Teurasjätteiden jalostaminen liikennepolttoaineiksi

Työssä määritettiin luokan 2 eläinperäisistä sivutuotteista liikennekäyttöön tuotettujen biodieselin ja biometaanin elinkaaren aikaiset kasvihuonekaasupäästöt ja tuotantoprosessien energiankulutukset perustuen kirjallisuuslähteistä saatuihin lähtötietoihin. Tätä kautta tutkittiin yhdistelmäprosessia, jossa tuotetaan molempia polttoaineita ja selvitettiin onko tällaisella tuotantotavalla mahdollista vähentää päästöjä ja parantaa polttoaineiden tuotannon energiatehokkuutta. Kasvihuone-kaasupäästöjen laskentamenetelmä pohjautuu direktiivissä 2009/28/EY annettuun ohjeistukseen ja eri kasvihuonekaasupäästöjen karakterisointi IPCC:n sadan vuoden tarkastelumalliin. Käytännön laskenta suoritettiin standardien SFS-EN ISO 14040 ja 14044 määrittelemän elinkaariarviointiselvityksen muodossa. Työssä käytetyn laskentamenetelmän ja tarkasteluun valittujen tuotanto-teknologioiden perusteella lasketut tulokset osoittavat, että yhdistelmäprosessilla ei saavuteta suurempia päästövähenemiä eikä parempaa energiatehokkuutta kuin nykyisin käytössä olevilla tuotantotavoilla. Tulokset ovat kuitenkin hyvin herkkiä laskennassa tehtyjen oletusten ja käytettyjen lähtötietojen vaihtelulle sekä valittujen laskentamenetelmien muutoksille. Suurin päästöjä ja energiankulutusta aiheuttava yksittäinen tekijä on kaikissa tuotejärjestelmissä luokan 2 sivutuotteiden esikäsittelyssä vaadittavaan steri-lointiin tarvittavan lämmön tuotanto. Tutkituissa tuotejärjestelmissä lämpö tuotetaan kokonaan tai osittain fossiilisilla polttoaineilla. Kasvihuone-kaasupäästöjä olisi mahdollista alentaa merkittävästi siirtymällä lämmön tuotannossa kokonaan uusiutuviin polttoaineisiin. Sterilointi on lain edellyttämä käsittelytapa ja siksi energiankulutusta on vallitsevissa olosuhteissa hyvin vaikea pienentää merkittävästi.

Liikennebiokaasun tuotannon ja käytön ulkoiset kustannukset Pohjois-Karjalassa

Työn tavoitteena oli selvittää liikennebiokaasuntuotannon ja käytön vaikutus liikenteen ulkoi-siin kustannuksiin Pohjois-Karjalassa. Biokaasua tuotetaan Joensuussa Kuhasalon jäteveden-puhdistamolla sekä Kontiosuon jäteasemalla, Kiteellä biokaasua tuotetaan BioKympin yh-teismädätyslaitoksessa. Lisäksi laskennassa huomioitiin yhden maatilakokoluokan biokaasun-tuotanto. Työssä selvitettiin kaksi skenaariota liikennebiokaasun tuotantomääräksi vuodelle 2015. Liikennebiokaasua voitaisiin tuottaa optimiskenaarion mukaan 3 426 MWh ja maksimi-tuotantoskenaarion mukaan 21 532 MWh. Liikennebiokaasun käytön vaikutukset liikenteen päästöihin laskettiin vuodelle 2015 ja vuo-delle 2020, jolloin liikennebiokaasua käytettäisiin 10 % liikenteen energiantarpeesta Pohjois-Karjalassa. Hiilidioksidipäästöt vähenevät vuoden 2020 tilanteessa samassa suhteessa kuin liikennebiokaasu korvaa fossiilisia polttoaineita. Muista päästöistä merkittävimmät päästövä-hennykset saatiin kun vuoden 2010 dieselautot muutettaisiin biokaasuautoiksi, tällöin hiuk-kaspäästöt alenisivat jopa 18 % vuoden 2010 päästöistä. Lisäksi selvitettiin liikenteen päästöjen pienenemisen vaikutus liikenteen aiheuttamiin ulkoisiin kustannuksiin. Laskettavat ulkoiset kustannukset olivat ilmastonmuutos, pakokaasupäästöt sekä energiariippuvuus. Pakokaasupäästöjen aiheuttamat ulkoiset kustannukset olivat vuonna 2010 noin 7 miljoonaa euroa. Liikennebiokaasua käyttävästä ajoneuvotyypistä riippuen ulkoiset kustannukset laskevat vuoden 2020 tilanteessa 10–16 % vuoden 2010 kustannuksista. Ilmastonmuutoksen ulkoiset kustannukset vuodelle 2010 olivat 9,5 miljoonaa euroa. Biokaasun käytön avulla kustannuksissa voitaisiin säästää 910 000 euroa vuonna 2020. Ener-giariippuvuuden hinta öljynkäytöstä oli vuonna 2010 noin 4,2 miljoonaa euroa ja vuonna 2020 kustannukset voisivat laskea 450 000 euroa.

Normal HC11 and ras-transformed mouse mammary cells are resistant to the antiproliferative effects of retinoic acid

The objective of the present study was to determine the effects of retinoic acid on the growth of the mouse mammary cells HC11 and HC11ras, which are a model for in vitro breast cancer progression. The expression of the two classes (RARs and RXRs) of retinoic acid receptor mRNAs was determined by Northern blot analysis. Receptor functional integrity was determined by testing whether RAR ß mRNA could be induced by retinoic acid. The effects of a 72-h exposure to 50 µM 13-cis retinoic acid on HC11 and HC11ras cell proliferation and HC11 cell differentiation were investigated by flow cytometric cell cycle analysis, and by determination of ß-casein mRNA expression, respectively. The possibility that retinoic acid would induce the expression of the vitamin D receptor and synergize with vitamin D, a known inhibitor of HC11 cell growth, was also investigated. HC11 cells expressed higher mRNA levels of both RAR a and RAR g when compared to HC11ras cells. In contrast, RAR ß, as well as RXR a, ß and g expression was low in both HC11 and HC11ras cells. In addition, RAR ß mRNA was induced by retinoic acid treatment in both cells. In spite of these observations, no effects were seen on cell proliferation or differentiation upon exposure to retinoic acid. Neither vitamin D receptor induction nor synergy with vitamin D on growth inhibition was observed. We conclude that the RAR expression profile could be related to the transformed state in HC11ras cells and that the retinoic acid resistance observed merits further investigation.

In vitro inhibitory effects of imatinib mesylate on stromal cells and hematopoietic progenitors from bone marrow

Imatinib mesylate (IM) is used to treat chronic myeloid leukemia (CML) because it selectively inhibits tyrosine kinase, which is a hallmark of CML oncogenesis. Recent studies have shown that IM inhibits the growth of several non-malignant hematopoietic and fibroblast cells from bone marrow (BM). The aim of the present study was to evaluate the effects of IM on stromal and hematopoietic progenitor cells, specifically in the colony-forming units of granulocyte/macrophage (CFU-GM), using BM cultures from 108 1.5- to 2-month-old healthy Swiss mice. The results showed that low concentrations of IM (1.25 µM) reduced the growth of CFU-GM in clonogenic assays. In culture assays with stromal cells, fibroblast proliferation and α-SMA expression by immunocytochemistry analysis were also reduced in a concentration-dependent manner, with a survival rate of approximately 50% with a dose of 2.5 µM. Cell viability and morphology were analyzed using MTT and staining with acrydine orange/ethidium bromide. Most cells were found to be viable after treatment with 5 µM IM, although there was gradual growth inhibition of fibroblastic cells while the number of round cells (macrophage-like cells) increased. At higher concentrations (15 µM), the majority of cells were apoptotic and cell growth ceased completely. Oil red staining revealed the presence of adipocytes only in untreated cells (control). Cell cycle analysis of stromal cells by flow cytometry showed a blockade at the G0/G1 phases in groups treated with 5-15 µM. These results suggest that IM differentially inhibits the survival of different types of BM cells since toxic effects were achieved.

Prosessiteollisuuden sekoittimen elinkaariarviointi

Työn tavoitteena oli tehdä elinkaariarviointi (LCA) prosessiteollisuuden sekoittimelle Helmix HF-80 ja analysoida LCA-tulokset, vaikutus ilmaston lämpenemisen potentiaalin (GWP) suhteen, sekä tutkia GWP-vaikutuksen pienentämisen mahdollisuuksia. Tutkittavan sekoittimen mahdolliset käyttökohteet ovat sellu- ja paperiteollisuus, raskaiden jätenesteiden käsittely, sekä muut teollisuusalueet, joissa käytetään tämän tyyppisiä laitteita. Työssä on muun muassa käsitelty sekoitusprosessit, sekoituslaitteiden tyypit, niiden rakenteiden ominaisuudet, käyttötarkoitus, toimintaperiaatteet, sekä sellu- ja paperi-teollisuudessa käytettävien sekoittimien yleiskatsaus. Työssä on myös kuvattu elinkaariarviointi (LCA) -menetelmä ja sen käyttötarkoitus. Tärkeimmistä tuloksista voi mainita sen, että sekoittimen (ilman säiliötä, sähkömoottoria ja vaihteistoa) kokoelinkaaren ilmastonlämpenemisen potentiaali (GWP) on noin 750 000 kg CO2-Equiv. Sekoittimen tuotanto- ja kierrätysaikana syntyy vain 1200 kg CO2-Equiv. ja suurin osa 748 000 kg CO2-Equiv. johtuu sähköenergian kulutuksesta käytön aikana. Käyttöajan vaikutusta voisi pienentää arvoksi 0 kg CO2-Equiv. käyttämällä pelkästään uusiutuvaa energiaa. Jos tuotantoaikana käytetty energia myös korvataan uusiutuvalla energialla, niin GWP-arvo koko elinkaaren aikana laskee arvoon 1006 kg CO2eqv., mikä on vain 0,13 % saaduista tuloksista. Suurin osa tästä arvosta liittyy sekoittimen materiaalin, tässä tapauksessa ruostumattoman teräksen tuotantoon.

Evaluación de la gestión de residuos derivados del proceso de impresión en Datapoint de Colombia S.A.S, y propuesta de mejora

Este trabajo se centra en el análisis de las actividades desarrolladas en torno a los servicios de procesos de impresión que ofrece la organización DATAPOINT de Colombia SAS para identificar los puntos críticos en la gestión de los residuos de impresión y las decisiones tomadas por parte de los involucrados durante todo el proceso (proveedores, clientes y la empresa), con el fin de revisar medidas y estrategias que permitan fortalecer la gestión integral de residuos de impresión a partir de una revisión y comparación de las mejores prácticas planteadas por los actores del sector. También se efectuaron recomendaciones con acciones de mejora que se podrían desarrollar con el fin de mitigar el impacto ambiental generado por estos residuos. Con la finalidad de cumplir con lo planteado se realizó inicialmente un estudio sobre la organización, sus clientes y proveedores para entender de manera integral la cadena de valor en torno a los tóner y su gestión inversa, (explicar) al igual que el entorno normativo tanto de manera nacional como internacional. Posteriormente, se identificaron los puntos de mejora comparando lo planteado por el proveedor versus lo ejecutado por los involucrados en el proceso, labor se realizó en campo con los clientes para entender la situación actual, sus necesidades y en que basan la toma de decisiones relacionada con el manejo de los residuos de impresión. Finalmente se listaran una serie de acciones de mejora y recomendaciones las cuales pueden ser incorporadas a los procesos críticos de DATAPOINT.

Methodology To model the energy and greenhouse gas emissions of electronic software distributions

A new electronic software distribution (ESD) life cycle analysis (LCA)methodology and model structure were constructed to calculate energy consumption and greenhouse gas (GHG) emissions. In order to counteract the use of high level, top-down modeling efforts, and to increase result accuracy, a focus upon device details and data routes was taken. In order to compare ESD to a relevant physical distribution alternative,physical model boundaries and variables were described. The methodology was compiled from the analysis and operational data of a major online store which provides ESD and physical distribution options. The ESD method included the calculation of power consumption of data center server and networking devices. An in-depth method to calculate server efficiency and utilization was also included to account for virtualization and server efficiency features. Internet transfer power consumption was analyzed taking into account the number of data hops and networking devices used. The power consumed by online browsing and downloading was also factored into the model. The embedded CO2e of server and networking devices was proportioned to each ESD process. Three U.K.-based ESD scenarios were analyzed using the model which revealed potential CO2e savings of 83% when ESD was used over physical distribution. Results also highlighted the importance of server efficiency and utilization methods.

In vitro inhibitory effect of trichostatin A on canine grade 3 mast cell tumor

Mast cell tumor (MCT) is one of the most prevalent neoplasms that affect skin and soft tissue in dogs. Because mast cell tumors present a great variety of clinical appearance and behavior, their treatment becomes a challenge. Trichostatin A (TSA), an antifungal antibiotic, has shown inhibitory effects on the proliferation and induction of apoptosis in various types of cancer cells. In order to evaluate the potential of trichostatin A as a therapeutic drug, cells of grade 3 MCT were cultured and treated with concentrations of 1 nM to 400 nM of TSA. MTT assay and trypan blue exclusion assays were performed to estimate cell growth and cell viability, and cell cycle analysis was evaluated. TSA treatment showed a reduction in numbers of viable cells and an increase of cell death by apoptosis. The cell cycle analysis showed an increase of hypodiploid cells and a reduction of G0/G1 and G2/M -phases. According to these results, trichostatin A may be an interesting potential chemotherapeutic agent for the treatment of canine MCT.

Effect of the anti-neoplastic drug doxorubicin on XPD-mutated DNA repair-deficient human cells

Doxorubicin (DOX), a member of the anthracycline group, is a widely used drug in cancer therapy. The mechanisms of DOX action include topoisomerase II-poisoning, free radical release, DNA adducts and interstrand cross-link (ICL) formation. Nucleotide excision repair(NER) is involved in the removal of helix-distorting lesions and chemical adducts, however, little is known about the response of NER-deficient cell lines to anti-tumoral drugs like DOX. Wild type and XPD-mutated cells, harbouring mutations in different regions of this gene and leading to XP-D, XP/CS or TTD diseases, were treated with this drug and analyzed for cell cycle arrest and DNA damage by comet assay. The formation of DSBs was also investigated by determination of gamma H2AX foci. Our results indicate that all three NER-deficient cell lines tested are more sensitive to DOX treatment, when compared to wild type cells or XP cells complemented by the wild type XPD cDNA, suggesting that NER is involved in the removal of DOX-induced lesions. The cell cycle analysis showed the characteristic G2 arrest in repair-proficient MRC5 cell line after DOX treatment, whereas the repair-deficient cell lines presented significant increase in sub-G1 fraction. The NER-deficient cell lines do not show different patterns of DNA damage formation as assayed by comet assay and phosphorylated H2AX foci formation. Knock-down of topoisomerase II alpha with siRNA leads to increased survival in both MRC5 and XP cells, however, XP cell line still remained significantly more sensitive to the treatment by DOX. Our study suggests that the enhanced sensitivity is due to DOX-induced DNA damage that is subject to NER, as we observed decreased unscheduled DNA synthesis in XP-deficient cells upon DOX treatment. Furthermore, the complementation of the XPD-function abolished the observed sensitivity at lower DOX concentrations, suggesting that the XPD helicase activity is involved in the repair of DOX-induced lesions. (C) 2009 Elsevier B.V. All rights reserved.

The value of a saved tree : The lifecycle of CO2-emissions associated with the combination of printing paper and electricity production of wood in the U.S. and Sweden

The pulp- and paper production is a very energy intensive industry sector. Both Sweden and the U.S. are major pulpandpaper producers. This report examines the energy and the CO2-emission connected with the pulp- and paperindustry for the two countries from a lifecycle perspective.New technologies make it possible to increase the electricity production in the integrated pulp- andpaper mill through black liquor gasification and a combined cycle (BLGCC). That way, the mill canproduce excess electricity, which can be sold and replace electricity produced in power plants. In thisprocess the by-products that are formed at the pulp-making process is used as fuel to produce electricity.In pulp- and paper mills today the technology for generating energy from the by-product in aTomlinson boiler is not as efficient as it could be compared to the BLGCC technology. Scenarios havebeen designed to investigate the results from using the BLGCC technique using a life cycle analysis.Two scenarios are being represented by a 1994 mill in the U.S. and a 1994 mill in Sweden.The scenariosare based on the average energy intensity of pulp- and paper mills as operating in 1994 in the U.S.and Sweden respectively. The two other scenarios are constituted by a »reference mill« in the U.S. andSweden using state-of-the-art technology. We investigate the impact of varying recycling rates and totalenergy use and CO2-emissions from the production of printing and writing paper. To economize withthe wood and that way save trees, we can use the trees that are replaced by recycling in a biomassgasification combined cycle (BIGCC) to produce electricity in a power station. This produces extra electricitywith a lower CO2 intensity than electricity generated by, for example, coal-fired power plants.The lifecycle analysis in this thesis also includes the use of waste treatment in the paper lifecycle. Both Sweden and theU.S. are countries that recycle paper. Still there is a lot of paper waste, this paper is a part of the countries municipalsolid waste (MSW). A lot of the MSW is landfilled, but parts of it are incinerated to extract electricity. The thesis hasdesigned special scenarios for the use of MSW in the lifecycle analysis.This report is studying and comparing two different countries and two different efficiencies on theBLGCC in four different scenarios. This gives a wide survey and points to essential parameters to specificallyreflect on, when making assumptions in a lifecycle analysis. The report shows that there arethree key parameters that have to be carefully considered when making a lifecycle analysis of wood inan energy and CO2-emission perspective in the pulp- and paper mill in the U.S. and in Sweden. First,there is the energy efficiency in the pulp- and paper mill, then the efficiency of the BLGCC and last theCO2 intensity of the electricity displaced by BIGCC or BLGCC generatedelectricity. It also show that with the current technology that we havetoday, it is possible to produce CO2 free paper with a waste paper amountup to 30%. The thesis discusses the system boundaries and the assumptions.Further and more detailed research, including amongst others thesystem boundaries and forestry, is recommended for more specificanswers.

Longevidade das micro e pequenas empresas prestadoras de serviços: um estudo das dimensões organizacionais e suas implicações

O presente estudo tem como objetivo fornecer subsídios para a discussão sobre a longevidade das micro e pequenas empresas (MPEs) prestadoras de serviços através da relação de parceria com a grande empresa. O enfoque delineia-se no espaço das dimensões organizacionais: estrutura e processos, comportamento, estratégias, tecnologia / inovação e ambiente. Tal escolha deu-se em função de duas razões: a primeira enquanto uma forma de recolocar na agenda de discussões brasileiras a questão da importância da vida longa às MPEs como agente dinamizador da economia; e a segunda, desmistificar a funcionalidade e indicadores de gestão a partir de sua longevidade. O estudo faz uma imersão diante dos desdobramentos das dimensões, com fatores que agregam um arcabouço de variáveis, discutidas à luz do referencial teórico do sistema sócio-técnico, estudos de ciclo de vida, na “lógica” da complexidade e nas evidências epistemológicas das dimensões organizacionais, creditando fatores tangíveis e intangíveis à margem de práticas da gestão das MPEs longevas. A estratégia metodológica sustentou-se em uma pesquisa quali-quantitativa, com investigação descritiva, explicativa e aplicada, com instrumentos de análise de dados primários e secundários e pesquisa de campo com aplicação de entrevistas e questionários. Foram pesquisadas 82 MPEs prestadoras de serviços com longevidade a partir de cinco anos e parceira da grande empresa. Após a análise dos dados evidenciou-se que as dimensões organizacionais alteram de forma sinérgica na vida dessas empresas. Os resultados comprovam ainda, que a parceria com a grande empresa explica significativamente na longevidade das MPEs. Ao final o estudo apresenta-se um quadro de indicadores de gestão das MPEs longevas, por dimensão organizacional.

Mutagênese induzida por flavonóides presentes do decocto das cascas da aroeira (Schinus terebinthifolius, Raddi)

The decoction of Brazilian pepper tree barks (Schinus terebinthifolius, Raddi), is used in medicine as wound healing and antiinflamatory. Once extracts from this plant are used for acceleration of scar s process, it is important to study their mutagenic and genotoxic potential. In previous works in our laboratory, it was observed mutagenicity caused by the decoction when in high concentrations. Among the chemical compounds of this plant that could be able to induce mutation, the flavonoids were the only group that was referred to have either an oxidant or antioxidant potential. The flavonoids were isolated, purified and quantified by adsorptive column chromatography under silica gel, bacterial and in vitro genotoxic tests were realized to determine if the flavonoids were the responsible agents for this mutagenicity found. The tests realized with plasmidial DNA were indicative that the flavonoids are probably genotoxic, due to the presence of correlation between increase of the flavonoid concentration and in plasmidial DNA double strand breakage visualized in agarose gel, as well as they were capable to generated abasic sites shown by the in vitro treatment with exonuclease III. The same tests with plasmidial DNA in the presence of copper [10 µM] and of a Tris-HCl pH 7.5 [10 µM] buffer were realized with the isolated flavonoids to determine if there would be or not participation of reactive oxygen species (ROS). The transformation of plasmidial DNA in different bacterial strains proficient and deficient in DNA repair enzymes in the presence or not of a Tris-HCl buffer, suggests that the enzymes that repair oxidative lesions are necessary to repair the lesions generated by the flavonoids and that ROS are generated and are necessary to promote the lesions. Bacterial tests with Escherichia coli strains of the CC collection (deficient or not for DNA repair enzymes), showed that the flavonoids are able to increase the frequency of mutations, mainly in strains mutated in repair enzymes (MutM, MutY-glicosylases and double mutant), suggesting that these agents are responsible for the enhancement in the mutation rate. In order to determine the mutation spectrum caused by the flavonoids of the Brazilian pepper tree stem bark, plasmidial DNA previously treated with the flavonoids were transformed in bacterial strains deficient and proficient in the DNA repair enzymes, followed by a blue-white selection with X-gal, DNA amplification by PCR and sequencing the positive mutant clones. Analysis of the mutants obtained from strains CC104, CC104mutM, CC104mutY, CC104mutMmutY, BW9101, BW9109 indicated a predominance of some mutations like G:C to C:G that can be correlated with the origin of 8-oxoG, due to oxidative lesions caused by the flavonoids. So it can concluded that the flavonoid isolated or in fractions enriched on them are genotoxic and mutagenic, and their mutations are predominantly oxidative, mediated by ROS, and the lesions are recognized by the BER system. In this way it is proposed that the flavonoids can act in two different ways to generate the DNA lesion: 1. in a Fenton-like reaction, when the flavonoid are in the presence of metal ions and that together with the water generate ROS that promotes the DNA lesions; 2. in another way the lesions can be generated by the formation of ROS due to the internal chemical structure of the flavonoid molecule due to the quantity and location of hydroxyl groups, and so producing the DNA lesions, those lesions can be directly (suggested by the in vitro experiments) or indirectly done (supported by the experiments using the CC bacterial strains)

Estudo in vitro dos efeitos da BMP-2 e do seu antagonista Noggin sobre a proliferação e migração celulares em carcinoma epidermóide de língua

Oral squamous cell carcinoma (OSCC) is the most prevalent malignancy in the oral cavity and reach a large number of individuals, has become an important public health problem. Studies have demonstrated changes in pathway components BMP in various types of cancers as prostate, colon, breast, gastric and OSCCs. Is the current knowledge that these proteins may exert pro-tumor effect in more advanced stages of neoplastic development coming to favor progression and invasion tumor. The inhibition of the signaling pathway BMP-2 through its antagonists, have shown positive results of antitumor activity and use of Noggin may be a novel therapeutic target for cancer. Given this evidence and the few studies with BMP-2, Noggin and OSCC, the objective of this research was to evaluate the effect of BMP-2 and its antagonist Noggin on proliferation and migration cell in line of cell cultures of human tongue squamous cell carcinoma (SCC25). The study was divided in three groups, a control group, where SCC25 cells suffered no treatment, a BMP-2 group, in which cells were treated with 100ng/ml of BMP-2 and a group of cells that were treated with 100ng/ml of Noggin. For the proliferation assay and cell cycle were established three time intervals (24, 48 and 72 hours). Proliferative activity was investigated by trypan blue and cell cycle analysis by staining with propidium iodide flow cytometry. The potential for migration / invasion of SCC25 cells was performing by a cell invasion assay using Matrigel in a 48-hour interval. The proliferation curve showed a higher proliferation in cells treated with BMP-2 in 72 hours (p < 0.05), and lower overgrowth and cell viability in Noggin group. Recombinant proteins favored a greater percentage of cells in cell cycle phase Go/G1 with a statistically significant difference in the interval of 24 hours (p < 0.05). BMP- 2 produced a greater invasion of cells studied as well as its antagonist Noggin inhibits invasion of cells (p < 0.05). Thus, these results indicate that BMP-2 promotes malignant phenotype, dues stimulates proliferation and invasion of SCC25 cells and, its antagonist Noggin may be an alternative treatment, due to inhibit the tumor progression

Atividade biológica de células-tronco da polpa de dentes decíduos humanos submetidas à criopreservação

Dental pulp stem cells have been widely investigated because of their ability to differentiate into both dental and non-dental cells, with potential use in therapies involving tissue engineering. The technique of cell cryopreservation represents a viable alternative for the conservation of these cells, since it stops reversibly, in a controlled manner, all of cell biological functions in an ultra low temperature. The present study aimed to evaluate, using in vitro experiments, the influence of a cryopreservation protocol on the biologic acti vity of stem cells from human exfoliated deciduous teeth (SHED). Cells obtained from the pulp of three deciduous teeth on end-stage exfoliation or with indicated extraction were expanded in α-MEM culture medium supplemented with antibiotics and 15% fetal bovine serum. At second subculture (P2), a group of cells were submitted to cryopreservation for 30 days in 10% DMSO diluted in fetal bovine serum, at -80º C, while the remind cells continued under normal conditions of cell culture. Cell proliferation was evaluated in both groups (not cryopreserved or cryopreserved) by Trypan blue stain essay at intervals of 24, 48 and 72h after plating. Cell cycle analysis of SHEDs submitted or not to the cryopreservation protocol was performed in the same intervals. Events related to cell death were studied by Annexyn V and PI expression under flow cytometry at the intervals of 24 and 72h. The presence of nuclear morphological changes was evaluated by DAPI staining at 72h interval. It was observed that both groups exhibited an upward cell proliferation curve, without considerable changes in cell viability throughout the experiment. The distribution of cell in the cell cycle phasis was consistent with cell proliferation in both groups. There were no nuclear morphological damages in the end range of the experiment. therefore, it is concluded that the proposed cryopreservation protocol is efficient for storing the studied cell type, allowing its use in future experimental studies