Appendix A. Supplementary data

Curcumin-loaded silica-based mesoporous materials: Synthesis, characterization and cytotoxic properties against cancer cells

Vishnu Sravan Bollu a,1, Ayan Kumar Barui a,1, Sujan Kumar Mondal a,1, Sanjiv Prashar b,

Mariano Fajardo b, David Briones c, Antonio Rodríguez-Diéguez c, Chitta Ranjan Patra a,1,*,

Santiago Gómez-Ruiz b,*

aBiomaterials Group, CSIR-Indian Institute of Chemical Technology, Uppal Road,

Tarnaka, Hyderabad - 500007, IndiabDepartamento de Biología y Geología, Física y Química Inorgánica, E.S.C.E.T., Universidad

Rey Juan Carlos, Calle Tulipán s/n, 28933, Móstoles (Madrid), SpaincDepartamento de Química Inorgánica, Universidad de Granada, Facultad de Ciencias, Fuente

Nueva s/n 18071 Granada, Spain1Academy of Scientific and Innovative Research (AcSIR), 2 Rafi Marg, New Delhi, India

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1. Dynamic light scattering study (DLS): Average particle size

Fig. S1(a-f). Size distribution of all mesoposours silica materials (V1-V6).

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2. Dynamic light scattering study (DLS): Charge (zeta potenial)

Fig. S2(a-f). Charge of all mesoporous silica materials (V1-V6).

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3. Electron microscopic images of MSU-2 and MCM-41

Fig. S3. SEM images of materials MSU-2 and MCM-41.

Fig. S4. TEM images of materials MSU-2 and MCM-41.

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4. Nitrogen adsorption-desorption isotherms

Fig. S5. Nitrogen adsorption-desorption isotherms of materials V4-V6 [y axis = Adsorbed volume (cm3/g); x axis = Relative presure P/P0].

5. Thermogravimetry analysis

Fig. S6. TG measurements of materials V1-V6. Exothermic processes take place when the heat flow increases.

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6. X-ray diffraction patterns

Fig. S7. X-ray diffraction patterns of materials V1-V6 [x axis = 2 Theta (º); y axis = Intensity (a. u.).

7. FT-IR measurements

Fig. S8. Example of FT-IR spectra of material MSU-2 and V3.

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8. Biological experiments

Table S1: Determination of cellular uptake of mesoporous silica materials in A549 and CHO cells using ICP-OES analysis.

MaterialSi (pg/cell)

A549 CHO

V1 4.3 2.56

V2 1.17 1.47

V3 22.5 12.4

V4 3.21 2.8

V5 3.33 0.84

V6 3.47 1.44

Fig. S9: Kinetic study for the internalization of curcumin loaded mesoporous (V3) material in A549 cells. a-a’’: partial enlarged images of (V3: 10 µM w.r.t curcumin, 12 h); b-b’’: partial enlarged images of (V6: 10 µM w.r.t curcumin, 12 h). Scale bar = 10 microns.

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Fig. S10 (a-b). Scratching assay in A549 cells with cyto-toxic dose of V3 and V6 materials. (a) This study reveals that the untreated control (UT) cells rapidly migrate toward the wound region. However, the wound area has remained unaltered in cells treated with V3 and V6 indicating their ability in inhibition of cancer cell migration. Row1: untreated control cells (a’-a’’’: UT); Row 2: cells treated with V3 (b’-b’’’: 10 µM w.r.t curcumin); Row 3: cells treated with V6 (c’-c’’’:10 µM w.r.t curcumin). (b) Quantification of wound area in a time span of 0-24 h using Image J Analysis software (NIH, Bethesda, MD, USA). Scale bar = 50 microns.

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Fig. S11 (a-b). Scratching assay in A549 cells with sub-toxic dose of V3 and V6 materials. (a) This study reveals that the untreated control cells (UT) rapidly migrate toward the wound region. However, the wound area has remained quite unaltered in cells treated with V3 and V6 indicating their ability in inhibition of cancer cell migration. Row 1: untreated control cells (a’-a’’’: UT); Row 2: cells treated with V3 (b’-b’’’: 5 µM w.r.t curcumin); Row 3: cells treated with V6 (c’-c’’’: 5 µM w.r.t curcumin). (b) Quantification of wound closure in a time span of 0-24 h using Image J Analysis software (NIH, Bethesda, MD, USA). Scale bar = 50 microns.

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Fig. S12. Determination of intracellular superoxide anion radical by fluorescence microscopy in A549 cells. The result shows that A549 cells treated with V1, V2, V4 and V5 show poor red fluorescence indicating the absence or presence of very lower level of super oxide ion radicals. (a-a’): untreated control cells; b-b’: cells treated with V1; c-c’: cells treated with V2; d-d’: cells treated with V4; e-e’: cells treated with V5.

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