TY  - JOUR
AU  - Lazarević, J. J.
AU  - Ralević, U.
AU  - Kukolj, Tamara
AU  - Bugarski, Diana
AU  - Lazarević, N.
AU  - Bugarski, Branko
AU  - Popović, Z., V
PY  - 2019
UR  - http://TechnoRep.tmf.bg.ac.rs/handle/123456789/4213
AB  - In investigation of (patho)physiological processes, cells represent frequently used analyte as an exceptional source of information. However, spectroscopic analysis of live cells is still very seldom in clinics, as well as in research studies. Among others, the reasons are long acquisition time during which autolysis process is activated, necessity of specified technical equipment, and inability to perform analysis in a moment of sample preparation. Hence, an optimal method of preserving cells in the existing state is of extreme importance, having in mind that selection of fixative is cell lineage dependent. In this study, two commonly used chemical fixatives, formaldehyde and methanol, are used for preserving primary mesenchymal stem cells extracted from periodontal ligament, which are valuable cell source for reconstructive dentistry. By means of Raman spectroscopy, cell samples were probed and the impact of these fixatives on their Raman response was analyzed and compared. Different chemical mechanisms are the core processes of formaldehyde and methanol fixation and certain Raman bands are shifted and/or of changed intensity when Raman spectra of cells fixed in that manner are compared. In order to get clearer picture, comprehensive statistical analysis was performed.
PB  - Pergamon-Elsevier Science Ltd, Oxford
T2  - Spectrochimica Acta Part A-Molecular and Biomolecular Spectroscopy
T1  - Influence of chemical fixation process on primary mesenchymal stem cells evidenced by Raman spectroscopy
EP  - 178
SP  - 173
VL  - 216
DO  - 10.1016/j.saa.2019.03.012
ER  - 

@article{
author = "Lazarević, J. J. and Ralević, U. and Kukolj, Tamara and Bugarski, Diana and Lazarević, N. and Bugarski, Branko and Popović, Z., V",
year = "2019",
abstract = "In investigation of (patho)physiological processes, cells represent frequently used analyte as an exceptional source of information. However, spectroscopic analysis of live cells is still very seldom in clinics, as well as in research studies. Among others, the reasons are long acquisition time during which autolysis process is activated, necessity of specified technical equipment, and inability to perform analysis in a moment of sample preparation. Hence, an optimal method of preserving cells in the existing state is of extreme importance, having in mind that selection of fixative is cell lineage dependent. In this study, two commonly used chemical fixatives, formaldehyde and methanol, are used for preserving primary mesenchymal stem cells extracted from periodontal ligament, which are valuable cell source for reconstructive dentistry. By means of Raman spectroscopy, cell samples were probed and the impact of these fixatives on their Raman response was analyzed and compared. Different chemical mechanisms are the core processes of formaldehyde and methanol fixation and certain Raman bands are shifted and/or of changed intensity when Raman spectra of cells fixed in that manner are compared. In order to get clearer picture, comprehensive statistical analysis was performed.",
publisher = "Pergamon-Elsevier Science Ltd, Oxford",
journal = "Spectrochimica Acta Part A-Molecular and Biomolecular Spectroscopy",
title = "Influence of chemical fixation process on primary mesenchymal stem cells evidenced by Raman spectroscopy",
pages = "178-173",
volume = "216",
doi = "10.1016/j.saa.2019.03.012"
}

Lazarević, J. J., Ralević, U., Kukolj, T., Bugarski, D., Lazarević, N., Bugarski, B.,& Popović, Z., V.. (2019). Influence of chemical fixation process on primary mesenchymal stem cells evidenced by Raman spectroscopy. in Spectrochimica Acta Part A-Molecular and Biomolecular Spectroscopy
Pergamon-Elsevier Science Ltd, Oxford., 216, 173-178.
https://doi.org/10.1016/j.saa.2019.03.012

Lazarević JJ, Ralević U, Kukolj T, Bugarski D, Lazarević N, Bugarski B, Popović ZV. Influence of chemical fixation process on primary mesenchymal stem cells evidenced by Raman spectroscopy. in Spectrochimica Acta Part A-Molecular and Biomolecular Spectroscopy. 2019;216:173-178.
doi:10.1016/j.saa.2019.03.012 .

Lazarević, J. J., Ralević, U., Kukolj, Tamara, Bugarski, Diana, Lazarević, N., Bugarski, Branko, Popović, Z., V, "Influence of chemical fixation process on primary mesenchymal stem cells evidenced by Raman spectroscopy" in Spectrochimica Acta Part A-Molecular and Biomolecular Spectroscopy, 216 (2019):173-178,
https://doi.org/10.1016/j.saa.2019.03.012 . .

TY  - JOUR
AU  - Bukara, Katarina
AU  - Jovanić, Svetlana
AU  - Drvenica, Ivana
AU  - Stančić, Ana
AU  - Ilić, Vesna Lj.
AU  - Rabasović, Mihailo D.
AU  - Pantelić, Dejan
AU  - Jelenković, Branislav
AU  - Bugarski, Branko
AU  - Krmpot, Aleksandar
PY  - 2017
UR  - http://TechnoRep.tmf.bg.ac.rs/handle/123456789/3747
AB  - The present study describes utilization of two photon excitation fluorescence (2PE) microscopy for visualization of the hemoglobin in human and porcine erythrocytes and their empty membranes (i.e., ghosts). High-quality, label-and fixation-free visualization of hemoglobin was achieved at excitation wavelength 730 nm by detecting visible autofluorescence. Localization in the suspension and spatial distribution (i.e., mapping) of residual hemoglobin in erythrocyte ghosts has been resolved by 2PE. Prior to the 2PE mapping, the presence of residual hemoglobin in the bulk suspension of erythrocyte ghosts was confirmed by cyanmethemoglobin assay. 2PE analysis revealed that the distribution of hemoglobin in intact erythrocytes follows the cells' shape. Two types of erythrocytes, human and porcine, characterized with discocyte and echinocyte morphology, respectively, showed significant differences in hemoglobin distribution. The 2PE images have revealed that despite an extensive washing out procedure after gradual hypotonic hemolysis, a certain amount of hemoglobin localized on the intracellular side always remains bound to the membrane and cannot be eliminated. The obtained results open the possibility to use 2PE microscopy to examine hemoglobin distribution in erythrocytes and estimate the purity level of erythrocyte ghosts in biotechnological processes.
PB  - Spie-Soc Photo-Optical Instrumentation Engineers, Bellingham
T2  - Journal of Biomedical Optics
T1  - Mapping of hemoglobin in erythrocytes and erythrocyte ghosts using two photon excitation fluorescence microscopy
IS  - 2
VL  - 22
DO  - 10.1117/1.JBO.22.2.026003
ER  - 

@article{
author = "Bukara, Katarina and Jovanić, Svetlana and Drvenica, Ivana and Stančić, Ana and Ilić, Vesna Lj. and Rabasović, Mihailo D. and Pantelić, Dejan and Jelenković, Branislav and Bugarski, Branko and Krmpot, Aleksandar",
year = "2017",
abstract = "The present study describes utilization of two photon excitation fluorescence (2PE) microscopy for visualization of the hemoglobin in human and porcine erythrocytes and their empty membranes (i.e., ghosts). High-quality, label-and fixation-free visualization of hemoglobin was achieved at excitation wavelength 730 nm by detecting visible autofluorescence. Localization in the suspension and spatial distribution (i.e., mapping) of residual hemoglobin in erythrocyte ghosts has been resolved by 2PE. Prior to the 2PE mapping, the presence of residual hemoglobin in the bulk suspension of erythrocyte ghosts was confirmed by cyanmethemoglobin assay. 2PE analysis revealed that the distribution of hemoglobin in intact erythrocytes follows the cells' shape. Two types of erythrocytes, human and porcine, characterized with discocyte and echinocyte morphology, respectively, showed significant differences in hemoglobin distribution. The 2PE images have revealed that despite an extensive washing out procedure after gradual hypotonic hemolysis, a certain amount of hemoglobin localized on the intracellular side always remains bound to the membrane and cannot be eliminated. The obtained results open the possibility to use 2PE microscopy to examine hemoglobin distribution in erythrocytes and estimate the purity level of erythrocyte ghosts in biotechnological processes.",
publisher = "Spie-Soc Photo-Optical Instrumentation Engineers, Bellingham",
journal = "Journal of Biomedical Optics",
title = "Mapping of hemoglobin in erythrocytes and erythrocyte ghosts using two photon excitation fluorescence microscopy",
number = "2",
volume = "22",
doi = "10.1117/1.JBO.22.2.026003"
}

Bukara, K., Jovanić, S., Drvenica, I., Stančić, A., Ilić, V. Lj., Rabasović, M. D., Pantelić, D., Jelenković, B., Bugarski, B.,& Krmpot, A.. (2017). Mapping of hemoglobin in erythrocytes and erythrocyte ghosts using two photon excitation fluorescence microscopy. in Journal of Biomedical Optics
Spie-Soc Photo-Optical Instrumentation Engineers, Bellingham., 22(2).
https://doi.org/10.1117/1.JBO.22.2.026003

Bukara K, Jovanić S, Drvenica I, Stančić A, Ilić VL, Rabasović MD, Pantelić D, Jelenković B, Bugarski B, Krmpot A. Mapping of hemoglobin in erythrocytes and erythrocyte ghosts using two photon excitation fluorescence microscopy. in Journal of Biomedical Optics. 2017;22(2).
doi:10.1117/1.JBO.22.2.026003 .

Bukara, Katarina, Jovanić, Svetlana, Drvenica, Ivana, Stančić, Ana, Ilić, Vesna Lj., Rabasović, Mihailo D., Pantelić, Dejan, Jelenković, Branislav, Bugarski, Branko, Krmpot, Aleksandar, "Mapping of hemoglobin in erythrocytes and erythrocyte ghosts using two photon excitation fluorescence microscopy" in Journal of Biomedical Optics, 22, no. 2 (2017),
https://doi.org/10.1117/1.JBO.22.2.026003 . .

TY  - JOUR
AU  - Drvenica, Ivana
AU  - Bukara, Katarina
AU  - Ilić, Vesna Lj.
AU  - Mišić, Danijela
AU  - Vasić, Borislav
AU  - Gajić, Radoš B.
AU  - Đorđević, Verica
AU  - Veljović, Đorđe
AU  - Belić, Aleksandar
AU  - Bugarski, Branko
PY  - 2016
UR  - http://TechnoRep.tmf.bg.ac.rs/handle/123456789/3329
AB  - The present study investigated preparation of bovine and porcine erythrocyte membranes from slaughterhouse blood as bio-derived materials for delivery of dexamethasone-sodium phosphate (DexP). The obtained biomembranes, i.e., ghosts were characterized in vitro in terms of morphological properties, loading parameters, and release behavior. For the last two, an UHPLC/-HESI-MS/MS based analytical procedure for absolute drug identification and quantification was developed. The results revealed that loading of DexP into both type of ghosts was directly proportional to the increase of drug concentration in the incubation medium, while incubation at 37 degrees C had statistically significant effect on loaded amount of DexP (P lt 0.05). The encapsulation efficiency was about fivefold higher in porcine compared to bovine ghosts. Insight into ghosts' surface morphology by field emission-scanning electron microscopy and atomic force microscopy confirmed that besides inevitable effects of osmosis, DexP inclusion itself had no observable additional effect on the morphology of the ghosts carriers. DexP release profiles were dependent on erythrocyte ghost type and amount of residual hemoglobin. However, sustained DexP release was achieved and shown over 3 days from porcine ghosts and 5 days from bovine erythrocyte ghosts.
PB  - Wiley, Hoboken
T2  - Biotechnology Progress
T1  - Biomembranes from slaughterhouse blood erythrocytes as prolonged release systems for dexamethasone sodium phosphate
EP  - 1055
IS  - 4
SP  - 1046
VL  - 32
DO  - 10.1002/btpr.2304
ER  - 

@article{
author = "Drvenica, Ivana and Bukara, Katarina and Ilić, Vesna Lj. and Mišić, Danijela and Vasić, Borislav and Gajić, Radoš B. and Đorđević, Verica and Veljović, Đorđe and Belić, Aleksandar and Bugarski, Branko",
year = "2016",
abstract = "The present study investigated preparation of bovine and porcine erythrocyte membranes from slaughterhouse blood as bio-derived materials for delivery of dexamethasone-sodium phosphate (DexP). The obtained biomembranes, i.e., ghosts were characterized in vitro in terms of morphological properties, loading parameters, and release behavior. For the last two, an UHPLC/-HESI-MS/MS based analytical procedure for absolute drug identification and quantification was developed. The results revealed that loading of DexP into both type of ghosts was directly proportional to the increase of drug concentration in the incubation medium, while incubation at 37 degrees C had statistically significant effect on loaded amount of DexP (P lt 0.05). The encapsulation efficiency was about fivefold higher in porcine compared to bovine ghosts. Insight into ghosts' surface morphology by field emission-scanning electron microscopy and atomic force microscopy confirmed that besides inevitable effects of osmosis, DexP inclusion itself had no observable additional effect on the morphology of the ghosts carriers. DexP release profiles were dependent on erythrocyte ghost type and amount of residual hemoglobin. However, sustained DexP release was achieved and shown over 3 days from porcine ghosts and 5 days from bovine erythrocyte ghosts.",
publisher = "Wiley, Hoboken",
journal = "Biotechnology Progress",
title = "Biomembranes from slaughterhouse blood erythrocytes as prolonged release systems for dexamethasone sodium phosphate",
pages = "1055-1046",
number = "4",
volume = "32",
doi = "10.1002/btpr.2304"
}

Drvenica, I., Bukara, K., Ilić, V. Lj., Mišić, D., Vasić, B., Gajić, R. B., Đorđević, V., Veljović, Đ., Belić, A.,& Bugarski, B.. (2016). Biomembranes from slaughterhouse blood erythrocytes as prolonged release systems for dexamethasone sodium phosphate. in Biotechnology Progress
Wiley, Hoboken., 32(4), 1046-1055.
https://doi.org/10.1002/btpr.2304

Drvenica I, Bukara K, Ilić VL, Mišić D, Vasić B, Gajić RB, Đorđević V, Veljović Đ, Belić A, Bugarski B. Biomembranes from slaughterhouse blood erythrocytes as prolonged release systems for dexamethasone sodium phosphate. in Biotechnology Progress. 2016;32(4):1046-1055.
doi:10.1002/btpr.2304 .

Drvenica, Ivana, Bukara, Katarina, Ilić, Vesna Lj., Mišić, Danijela, Vasić, Borislav, Gajić, Radoš B., Đorđević, Verica, Veljović, Đorđe, Belić, Aleksandar, Bugarski, Branko, "Biomembranes from slaughterhouse blood erythrocytes as prolonged release systems for dexamethasone sodium phosphate" in Biotechnology Progress, 32, no. 4 (2016):1046-1055,
https://doi.org/10.1002/btpr.2304 . .

TY  - JOUR
AU  - Bukara, Katarina
AU  - Drvenica, Ivana
AU  - Ilić, Vesna Lj.
AU  - Stančić, Ana
AU  - Mišić, Danijela
AU  - Vasić, Borislav
AU  - Gajić, Radoš
AU  - Vučetić, Dušan
AU  - Kiekens, Filip
AU  - Bugarski, Branko
PY  - 2016
UR  - http://TechnoRep.tmf.bg.ac.rs/handle/123456789/3318
AB  - The objective of our study was to develop controlled drug delivery system based on erythrocyte ghosts for amphiphilic compound sodium diclofenac considering the differences between erythrocytes derived from two readily available materials - porcine slaughterhouse and outdated transfusion human blood. Starting erythrocytes, empty erythrocyte ghosts and diclofenac loaded ghosts were compared in terms of the encapsulation efficiency, drug releasing profiles, size distribution, surface charge, conductivity, surface roughness and morphology. The encapsulation of sodium diclofenac was performed by an osmosis based process - gradual hemolysis. During this process sodium diclofenac exerted mild and delayed antihemolytic effect and increased potassium efflux in porcine but not in outdated human erythrocytes. FTIR spectra revealed lack of any membrane lipid disorder and chemical reaction with sodium diclofenac in encapsulated ghosts. Outdated human erythrocyte ghosts with detected nanoscale damages and reduced ability to shrink had encapsulation efficiency of only 8%. On the other hand, porcine erythrocyte ghosts had encapsulation efficiency of 37% and relatively slow drug release rate. More preserved structure and functional properties of porcine erythrocytes related to their superior encapsulation and release performances, define them as more appropriate for the usage in sodium diclofenac encapsulation process.
PB  - Elsevier Science Bv, Amsterdam
T2  - Journal of Biotechnology
T1  - Comparative studies on osmosis based encapsulation of sodium diclofenac in porcine and outdated human erythrocyte ghosts
EP  - 22
SP  - 14
VL  - 240
DO  - 10.1016/j.jbiotec.2016.10.017
ER  - 

@article{
author = "Bukara, Katarina and Drvenica, Ivana and Ilić, Vesna Lj. and Stančić, Ana and Mišić, Danijela and Vasić, Borislav and Gajić, Radoš and Vučetić, Dušan and Kiekens, Filip and Bugarski, Branko",
year = "2016",
abstract = "The objective of our study was to develop controlled drug delivery system based on erythrocyte ghosts for amphiphilic compound sodium diclofenac considering the differences between erythrocytes derived from two readily available materials - porcine slaughterhouse and outdated transfusion human blood. Starting erythrocytes, empty erythrocyte ghosts and diclofenac loaded ghosts were compared in terms of the encapsulation efficiency, drug releasing profiles, size distribution, surface charge, conductivity, surface roughness and morphology. The encapsulation of sodium diclofenac was performed by an osmosis based process - gradual hemolysis. During this process sodium diclofenac exerted mild and delayed antihemolytic effect and increased potassium efflux in porcine but not in outdated human erythrocytes. FTIR spectra revealed lack of any membrane lipid disorder and chemical reaction with sodium diclofenac in encapsulated ghosts. Outdated human erythrocyte ghosts with detected nanoscale damages and reduced ability to shrink had encapsulation efficiency of only 8%. On the other hand, porcine erythrocyte ghosts had encapsulation efficiency of 37% and relatively slow drug release rate. More preserved structure and functional properties of porcine erythrocytes related to their superior encapsulation and release performances, define them as more appropriate for the usage in sodium diclofenac encapsulation process.",
publisher = "Elsevier Science Bv, Amsterdam",
journal = "Journal of Biotechnology",
title = "Comparative studies on osmosis based encapsulation of sodium diclofenac in porcine and outdated human erythrocyte ghosts",
pages = "22-14",
volume = "240",
doi = "10.1016/j.jbiotec.2016.10.017"
}

Bukara, K., Drvenica, I., Ilić, V. Lj., Stančić, A., Mišić, D., Vasić, B., Gajić, R., Vučetić, D., Kiekens, F.,& Bugarski, B.. (2016). Comparative studies on osmosis based encapsulation of sodium diclofenac in porcine and outdated human erythrocyte ghosts. in Journal of Biotechnology
Elsevier Science Bv, Amsterdam., 240, 14-22.
https://doi.org/10.1016/j.jbiotec.2016.10.017

Bukara K, Drvenica I, Ilić VL, Stančić A, Mišić D, Vasić B, Gajić R, Vučetić D, Kiekens F, Bugarski B. Comparative studies on osmosis based encapsulation of sodium diclofenac in porcine and outdated human erythrocyte ghosts. in Journal of Biotechnology. 2016;240:14-22.
doi:10.1016/j.jbiotec.2016.10.017 .

Bukara, Katarina, Drvenica, Ivana, Ilić, Vesna Lj., Stančić, Ana, Mišić, Danijela, Vasić, Borislav, Gajić, Radoš, Vučetić, Dušan, Kiekens, Filip, Bugarski, Branko, "Comparative studies on osmosis based encapsulation of sodium diclofenac in porcine and outdated human erythrocyte ghosts" in Journal of Biotechnology, 240 (2016):14-22,
https://doi.org/10.1016/j.jbiotec.2016.10.017 . .