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Inorganic phosphate nanorods are a novel fluorescent label in cell biology.

Patra CR, Bhattacharya R, Patra S, Basu S, Mukherjee P, Mukhopadhyay D - J Nanobiotechnology (2006)

Bottom Line: We report the first use of inorganic fluorescent lanthanide (europium and terbium) ortho phosphate [LnPO4.H2O, Ln = Eu and Tb] nanorods as a novel fluorescent label in cell biology.These nanorods, synthesized by the microwave technique, retain their fluorescent properties after internalization into human umbilical vein endothelial cells (HUVEC), 786-O cells, or renal carcinoma cells (RCC).The cellular internalization of these nanorods and their fluorescence properties were characterized by fluorescence spectroscopy (FS), differential interference contrast (DIC) microscopy, confocal microscopy, and transmission electron microscopy (TEM).

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Biochemistry and Molecular Biology, Mayo Clinic Cancer Center, Mayo Clinic, Rochester, Minnesota, USA. patra.chittaranjan@mayo.edu

ABSTRACT
We report the first use of inorganic fluorescent lanthanide (europium and terbium) ortho phosphate [LnPO4.H2O, Ln = Eu and Tb] nanorods as a novel fluorescent label in cell biology. These nanorods, synthesized by the microwave technique, retain their fluorescent properties after internalization into human umbilical vein endothelial cells (HUVEC), 786-O cells, or renal carcinoma cells (RCC). The cellular internalization of these nanorods and their fluorescence properties were characterized by fluorescence spectroscopy (FS), differential interference contrast (DIC) microscopy, confocal microscopy, and transmission electron microscopy (TEM). At concentrations up to 50 microg/ml, the use of [3H]-thymidine incorporation assays, apoptosis assays (TUNEL), and trypan blue exclusion illustrated the non-toxic nature of these nanorods, a major advantage over traditional organic dyes.

No MeSH data available.


Related in: MedlinePlus

Effect of fluorescent nanorods (EuPO4 and TbPO4) with different concentrations to normal HUVEC was observed by [3H]thymidine incorporation asssay. A serum-starved HUVEC was treated with (A) EuPO4·H2O nanorods and (B) TbPO4·H2O nanorods at the concentration range of 1–100 μg/mL [Eu1 = 1 μg/ml, Eu50 = 50 μg/ml, Eu100 = 100 μg/ml. Similarly, Tb1 = 1 μg/ml, Tb50 = 50 μg/ml, Tb100 = 100 μg/ml]. Average of three independent experiments, each was done in triplicate.
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Figure 8: Effect of fluorescent nanorods (EuPO4 and TbPO4) with different concentrations to normal HUVEC was observed by [3H]thymidine incorporation asssay. A serum-starved HUVEC was treated with (A) EuPO4·H2O nanorods and (B) TbPO4·H2O nanorods at the concentration range of 1–100 μg/mL [Eu1 = 1 μg/ml, Eu50 = 50 μg/ml, Eu100 = 100 μg/ml. Similarly, Tb1 = 1 μg/ml, Tb50 = 50 μg/ml, Tb100 = 100 μg/ml]. Average of three independent experiments, each was done in triplicate.

Mentions: Since these inorganic nanorods show distinct fluorescence activity upon cellular internalization, we have decided to use these materials as a fluorescent label for HUVEC and 786-O cells. We examined their in vitro toxicity with [3H]-thymidine incorporation assays [29] on normal endothelial cells (HUVEC) and found them to be non-toxic (Fig. 8A–B). Although there were indications that exposure to certain nanomaterials might lead to adverse biological effects, this appears to dependent upon the chemical and physical properties of the material [4,27,28]. The potential toxicity of inorganic fluorescent nanoparticles has recently become a topic of considerable importance and discussion, especially since in vivo toxicity is likely to be a key factor in determining whether fluorescent probes will be approved by regulatory agencies for human clinical use. HUVEC proliferation [29] was clearly not affected from internalization of materials up to 50 mg/ml compared to control samples (Fig. 8A–B); however, at concentrations greater than 50 mg/ml, nanorods were detected to be toxic. Experiments were repeated in triplicate and results were reproducible.


Inorganic phosphate nanorods are a novel fluorescent label in cell biology.

Patra CR, Bhattacharya R, Patra S, Basu S, Mukherjee P, Mukhopadhyay D - J Nanobiotechnology (2006)

Effect of fluorescent nanorods (EuPO4 and TbPO4) with different concentrations to normal HUVEC was observed by [3H]thymidine incorporation asssay. A serum-starved HUVEC was treated with (A) EuPO4·H2O nanorods and (B) TbPO4·H2O nanorods at the concentration range of 1–100 μg/mL [Eu1 = 1 μg/ml, Eu50 = 50 μg/ml, Eu100 = 100 μg/ml. Similarly, Tb1 = 1 μg/ml, Tb50 = 50 μg/ml, Tb100 = 100 μg/ml]. Average of three independent experiments, each was done in triplicate.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC1636656&req=5

Figure 8: Effect of fluorescent nanorods (EuPO4 and TbPO4) with different concentrations to normal HUVEC was observed by [3H]thymidine incorporation asssay. A serum-starved HUVEC was treated with (A) EuPO4·H2O nanorods and (B) TbPO4·H2O nanorods at the concentration range of 1–100 μg/mL [Eu1 = 1 μg/ml, Eu50 = 50 μg/ml, Eu100 = 100 μg/ml. Similarly, Tb1 = 1 μg/ml, Tb50 = 50 μg/ml, Tb100 = 100 μg/ml]. Average of three independent experiments, each was done in triplicate.
Mentions: Since these inorganic nanorods show distinct fluorescence activity upon cellular internalization, we have decided to use these materials as a fluorescent label for HUVEC and 786-O cells. We examined their in vitro toxicity with [3H]-thymidine incorporation assays [29] on normal endothelial cells (HUVEC) and found them to be non-toxic (Fig. 8A–B). Although there were indications that exposure to certain nanomaterials might lead to adverse biological effects, this appears to dependent upon the chemical and physical properties of the material [4,27,28]. The potential toxicity of inorganic fluorescent nanoparticles has recently become a topic of considerable importance and discussion, especially since in vivo toxicity is likely to be a key factor in determining whether fluorescent probes will be approved by regulatory agencies for human clinical use. HUVEC proliferation [29] was clearly not affected from internalization of materials up to 50 mg/ml compared to control samples (Fig. 8A–B); however, at concentrations greater than 50 mg/ml, nanorods were detected to be toxic. Experiments were repeated in triplicate and results were reproducible.

Bottom Line: We report the first use of inorganic fluorescent lanthanide (europium and terbium) ortho phosphate [LnPO4.H2O, Ln = Eu and Tb] nanorods as a novel fluorescent label in cell biology.These nanorods, synthesized by the microwave technique, retain their fluorescent properties after internalization into human umbilical vein endothelial cells (HUVEC), 786-O cells, or renal carcinoma cells (RCC).The cellular internalization of these nanorods and their fluorescence properties were characterized by fluorescence spectroscopy (FS), differential interference contrast (DIC) microscopy, confocal microscopy, and transmission electron microscopy (TEM).

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Biochemistry and Molecular Biology, Mayo Clinic Cancer Center, Mayo Clinic, Rochester, Minnesota, USA. patra.chittaranjan@mayo.edu

ABSTRACT
We report the first use of inorganic fluorescent lanthanide (europium and terbium) ortho phosphate [LnPO4.H2O, Ln = Eu and Tb] nanorods as a novel fluorescent label in cell biology. These nanorods, synthesized by the microwave technique, retain their fluorescent properties after internalization into human umbilical vein endothelial cells (HUVEC), 786-O cells, or renal carcinoma cells (RCC). The cellular internalization of these nanorods and their fluorescence properties were characterized by fluorescence spectroscopy (FS), differential interference contrast (DIC) microscopy, confocal microscopy, and transmission electron microscopy (TEM). At concentrations up to 50 microg/ml, the use of [3H]-thymidine incorporation assays, apoptosis assays (TUNEL), and trypan blue exclusion illustrated the non-toxic nature of these nanorods, a major advantage over traditional organic dyes.

No MeSH data available.


Related in: MedlinePlus