scholarly journals Annexin V Release and Transmembrane Mitochondrial Potential during Storage of Apheresis-Derived Platelets Treated for Pathogen Reduction

2010 ◽  
Vol 37 (1) ◽  
pp. 7-12 ◽  
Author(s):  
Susanne M. Picker ◽  
Larissa Oustianskaia ◽  
Volker Schneider ◽  
Birgit S. Gathof
Keyword(s):  
Annexin V ◽  
Mitochondrial Potential ◽  
Pathogen Reduction ◽  
Transmembrane Mitochondrial Potential

scholarly journals Evaluation of viability indicators of bovine spermatozoa after exposure to silicon dimethylglycerolate using flow cytofluorimetry

2021 ◽  
Vol 209 (06) ◽  
pp. 53-60
Author(s):  
Alena Nakidkina ◽  
T. I. KUZMINA
Keyword(s):  
Transmembrane Potential ◽  
Membrane Integrity ◽  
Chemical Properties ◽  
Annexin V ◽  
Mitochondrial Potential ◽  
Male Gametes ◽  
Biological Compatibility ◽  
Wide Range ◽  
Bovine Spermatozoa ◽  
Transmembrane Mitochondrial Potential

Abstract. Silicon and its dioxide (silica) demonstrate good biological compatibility and a wide range of physical and chemical properties, depending on the production and processing method. In particular, silicon dimethylglycerolate (SDMG) has transmucous and transcutaneous drug conductivity, and, as a hydrogel, may be of interest for the oocytes and embryos cultivation medium structuring and/or media for cryopreservation/thawing of gametes. The aim of this study was to examine the effect of SDMG at concentrations of 0.2 % and 0.02 % on the transmembrane potential of mitochondria and cell viability of bovine spermatozoa. Methods. Sperm subpopulations were assessed for (non)viability indicators (disrupted transmembrane potential of mitochondria, externalization of phosphatidylserine and plasma membrane integrity loss) by flow cytometry with two sets of fluorescent probes. Mitochondrial transmembrane potential was measured using 3,3'-dihexyloxacarbocyanine iodide (DiOC6(3))/ethidium bromide, and externalization of phosphatidylserine – using Annexin V-FITC/propidium iodide pair. The results of this work indicate that SDMG in concentrations of 0.2 % and 0.02 % does not affect the transmembrane mitochondrial potential, externalization of phosphatidylserine or necrotic processes in the population of bovine spermatozoa. The scientific novelty. The data is obtained for the first time on the absence of cytotoxicity of SDMG for male gametes. Together with the shown positive effect of this compound on the morphological parameters and the state of nuclear chromatin of porcine oocytes after intrafollicular vitrification, it should be concluded that silicon-containing glycerohydrogels are of interest as a component of sperm cryopreservation/thawing media.

Hypomethylation Therapy of Decitabine in Patients with Myelodysplastic Syndromes (MDS) Induces Apoptosis and Reduces Proliferation.

Blood ◽  
2005 ◽  
Vol 106 (11) ◽  
pp. 371-371
Author(s):  
I. Jilani ◽  
H. Kantarjian ◽  
M. Gorre ◽  
J.-P. Issa ◽  
J. Bennett ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Annexin V ◽  
Wilcoxon Test ◽  
Brdu Incorporation ◽  
Direct Result ◽  
Multiparameter Flow Cytometry ◽  
Aberrant Methylation ◽  
Mitochondrial Potential ◽  
Cd34 Cells ◽  
Significant Difference

Abstract Background: MDS is characterized by cytopenias believed to be the direct result of increased apoptosis of the hematopoietic cells in the bone marrow despite an increase in proliferation. Aberrant methylation resulting in leukemogenesis is frequent in MDS and is a potential target for pharmacologic therapy. Decitabine (Dacogen™; DAC), which has been shown to be effective in treating patients with MDS (Saba et al, Kantarjian et al), indirectly depletes methylcytosine, resulting in hypomethylation of target genes. However, it is not known whether this correction of methylation leads to reducing the apoptosis, allowing normal cells to grow, or by increasing the apoptosis killing off the tumor cells. Methods: We studied apoptosis and proliferation in patients with MDS treated on a randomized protocol to receive either DAC or best supportive care (SC). Apoptosis as measured by annexin V and mitochondrial potential was studied in various subpopulations of cells using multiparameter flow cytometry. Proliferation was also measured in a similar fashion using BrdU incorporation. Bone marrow (BM) and peripheral blood (PB) samples were collected from patients at baseline and at various times on therapy. Results: At baseline, the DAC group showed no significant difference in apoptosis, proliferation or percent of CD34+ cells from the SC group in BM (n = 39 and 33 patients, respectively) or PB (n = 51 and 50, respectively). After three months on treatment a significant increase in apoptosis (annexin V) in CD34+ cells was noted in the DAC arm but not in the SC arm (Wilcoxon test, P=0.01). Similar results were obtained when disturbance in mitochondrial potential was measured in the blast population (P=0.02). Interestingly, a similar increase in apoptosis was observed in CD8+ cells in the DAC arm (P=0.02). The increase in apoptosis was augmented with additional courses of DAC therapy. A greater reduction in proliferation (BrdU incorporation) in the CD34+ cells in the decitabine arm compared to SC (P=0.01) was also observed. Those DAC-treated patients who had a higher proliferation rate (BrdU incorporation) at diagnosis were more likely to achieve a CR than those with low level of proliferation (P=0.03). Conclusions: Decitabine therapy in patients with MDS leads to hypomethylation, but the net effects include high levels of apoptosis and the death of the neoplastic cells and a complimentary reduction of proliferation of the leukemic cells.

scholarly journals Targeting Lon Protease Activity Promotes Growth Inhibition and Apoptosis in Acute Myeloid Leukemia Cell Lines

Blood ◽  
2014 ◽  
Vol 124 (21) ◽  
pp. 5130-5130
Author(s):  
Serena De Matteis ◽  
Silvia Carloni ◽  
Roberta Napolitano ◽  
Michele Zanoni ◽  
Lara Gibellini ◽  
...  
Keyword(s):  
Acute Myeloid Leukemia ◽  
Cell Lines ◽  
Protease Activity ◽  
Myeloid Leukemia ◽  
Annexin V ◽  
Time Dependent ◽  
Lon Protease ◽  
Mitochondrial Potential ◽  
Induction Of Apoptosis ◽  
Acute Myeloid

Abstract INTRODUCTION Acute myeloid leukemia (AML) is a progressive malignant disorder of the myeloid lineage of hematopoietic cells, where overall three-year survival is below 20% for patients above 65 years (1). Therapy resistance or relapsed are the main causes of mortality among these patients. The functional changes that affect mitochondria may contribute to the development of cancer (2). The mitochondrial Lon, ubiquitously expressed in human cells and tissues, seems to be involved in protein quality control and stress response pathways in mitochondria. Several stress conditions induce an upregulation of Lon that may promote tumorigenesis by inhibiting apoptosis and promoting cell proliferation, transformation and angiogenesis (3). The role of Lon in AML has not been described yet. Our study aims to investigate if the activity of the protease may be linked to AML leukemogenesis and drug resistance. In particular we tested Lon protease activity inhibitors to clarify a new pathogenetic mechanism in controlling drug responsiveness in this haematological disorder. METHODS AML cell lines Kasumi-1 (acute myeloblastic leukemia), MOLM-13 (acute monocytic leukemia) and HL-60 (acute promyelocytic leukemia) were used to perform the experiments. Lon expression was measured by RT-PCR and immunoblotting. Cell-growth was quantified by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Reactive oxygen species (ROS) were evaluated by Nitro Blue Tetrazolium (NBT) assay. For cell cycle analysis, cells were fixed with ethanol, resuspended in phosphate-buffered saline (PBS) containing 0.1mg/mL RNase A, NP40 0,15% and 100mg/mL of propidium iodide (PI) and analysed by using flow cytometry and Modfit software. The early and late apoptosis was assessed with Annexin V-FITC Apoptosis Detection kit. To measure the mitochondrial potential, JC-1 Mitochondrial Membrane Potential assay kit was used. RESULTS Lon protease was expressed at high levels in AML cell lines compared with resting or activated peripheral blood mononuclear cells and monocytes, but less than other cancer cell lines such as K562 (human erythromyeloblastoid leukemia), HDLM-2 (Hodgkin lymphoma, nuclear sclerosing), SHSY5Y (neuroblastoma), MCF7 (breast carcinoma). Based on previous works (4), which show that 2-Cyano-3,12-dioxooleana-1,9-dien-28-oic acid (CDDO) and its derivates inhibit the Lon protease activity, we evaluated the effect of two inhibitors on cell growth and the induction of apoptosis. The leukemic cell lines were treated with CDDO and CDDO-Me (CDDO methyl-ester) at different concentrations for 24, 48 and 72h. Both compounds inhibited the cell proliferation in a dose- and time-dependent manner and induced an increase of ROS. We observed a time-dependent increase in the percentage of Annexin V+ cells after treatment with CDDO (1,25µM) and CDDO-Me (0,25µM). The induction of apoptosis after CDDO exposure was important only at 72h, while we observed a percentage of apoptosis over 60% starting from 24h of CDDO-Me treatment. Interestingly, CDDO induced a time-dependent arrest in G2/M phase in MOLM-13 cells and in G0/G1 phase in Kasumi-1 and HL-60 cells. Instead the CDDO-Me induced an alteration of all cell cycle phases, probably for its cytotoxicity. Moreover CDDO-Me caused a higher decrease in mitochondrial potential (Dψm) than CDDO in all cell lines. CONCLUSION In conclusion, these findings show that Lon protease inhibitors promote growth inhibition and apoptosis and support the concept that mitochondrial Lon may represent a novel anticancer drug target. REFERENCES 1. Burnett A, Wetzler M, Lowenberg B. Therapeutic advances in acute myeloid leukemia. J Clin Oncol. 2011, 29:487-494. 2. Gogvadze V, Orrenius S, Zhivotovsky B. Mitochondria in cancer cells: what is so special about them? Trends Cell Biol. 2008, 18(4):165-73. 3. Ngo JK, Pomatto LC, Davies KJ. Upregulation of the mitochondrial Lon Protease allows adaptation to acute oxidative stress but dysregulation is associated with chronic stress, disease, and aging. Redox Biol. 2013, 1(1):258-64. 4. Bernstein SH, Venkatesh S, Li M, Lee J, Lu B, Hilchey SP, Morse KM, Metcalfe HM, Skalska J, Andreeff M, Brookes PS, Suzuki CK. The mitochondrial ATP-dependent Lon protease: a novel target in lymphoma death mediated by the synthetic triterpenoid CDDO and its derivatives. Blood. 2012, 119(14):3321-9. Disclosures Martinelli: Novartis: Consultancy, Speakers Bureau; BMS: Consultancy, Speakers Bureau; Pfizer: Consultancy; ARIAD: Consultancy.

scholarly journals Insight into Factors Influencing Wound Healing Using Phosphorylated Cellulose-Filled-Chitosan Nanocomposite Films

2021 ◽  
Vol 22 (21) ◽  
pp. 11386
Author(s):  
Marta Kędzierska ◽  
Sara Blilid ◽  
Katarzyna Miłowska ◽  
Joanna Kołodziejczyk-Czepas ◽  
Nadia Katir ◽  
...  
Keyword(s):  
Wound Healing ◽  
Antioxidant Properties ◽  
Nanocomposite Films ◽  
Coagulation System ◽  
Mitochondrial Potential ◽  
Transparent Films ◽  
Accelerate Wound Healing ◽  
Hemostatic Activity ◽  
Potential Factors ◽  
Transmembrane Mitochondrial Potential

Marine polysaccharides are believed to be promising wound-dressing nanomaterials because of their biocompatibility, antibacterial and hemostatic activity, and ability to easily shape into transparent films, hydrogels, and porous foams that can provide a moist micro-environment and adsorb exudates. Current efforts are firmly focused on the preparation of novel polysaccharide-derived nanomaterials functionalized with chemical objects to meet the mechanical and biological requirements of ideal wound healing systems. In this contribution, we investigated the characteristics of six different cellulose-filled chitosan transparent films as potential factors that could help to accelerate wound healing. Both microcrystalline and nano-sized cellulose, as well as native and phosphorylated cellulose, were used as fillers to simultaneously elucidate the roles of size and functionalization. The assessment of their influences on hemostatic properties indicated that the tested nanocomposites shorten clotting times by affecting both the extrinsic and intrinsic pathways of the blood coagulation system. We also showed that all biocomposites have antioxidant capacity. Moreover, the cytotoxicity and genotoxicity of the materials against two cell lines, human BJ fibroblasts and human KERTr keratinocytes, was investigated. The nature of the cellulose used as a filler was found to influence their cytotoxicity at a relatively low level. Potential mechanisms of cytotoxicity were also investigated; only one (phosphorylated microcellulose-filled chitosan films) of the compounds tested produced reactive oxygen species (ROS) to a small extent, and some films reduced the level of ROS, probably due to their antioxidant properties. The transmembrane mitochondrial potential was very slightly lowered. These biocompatible films showed no genotoxicity, and very importantly for wound healing, most of them significantly accelerated migration of both fibroblasts and keratinocytes.

scholarly journals Efavirenz Induced Suicidal Death of Human Erythrocytes

2015 ◽  
Vol 37 (6) ◽  
pp. 2496-2507 ◽  
Author(s):  
Rosi Bissinger ◽  
Ghada Bouguerra ◽  
Abdulla Al Mamun Bhuyan ◽  
Sabrina Waibel ◽  
Salem Abbès ◽  
...  
Keyword(s):  
Kinase Inhibitor ◽  
Annexin V ◽  
Casein Kinase ◽  
Human Erythrocytes ◽  
Cell Shrinkage ◽  
P38 Kinase ◽  
Mitochondrial Potential ◽  
Forward Scatter ◽  
Erythrocyte Surface ◽  
Phosphatidylserine Translocation

Background/Aims: The reverse transcriptase inhibitor efavirenz utilized for the treatment of human immunodeficiency virus (HIV)-1 infection, triggers suicidal cell death or apoptosis, an effect in part due to interference with mitochondrial potential. Side effects of efavirenz include anemia. Causes of anemia include accelerated clearance of circulating erythrocytes. Even though lacking mitochondria, erythrocytes may enter suicidal erythrocyte death or eryptosis, which is characterized by cell shrinkage and cell membrane scrambling with phosphatidylserine translocation to the erythrocyte surface. Triggers of eryptosis include Ca2+ entry and increase of cytosolic Ca2+ activity ([Ca2+]i), oxidative stress, ceramide, as well as activation of p38 kinase, casein kinase 1α and/or cyclooxygenase. The present study explored, whether and how efavirenz induces eryptosis. Methods: Phosphatidylserine exposure at the cell surface was estimated from annexin V binding, cell volume from forward scatter, [Ca2+]i from Fluo3-fluorescence, ROS formation from DCFDA dependent fluorescence, and ceramide abundance utilizing selective antibodies. Results: A 48 hours exposure of human erythrocytes to efavirenz (≥ 2 µg/ml) significantly increased the percentage of annexin-V-binding cells, significantly decreased forward scatter (2 µg/ml), significantly increased Fluo3-fluorescence (≥ 2 µg/ml), but did not significantly modify DCFDA fluorescence or ceramide abundance. The effect of efavirenz on annexin-V-binding was significantly blunted, but not abolished by removal of extracellular Ca2+. The effect of efavirenz on annexin-V-binding was further significantly blunted by p38 kinase inhibitor SB203580 (2 µM) and casein kinase 1α inhibitor D4476 (10 µM), but not by cyclooxygenase inhibitor aspirin (50 µM). Conclusions: Efavirenz triggers cell shrinkage and phosphatidylserine translocation to the erythrocyte surface, an effect in part due to stimulation of Ca2+ entry as well as activation of p38 kinase and casein kinase 1α.

Platelet Mitochondrial Potential and Function Is Altered During Severe Septic Shock.

Blood ◽  
2009 ◽  
Vol 114 (22) ◽  
pp. 5074-5074
Author(s):  
Andrea Artoni ◽  
Alessandro Protti ◽  
Anna Lecchi ◽  
Giovanna Motta ◽  
Francesca Rosini ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Septic Shock ◽  
Platelet Aggregation ◽  
Platelet Function ◽  
Sofa Score ◽  
Conflicts Of Interest ◽  
Platelet Rich Plasma ◽  
Annexin V ◽  
Mitochondrial Potential ◽  
Serotonin Content

Abstract Abstract 5074 Background Septic shock is a condition characterized by a systemic inflammatory response associated to organ dysfunction. Primary hemostasis is impaired in sepsis, and multiple causative mechanisms have been proposed. Methods In this study we evaluated platelet function in patients with septic shock. In particular platelet mitochondrial potential was studied as a possible cause of platelet function impairment in septic shock. To test the hypothesis we enrolled 21 consecutive patients admitted to our intensive care unit (ICU) with diagnosis of septic shock. All patients were analysed within 24 hours from admission. 30 ml of citrated blood were withdrawn from each patient and the platelet function tests were performed. Platelet mitochondrial potential (Dy) was assessed by flow cytometry through JC1 staining, espressed as the ratio of FL-1 and FL-2 fluorescence. Platelet aggregation was performed on platelet rich plasma in 5 patients not assuming any drug known to influence platelet function with the following aggregating agents: ADP (final concentration 4 μM), collagen (2 μg/ml), U46619 (10 μM) and TRAP (10 μM). Platelet ATP secretion was quantified and intraplatelet δ-granules content dosage was performed. Expression of GpIb, of GpIIb/IIIa, and of annexin V binding were evaluated by flow cytometry. The study was approved by the Hospital Ethical Committee. Results at admission platelet mitochondrial potential was reduced in septic patients in comparison to normal controls (2.6±1.4 vs 3.4±0.3 p=0.06). At day one there was a good correlation between platelet mitochondrial impairment and the Sepsis-related Organ Failure Assessment (SOFA) score, an indicator of sepsis severity (R2=0,39 p<0.001 n=18) with higher values indicating higher degree of severity. Platelets taken from patients with a SOFA score above the group median value (>9, n=8) had Dy values significantly lower than those taken from less severely ill patients (SOFA score ' 9, n=10) and healthy volunteers (1.6±0.6 vs. 3.3±1.4 vs. 3.4±0.3; p<0.01 one way analysis of variance). At day one platelet aggregation was severely impaired, especially with strong aggregating agents as collagen (3/5 of patients with maximum platelet aggregation below 25%). The maximum aggregation caused by collagen seemed to correlate to platelet mitochondrial potential (R2=0,72 n=5 p=0.07). Platelet secretion was equally defective at admission, with pathological secretion to collagen stimulus in 40% of tested patients. Intraplatelet ADP content resulted low in 66% tested (n= 12), while serotonin content was pathological in 58% of patients (n= 12). Platelet GpIIb/IIIa and GpIb expression and annexin V binding were within normal limits. Conclusion we demonstrated in a very well selected group of patients that during the early phase of a septic shock a mitochondrial dysfunction occurs, and this may have an impact on platelets dysfunction observed in this condition. Disclosures No relevant conflicts of interest to declare.

scholarly journals Cytotoxicity of PP(Arg)2- and PP(Ala)2(Arg)2-based photodynamic therapy and early stage of apoptosis induction in human breast cancers in vitro.

2012 ◽  
Vol 59 (4) ◽  
Author(s):  
Agata Nowak-Stępniowska ◽  
Katarzyna Wiktorska ◽  
Maciej Małecki ◽  
Małgorzata Milczarek ◽  
Katarzyna Lubelska ◽  
...  
Keyword(s):  
Photodynamic Therapy ◽  
Apoptotic Cell ◽  
Early Stage ◽  
Protoporphyrin Ix ◽  
Membrane Receptors ◽  
Breast Cancers ◽  
Apoptosis Induction ◽  
Mitochondrial Potential ◽  
Transmembrane Mitochondrial Potential

Mitochondria are cell energetic centers where ATP is produced. They also play a very important role in the PDT as intracellular sites of photosensitizer localization. Photosensitizers gathering in mitochondria (like porphyrin derivatives used in this work) are more effective in tumor cell destruction. Moreover, it was assumed that di-amino acid substituents attached to porphyrin ring will strengthen the effectivity of interaction with membrane receptors of examined cells. MTT assay was performed to investigate the influence of PP(Arg)(2) and PP(Ala)(2)(Arg)(2)-based PDT on breast cancer cell viability for 24 h, 48 h and 120 h after cell irradiation. Then the influence of PP(Ala)(2)(Arg)(2)- and PP(Arg)(2)-mediated PDT on early mitochondrial apoptosis induction via measurements of the transmembrane mitochondrial potential changes was examined. Results showed that lower energy doses and maximal nontoxic photosensitizer doses of PP(Ala)(2)(Arg)(2) and PP(Arg)(2) applied in PDT can imply apoptotic cell death. It was confirmed that modification of the protoporphyrin IX by attaching two alanine substituents raised the efficiency of photodynamic therapy.

scholarly journals Cell cycle and transmembrane mitochondrial potential analysis after treatment with chromium(iii), iron(iii), molybdenum(iii) or nickel(ii) and their mixtures

2019 ◽  
Vol 8 (2) ◽  
pp. 188-195 ◽  
Author(s):  
Sylwia Terpilowska ◽  
Andrzej K. Siwicki
Keyword(s):  
Cell Cycle ◽  
Hepg2 Cells ◽  
Transmembrane Potential ◽  
Mitochondrial Transmembrane Potential ◽  
Mitochondrial Potential ◽  
Potential Analysis ◽  
Transmembrane Mitochondrial Potential

The aim of this study was to examine the effect of chromium(iii), iron(iii), molybdenum(iii) and nickel(ii) and their combinations on the cell cycle and mitochondrial transmembrane potential (MTP) in BALB/3T3 and HepG2 cells.

scholarly journals Ruthenium(II) and Platinum(II) Complexes with Biologically Active Aminoflavone Ligands Exhibit In Vitro Anticancer Activity

2021 ◽  
Vol 22 (14) ◽  
pp. 7568
Author(s):  
Małgorzata Fabijańska ◽  
Maria M. Kasprzak ◽  
Justyn Ochocki
Keyword(s):  
Anticancer Activity ◽  
Cell Lines ◽  
Cisplatin Resistance ◽  
Annexin V ◽  
Biologically Active ◽  
Caspase 8 ◽  
Mitochondrial Potential ◽  
Apoptosis Pathway ◽  
Ruthenium Compounds

Continuing our studies on the mechanisms underlying the cytotoxicity of potential drugs, we have described several aspects of the in vitro anticancer activity of ruthenium(II) and platinum(II) complexes with bioactive, synthetic aminoflavone ligands. We examined the mechanism of proapoptotic activity of cis-dichlorobis(3-imino-2-methoxyflavanone)ruthenium(II), cis-dichlorobis(3-imino-2-ethoxyflavanone)ruthenium(II), and trans-dichlorobis(3-aminoflavone)platinum(II). Cisplatin was used as a reference compound. The cytotoxicity was investigated by MTT assay. The mechanism of proapoptotic activity of the tested compounds was investigated by evaluation of caspase-8 activity, cytometric analysis of annexin-V positive cells, and mitochondrial potential loss measurement. The results showed that ruthenium compounds break partially or completely the cisplatin resistance by activating the caspase 8-dependent apoptosis pathway and loss of mitochondrial membrane potential. Platinum compounds also have a cytostatic effect, but their action requires more exposure time. Potential mechanisms underlying drug resistance in the two pairs of cancer cell lines were investigated: total glutathione content, P-glycoprotein activity, and differences in the activity of DNA repair induced by nucleotide excision. Results showed that cisplatin-resistant cells have elevated glutathione levels relative to sensitive cells. Moreover, they indicated the mechanisms enabling cells to avoid apoptosis caused by DNA damage. Pg-P activity has no effect on the development of cisplatin resistance in the cell lines described.

scholarly journals Cytotoxicity of PP(Arg)(2)- and Hp(Arg)(2)-mediated photodynamic therapy and early stage of apoptosis induction in prostate carcinoma in vitro.

2011 ◽  
Vol 58 (4) ◽  
Author(s):  
Agata Nowak-Stępniowska ◽  
Katarzyna Wiktorska ◽  
Maciej Małecki ◽  
Anna Romiszewska ◽  
Alfreda Padzik-Graczyk
Keyword(s):  
Prostate Carcinoma ◽  
Mitochondrial Membrane ◽  
Laser Scanning ◽  
Early Stage ◽  
Mitochondrial Potential ◽  
Mitochondrial Membrane Depolarization ◽  
Porphyrin Photosensitizers ◽  
Transmembrane Mitochondrial Potential ◽  
Energy Dose

Porphyrin photosensitizers tend to localize in mitochondria. The depolarization of mitochondrial membrane is one of the early stages of apoptosis and Laser Scanning Fluorescence Microscopy allows to determine changes in transmembrane mitochondrial potential under influence of PDT depending on the kind of photosensitizer (PP(Arg)(2), Hp(Arg)(2)), the energy dose (5, 10, 30 and 50 J/cm(2)) and time periods (24 and 48 hours after irradiation) in the LNCaP (lymphonodal metastasis of prostate carcinoma, the androgen dependent cell line). Cyototoxicity induced by PP(Arg)(2)- and Hp(Arg)(2)-based PDT depending on energy dose and time after irradiation in prostate carcinoma is determined with MTT. Generally, it was shown that lower energy doses induce greater changes in transmembrane mitochondrial potential. Hp(Arg)(2)-based PDT was more effective causing greater mitochondrial membrane depolarization and cell viability decrease in comparison to PP(Arg)(2)-mediated PDT (in the case of maximal nontoxic photosensitizer doses used).

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