The use of size-defined DNA-functionalized calcium phosphate nanoparticles to minimise intracellular calcium disturbance during transfection

Biomaterials ◽  
2009 ◽  
Vol 30 (35) ◽  
pp. 6794-6802 ◽  
Author(s):  
Sebastian Neumann ◽  
Anna Kovtun ◽  
Irmgard D. Dietzel ◽  
Matthias Epple ◽  
Rolf Heumann
2010 ◽  
Vol 441 ◽  
pp. 299-305 ◽  
Author(s):  
Matthias Epple ◽  
Anna Kovtun

Calcium phosphate is a natural biomineral and therefore possesses an excellent biocompatibility due to its chemical similarity to human hard tissue (bone and teeth). Calcium phosphate nanoparticles can be precipitated under controlled conditions and used as carrier in biological systems, e.g. to transfer nucleic acids or drugs. Such nanoparticles can also be suitably functionalized with fluorescing dyes, polymeric agents, pro-drugs or activators. The small monodisperse nanoparticles only mildly influence the intracellular calcium level and therefore are not toxic for cells.


Author(s):  
Manuel Rivas ◽  
Pau Turon ◽  
Carlos Alemán ◽  
Jordi Puiggalí ◽  
Luis J. del Valle

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.


2014 ◽  
Vol 70 (a1) ◽  
pp. C67-C67
Author(s):  
Babak Mostaghaci ◽  
Brigitta Loretz ◽  
Robert Haberkorn ◽  
Guido Kickelbick ◽  
Claus-Michael Lehr

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.


2004 ◽  
Vol 14 (14) ◽  
pp. 2213 ◽  
Author(s):  
T. Welzel ◽  
I. Radtke ◽  
W. Meyer-Zaika ◽  
R. Heumann ◽  
M. Epple

2012 ◽  
Vol 109 (35) ◽  
pp. 14170-14175 ◽  
Author(s):  
S. Boonrungsiman ◽  
E. Gentleman ◽  
R. Carzaniga ◽  
N. D. Evans ◽  
D. W. McComb ◽  
...  

Agronomy ◽  
2021 ◽  
Vol 11 (6) ◽  
pp. 1026
Author(s):  
Federica Gaiotti ◽  
Marco Lucchetta ◽  
Giacomo Rodegher ◽  
Daniel Lorenzoni ◽  
Edoardo Longo ◽  
...  

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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