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. 2023 Jun 9;9(6):475.
doi: 10.3390/gels9060475.

Collagen Hydrolysate Effects on Photodynamic Efficiency of Gallium (III) Phthalocyanine on Pigmented Melanoma Cells

Vanya Mantareva  1 , Ivan Iliev  2 , Inna Sulikovska  2 , Mahmut Durmuş  3 , Tsanislava Genova  4
Affiliations

Collagen Hydrolysate Effects on Photodynamic Efficiency of Gallium (III) Phthalocyanine on Pigmented Melanoma Cells

Vanya Mantareva et al. Gels. .

Abstract

The conjugation of photosensitizer with collagen seems to be a very promising approach for innovative topical photodynamic therapy (PDT). The study aims to evaluate the effects of bovine collagen hydrolysate (Clg) on the properties of gallium (III) phthalocyanine (GaPc) on pigmented melanoma. The interaction of GaPc with Clg to form a conjugate (GaPc-Clg) showed a reduction of the intensive absorption Q-band (681 nm) with a blue shift of the maximum (678 nm) and a loss of shape of the UV-band (354 nm). The fluorescence of GaPc, with a strong emission peak at 694 nm was blue shifted due to the conjugation which lower intensity owing to reduce quantum yield (0.012 vs. 0.23, GaPc). The photo- and dark cytotoxicity of GaPc, Glg and GaPc-Clg on pigmented melanoma cells (SH-4) and two normal cell lines (BJ and HaCaT) showed a slight decrease of cytotoxicity for a conjugate, with low selectivity index (0.71 vs. 1.49 for GaPc). The present study suggests that the ability of collagen hydrolysate to form gels minimizes the high dark toxicity of GaPc. Collagen used for conjugation of a photosensitizer might be an essential step in advanced topical PDT.

Keywords: bovine collagen hydrolysate and gels; gallium phthalocyanine; mouse embryonal fibroblasts; normal keratocytes and fibroblasts; photodynamic therapy; pigmented melanoma.

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Conflict of interest statement

The authors declare no conflict of interest. The funder 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

Scheme 1
Scheme 1
Synthetic pathways for synthesis of Gallium (III) phthalocyanine (GaPc, 3).
Figure 1
Figure 1
Absorption spectra of Ga(III)-phthalocyanine (GaPc) in: dimethylsulfoxide (DMSO) and phosphate-buffered saline (PBS) for 6 µM GaPc (a) and in PBS with hydrolyzed collagen (Clg) for a range of concentrations for a constant concentration of 6 µM GaPc (b).
Figure 2
Figure 2
Fluorescence spectra: (a) collagen hydrolysate (Clg) in acetic acid (0.05% acetic acid) and phosphate-buffered saline (PBS, pH 7.2) at exc: 226 nm; and (b) conjugate with Clg (GaPc–Clg) at exc: 615 nm.
Figure 3
Figure 3
Dark and photo-cytotoxicity: (a) Ga (III) phthalocyanine (GaPc), bovine collagen hydrolysate (Clg) and the conjugate of both (GaPc–Clg), and (b) Clg, all studied on embryonal BALB 3T3 cell line (exposure with solar light-emitting diode, LED 360 nm–850 nm).
Figure 4
Figure 4
Dark and photo-cytotoxicity of Gallium(III) phthalocyanine (GaPc), and the conjugate GaPc–Clg on melanoma cell line (SH-4) versus normal cell lines BJ (a); and HaCaT (b). Light from a light-emitting diode (LED) at 660 nm with a light dose of 50 J/cm2.
Figure 5
Figure 5
Dark and photo-cytotoxicity of bovine collagen hydrolysate (Clg) on SH-4 vs. BJ and HaCaT. Irradiation from a light-emitting diode (LED) at 660 nm with a light dose of 50 J/cm2.
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