abstract

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ABSTRACT Fruits of Romanian native plants from Adoxaceae family (European Cranberrybush - Viburnum opulus L. –E1 and European Black Elderberry - Sambucus nigra L.-E2) have biological activity mainly due to their high content of anthocyanins and other polyphenols. They were used to obtain new nanomaterials based on gold nanoparticles. We have controled the size, shape and surface morphology of these nanomaterials (AuNPs-E1, AuNPs-E2) prepared by using a green method; the new nanomaterials have been used in medicine to treat some skin diseases. We have evaluated their in vitro and in vivo effect and we have identified the effectiveness of new topic therapies based on nanotechnology using high frequency ultrasound. THE NON-INVASIVE ASSESSMENT OF THE EFFICACY OF GOLD NANOPARTICLES-ANTHOCYANINS NANOMATERIALS ON CUTANEOUS PATHOLOGY Liliana Olenic 1, Adriana Vulcu 1 , Camelia Grosan 1 , Luminita David 2 , Bianca Moldovan 2 , Maria Perde-Schrepler 3 , Adriana Gabriela Filip 4 , Maria Crisan 4 , Ioana Chiorean 5 1 National Institute for Research and Development of Isotopic and Molecular Technologies, 65-103 Donath Street, Cluj-Napoca, Romania 2 Faculty of Chemistry and Chemical Engineering, 11 Arany Janos Street RO 400028, Cluj-Napoca, Romania 3 “Ion Chiricuţă” Oncology Institute, 34-36 Republicii Street Cluj-Napoca, Romania 4 Faculty of General Medicine, Iuliu Haţieganu University of Medicine and Pharmacy Strada Emil Isac 13, Cluj-Napoca, Romania 5 Faculty of Mathematics and Informatics, , Babeş-Bolyai –University, Kogălniceanu Street, 1, Cluj-Napoca, Romania 400084 EXPERIMENTAL RESULTS AND DISCUSSIONS CONCLUSION Gold nanoparticles functionalized with anthocyanins extracted from Viburnum opulus L. and Sambucus nigra L. (AuNPs-E1 and AuNPs-E2) were prepared and characterized by UV-Vis and FTIR spectroscopy and by Transmission Electron Spectroscopy (TEM) (diameters between 25-100 nm) There were assessed cytotoxicity tests of the extract alone and of the functionalized nanoparticles; To study the antiinflammatory effect, keratinocytes treated with non- toxic doses of anthocyanins and nanoparticles were exposed to increasing doses of UVB irradiation Figure 1. (a) Viburnum opulus L., berries; (b) Sambucus Nigra L., berries;(c) And (d) TEM images of AuNPs with E1 respectively E2 INCDTIM Figure 2. UV-Vis spectra of (a) AuNPs- E1; (b) AuNPs-E2 Acknowledgement This work was elaborated in the frame project no 147/2012, through the program “Partenerships in priority areas-PN II”, developed with the support of ANCS, CNDI- UEFISCDI 300 400 500 600 700 800 900 0.1 0.2 0.3 0.4 0.5 0.6 0.7 0.8 A uN P s-E 1 pH 7 .5 cen trif A b s o rb a n ce (a.u .) nm ) A uN P s-E 1 pH 7.5 (a) 400 500 600 700 800 900 0.10 0.15 0.20 0.25 0.30 0.35 0.40 A uN P S -E 2pH 9.5 centrif A b so rb an c e (a .u .) (nm ) A uN P S -E 2pH 9.5 (b ) . In vitro toxicity studies H aC aT 0 7. 5 15 37 .5 75 15 0 37 5 750 0.00 0.05 0.10 0.15 0.20 *** *** *** *** ** * AuE1 NP conc.( g/m l) opticaldensity (492nm ) A431 0 7.5 15 37.5 75 150 375 750 0.0 0.1 0.2 0.3 0.4 ** *** * AuE1 NP conc.( g/m l) opticaldensity (492nm ) (a) (b) H aC aT 0.00 0.05 0.10 0.15 0.20 *** *** *** *** * AuE2 NP conc.( g/m l) opticaldensity (492nm ) A431 0 7.5 15 37.5 75 150 375 750 0.0 0.1 0.2 0.3 0.4 ** *** *** AuE2 NP conc.( g/m l) opticaldensity (492nm ) (d) (c) Figure 3. The viability of HaCaTcell and A431 cells function of nanomaterials concentration H aC aT 0.0 0.1 0.2 AuNPE2 AuNPE1 Au NP conc. opticaldensity (492nm ) A431 0.0 0.1 0.2 0.3 0.4 AuE1NP AuE2 NP Au N P conc. opticaldensity (492nm ) Figure 4. Compared of cell survival between the two line of cells with the two types of nanoparticles 2. In vivo toxicity studies MDA ctrl A T 1 A T 2 diclofenac Au AT A g A T 0 1 2 3 ** ** * nm oli/m g P GSH ctrl A T 1 A T 2 diclofenac A u A T Ag AT 0 10 20 30 40 nm oli/m g P Figure 5. Level of malondialdehyde for different lots Figure 6.Glutatione level for different lots Edem a 2h ctrl diclofenac A T 1 A T 2 A u A T Ag AT 0.0 0.5 1.0 1.5 ** ** * Figure 7. Edema of foot after 2 h treatment 3. Evaluation of new prepared nanomaterials on skin diseases Figure 8. Images token with high-frequency ultrasound before and after topical application of new nanomaterials for 2 weeks Figure 9. The histogram of skin thickness for 8 patients before and after treatment with nanomaterials

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INCDTIM. (b). (a). (c). (d). THE NON-INVASIVE ASSESSMENT OF THE EFFICACY OF GOLD NANOPARTICLES-ANTHOCYANINS NANOMATERIALS ON CUTANEOUS PATHOLOGY. Liliana Olenic 1, Adriana Vulcu 1 , Camelia Grosan 1 , Luminita David 2 , Bianca Moldovan 2 , - PowerPoint PPT Presentation

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Page 1: ABSTRACT

ABSTRACT Fruits of Romanian native plants from Adoxaceae family (European Cranberrybush - Viburnum opulus L. –E1 and European Black Elderberry -

Sambucus nigra L.-E2) have biological activity mainly due to their high content of anthocyanins and other polyphenols. They were used to obtain new nanomaterials based on gold nanoparticles. We have controled the size, shape and surface morphology of these nanomaterials (AuNPs-E1, AuNPs-E2) prepared by using a green method; the new nanomaterials have been used in medicine to treat some skin diseases.

We have evaluated their in vitro and in vivo effect and we have identified the effectiveness of new topic therapies based on nanotechnology using high frequency ultrasound.

THE NON-INVASIVE ASSESSMENT OF THE EFFICACY OF GOLD NANOPARTICLES-ANTHOCYANINS NANOMATERIALS ON CUTANEOUS

PATHOLOGY Liliana Olenic1, Adriana Vulcu1, Camelia Grosan1, Luminita David2, Bianca Moldovan2,

Maria Perde-Schrepler3, Adriana Gabriela Filip4 , Maria Crisan4, Ioana Chiorean5

1National Institute for Research and Development of Isotopic and Molecular Technologies, 65-103 Donath Street, Cluj-Napoca, Romania2Faculty of Chemistry and Chemical Engineering, 11 Arany Janos Street RO 400028, Cluj-Napoca, Romania

3“Ion Chiricuţă” Oncology Institute, 34-36 Republicii Street Cluj-Napoca, Romania 4Faculty of General Medicine, Iuliu Haţieganu University of Medicine and Pharmacy Strada Emil Isac 13, Cluj-Napoca, Romania

5Faculty of Mathematics and Informatics, , Babeş-Bolyai –University, Kogălniceanu Street, 1, Cluj-Napoca, Romania 400084

EXPERIMENTAL

RESULTS AND DISCUSSIONS

CONCLUSIONGold nanoparticles functionalized with anthocyanins extracted from Viburnum opulus L. and Sambucus nigra L. (AuNPs-E1 and AuNPs-E2) were prepared and characterized by UV-Vis and FTIR spectroscopy and by Transmission Electron Spectroscopy (TEM) (diameters between 25-100 nm) There were assessed cytotoxicity tests of the extract alone and of the functionalized nanoparticles;To study the antiinflammatory effect, keratinocytes treated with non-toxic doses of anthocyanins and nanoparticles were exposed to increasing doses of UVB irradiation The histograms of the skin thickness for 8 patients presented an involution of diseases after treatment, fact that indicates the very good antiinflammatory effect of our nanomaterials.

Figure 1. (a) Viburnum opulus L., berries; (b) Sambucus Nigra L., berries;(c) And (d) TEM images of AuNPs with E1 respectively E2

INCDTIM

Figure 2. UV-Vis spectra of (a) AuNPs-E1; (b) AuNPs-E2

AcknowledgementThis work was elaborated in the frame project no 147/2012, through the program “Partenerships in priority areas-PN II”, developed with the support of ANCS, CNDI-UEFISCDI

300 400 500 600 700 800 900

0.1

0.2

0.3

0.4

0.5

0.6

0.7

0.8

AuNPs-E1 pH 7.5 centrif

Abs

orba

nce

(a.u

.)

nm)

AuNPs-E1 pH 7.5

(a)

400 500 600 700 800 900

0.10

0.15

0.20

0.25

0.30

0.35

0.40

AuNPS-E2pH 9.5 centrif

Abs

orba

nce

(a.u

.)

(nm)

AuNPS-E2pH 9.5

(b)

1. In vitro toxicity studies

HaCaT

0 7.5 15 37.5 75 15

037

575

00.00

0.05

0.10

0.15

0.20

***************

AuE1 NP conc. (g/ml)

optic

al d

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ty (4

92nm

)

A431

0 7.5 15 37.5 75 15

037

575

00.0

0.1

0.2

0.3

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**

***

*

AuE1 NP conc. (g/ml)

optic

al d

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ty (4

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)

(a) (b)

HaCaT

0.00

0.05

0.10

0.15

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***

****** ***

*

AuE2 NP conc. (g/ml)

optic

al d

ensi

ty (4

92nm

)

A431

0 7.5 15 37.5 75 15

037

575

00.0

0.1

0.2

0.3

0.4

**

*** ***

AuE2 NP conc. (g/ml)

optic

al d

ensi

ty (4

92nm

)

(d)(c)

Figure 3. The viability of HaCaTcell and A431 cells function of nanomaterials concentration

HaCaT

0.0

0.1

0.2AuNPE2AuNPE1

Au NP conc.

optic

al d

ensi

ty (4

92nm

)

A431

0.0

0.1

0.2

0.3

0.4AuE1NPAuE2 NP

Au NP conc.

optic

al d

ensi

ty (4

92nm

)

Figure 4. Compared of cell survival between the two line of cells with the two types of nanoparticles

2. In vivo toxicity studiesMDA

ctrl

AT 1AT 2

diclofen

acAu A

TAg A

T0

1

2

3

** ** *

nmol

i/mg

P

GSH

ctrl

AT 1AT 2

diclofen

acAu A

TAg A

T0

10

20

30

40

nmol

i/mg

P

Figure 5. Level of malondialdehyde for different lots

Figure 6.Glutatione level for different lots

Edema 2h

ctrl

diclofen

acAT 1

AT 2Au A

T

Ag AT

0.0

0.5

1.0

1.5

**

***

Figure 7. Edema of foot after 2 h treatment

3. Evaluation of new prepared nanomaterials on skin diseases

Figure 8. Images token with high-frequency ultrasound before and after topical application of new nanomaterials for

2 weeks

Figure 9. The histogram of skin thickness for 8 patients before and after treatment with

nanomaterials