Publications by authors named "Andrei Moroz"

23 Publications

  • Page 1 of 1

Exposure to Bacteriophages T4 and M13 Increases Integrin Gene Expression and Impairs Migration of Human PC-3 Prostate Cancer Cells.

Antibiotics (Basel) 2021 Oct 3;10(10). Epub 2021 Oct 3.

Laboratory of Extracellular Matrix Biology, Department of Structural and Functional Biology, Institute of Biosciences of Botucatu, Sao Paulo State University (UNESP), Botucatu 18618-689, SP, Brazil.

The interaction between bacteriophages and integrins has been reported in different cancer cell lines, and efforts have been undertaken to understand these interactions in tumor cells along with their possible role in gene alterations, with the aim to develop new cancer therapies. Here, we report that the non-specific interaction of T4 and M13 bacteriophages with human PC-3 cells results in differential migration and varied expression of different integrins. PC-3 tumor cells (at 70% confluence) were exposed to 1 × 10 pfu/mL of either lytic T4 bacteriophage or filamentous M13 bacteriophage. After 24 h of exposure, cells were processed for a histochemical analysis, wound-healing migration assay, and gene expression profile using quantitative real-time PCR (qPCR). qPCR was performed to analyze the expression profiles of integrins , , , , and . Our findings revealed that PC-3 cells interacted with T4 and M13 bacteriophages, with significant upregulation of , , , genes after phage exposure. PC-3 cells also exhibited reduced migration activity when exposed to either T4 or M13 phages. These results suggest that wildtype bacteriophages interact non-specifically with PC-3 cells, thereby modulating the expression of integrin genes and affecting cell migration. Therefore, bacteriophages have future potential applications in anticancer therapies.
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http://dx.doi.org/10.3390/antibiotics10101202DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8532711PMC
October 2021

Thinking Outside the Box to Obtain the Best Antibodies!

Authors:
Andrei Moroz

Monoclon Antib Immunodiagn Immunother 2021 10;40(5):201-202

Monoclonal Antibodies Laboratory, Department of Clinical Analysis, School of Pharmaceutical Sciences, São Paulo State University (UNESP), Araraquara, Brazil.

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http://dx.doi.org/10.1089/mab.2021.0041DOI Listing
October 2021

Detection of factor VIII and D-dimer biomarkers for venous thromboembolism diagnosis using electrochemistry immunosensor.

Talanta 2020 Nov 25;219:121241. Epub 2020 Jun 25.

Universidade Federal de São Paulo, Instituto de Ciência e Tecnologia, São José Dos Campos, SP, Brazil. Electronic address:

Venous thromboembolism (VTE) is a serious clinical condition which early and accurate diagnosis may contribute to the reduction of associated morbidity and mortality. VTE occurs when a blood clot (thrombus) blocks the vein blood flow causing deep vein thrombosis (DVT) and, when it migrates to the lungs, it may clog the pulmonary arteries characterizing pulmonary embolism (PE). Analysis using fibrin degradation products or D-dimer and coagulation factor VIII may assist early diagnosis of VTE. Thus, two immunosensors were built using layer-by-layer (LbL) films technique, one containing the anti-D-dimer immobilized on polyethylene imine (PEI) and another the anti-FVIII on silk fibroin (SF). Immunosensor response, the antigen-antibody specific interaction, was investigated using cyclic voltammetry. When immunosensors, PEI/anti-D-dimer and SF/anti-FVIII, were exposed to antigens, D-dimer and Factor VIII, the voltammograms area and current were significantly increased with increasing specific antigen concentration. The specific interaction was confirmed with control experiments, electrodes containing only PEI or SF, that no significant changes in the voltammogram responses were observed and principal component analysis confirmed these results. The films formation and response were verified using scanning electronic microscopy (SEM). The developed immunosensor seems to be a promising and effective early complementary exam to assist in the VTE diagnosis, through the combined response of two biomarkers very sensible.
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http://dx.doi.org/10.1016/j.talanta.2020.121241DOI Listing
November 2020

Mesenchymal stem cell (MSc) secretome: A possible therapeutic strategy for intensive-care COVID-19 patients.

Med Hypotheses 2020 Sep 25;142:109769. Epub 2020 Apr 25.

São Paulo State Univ UNESP, Botucatu Medical School, Blood Transfusion Center, Cell Engineering Lab, Botucatu, SP, Brazil; São Paulo State Univ UNESP, Araraquara School of Pharmaceutical Sciences, Department of Clinical Analysis Laboratory of Monoclonal Antibodies, Araraquara, SP, Brazil.

As an emerging global health challenge, COVID-19 requires international knowledge to reach novel possible therapeutic strategies, especially for intensive-care patients. During the early stages of infection, pneumocytes II are the primary infected cells, harming the respiratory system. We have previous evidence in murine models that MSc's secretome can be used to treat pulmonary injuries induced with bleomycin, due to its content: growth factors, extracellular vesicles, and exosomes. We hypothesize and strongly recommend MSc secretome testing and production, in xenofree conditions, to be used as an alternative approach in SARS-Cov-2 patients in critical conditions.
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http://dx.doi.org/10.1016/j.mehy.2020.109769DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7252028PMC
September 2020

Correction to: Culture of mesenchymal stem cells derived from equine synovial membrane in alginate hydrogel microcapsules.

Stem Cell Res Ther 2018 10 7;9(1):259. Epub 2018 Oct 7.

Department of Veterinary Surgery and Anesthesiology, University of Veterinary Medicine and Animal Science UNESP, District of Rubião Júnior, s / n, Botucatu, São Paulo, Brazil.

The original article [1] contained a minor error regarding the mean diameter of the alginate microcapsules described in relation to Fig. 4 in the Results section. The microcapsules had an actual mean diameter of 3000 μm instead of 1000 μm as mistakenly mentioned in the original article.
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http://dx.doi.org/10.1186/s13287-018-0999-6DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6174064PMC
October 2018

Multiple Tolerization Reduces Antibody Binding Against Tolerogen Cells: Implications for the Production of Monoclonal Antibodies.

Monoclon Antib Immunodiagn Immunother 2018 Apr;37(2):100-104

1 Monoclonal Antibody Laboratory, Proteomics Center , Univ Estadual Paulista-UNESP, School of Pharmaceutical Sciences, Araraquara, Brazil .

We report an immunization technique that can update the production of monoclonal antibodies (mAbs): the multiple tolerization subtractive immunization (MTSI). A total of 10 BALB/C mice were used. Animals in group 1 received one inoculation of RWPE-1 cells (nontumoral), followed by cyclophosphamide, and then received serial inoculations of nonirradiated PC3 cells (tumoral). Animals in group 2 received our MTSI protocol, as follows: one inoculation of RWPE-1 cells, followed by cyclophosphamide (Cy). This whole tolerization step was repeated three other times, with 14-day intervals between the last Cy exposure and the next RWPE-1 cell inoculation. Finally, the animals received the same nonirradiated PC3 cell exposure as group 1. Blood was taken from each animal, and their polyclonal sera individually tested against the nontumoral RWPE-1 cells in flow cytometry. We found out that, after the MTSI was employed, the serum of the immunized animals, in group 2, contained considerably less antibodies that reacted against the tolerogenic cells, compared with the serum of the animals that underwent regular subtractive immunization. We showed that, by repeating the tolerization cycles, the polyclonal antibodies produced by mice have a reduced specificity toward common/immunodominant epitopes present at nontumoral cells, and thus this technique can be readily used by others in studies involving murine mAb protocols.
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http://dx.doi.org/10.1089/mab.2017.0055DOI Listing
April 2018

Culture of mesenchymal stem cells derived from equine synovial membrane in alginate hydrogel microcapsules.

BMC Vet Res 2018 Mar 27;14(1):114. Epub 2018 Mar 27.

Department of Veterinary Surgery and Anesthesiology, University of Veterinary Medicine and Animal Science UNESP, District of Rubião Júnior, s / n, Botucatu, São Paulo, Brazil.

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http://dx.doi.org/10.1186/s12917-018-1425-0DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5870504PMC
March 2018

Ultrastructural analysis and residual DNA evaluation of rabbit vein scaffold.

Acta Cir Bras 2017 Sep;32(9):706-711

Associate Professor, Department of Urology, Botucatu Medical School, UNESP, Botucatu-SP, Brazil. Conception and design of the study, manuscript writing, critical revision.

Purpose: To investigate the ultrastructural characteristics and analysis of residual DNA in scaffold models, produced with decellularized vena cava in an experimental model with rabbits.

Methods: Three groups were created for ultrastructural and residual DNA analysis: group 1 - control, consisting of samples of vena cava in natura; group 2 - SD, consisting of vein fragments submitted to 2% sodium deoxycholate decellularization by shaking (160rpm - Shaker News Brunswick Scientific®) for 1 hour at controlled temperature shaker at 37°C; group 3 - SDS, consisting of vein fragments submitted to 1% sodium dodecyl sulfate decellularization under the same previous condition, for 2 hours.

Results: The ultrastructural matrix of the blood vessel maintained its vintegrity after either decellularization models. The results of the two quantification methods demonstrated a significant decrease in the DNA content of the decellularized vena cava samples as compared to the control samples and, differed statistically from each other, p <0.05.

Conclusion: The 2% DS protocol for vein decellularization, in this experimental model, was considered the best protocol because it presented less amount of residual DNA without causing substantial destruction of the extracellular matrix.
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http://dx.doi.org/10.1590/s0102-865020170090000003DOI Listing
September 2017

Enhanced immunization techniques to obtain highly specific monoclonal antibodies.

MAbs 2018 01 6;10(1):46-54. Epub 2017 Oct 6.

a Universidade Estadual Paulista (Unesp), Faculdade de Ciências Farmacêuticas, Araraquara, Proteomics Center, Monoclonal Antibody Lab. , Araraquara , Brazil.

Despite fast advances in genomics and proteomics, monoclonal antibodies (mAbs) are still a valuable tool for areas such as the evolution of basic research in stem cells and cancer, for immunophenotyping cell populations, diagnosing and prognosis of diseases, and for immunotherapy. To summarize different subtractive immunization approaches successfully used for the production of highly specific antibodies, we identified scientific articles in NCBI PubMed using the following search terms: subtractive immunization, monoclonal antibody, tolerization, neonatal, high-zone tolerance, masking immunization. Patent records were also consulted. From the list of results, we included all available reports, from 1985 to present, that used any enhanced immunization technique to produce either polyclonal or monoclonal antibodies. Our examination yielded direct evidence that these enhanced immunization techniques are efficient in obtaining specific antibodies to rare epitopes, with different applications, such as to identify food contaminants or tumor cells.
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http://dx.doi.org/10.1080/19420862.2017.1331804DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5800380PMC
January 2018

Cadmium exposure inhibits MMP2 and MMP9 activities in the prostate and testis.

Biochem Biophys Res Commun 2015 Feb 16;457(4):538-41. Epub 2015 Jan 16.

Univ Estadual Paulista - UNESP, Institute of Biosciences, Department of Morphology, Extracellular Matrix Laboratory, Botucatu, SP, Brazil. Electronic address:

Matrix metalloproteinases (MMPs) are zinc (Zn(2+)) and calcium (Ca(2+)) dependant endopeptidases, capable of degradation of numerous components of the extracellular matrix. Cadmium (Cd(2+)) is a well known environmental contaminant which could impair the activity of MMPs. In this sense, this study was conducted to evaluate if Cd(2+) intake inhibits these endopeptidases activities at the rat prostate and testicles and if it directly inhibits the activity of MMP2 and MMP9 at gelatinolytic assays when present in the incubation buffer. To investigate this hypothesis, Wistar rats (5 weeks old), were given tap water (untreated, n = 9), or 15 ppm CdCl2 diluted in drinking water, during 10 weeks (n = 9) and 20 weeks (n = 9). The animals were euthanized and their ventral prostate, dorsal prostate, and testicles were removed. These tissue samples were processed for protein extraction and subjected to gelatin zymography evaluation. Additionally, we performed an experiment of gelatin zymography in which 5 μM or 2 mM cadmium chloride (CdCl2) was directly dissolved at the incubation buffer, using the prostatic tissue samples from untreated animals that exhibited the highest MMP2 and MMP9 activities in the previous experiment. We have found that CdCl2 intake in the drinking water led to the inhibition of 35% and 30% of MMP2 and MMP9 (p < 0.05) at the ventral prostate and testis, respectively, in Cd(2+) treated animals when compared to controls. Moreover, the activities of the referred enzymes were 80% and 100% inhibited by 5 μM and 2 mM of CdCl2, respectively, even in the presence of 10 mM of CaCl2 within the incubation buffer solution. These important findings demonstrate that environmental cadmium contamination may deregulate the natural balance in the extracellular matrix turnover, through MMPs downregulation, which could contribute to the toxic effects observed in prostatic and testicular tissue after its exposure.
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http://dx.doi.org/10.1016/j.bbrc.2015.01.019DOI Listing
February 2015

Polidocanol versus hypertonic glucose for sclerotherapy treatment of reticular veins of the lower limbs: study protocol for a randomized controlled trial.

Trials 2014 Dec 19;15:497. Epub 2014 Dec 19.

Department of Surgery and Orthopedics, School of Medicine, São Paulo State University (UNESP), Rubião Junior s/n, CEP 18,618-970 Botucatu, SP, Brazil.

Background: The prevalence of chronic venous disease is high and occurs more frequently in females. According to the clinical, etiological, anatomical, and pathological classification (CEAP) definition, the reticular veins are included in the C1 class and are mainly associated with aesthetic complaints. Several invasive techniques are used for treatment, including mini phlebectomy, laser ablation, and radiofrequency ablation. However, a wide range of sclerosing agents may serve as minimally invasive alternatives, promoting chemical sclerosis of the vein wall. Although this technique is routinely performed around the world, there is no consensus on the most efficacious and safe chemical agent to be used.

Methods/design: Inclusion criteria are women between 18 and 69 years old with at least 10 cm long reticular veins in the lower limbs, on the outer side of the leg/thigh. Patients with CEAP 2 to 6, or with allergies, pregnancy, performing breastfeeding, or with any dermatologic or clinical problems will be excluded. Patients with venous ultrasound mapping showing involvement of saphenous trunks and/or a deep venous system will also be excluded. Patients will be randomized into two groups, one receiving 75% pure glucose and the other group receiving 0.2% polidocanol diluted in 70% glucose. Just one limb and one session per patient will be performed. The sclerosing agent volume will not exceed 5 mL. Clinical follow-up will include visits on days 7 and 60, always with photographic documentation.

Discussion: This project aims to enroll 96 patients and subject them to a double-blind treatment after the randomization process. The design is intended to evaluate efficacy through a primary end point and safety through a secondary end point. Forty-eight patients have currently been enrolled. Preliminary results for these patients showed that 25 received treatment, 2 were excluded, and 22 returned after 7 days and showed no greater adverse events. To date, establishing efficacy criteria has not been possible, and no patients have reached the 60-day return point. These data may help doctors choose the best chemical agent for the treatment of reticular veins.

Trial Registration: ClinicalTrials.gov Identifier: NCT02054325, 3/02/2014.
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http://dx.doi.org/10.1186/1745-6215-15-497DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4301449PMC
December 2014

Light-emitting diode effects on combined decellularization of tracheae. A novel approach to obtain biological scaffolds.

Acta Cir Bras 2014 Aug;29(8):485-92

Department of Urology, Medical School, UNESP, Botucatu, SP, Brazil.

Purpose: To obtain a decellularized tracheal scaffold associating traditional approaches with the novel light-emitting diode (LED) proposal.

Methods: This study was performed with New Zealand adult rabbits weighing 3.0 - 4.0 kg. Different protocols (22) were used combining physical (agitation and LED irradiation), chemical (SDS and Triton X-100 detergents), and enzymatic methods (DNase and RNase).

Results: Generally, the cells surrounding soft tissues were successfully removed, but none protocol removed cells from the tracheal cartilage. However, longer protocols were more effective. The cost-benefits relation of the enzymatic processes was not favorable. It was possible to find out that the cartilaginous tissue submitted to the irradiation with LED 630nm and 475 nm showed an increased number of gaps without cells, but several cells were observed to be still present.

Conclusion: The light-emitting diode is a promising tool for decellularization of soft tissues.
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http://dx.doi.org/10.1590/s0102-86502014000800002DOI Listing
August 2014

Morphofunctional characterization of decellularized vena cava as tissue engineering scaffolds.

Exp Cell Res 2014 Aug 12;326(1):103-11. Epub 2014 Jun 12.

Blood Transfusion Center, Cell Engineering Laboratory, Botucatu Medical School, São Paulo State University (UNESP), Botucatu, SP, Brazil; Department of Urology, Botucatu Medical School, São Paulo State University (UNESP), Botucatu, SP, Brazil.

Clinical experience for peripheral arterial disease treatment shows poor results when synthetic grafts are used to approach infrapopliteal arterial segments. However, tissue engineering may be an option to yield surrogate biocompatible neovessels. Thus, biological decellularized scaffolds could provide natural tissue architecture to use in tissue engineering, when the absence of ideal autologous veins reduces surgical options. The goal of this study was to evaluate different chemical induced decellularization protocols of the inferior vena cava of rabbits. They were decellularized with Triton X100 (TX100), sodium dodecyl sulfate (SDS) or sodium deoxycholate (DS). Afterwards, we assessed the remaining extracellular matrix (ECM) integrity, residual toxicity and the biomechanical resistance of the scaffolds. Our results showed that TX100 was not effective to remove the cells, while protocols using SDS 1% for 2h and DS 2% for 1h, efficiently removed the cells and were better characterized. These scaffolds preserved the original organization of ECM. In addition, the residual toxicity assessment did not reveal statistically significant changes while decellularized scaffolds retained the equivalent biomechanical properties when compared with the control. Our results concluded that protocols using SDS and DS were effective at obtaining decellularized scaffolds, which may be useful for blood vessel tissue engineering.
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http://dx.doi.org/10.1016/j.yexcr.2014.05.023DOI Listing
August 2014

Finasteride inhibits human prostate cancer cell invasion through MMP2 and MMP9 downregulation.

PLoS One 2013 30;8(12):e84757. Epub 2013 Dec 30.

Univ Estadual Paulista - UNESP, Institute of Biosciences, Department of Morphology, Extracellular Matrix Lab, Botucatu, São Paulo, Brazil.

Introduction: The use of the 5-alpha reductase inhibitors (5-ARIs) finasteride and dutasteride for prostate cancer prevention is still under debate. The FDA recently concluded that the increased prevalence of high-grade tumors among 5-ARI-treated patients must not be neglected, and they decided to disallow the use of 5-ARIs for prostate cancer prevention. This study was conducted to verify the effects of finasteride on prostate cell migration and invasion and the related enzymes/proteins in normal human and tumoral prostatic cell lines.

Materials And Methods: RWPE-1, LNCaP, PC3 and DU145 cells were cultivated to 60% confluence and exposed for different periods to either 10 µM or 50 µM finasteride that was diluted in culture medium. The conditioned media were collected and concentrated, and MMP2 and MMP9 activities and TIMP-1 and TIMP-2 protein expression were determined. Cell viability, migration and invasion were analyzed, and the remaining cell extracts were submitted to androgen receptor (AR) detection by western blotting techniques. Experiments were carried out in triplicate.

Results: Cell viability was not significantly affected by finasteride exposure. Finasteride significantly downregulated MMP2 and MMP9 activities in RWPE-1 and PC3 cells and MMP2 in DU145 cells. TIMP-2 expression in RWPE-1 cells was upregulated after exposure. The cell invasion of all four tested cell lines was inhibited by exposure to 50 µM of finasteride, and migration inhibition only occurred for RWPE-1 and LNCaP cells. AR was expressed by LNCaP, RWPE-1 and PC3 cells.

Conclusions: Although the debate on the higher incidence of high-grade prostate cancer among 5-ARI-treated patients remains, our findings indicate that finasteride may attenuate tumor aggressiveness and invasion, which could vary depending on the androgen responsiveness of a patient's prostate cells.
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http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0084757PLOS
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3875555PMC
February 2015

Platelet and plasma bioactive scaffolds for stem cell differentiation: what are we missing?

Platelets 2014 31;25(7):556-7. Epub 2013 Oct 31.

Cell Engineering Lab, Blood Transfusion Center, Botucatu Medical School, Univ Estadual Paulista , UNESP, Botucatu, SP , Brazil and.

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http://dx.doi.org/10.3109/09537104.2013.836748DOI Listing
June 2015

Platelet-rich plasma and chronic wounds: remaining fibronectin may influence matrix remodeling and regeneration success.

Cytotherapy 2013 Nov 30;15(11):1436-9. Epub 2013 Aug 30.

Blood Transfusion Center, Cell Engineering Laboratory, Botucatu Medical School, Universidade Estadual Paulista-UNESP, Botucatu, SP, Brazil; Department of Morphology, Extracellular Matrix Laboratory, Botucatu Biosciences Institute, Universidade Estadual Paulista-UNESP, Botucatu, SP, Brazil. Electronic address:

Background: Platelet-rich plasma has been largely used as a therapeutic option for the treatment of chronic wounds of different etiologies. The enhanced regeneration observed after the use of platelet-rich plasma has been systematically attributed to the growth factors that are present inside platelets' granules.

Aim: We hypothesize that the remaining plasma and platelet-bound fibronectin may act as a further bioactive protein in platelet-rich plasma preparations.

Methods: Recent reports were analyzed and presented as direct evidences of this hypotheses.

Results: Fibronectin may directly influence the extracellular matrix remodeling during wound repair. This effect is probably through matrix metalloproteinase expression, thus exerting an extra effect on chronic wound regeneration.

Conclusions: Physicians should be well aware of the possible fibronectin-induced effects in their future endeavors with PRP in chronic wound treatment.
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http://dx.doi.org/10.1016/j.jcyt.2013.05.019DOI Listing
November 2013

Comment on: fibronectin in tissue regeneration: timely disassembly of the scaffold is necessary to complete the build.

Authors:
Andrei Moroz

Cell Mol Life Sci 2013 Nov 22;70(22):4255-6. Epub 2013 Aug 22.

Cell Engineering Lab, Blood Transfusion Center, Botucatu Medical School, Univ Estadual Paulista, UNESP, District of Rubião Júnior S/N, Botucatu, SP, 18618-970, Brazil,

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http://dx.doi.org/10.1007/s00018-013-1453-7DOI Listing
November 2013

Tissue-engineered blood vessel substitute by reconstruction of endothelium using mesenchymal stem cells induced by platelet growth factors.

J Vasc Surg 2014 Jun 2;59(6):1677-85. Epub 2013 Jul 2.

Cell Engineering Laboratory, Blood Transfusion Center, Botucatu Medical School, UNESP-Paulista State University, Botucatu, Brazil; Department of Urology, Botucatu Medical School, UNESP-Paulista State University, Botucatu, Brazil.

Background: Cardiovascular diseases remain leaders as the major causes of mortality in Western society. Restoration of the circulation through construction of bypass surgical treatment is regarded as the gold standard treatment of peripheral vascular diseases, and grafts are necessary for this purpose. The great saphenous vein is often not available and synthetic grafts have their limitations. Therefore, new techniques to produce alternative grafts have been developed and, in this sense, tissue engineering is a promising alternative to provide biocompatible grafts. This study objective was to reconstruct the endothelium layer of decellularized vein scaffolds, using mesenchymal stem cells (MSCs) and growth factors obtained from platelets.

Methods: Fifteen nonpregnant female adult rabbits were used for all experiments. Adipose tissue and vena cava were obtained and subjected to MSCs isolation and tissue decellularization, respectively. MSCs were subjected to differentiation using endothelial inductor growth factor (EIGF) obtained from human platelet lysates. Immunofluorescence, histological and immunohistochemical analyses were employed for the final characterization of the obtained blood vessel substitute.

Results: The scaffolds were successfully decellularized with sodium dodecyl sulfate. MSCs actively adhered at the scaffolds, and through stimulation with EIGF were differentiated into functional endothelial cells, secreting significantly higher quantities of von Willebrand factor (0.85 μg/mL; P < .05) than cells cultivated under the same conditions, without EIGF (0.085 μg/mL). Cells with evident morphologic characteristics of endothelium were seen at the lumen of the scaffolds. These cells also stained positive for fascin protein, which is highly expressed by differentiated endothelial cells.

Conclusions: Taken together, the use of decellularized bioscaffold and subcutaneous MSCs seems to be a potential approach to obtain bioengineered blood vessels, in the presence of EIGF supplementation.
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http://dx.doi.org/10.1016/j.jvs.2013.05.032DOI Listing
June 2014

Fibronectin induces MMP2 expression in human prostate cancer cells.

Biochem Biophys Res Commun 2013 Jan 19;430(4):1319-21. Epub 2012 Dec 19.

Univ Estadual Paulista - UNESP, Institute of Biosciences, Department of Morphology, Botucatu, SP, Brazil.

High-grade prostate cancers express high levels of matrix metalloproteinases (MMPs), major enzymes involved in tumor invasion and metastasis. However, the tumor cell lines commonly employed for prostate cancer research express only small amounts of MMPs when cultivated as monolayer cultures, in common culture media. The present study was conducted to ascertain whether culture conditions that include fibronectin can alter MMP2 and MMP9 expression by the human prostatic epithelial cell lines RWPE-1, LNCaP and PC-3. These cells were individually seeded at 2×10(4) cells/cm(2), cultivated until they reached 80% confluence, and then exposed for 4h to fibronectin, after which the conditioned medium was analyzed by gelatin zymography. Untreated cells were given common medium. Only RWPE-1 cells express detectable amounts of MMP9 when cultivated in common medium, whereas the addition of fibronectin induced high expression levels of pro and active forms of MMP2 in all tested cell lines. Our findings demonstrate that normal and tumor prostate cell lines express MMP2 activity when in contact with extracellular matrix components or blood plasma proteins such as fibronectin. Future studies of transcriptomes and proteomes in prostate cancer research using these cell lines should not neglect these important conclusions.
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http://dx.doi.org/10.1016/j.bbrc.2012.12.031DOI Listing
January 2013

Chronic caffeine intake increases androgenic stimuli, epithelial cell proliferation and hyperplasia in rat ventral prostate.

Int J Exp Pathol 2012 Dec;93(6):429-37

Department of Morphology, Institute of Biosciences, Univ Estadual Paulista (UNESP), Botucatu, SP, Brazil.

Coffee intake has been associated with a low risk of developing cancer, including prostate cancer, which is one of the most commonly diagnosed cancer in men. However, few studies have evaluated the chronic effects of caffeine, which is the most abundant methylxanthine in coffee, on prostate morphology and physiology. In the present study, we investigated the effects of chronic, low-dose caffeine intake on rat prostate morphology from puberty to adulthood. Five-week-old male Wistar rats were randomized into two experimental groups: caffeine-treated (20 ppm in drinking water, n = 12) and control (n = 12). The ventral and dorsolateral prostates were dissected, weighted and submitted to morphological, morphometrical and immunohistochemical analysis of cellular proliferation, apoptosis and androgen receptor (AR) tissue expression. The testosterone (T) and dihydrotestosterone (DHT) concentrations were measured in the plasma. Our results show that caffeine intake increased the concentrations of T and DHT, organ weight, epithelial cell proliferation and AR tissue expression in the ventral prostatic lobe. All the ventral prostates from the caffeine-treated animals presented various degrees of epithelial and stromal hyperplasia. Our results suggest that chronic caffeine intake from puberty increases androgenic signalling and cell proliferation in the rat prostate gland and can be related to the development of benign prostatic hyperplasia.
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http://dx.doi.org/10.1111/j.1365-2613.2012.00843.xDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3521899PMC
December 2012

Caffeine reduces cadmium accumulation in the organism and enhances the levels of antioxidant protein expression in the epididymis.

Reprod Toxicol 2013 Jan 23;35:137-43. Epub 2012 Oct 23.

Department of Morphology, Institute of Biosciences-University Estadual Paulista, Botucatu, Sao Paulo, Brazil.

The aim of this study was to investigate the effects of caffeine (20 mg/L) intake on cadmium (15 mg/L) accumulation in the rat blood, testes, epididymis and prostate as well as cadmium-induced changes to the antioxidant defense system of the epididymis. Caffeine reduced the cadmium concentration in all tissues analyzed. Meanwhile, cadmium reduced catalase activity and increased superoxide dismutase (SOD) activity in the epididymis. Caffeine increased SOD activity, catalase and glutathione tissue expression and sustains the cadmium's effect on catalase and GSP-Px activity. No differences in the expression of metallothionein and lipid peroxidation were observed among the different treatments in the epididymis. In conclusion, low doses of cadmium alter the antioxidant enzymatic profile of the epididymis, but not induced oxidative lipid damage. Caffeine intake reduces overall cadmium accumulation in the organism and enhances the levels of antioxidant protein expression in the epididymis, thus exerting a protective effect against this metal.
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http://dx.doi.org/10.1016/j.reprotox.2012.10.009DOI Listing
January 2013

Platelet lysate 3D scaffold supports mesenchymal stem cell chondrogenesis: an improved approach in cartilage tissue engineering.

Platelets 2013 30;24(3):219-25. Epub 2012 May 30.

Extracellular Matrix Laboratory, Department of Morphology, Institute of Biosciences, Universidade Estadual Paulista-UNESP, District of Rubião Júnior S/N, 18618-970, Botucatu, SP, Brazil.

Articular lesions are still a major challenge in orthopedics because of cartilage's poor healing properties. A major improvement in therapeutics was the development of autologous chondrocytes implantation (ACI), a biotechnology-derived technique that delivers healthy autologous chondrocytes after in vitro expansion. To obtain cartilage-like tissue, 3D scaffolds are essential to maintain chondrocyte differentiated status. Currently, bioactive 3D scaffolds are promising as they can deliver growth factors, cytokines, and hormones to the cells, giving them a boost to attach, proliferate, induce protein synthesis, and differentiate. Using mesenchymal stem cells (MSCs) differentiated into chondrocytes, one can avoid cartilage harvesting. Thus, we investigated the potential use of a platelet-lysate-based 3D bioactive scaffold to support chondrogenic differentiation and maintenance of MSCs. The MSCs from adult rabbit bone marrow (n = 5) were cultivated and characterized using three antibodies by flow cytometry. MSCs (1 × 10(5)) were than encapsulated inside 60 µl of a rabbit platelet-lysate clot scaffold and maintained in Dulbecco's Modified Eagle Medium Nutrient Mixture F-12 supplemented with chondrogenic inductors. After 21 days, the MSCs-seeded scaffolds were processed for histological analysis and stained with toluidine blue. This scaffold was able to maintain round-shaped cells, typical chondrocyte metachromatic extracellular matrix deposition, and isogenous group formation. Cells accumulated inside lacunae and cytoplasm lipid droplets were other observed typical chondrocyte features. In conclusion, the usage of a platelet-lysate bioactive scaffold, associated with a suitable chondrogenic culture medium, supports MSCs chondrogenesis. As such, it offers an alternative tool for cartilage engineering research and ACI.
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http://dx.doi.org/10.3109/09537104.2012.686255DOI Listing
September 2013

Isolation and immunophenotypic characterization of mesenchymal stem cells derived from equine species adipose tissue.

Vet Immunol Immunopathol 2009 Dec 30;132(2-4):303-6. Epub 2009 Jun 30.

Department of Veterinary Surgery and Anesthesiology, School of Veterinary Medicine and Animal Science, São Paulo State University, Botucatu, São Paulo, Brazil.

The purpose of this work was to isolate and cultivate mesenchymal stem cells (MSC) derived from equine adipose tissue and conduct cellular characterization with the following markers: CD90, CD44 and CD13. Adipose tissue collection was performed at the base of the horses' tails, followed by immediate isolation and cultivation of the MSC and posterior characterization by flow cytometry for the interspecies reaction test using mouse anti-rat CD90 monoclonal antibody (mAb), fluorescein isothiocyanate (FITC), and tests with specific mAb mouse anti-horse CD13 and mouse anti-horse CD44. The technique used for isolation and cell cultivation proved to be safe and viable. The CD90 mAb expressed cross-reaction with MSC derived from equine adipose tissue and CD44 showed greater expression in cells as the number of culture passages increased. Although marker CD13 expresses reaction in other studies involving MSC in different species, it presented no expression in the experiment realized. The results obtained revealed the immunophenotypic characterization of the surface of isolated and cultivated MSC, classifying these cells as a promising type of progenitor cells that can be applied in equine cellular therapy.
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http://dx.doi.org/10.1016/j.vetimm.2009.06.014DOI Listing
December 2009
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