Porphyrin Derivative Conjugated with Gold Nanoparticles for Dual-Modality Photodynamic and Photothermal Therapies In Vitro

Gold nanoparticles (Au NPs) have been confirmed to show excellent photothermal conversion property for tumor theranostic applications. To improve the antitumor efficacy, a novel nanoplatform system composed of porphyrin derivative and Au NPs was fabricated to study the dual-modality photodynamic and photothermal therapy with laser irradiation. Modified chitosan was coated on the Au NPs surface via ligand exchange between thiol groups and Au. The chitosan-coated Au NPs (QCS-SH/Au NPs) were further conjugated with meso-tetrakis­(4-sulphonatophenyl)­porphyrin (TPPS) via electrostatic interaction to obtain the porphyrin-conjugated Au hybrid nanoparticles (TPPS/QCS-SH/Au NPs). Size, morphology, and properties of the prepared nanoparticles were confirmed by Zeta potential, nanoparticle size analyzer, transmission electron microscopy (TEM), and UV–vis spectroscopy. Moreover, both photothermal therapy (PTT) and photodynamic therapy (PDT) were investigated. Compared with alone Au NPs or TPPS, the hybrid TPPS/QCS-SH/Au NPs with lower cytotoxicity showed durable elevated temperature to around 56 °C and large amount of singlet oxygen (¹O₂) produced from TPPS. Thus, the hybrid nanoparticles showed a more significant synergistic therapy effect of hyperthermia from PTT as well as ¹O₂ from PDT, which has potential applications in the tumor therapy fields

Key findings.

<p>While ROS activation by IS and PCS have been widely studied, we have shown in this study that IS and PCS activates ASK1, a ROS-driven protein kinase, and its downstream MAPKs (p38MAPK and ERK1/2) as well as NF-κB, leading to the upregulation of fetal genes (α-SkM-Ac and β-MHC) to promote cardiac hypertrophy and pro-fibrotic genes (TGF-β1 and <i>ctgf</i>) to cause cardiac and renal fibrosis.</p

Le Courrier

27 juillet 18331833/07/27 (N208)

Signaling pathway activation in <i>p</i>-cresol sulfate-stimulated renal mesangial cells.

<p>(A) Representative images of the Western blot analyses of RMCs stimulated with PCS and co-treated with (A) 100 μM Probenecid and 1.0 μM G226 for (i) phospho-ASK1, (ii) phospho-p38 (*<i>p</i> = 0.012 IS [10 μM] vs control, unpaired t-test), (iii) phospho-ERK1/2, (iv) phospho-NF-κB; and (B) 3.0 μM RWJ-67657 and 1.0 μM U0126 for (i) phospho-p38, (ii) phospho-ERK1/2, (iii) phospho-NF-κB and (iv) Pan Actin. Data are presented as mean ± SEM (n = 3). *<i>p</i><0.05, **<i>p</i><0.01 vs control, ^#<i>p</i><0.05, ^##<i>p</i><0.01 vs PCS [100 μM], One-way Anova.</p

Effects of <i>p</i>-cresol sulfate on cardiac cell viability.

<p>PCS (0.0001 to 100 μM) did not alter NCF viability as determined by MTT assay. Data are presented as mean ± SEM, each with quadruplets from three independent experiments and analyzed with One-way Anova.</p

Effects of indoxyl sulfate and <i>p</i>-cresol sulfate stimulation with and without selective ASK1 inhibitor on HK2 cell and neonatal cardiac fibroblast viability.

<p>MTT assay showed both IS and PCS stimulations (10 and 100 μM, respectively) (A) do not alter cellular viability of HK2 cell and NCFs in the presence and absence of G226 (HK2: 0.1 to 3.0 μM; NCF: 0.03 to 1.0 μM). Data are presented as mean ± SEM of quadruplets from three independent experiments and analyzed with One-way Anova.</p

Effects of <i>p</i>-cresol sulfate on cardiac myocyte hypertrophy and fibroblast collagen synthesis.

<p> PCS significantly induced NCM hypertrophy starting from the lowest dose (0.001 μM, <i>p</i><0.01 vs control) (A) and NCF collagen synthesis (B) starting at 0.003 (<i>p</i><0.01 vs control). Ang-II has been included as a positive control. Data are presented as mean ± SEM of triplicates from three experiments. *<i>p</i><0.05, **<i>p</i><0.01, ***<i>p</i><0.001, ****<i>p</i><0.0001 vs control, One-way Anova.</p

Signaling pathway activation in <i>p</i>-cresol sulfate-stimulated neonatal cardiac myocytes.

<p>Representative images and quantification of Western blot analyses of NCMs stimulated with PCS in the absence and presence of (A) 100 μM Probenecid and 1.0 μM G226 for (i) phospho-ASK1, (ii) phospho-p38, (iii) phospho-ERK1/2, (iv) phospho-NF-κB; and (B) 3.0 μM RWJ-67657 and 1.0 μM U0126 for (i) phospho-p38, (ii) phospho-ERK1/2, (iii) phospho-NF-κB and (iv) Pan Actin. Data are presented as mean ± SEM (n = 3). *<i>p</i><0.05, **<i>p</i><0.01, ***<i>p</i><0.001 vs control, ^#<i>p</i><0.05, ^##<i>p</i><0.01, ^###<i>p</i><0.001 vs PCS [100 μM], One-way Anova.</p

Effects of indoxyl sulfate and <i>p</i>-cresol sulfate stimulated on pro-hypertrophic gene expression of NCMs and pro-fibrotic gene expression of NCFs and HK2 cells.

<p>IS (10 μM) and PCS (100 μM) upregulated gene expression of α-SkM-Ac and β-MHC in NCMs (A, i-iv) and TGF-β1 and <i>ctgf</i> in NCFs (B, i-iv) and HK2 cells (C, i-iv), which were suppressed by G226, Probenecid, RWJ-67657 and U0126. *<i>p</i><0.05, **<i>p</i><0.01, ***p<0.001 IS [10 μM] or PCS [100 μM] vs control, ^#<i>p</i><0.05, ^##<i>p</i><0.01, ^###<i>p</i><0.001^####<i>p</i><0.0001 vs IS [10 μM] or PCS [100 μM], One-way Anova. These results are representatives of two independent experiments in triplicates.</p

Signaling pathway activation in indoxyl sulfate-stimulated renal mesangial cells.

<p>Representative images and quantification of Western blot analyses of RMCs stimulated with IS and co-treated with (A) 100 μM Probenecid and 1.0 μM G226 for (i) phospho-ASK1, (ii) phospho-p38 (^##<i>p</i> = 0.0014 G226 [1.0 μM] vs IS [10 μM], unpaired t-test), (iii) phospho-ERK1/2, (iv) phospho-NF-κB (^##<i>p</i> = 0.0014 G226 [1.0 μM] vs IS [10 μM], unpaired t-test); and (B) 3.0 μM RWJ-67657 and 1.0 μM U0126 for (i) phospho-p38, (ii) phospho-ERK1/2 (**<i>p</i> = 0.0063 IS [10 μM] vs control, ^##<i>p</i> = 0.0136 U0126 [1.0 μM] vs IS [10 μM], unpaired t-test), (iii) phospho-NF-κB and (iv) Pan Actin. Data are presented as mean ± SEM (n = 3). **<i>p</i><0.01, ****<i>p</i><0.0001 vs control, ^#<i>p</i><0.05, ^##<i>p</i><0.01, ^####<i>p</i><0.0001 vs IS [10 μM], One-way Anova.</p