-κB pathway. subunits, while 50% of LW signiﬁcantly reduced the expression of
for 4 h. As described in the Materials and Methods section, p50 NF- cells with 50% of LW signiﬁcantly limited this trend (P < 0.01);
κB labeling was categorized as weak, medium, or strong (Table 1) 25% of LW reduced the eﬀects of IL-1β, even if not in a signiﬁcant
(Table 1, Figure 7B). No relevant changes in signal intensity were Based on the previous reported results, and to conﬁrm the
observed following treatment with 25% LW alone (Figure 7C) or possible role of NF-κB pathway in mediating LW induced activity,
quantitative real time PCR. The data analysis was calculated as fold change to control culture (CTRL = equal to 1). Data were represented as mean ±
1). Data were represented as mean ± standard deviation. **P < 0.01 versus CTRL; P < 0.05 versus IL-1β.
1). Data were represented as mean ± standard deviation. *P < 0.05, **P < 0.01 versus CTRL; P < 0.05, P < 0.01 versus IL-1β.
TABLE 1 Immunoﬂuorescence for p50 nuclear factor (NF)-κB was analyzed and classiﬁed into
intensity signal % signal % signal % 4 Discussion
25% of LW+ IL-1β 5 10 85 and in part, to its chemical composition (2–6).
50% of LW 85 10 5 In this preliminary study, we examined the potential biological
0% of LW+ IL-1β 10 55 35 properties of LW, tested as a whole, in human OA chondrocyte
(BAY 11–7082, IKKα/β), and then, incubated with 50% of LW for results in terms of viability of chondrocytes and was performed

Methods (brief)

PUBLISHED 20 April 2026 Giulia Collodel4 , Elena Moretti4 , Seraﬁno Carta5 and
Collodel G, Moretti E, Carta S and Independent Researcher, Siena, Italy, 2 Health Resort of Levico and Vetriolo, Levico Terme, Trento,
Collodel, Moretti, Carta and Cheleschi.
the biological mechanisms responsible for the activity of mineral 2.1 Sample collection and isolation of the
water (7–9). Human OA cartilage samples were collected from femoral
starting density of 1 × 105 cells/well, until 85% conﬂuence, in Cells were then harvested, collected into cytometry tubes, and
Health Resort (Levico Terme, Italy), directly collected from the ﬂow cytometry.
the samples arrived at our laboratory, they were analyzed for their assessment. For the detection of apoptotic cells, the analysis was
the samples were accurately ﬁltered using a 0.2 μm ﬁlter. Chemical to each dye, and the results were reported as percentage of
Following treatment procedure, cells were collected, and total
The Ct values of each sample were calculated and converted
sample/absorbance of control × 100. Results were referred to as
collected into cytometry tubes, and centrifuged. The pellet Biotechnology, Birmingham, AL, USA). Then, the coverslips were
for each of the samples collected and the results were expressed following experiments, in agreement with our previous study (35)
using 2CT values for each sample. All tests were provided treatment of the cells with LW at 50% (P < 0.05) (Figures 1a–c,
vitro studies have been performed on human cutaneous samples provided their written informed consent to participate in this study.
samples. The studies were conducted in accordance with the of the authors and do not necessarily represent those of
Evidence from in vitro studies performed on human and animal samples. Int J

Implications

This page makes the source discoverable for category-level evidence routing. Values remain source-native and should be used only with the stated matrix, species, basis, geography, and censoring context from the paper. The page does not convert total mercury to methylmercury or use total arsenic as inorganic arsenic.

Wiki pages this source may touch

Verification notes

Identity check: DOI, raw handle, candidate cite-key, and SHA-256 were compared against existing wiki/sources/ pages before creation.
Full-PDF read: pdftotext -layout was run on the full PDF twice; extracted text hashes matched before the page was written.
Numeric verification: numeric/table-bearing lines were selected mechanically from the verified extraction and preserved without unit conversion or rounding.
Brand firewall: the worker skips PDFs when extracted numeric lines appear brand/manufacturer-sensitive; this page contains category-level or species-level evidence only.
HMTc firewall: no threshold, percentile, pass/fail, clean/dirty, or certification math is stated.

Update history

The five most recent substantive edits to this page, classified major (evidence or structure moved), correction (a published value or statement was wrong and has been fixed), or minor (narrative rewritten without changing the underlying evidence). Each description is derived from what the edit did to this page; the linked commit is the authoritative record, routine regeneration passes are excluded, and the full version history lives in git. When DOI minting comes online (see schema docs), each entry below will also link to a version-pinned DataCite DOI.

Commit	Date	Change	Description
b01ec52c	2026-08-04	major	2 sections added
d49e450f	2026-08-03	major	5 sections added; narrative text revised