Cytotoxic effects of cetuximab-conjugated calcium phosphate-nanoparticles on epithelial and mesenchymal cells

AbstractIntracellular calcium (Ca2+) is a key signaling element that is involved in a great variety of fundamental biological processes. Thus, Ca2+ deregulation would be involved in the cancer cell progression and damage of mitochondrial membrane and DNA, which lead to apoptosis and necrosis. In this study, we have prepared amorphous calcium phosphate nanoparticles (ACP NPs) for studied their incorporation by endocytosis or electroporation to epithelial, endothelial and fibroblast cells (MCF-7, HUVEC and COS-1 cells, respectively). Our results showed that internalized ACP NPs have cytotoxic effects as a consequence of the increase of the intracellular calcium content. The endocytosis pathways showed a greater cytotoxic effect since calcium ions could easily be released from the nanoparticles and be accumulated in the lysosomes and mitochondria. In addition, the cytotoxic effect could be reversed when calcium ion was chelated with ethylene glycol-bis(2-aminoethylether)-N,N,N′,N′-tetraacetic acid (EGTA). Modification of ACP NPs by coating with different compounds based on phosphates was also evaluated. The results indicated a reduction of the cytotoxic effect, in the order polyphosphate < phosphonic acid < orthophosphate. A differential cytotoxic effect of ACP-NPs was observed in function of the cell type; the cytotoxic effect can be ordered as i.e., HUVEC > COS-1 > MCF-7. The greater cytotoxic effect caused by the increase of intracellular calcium that is observed in normal cells and the greater resistance of cancer cells suggests new perspectives for cancer research.

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TOXICITY OF SPHERICAL AND NEEDLE-SHAPED CALCIUM PHOSPHATE BIONS TO INJURED INTIMA OF THE RAT ABDOMINAL AORTA

ZHurnal «Patologicheskaia fiziologiia i eksperimental`naia terapiia» ◽

10.25557/0031-2991.2019.02.80-88 ◽

2019 ◽

pp. 80-88

Author(s):

А.Г. Кутихин ◽

Д.К. Шишкова ◽

Е.А. Великанова ◽

А.В. Миронов ◽

Е.О. Кривкина ◽

...

Keyword(s):

Smooth Muscle ◽

Calcium Phosphate ◽

Systemic Inflammation ◽

Intravenous Administration ◽

Intimal Hyperplasia ◽

Mesenchymal Cells ◽

Enzyme Linked Immunosorbent Assay ◽

Cytotoxic Action ◽

Alizarin Red ◽

Synthetic Phenotype

Цель исследования - оценка токсического действия сферических кальций-фосфатных бионов и игольчатых кальций-фосфатных бионов на предварительно поврежденную интиму аорты крыс. Методика. Токсическое действие сферических кальций-фосфатных бионов и игольчатых кальций-фосфатных бионов на поврежденную интиму брюшной аорты крыс линии Wistar (n = 10 на группу) оценивали путем их однократного внутривенного введения после баллонной ангиопластики с эксплантацией поврежденного участка аорты через 5 нед. Биоптаты анализировали: 1) классическими гистологическими методами (окрашивание гематоксилин-эозином, ализариновым красным, по Вейгерту-ван Гизону и по Расселлу-Мовату); 2) иммунофлюоресцентным окрашиванием криосрезов (сочетанное окрашивание на CD31 и CD34, на CD31 и α-гладкомышечный актин (α-ГМА), на виментин и α-ГМА, на коллаген IV типа и α-ГМА). Для оценки влияния системного воспаления на КФБ-индуцированную эндотелиотоксичность определяли содержание моноцитарного хемоаттрактантного белка (МСР-1/CCL2) и церулоплазмина в сыворотке крови прооперированных крыс посредством иммуноферментного анализа. Результаты. Сферические кальций-фосфатные бионы и игольчатые кальций-фосфатные бионы вызывали выраженную гипертрофию интимы брюшной аорты в 90% (9 из 10 крыс) и 80% случаев (8 из 10 крыс) соответственно, в то время как частота гипертрофии в группе контрольных крыс составила лишь 10% (1 из 10 крыс). Неоинтима при экспозиции интимы брюшной аорты обоим типам бионов характеризовалась переходом фенотипа клеток мезенхимального ряда с контрактильного (α-ГМА-положительные и виментин-отрицательные гладкомышечные клетки) и неактивного (α-ГМА-отрицательные и виментин-положительные фибробласты) на активный синтетический (α-ГМА- и виментин-положительные клетки), что приводило к формированию значительных количеств экстрацеллюлярного матрикса. Внутривенное введение сферических кальций-фосфатных бионов и игольчатых кальций-фосфатных бионов не приводило к изменению уровней МСР-1/CCL2 и церулоплазмина в сыворотке крови, что свидетельствовало об отсутствии их возможного влияния на развитие системного воспалительного ответа. Заключение. Внутривенное введение кальций-фосфатных бионов после повреждения интимы брюшной аорты крыс путем баллонной ангиопластики вызывает развитие гипертрофии интимы, частота и выраженность которой не зависит от формы кальций-фосфатных бионов и которая характеризуется переходом фенотипа клеток мезенхимального ряда из контрактильного/неактивного на активный синтетический. При этом эндотелиотоксическое действие кальций-фосфатных бионов обусловлено их непосредственным воздействием на эндотелий, а не развитием системного воспаления. Purpose. To compare toxicity of spherical calcium phosphate bions (SCPB) and needle-shaped calcium phosphate bions (NCPB) to injured intima of rat aortas. Methods. Toxicity of SCPB and NCPB to injured abdominal aortas of Wistar rats (n = 10 per group) was evaluated using intravenous administration of the bions after balloon angioplasty. Rats were sacrificed five weeks postoperation, and an injured aortic segment was excised. Tissue preparations were stained with hematoxylin and eosin, alizarin red S, Weigert-van Gieson, and Movat’s pentachrome stains. Selected tissue samples were then examined using combined immunofluorescence staining (CD31/CD34, CD31/α-smooth muscle actin (α-SMA), α-SMA/vimentin, and α-SMA/collagen IV). Possible influence of systemic inflammation on CPB-induced endothelial toxicity was assessed by measuring monocyte chemoattractant protein-1 and ceruloplasmin in rat serum using the enzyme-linked immunosorbent assay. Results. Intravenous administration of SCPB or NCPB provoked intimal hyperplasia in 90% (9 of 10) and 80% (8 of 10) of rats vs. 10% (1 of 10) in the control group. The neointima was characterized by a phenotypic switch of mesenchymal cells, i.e. transition of a contractile (α-SMA-positive, vimentin-negative vascular smooth muscle cells) and quiescent (α-SMA-negative vimentin-positive fibroblasts) to an active synthetic phenotype (double-positive cells), which resulted in deposition of the extracellular matrix. Neither SCPB nor NCPB changed serum levels of pro-inflammatory molecules, МСР-1/CCL2, and ceruloplasmin. Conclusions. Intravenous administration of CPB upon balloon-induced vascular injury caused intimal hyperplasia regardless of the CPB shape. Hyperplasia foci were characterized by a switch of mesenchymal cells from a contractile/quiescent to an active synthetic phenotype. Endothelial toxicity of CPBs was defined by their direct cytotoxic action rather than induction of systemic inflammation.

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One-pot synthesis of injectable methylcellulose hydrogel containing calcium phosphate nanoparticles

Carbohydrate Polymers ◽

10.1016/j.carbpol.2016.10.055 ◽

2017 ◽

Vol 157 ◽

pp. 775-783 ◽

Cited By ~ 22

Author(s):

Hanna Park ◽

Min Hee Kim ◽

Young Il Yoon ◽

Won Ho Park

Keyword(s):

Calcium Phosphate ◽

One Pot ◽

One Pot Synthesis ◽

Calcium Phosphate Nanoparticles

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Calcium phosphate phase transition in the presence of Aminosilane

Acta Crystallographica Section A Foundations and Advances ◽

10.1107/s205327331409932x ◽

2014 ◽

Vol 70 (a1) ◽

pp. C67-C67

Author(s):

Babak Mostaghaci ◽

Brigitta Loretz ◽

Robert Haberkorn ◽

Guido Kickelbick ◽

Claus-Michael Lehr

Keyword(s):

Phase Transition ◽

Calcium Phosphate ◽

Blood Compatibility ◽

Transfection Efficiency ◽

Hek293 Cells ◽

Step Method ◽

Calcium Phosphate Nanoparticles ◽

Amine Groups ◽

The Impact

Calcium phosphate has been the point of interest for in vitro gene delivery for many years because of its biocompatibility and straight forward application. However, there are some limitations regarding in vivo administration of these particles mostly because of vast agglomeration of the particles and lack of strong bond between the particles and pDNA. We introduced a simple single step method to functionalize calcium phosphate nanoparticles with Aminosilanes having a different number of amine groups. The nanoparticles were characterized chemically and structurally and their toxicity and interaction with pDNA were studied as well. Results revealed that different crystalline phase of calcium phosphate nanoparticles (Brushite and Hydroxyapatite) with a size below 150 nm were prepared, depending on conditions of synthesis and phase, each with a narrow size distribution. The aminosilane agents caused oriented nucleation and growth of crystallites and can decrease the pH for producing hydroxyapatite phase. The phenomenon could be revealed with the presence of anisotropy in the structure of synthesized hydroxyapatite. The number of amine groups in the Aminosilane agent could change the phase transition pH. Brushite particles revealed to have stronger interaction with pDNA mostly because of their higher positive surface charge. Both particles showed blood compatibility and negligible toxicity. Transfection experiment revealed the capability of both brushite and hydroxyapatite particles to transfect A549 and HEK293 cells. The new modified nanoparticles can be stored in a dried state and re-dispersed easily at the time of administration. Moreover, the transfection efficiency is higher in comparison with conventional calcium phosphate. This study showed the impact of presence and type of the modifying agent on the crystal structure and the amount of surface functionalization of nanoparticles, which in consequence influenced their interaction with cells.

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Transfection of cells with custom-made calcium phosphate nanoparticles coated with DNA

Journal of Materials Chemistry ◽

10.1039/b401644k ◽

2004 ◽

Vol 14 (14) ◽

pp. 2213 ◽

Cited By ~ 104

Author(s):

T. Welzel ◽

I. Radtke ◽

W. Meyer-Zaika ◽

R. Heumann ◽

M. Epple

Keyword(s):

Calcium Phosphate ◽

Custom Made ◽

Calcium Phosphate Nanoparticles

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Urea-Doped Calcium Phosphate Nanoparticles as Sustainable Nitrogen Nanofertilizers for Viticulture: Implications on Yield and Quality of Pinot Gris Grapevines

Agronomy ◽

10.3390/agronomy11061026 ◽

2021 ◽

Vol 11 (6) ◽

pp. 1026

Author(s):

Federica Gaiotti ◽

Marco Lucchetta ◽

Giacomo Rodegher ◽

Daniel Lorenzoni ◽

Edoardo Longo ◽

...

Keyword(s):

Calcium Phosphate ◽

Nitrogen Source ◽

Foliar Application ◽

N Fertilization ◽

Plant Nitrogen ◽

Yield And Quality ◽

Berry Quality ◽

Potential Benefits ◽

Calcium Phosphate Nanoparticles

In recent years, the application of nanotechnology for the development of new “smart fertilizers” is regarded as one of the most promising solutions for boosting a more sustainable and modern grapevine cultivation. Despite showing interesting potential benefits over conventional fertilization practices, the use of nanofertilizers in viticulture is still underexplored. In this work, we investigated the effectiveness of non-toxic calcium phosphate nanoparticles (Ca3(PO4)2∙nH2O) doped with urea (U-ACP) as a nitrogen source for grapevine fertilization. Plant tests were performed for two years (2019–2020) on potted adult Pinot gris cv. vines grown under semi-controlled conditions. Four fertilization treatments were compared: N1: commercial granular fertilization (45 kg N ha−1); N2: U-ACP applied in fertigation (36 kg N ha−1); N3: foliar application of U-ACP (36 kg N ha−1); C: control, receiving no N fertilization. Plant nitrogen status (SPAD), yield parameters as well as those of berry quality were analyzed. Results here presented clearly show the capability of vine plants to recognize and use the nitrogen supplied with U-ACP nanoparticles either when applied foliarly or to the soil. Moreover, all of the quali–quantitative parameters measured in vine plants fed with nanoparticles were perfectly comparable to those of plants grown in conventional condition, despite the restrained dosage of nitrogen applied with the nanoparticles. Therefore, these results provide both clear evidence of the efficacy of U-ACP nanoparticles as a nitrogen source and the basis for the development of alternative nitrogen fertilization strategies, optimizing the dosage/benefit ratio and being particularly interesting in a context of a more sustainable and modern viticulture.

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