Antitumor efficacy of silver nanoparticles reduced with β-D-glucose as neoadjuvant therapy to prevent tumor relapse in a mouse model of breast cancer

Introduction: Neoadjuvant therapy constitutes a valuable modality for diminishing tumor volume prior to surgical resection. Nonetheless, its application encounters limitations in the context of recurrent tumors, which manifest resistance to conventional treatments. Silver nanoparticles (AgNPs) have emerged as a promising alternative for cancer treatment owing to their cytotoxic effects.Methods: Cellular viability was assessed by Alamar blue assay in 4T1 breast cancer cell line. Silver biodistribution was detected by an inductively coupled plasma optical emission spectrometer in an in vivo mice model. For neoadjuvant evaluation, mice were randomized and treated intratumoral with AgNPs-G or intraperitoneally with doxorubicin (DOX) as a control. Recurrence was determined after 170 days by counting lung metastatic nodules (dyed with Bouin solution) with histological confirmation by H&E. Masson’s stain, Ki67 immunohistochemistry, and a TUNEL assay were performed in lungs from treated mice.Results: AgNPs-G reduced 4T1 cell viability and in an ex vivo assay the AgNPs-G decreased the tumor cell viability. After intravenous administration of AgNPs-G were detected in different organs. After intratumor administration, AgNPs-G are retained. The AgNPs-G treatment significantly reduced tumor volume before its surgical resection. AgNPs-G reduced the development of lung metastatic nodules and the expression of Ki67. TUNEL assay indicated that AgNPs-G didn’t induce apoptosis.Conclusions: We concluded that intratumor administration of AgNPs-G reduced tumor volume before surgical resection, alongside a reduction in lung metastatic nodules, and Ki67 expression. These findings provide valuable insights into the AgNPs-G potential for intratumor and neoadjuvant cancer therapies. However, further research is needed to explore their full potential and optimize their use in clinical settings

Development of a new generation of miniemulsion based on cottonseed oil with α-tocopherol and ZnO and evaluation of its adjuvant activity

Background Emulsions have been widely used as immunological adjuvants. But the use of materials derived from plants such as cottonseed oil, alpha-tocopherol, or minerals such as zinc, as well as their use at the nanometric scale has been little explored. In this study, we develop a new miniemulsion and evaluated its antioxidant and phagocytic capacity, as well as parameters related to immune response stimulation by cytokine expression and antibodies production in a mice model. Methods Formulated CN (cottonseed oil miniemulsion) and CNZ (cottonseed oil miniemulsion whit zinc oxide nanoparticles) miniemulsions were characterized by scanning electronic microscopy SEM, DLS and FT-IR. In murine macrophages, splenocytes and thymocytes primary cultures safety and cytotoxicity were determined by MTT. In macrophages the antioxidant and phagocytic capacity was evaluated. In BALB/c mice, the stimulation of the immune system was determined by the expression of cytokines and the production of antibodies. Results The CN and CNZ presented stability for 90 days. Immediately after preparation, the CN presented a higher particle size (543.1 nm) than CNZ (320 nm). FT-IR demonstrated the correct nanoparticle synthesis by the absence of sulfate groups. CN and CNZ (1.25 to 10 µL/mL) had no toxic effect on macrophages (p = 0.108), splenocytes (p = 0.413), and thymocytes (p = 0.923). All CN and CNZ doses tested induced nitric oxide and antioxidants production in dose dependent manner when compared with control. CN-ovalbumin and CNZ-ovalbumin treatments in femoral subcutaneous tissue area showed inflammation with higher leukocyte infiltration compared with FCA. The intraperitoneal administration with CN, CNZ, and FCA showed a higher total intraperitoneal cells recruitment (CD14+) after 24 h of inoculation than control (p = 0.0001). CN and CNZ increased the phagocyte capacity with respect to untreated macrophages in the Candida albicans-phagocytosis assay. The evaluation of residual CFU indicated that only CN significantly decreased (p = 0.004) this value at 3 h. By other side, only CN increased (p = 0.002) the nitric oxide production. CNZ stimulated a major INFγ secretion compared with FCA at day 7. A major IL-2 secretion was observed at days 7 and 14, stimulated with CN and CNZ. Both miniemulsions did not affect the antibody isotypes production (IgG1, IgG2a, IgG3, IgA and IgM) at days 7, 14, 28, and 42. CN induced a significant IgG production against OVA, but lesser than FCA. Conclusions The two new miniemulsions with adjuvant and antioxidant capacity, were capable of generating leukocyte infiltration and increased cytokines and antibodies production

Values for the categorical (sex, IgG) and continuous variables (glucose, urea, creatinine, uric acid, BMI, WHR, MAP BR, HR and temperature) of the cohort.

Values for the categorical (sex, IgG) and continuous variables (glucose, urea, creatinine, uric acid, BMI, WHR, MAP BR, HR and temperature) of the cohort.</p

Fig 2 -

Box-whisker charts of MANOVA scores, for (a) the factor Susceptibility (p>0.05), (b) the factor Sex (p<0.001) and (c) the interaction Susceptibility × Sex (p = 0.033). Baseline values are shown for male and female participants who remained seronegative (M- and F-), or who were found to have been infected (M+ and F+), during a follow-up study six months later. Outliers are shown explicitly (black dots).</p

Composition of the cohort in terms of male (M) and female participants (F), and post-hoc defined groups of participants that remain seronegative (IgG-) or become seropositive (IgG+) over the course of 6 months.

Composition of the cohort in terms of male (M) and female participants (F), and post-hoc defined groups of participants that remain seronegative (IgG-) or become seropositive (IgG+) over the course of 6 months.</p

Baseline values for anthropometric, biochemical, and physiological variables of the groups of male and female participants that remain seronegative (M- and F-, respectively) and become seropositive over the course of 6 months (M+ and F+, respectively).

All variables, except cholesterol, are non-normal for at least one of the groups, and therefore are reported in standard units as Q2(Q1, Q3) (upper half of the table), where Q2 is second quartile, i.e., the median, Q1 y Q3 are first and third quartiles respectively. Non-normal data is log transformed and represented as mean (SD) (bottom half of the table), where SD is standard deviation. For completeness, cholesterol is given in both formats without log transformation. Statistically significant factors and/or interaction using 2-way ANOVA and statistically significant pairwise group comparisons using the Mann-Whitney U test are also indicated for individual variables.</p

Fig 3 -

Box-whisker charts of values of the continuous variables that show significant pairwise differences between groups using the Mann-Whitney U test, (a) waist-to-hip ratio (WHR), (b) mean arterial pressure (MAP), (c) creatinine, (d) uric acid, (e) urea, and (f) breathing rate (BR). Baseline values are shown for male and female participants who remained seronegative (M- and F-), or who were found to have been infected (M+ and F+), during a follow-up study six months later. Normative ranges of reference values are indicated for male (blue-shaded background) and female participants (pink-shaded background). Outliers are shown explicitly (black dots).</p

Flowchart of the selection process of the participants based on the inclusion and exclusion.

Flowchart of the selection process of the participants based on the inclusion and exclusion.</p