Aggregation Behavior of Protoporphyrin IX in Aqueous Solutions:  Clear Evidence of Vesicle Formation

Scolaro, Luigi Monsù; Castriciano, Maria Angela; Romeo, Andrea; Patanè, Salvatore; Cefalì, Eugenio; Allegrini, M.

doi:10.1021/jp013155h

Cited by 155 publications

(163 citation statements)

References 35 publications

Supporting

Mentioning

150

Contrasting

Unclassified

Order By: Relevance

“…According to Scolaro et al (2002), porphyrins exhibit aggregation, a π-π stacking interaction in water while spectroscopic studies performed in water as a function of pH demonstrated that they are monomer between pH 0-3; dimmer in pH above 8 and higher aggregates in the pH 3-7. At pH 1, PpIX absorption spectra exhibits a strong absorption band (B-band) at λ (nm) = 406 (1.7) and four weaker absorption bands (Q-bands) at λ (nm) = 516 (9.6); 554 (17.2); 600 (11.6) and 626 (8.0).…”

Section: Resultsmentioning

confidence: 99%

Fluorometric quantification of protoporphyrin IX in biological skin samples from in vitro penetration/permeation studies

Rossetti

Depieri

Tedesco

et al. 2010

Braz. J. Pharm. Sci.

View full text Add to dashboard Cite

A fluorometric analytical method was developed for quantification of protoporphyrin IX (PpIX) in skin samples and receptor phase solution after in vitro cutaneous penetration/permeation studies. Analytical conditions used were: excitation and emission wavelengths: 400 nm and 632 nm; bandwidth: 0.5 nm; excitation and emission slits: 10/10. PpIX was recovered from two different layers of skin, the stratum corneum (SC) and the epidermis plus dermis ([E+D]), by vortex homogenization, probe and bath sonication, using DMSO as an extraction solvent. The detection and quantification limits were 0.002 and 0.005 µg/mL, respectively. The assay was linear from 0.005 -0.5 µg/mL. The within-day and betweenday assay precision and accuracy in DMSO and receptor phase solution were each studied at the two concentration levels 0.04 and 0.2 µg/mL, and 0.01 and 0.08 µg/mL, respectively. The coefficients of variation and deviation from the theoretical values were lower than 5%. The skin recovery of PpIX from SC and [E+D] layers using two different concentrations (0.5 and 1.0 µg/mL) were all above 90.0%. The method described has potential application to in vitro penetration/permeation studies of PpIX using porcine skin as a biological membrane model. Uniterms: Fluorometric quantification. Protoporphyrin IX/determination. Photodynamic therapy. Porcine skin. Cutaneous penetration/in vitro study. Cutaneous permeation/in vitro study.Um método analítico por espectrofluorimetria foi desenvolvido para quantificar a protoporfirina IX (Pp IX) em amostras de pele e fase receptora após a realização de testes in vitro de penetração/permeação cutâneas. As condições analíticas utilizadas foram: comprimentos de onda de excitação e emissão: 400 nm e 632 nm; largura de banda: 0,5 nm; fendas de excitação e emissão: 10/10. A PpIX foi extraída de amostras de estrato córneo (EC) e da epiderme sem estrato córneo + derme ([E+D]) através da agitação em vórtex e sonicação por haste e banho, utilizando-se o DMSO como solvente extrator. O limite de detecção e quantificação foram, respectivamente, de 0,002 e 0,005 µg/mL. O método mostrou-se linear da faixa de 0,005 -0,5 µg/mL. A precisão e exatidão intra e inter-ensaio em DMSO e na fase receptora foram validadas utilizando-se duas concentrações distintas, respectivamente, de 0,004 e 0,2 µg/mL, e 0,01 e 0,08 µg/mL. Os valores de coeficiente de variação e o desvio do valor teórico foram inferiores a 5%. A recuperação da PpIX das camadas da pele (EC e [E+D]) utilizando-se duas concentrações distintas (0,5 e 1,0 µg/mL) foram todas acima de 90,0%. O método descrito pode ser utilizado para determinação da PpIX após estudos de penetração/permeação cutânea in vitro utilizando pele de porco como modelo de membrana.Unitermos: Espectrofluorimetria/análise quantitativa. Quantificação fluorimétrica. Protoporfirina IX/ determinação. Terapia fotodinâmica. Pele porcina. Penetração cutânea/estudo in vitro. Permeação cutânea/estudo in vitro.

show abstract

Section: Resultsmentioning

confidence: 99%

Fluorometric quantification of protoporphyrin IX in biological skin samples from in vitro penetration/permeation studies

Rossetti

Depieri

Tedesco

et al. 2010

Braz. J. Pharm. Sci.

View full text Add to dashboard Cite

show abstract

“…Surprisingly, the fluorescence intensity of free PPIX increased nonlinearly with a rising concentration of PPIX. This may be caused by its low solubility and self-aggregation (37)(38) and may affect the PPIX concentration determination based on fluorescence spectroscopy. The EfeB-PPIX complex has a much higher fluorescence intensity compared with that of free PPIX at a corresponding concentration.…”

Section: Efeb Binds Ppix and Enhances Its Fluorescence Intensity-mentioning

confidence: 99%

“…The EfeB-PPIX complex can also be purified from cell lysate containing EfeB and additive PPIX. However, the ITC run for PPIX and EfeB does not provide a significant result because of its low solubility and self-stacking (37)(38). Titrating heme into the EfeB-PPIX complex gives a K d of 2.14 (Ϯ0.5) ϫ 10 Ϫ7 M (supplemental Fig.…”

Section: Efeb Binds Ppix and Enhances Its Fluorescence Intensity-mentioning

confidence: 99%

Crystal Structure and Biochemical Features of EfeB/YcdB from Escherichia coli O157

Wang

et al. 2011

Journal of Biological Chemistry

View full text Add to dashboard Cite

EfeB/YcdB is a member of the dye-decolorizing peroxidase (DyP) protein family. A recent study has shown that this protein can extract iron from heme without breaking the tetrapyrrole ring. We report the crystal structure of EfeB from Escherichia coli O157 bound to heme at 1.95 Å resolution. The EfeB monomer contains two domains. The heme molecule is located in a large hydrophobic pocket in the C-terminal domain. A long loop connecting the two domains extensively interacts with the heme, which is a distinctive structural feature of EfeB homologues. A large tunnel formed by this loop and the ␤-sheet of C-terminal domain provides a potential cofactor/substrate binding site. Biochemical data show that the production of protoporphyrin IX (PPIX) is closely related to the peroxidation activity. The mutant D235N keeps nearly the same activity of guaiacol peroxidase as the wild-type protein, whereas the corresponding mutation in the classic DyP protein family completely abolished the peroxidation activity. These results suggest that EfeB is a unique member of the DyP protein family. In addition, dramatically enhanced fluorescence excitation and emission of EfeB-PPIX was observed, implying this protein may be used as a red color fluorescence marker.

show abstract

“…The LED downlight was chosen because PpIX exhibits maximum absorbance at 409 nm light but also absorbs components at longer wavelengths in the visible light range [21], and white light contains all visible light, ranging from 400-700 nm. Furthermore, when considering the LED downlight properties and size, the heat generated by the light is very low and it is useful for the exposure of an entire 96-well plate.…”

Section: In Vitro Pdt Studymentioning

confidence: 99%

“…PpIX exists as a monomer in the pH range 0-3, as an aggregate in the pH range 3-8 and as a dimer in any pH greater than 8 [21]. Aggregation behavior of PpIX in neutral water makes it inappropriate for use in physiological studies.…”

Section: Introductionmentioning

confidence: 99%

Protoporphyrin IX-loaded magnetoliposomes as a potential drug delivery system for photodynamic therapy: Fabrication, characterization and in vitro study

Başoğlu

Bilgin

Demir

2016

Photodiagnosis and Photodynamic Therapy

View full text Add to dashboard Cite

Graphical abstractHighlights  Protoporphyrin IX loaded Magnetoliposomes were produced small enough for intravenous application. Protoporphyrin IX loaded Magnetoliposomes indicated its efficacy in nano-molar concentration for the in vitro photodynamic therapy application.  White LED light source provides sufficient energy to stimulate protoporphyrin molecules in cell culture. AbstractBackground: Protoporphyrin IX (PpIX) is a well-known photosensitizer that has great potential

show abstract

Aggregation Behavior of Protoporphyrin IX in Aqueous Solutions: Clear Evidence of Vesicle Formation

Cited by 155 publications

References 35 publications

Fluorometric quantification of protoporphyrin IX in biological skin samples from in vitro penetration/permeation studies

Fluorometric quantification of protoporphyrin IX in biological skin samples from in vitro penetration/permeation studies

Crystal Structure and Biochemical Features of EfeB/YcdB from Escherichia coli O157

Protoporphyrin IX-loaded magnetoliposomes as a potential drug delivery system for photodynamic therapy: Fabrication, characterization and in vitro study

Contact Info

Product

Resources

About