Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2023 Sep 19;15(9):2340.
doi: 10.3390/pharmaceutics15092340.

A New Paclitaxel Formulation Based on Secretome Isolated from Mesenchymal Stem Cells Shows a Significant Cytotoxic Effect on Osteosarcoma Cell Lines

Alessia Giovanna Santa Banche Niclot  1 , Elena Marini  1 , Ivana Ferrero  2 , Francesco Barbero  3 , Elena Rosso  4 , Ivana Fenoglio  3 , Alessandro Barge  4 , Augusto Pessina  5 , Valentina Coccè  5 , Francesca Paino  5 , Katia Mareschi  1   2 , Franca Fagioli  1   2
Affiliations

A New Paclitaxel Formulation Based on Secretome Isolated from Mesenchymal Stem Cells Shows a Significant Cytotoxic Effect on Osteosarcoma Cell Lines

Alessia Giovanna Santa Banche Niclot et al. Pharmaceutics. .

Abstract

Background: Osteosarcoma (OS) represents a rare cancer with an unfavorable prognosis that needs innovative treatment. The aim was to isolate a secretome from mesenchymal stem cells (MSCs) that are treated with paclitaxel (PTX)-containing microvesicles as a drug delivery system and analyze its cytotoxic effects on OS cell lines (SJSA, MG63, and HOS).

Methods: Three batches of secretome (SECR-1, SECR-2, and SECR-3) were produced from three bone marrow (BM) MSCs samples treated for 24 h with 15 µg/mL of PTX or with a standard medium. The viability of the OS cell lines after 5 days of exposure to SECR-1-2-3 (pure and diluted to 1:2 and 1:4) was analyzed with an MTT assay. The same SECR batches were analyzed with high-performance liquid chromatography (HPLC) and with a nanoparticle tracking assay (NTA).

Results: A statistically significant decrease in the viability of all OS cell lines was observed after treatment with SECR-PTX 1-2-3 in a dose-response manner. The NTA analyses showed the presence of nanoparticles (NPs) with a mean size comparable to that of extracellular vesicles (EVs). The HPLC analyses detected the presence of PTX in minimal doses in all SECR batches.

Conclusions: This proof-of-concept study showed that the conditioned medium isolated from MSCs loaded with PTX had a strong cytotoxic effect on OS cell lines, due to the presence of EV and PTX.

Keywords: drug delivery system; mesenchymal stem cells; osteosarcoma; paclitaxel; secretome.

PubMed Disclaimer

Conflict of interest statement

The authors declare no conflict of interest. The funders had no role in the design of the study; in the collection, analyses, or interpretation of data; in the writing of the manuscript, or in the decision to publish the results.

Figures

Figure 1
Figure 1
Immunophenotypic analysis of three batches of BM-MSCs, before and after PTX treatment.
Figure 2
Figure 2
The means of three experiments to perform a viability test with an MTT assay on three OS cell lines: (A) SJSA; (B) MG63; (C) HOS, n = 3. CTRL Viability: OS cell lines treated with standard medium; CTRL Mortality: cells treated with 30 µg/mL of PTX; SECR-CTRL: the condition after treatment of three batches of secretome, isolated from untreated MSCs (SECR-CTR-1,2,3), and after the loading of PTX (SECR-PTX-1,2,3), either pure or diluted to 1:2 and 1:4. The symbols **, ***, and **** indicate a significant difference, with p-values of <0.01, <0.001, and <0.0001, respectively.
Figure 2
Figure 2
The means of three experiments to perform a viability test with an MTT assay on three OS cell lines: (A) SJSA; (B) MG63; (C) HOS, n = 3. CTRL Viability: OS cell lines treated with standard medium; CTRL Mortality: cells treated with 30 µg/mL of PTX; SECR-CTRL: the condition after treatment of three batches of secretome, isolated from untreated MSCs (SECR-CTR-1,2,3), and after the loading of PTX (SECR-PTX-1,2,3), either pure or diluted to 1:2 and 1:4. The symbols **, ***, and **** indicate a significant difference, with p-values of <0.01, <0.001, and <0.0001, respectively.
Figure 3
Figure 3
Mean size and size distribution of SECR, obtained via nanoparticle tracking analysis (NTA). (A) Mean size distribution of SECR-CTRL-1/2/3 (orange) and SECR-PTX-1/2/3 (green). (B) Size distribution of SECR-CTRL-1, not labeled (red) and labeled (blue) with the fluorescent membrane stain, CMG. (C) Size distribution of SECR-CTRL-1, both not labeled (red) and labeled (blue) with the fluorescent membrane stain, CMG. The symbols **** indicate a significant difference with a p-value of <0.0001.
Figure 4
Figure 4
The linear relationship between paclitaxel concentration in EVs (PTX/EVs Tot ng/mL) and viability (%) in three OS cell lines (SJSA, MG63, and HOS), which were treated with three batches of secretome (SECR-1, SECR-2, and SECR-3). The dotted line refers to the quantity of PTX/EVs and the straight line is the trend line.
See this image and copyright information in PMC

References

    1. Hingorani P., Janeway K., Crompton B.D., Kadoch C., Mackall C.L., Khan J., Shern J.F., Schiffman J., Mirabello L., Savage S.A., et al. Current State of Pediatric Sarcoma Biology and Opportunities for Future Discovery: A Report from the Sarcoma Translational Research Workshop. Cancer Genet. 2016;209:182–194. doi: 10.1016/j.cancergen.2016.03.004. - DOI - PMC - PubMed
    1. Anderson M.E. Update on Survival in Osteosarcoma. Orthop. Clin. N. Am. 2016;47:283–292. doi: 10.1016/j.ocl.2015.08.022. - DOI - PubMed
    1. Ingley K.M., Maleddu A., Grange F.L., Gerrand C., Bleyer A., Yasmin E., Whelan J., Strauss S.J. Current Approaches to Management of Bone Sarcoma in Adolescent and Young Adult Patients. Pediatr. Blood Cancer. 2022;69:e29442. doi: 10.1002/pbc.29442. - DOI - PubMed
    1. Lenna S., Bellotti C., Duchi S., Martella E., Columbaro M., Dozza B., Ballestri M., Guerrini A., Sotgiu G., Frisoni T., et al. Mesenchymal Stromal Cells Mediated Delivery of Photoactive Nanoparticles Inhibits Osteosarcoma Growth in Vitro and in a Murine in Vivo Ectopic Model. J. Exp. Clin. Cancer Res. 2020;39:40. doi: 10.1186/s13046-020-01548-4. - DOI - PMC - PubMed
    1. Yoon S.J., Moon Y.J., Chun H.J., Yang D.H. Doxorubicin·Hydrochloride/Cisplatin-Loaded Hydrogel/Nanosized (2-Hydroxypropyl)-Beta-Cyclodextrin Local Drug-Delivery System for Osteosarcoma Treatment In Vivo. Nanomaterials. 2019;9:E1652. doi: 10.3390/nano9121652. - DOI - PMC - PubMed