Abstract The use of platelet-rich plasma (PRP) and adipose-derived stromal cells (ADSC) have been investigated as a form of wound healing enhancement. The objective of this work was to evaluate the association of red propolis (RP) and PRP as inducers of ADSC for application in tissue regeneration. Adipose tissue post-collection and post-cryopreservation was isolated with type II collagenase, characterized by flow cytometry, and differentiated into osteogenic, chondrogenic and adipose cell. The viability of ADSC was evaluated when exposed to different concentrations of RP using the MTT and trypan blue assay. Acridine orange and ethidium bromide (AO/EB) was performed to evaluate cell death events. Horizontal migration methods were investigated in ADSC using autologous and homologous PRP associated with RP (PRP/RP). All assays were processed in triplicate. Flow cytometry and cellular differentiation showed that type II collagenase was effective for isolating ADSC post-collection and post-cryopreservation. RP extracts at concentrations of up to 50 μg.mL-1 presented no cytotoxic effects. Association of PRP and RP at 25 and 50 μg.ml-1 influenced ADSC migration, with total closure on the seventh day after exposition. The results here presented could stimulate proliferation of ADSC cells that may contribute directly or indirectly to the reconstructive process of tissue regeneration. plateletrich platelet rich (PRP adiposederived derived (ADSC enhancement (RP regeneration postcollection post collection postcryopreservation cryopreservation osteogenic assay AO/EB AOEB AO EB (AO/EB events PRP/RP. PRPRP PRP/RP . (PRP/RP) triplicate postcryopreservation. cryopreservation. 5 μg.mL1 μgmL1 μgmL μg.mL 1 μg mL μg.mL- effects 2 μg.ml1 μgml1 μgml μg.ml ml μg.ml- exposition (PRP/RP mL1 ml1

Abstract In this work, NRL films were enriched with red propolis and silver nanoparticles with the aim of obtaining dressings with bactericide properties. To improve the properties of these materials, plasma treatment was also applied to the films. Cytotoxic effects were evaluated by MTT and agar diffusion assays on secondary and stem cells. Determination of cell behavior on the films were assessed by EDS and FEG-SEM. In general, the plasma modified NRL films presented hydrophilic character, favorable condition for biomedical applications. RBS results showed that silver is probably in nanoparticulate form. MTT assays revealed that non-cytotoxic effects were observed for all the samples extracts. Either the plasma treatment or silver deposition were effective in sterilizing the samples and modifying their surfaces. It was demonstrated in this in vitro study that ADSCs associated with NRL films enriched with propolis should be considered promising for application in the healing of occlusive wounds.

This paper reports the DNA interaction and cytotoxicity of binary and ternary copper(II) complexes of valproic acid with phenanthroline or bipyridine and the complementary characterization of Cu(Valp)2Bipy by crystallography. Circular dichroism, plasmid and oligonucleotide assays have shown that the complexes interact with DNA in different manners and are able to cleave plasmid DNA and oligonucleotides at concentrations from 10 to 1000 mol L-1. The plasmid DNA cleavage is enhanced 650, 375 and 285 times by photo activation with UVB light for the complexes Cu(Valp)2Phen, Cu(Valp)2Bipy and Cu2(Valp)4, respectively. Cytotoxicity assays demonstrated that V79 cells were more sensitive to CuII derivatives than sodium valproate as evaluated by 3-(4,5-dimethylthiazol-2yl)-2,5-diphenyl tetrazolium bromide (MTT) and clonogenic assays, where the cytotoxic profile of the compounds was: Cu(Valp)2Phen higher than Cu(Valp)2Bipy, higher than Cu2(Valp)4. Therefore, we report two copper complexes, which interact with DNA and promote its cleavage, leading to an enhanced cytotoxicity when compared to valproic acid.

RESUMO Objetivo: Definir escores histológicos de degeneração do disco intervertebral que permitam a identificação de características morfológicas da doença, além de melhorar o conhecimento sobre a discopatia degenerativa lombar por meio de marcadores imuno-histoquímicos. Métodos: As colorações histológicas de hematoxilina e eosina, azul de alcian/PAS, tricrômica de Masson e safranina O/FCF foram utilizadas em cortes de disco intervertebral degenerado de pacientes com discopatia degenerativa lombar. Os marcadores proteicos definidos na imuno-histoquímica permitiram a avaliação da proliferação celular (Ki-67) e da apoptose (p53). Resultados: Os dados do estudo permitiram a determinação da coloração de safranina O/FCF como a mais eficaz para avaliar os parâmetros tais como a área, o diâmetro e o número de agrupamentos de condrócitos. Também se determinou a importância do uso das colorações histológicas de forma associada, como safranina O/FCF, tricrômica de Masson, azul de alcian/PAS e hematoxilina e eosina, uma vez que elas são complementares para a verificação histopatológica da degeneração do disco intervertebral. Pela técnica da expressão de proteínas com técnica imuno-histoquímica, foi possível considerar dois estágios de degeneração do disco: proliferação de células com a formação de agrupamentos de condrócitos, seguida pela indução de apoptose. Conclusão: Este estudo permitiu definir a caracterização histológica e imuno-histoquímica em discopatia degenerativa lombar e seus graus de evolução, determinando novos escores de degeneração discal.

ABSTRACT Objective: To define histological scores for intervertebral disc degeneration that would enable the definition of morphological characteristics of disease, besides improving knowledge of the lumbar degenerative disc disease by means of immunohistochemical markers. Methods: Hematoxylin and Eosin, Alcian/PAS, Masson Trichrome and Safranin O/FCF staining was used on the intervertebral disc degeneration sections of patients with lumbar degenerative disc disease. The protein markers defined in immunohistochemistry were cell proliferation (Ki-67) and apoptosis (p53). Results: The study data enabled the determination of Safranin O/FCF stain as the most effective one for evaluating parameters such as area, diameter, and number of chondrocyte clusters. The importance of using stains in association, such as Safranin O/FCF, Masson Trichrome, Alcian/PAS and Hematoxylin and Eosin, was also determined, as they are complementary for the histopathological verification of intervertebral disc degeneration. By expressing proteins using the immunohistochemistry technique, it was possible to consider two stages of disc degeneration: cell proliferation with chondrocyte cluster formation, and induction of apoptosis. Conclusion: This study enabled the histological and immunohistochemical characterization to be determined for lumbar degenerative disc disease, and its degrees of evolution, by determining new disc degeneration scores.

RESUMEN Objetivo: Definir valores histológicos de degeneración del disco intervertebral que permitan la definición de las características morfológicas de la enfermedad y mejorar el conocimiento de la enfermedad degenerativa del disco lumbar mediante marcadores inmunohistoquímicos. Métodos: Las coloraciones histológicas con hematoxilina y eosina, azul alcián/PAS, tricrómico de Masson y safranina O/FCF se utilizaron en secciones de los discos intervertebrales degenerados de pacientes con enfermedad degenerativa del disco lumbar. Los marcadores de proteínas definidos por inmunohistoquímica permitieron la evaluación de la proliferación celular (Ki-67), y la apoptosis (p53) . Resultados: Los datos de la determinación permitieron establecer la tinción con safranina O/FCF como la más eficaz para evaluar parámetros tales como el área, diámetro y número de agrupaciones de condrocitos. Se determinó también la importancia del uso asociado de diversas tinciones, como safranina O/FCF, tricrómico de Masson, azul de alcián/PAS y hematoxilina y eosina, ya que son complementarias para la verificación histopatológica de la degeneración del disco intervertebral. Por la técnica de la expresión de proteína con análisis inmunohistoquímica, fue posible establecer dos etapas de la degeneración del disco: la proliferación de células con la formación de agrupaciones de condrocitos y la inducción de la apoptosis. Conclusión: Este estudio permitió definir la caracterización histológica e inmunohistoquímica en la enfermedad degenerativa del disco lumbar y sus grados de evolución, mediante la determinación de nuevas puntuaciones de degeneración del disco.

RESUMO Salvia officinalis (Lamiaceae) é muito utilizado no sul do Brasil como condimento, tempeiro e chá para o tratamento de várias doenças. O objetivo deste estudo foi caracterizar compostos químicos no extrato hidroalcoólica (ExtHS) e aquosas (ExtAS) de Salvia officinalis (L.) por cromatografia gasosa acoplado a espectrometria de massa (GC-MS) e pela espectrometria de ionização por electrospray de alta resolução acoplado a massas (ESI-QTOF MS/MS), avaliar a capacidade in vitro para eliminar o radical livre 2,2-difenil-1-picrilhidrazil (DPPH•) e 2,2'-azino-bis (6-3-etilbenzotiazolina-sulfónico) (ABTS•+), atividade da catalase (CAT-like) e superóxido dismutase (SOD-like), além disso, atividade citotóxica por ensaio MTT, alterações na morfologia celular por Giemsa e mecanismo de apoptose induzida por anexina V/iodeto de propídio. A identificação química revelou a presença de ácidos e compostos fenólicos nos extratos de sálvia. Na análise antioxidante in vitro ambos os extratos indicaram atividades promissoras. O ensaio citotóxico em células tumorais (HEp-2, HeLa, A-549, HT-29 e A-375) e em não-tumoral (células HEK-293 e MRC-5), mostraram seletividade para celulas tumorais. A imunicitoquímica apresentou a maioria das células tumorais nos estágios tardios de processo de apoptose e necrose. Tendo em conta os resultados aqui apresentados, Salvia officinalis brasileira (L.) usada como condimento e chá pode proteger o corpo de algumas doenças, em particular aquelas que envolvem estresse oxidativo, como desordens neurodegenerativas, inflamação e câncer.

ABSTRACT Salvia officinalis (Lamiaceae) has been used in south of Brazil as a diary homemade, in food condiment and tea-beverage used for the treatment of several disorders. The objective of this study was to characterize chemical compounds in the hydroalcoholic (ExtHS) and aqueous (ExtAS) extract from Salvia officinalis (L.) by gas chromatography-mass spectrometry (GC-MS) and by high-resolution electrospray ionization mass spectrometry (ESI-QTOF MS/MS), evaluate in vitro ability to scavenge the free radical 2,2-diphenyl-1-picrylhydrazyl (DPPH•) and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS•+), catalase (CAT-like) and superoxide dismutase (SOD-like) activity, moreover cytotoxic by MTT assay, alterations on cell morphology by giemsa and apoptotic-induced mechanism for annexin V/propidium iodide. Chemical identification sage extracts revealed the presence of acids and phenolic compounds. In vitro antioxidant analysis for both extracts indicated promising activities. The cytotoxic assays using tumor (Hep-2, HeLa, A-549, HT-29 and A-375) and in non-tumor (HEK-293 and MRC-5), showed selectivity for tumor cell lines. Immunocytochemistry presenting a majority of tumor cells at late stages of the apoptotic process and necrosis. Given the results presented here, Brazilian Salvia officinalis (L.) used as condiment and tea, may protect the body against some disease, in particularly those where oxidative stress is involved, like neurodegenerative disorders, inflammation and cancer.

A biflorina é uma o-naftoquinona com comprovados efeitos citotóxicos frente a células tumorais além de atividades antimicrobiana, antitumoral e antimutagênica. A biflorina é um composto isolado das raízes da planta Capraria biflora L.(Schrophulariaceae), originária das Antilhas e da América do Sul, que está localizada em zonas temperadas e tropicais. Este composto mostrou uma potente atividade contra microorganismos gram-positivos e bactérias álcool-ácido resistentes. Tem mostrado eficiência em inibir a proliferação celular em linhagens tumorais CEM, HL-60, B16, HCT-8, MCF-7. Recentemente, a linhagem tumoral SK-Br3 foi tratada com biflorina e demonstrou importante efeito citotóxico. Neste artigo, relatamos informações relacionadas com os primeiros estudos de caracterização estrutural, bem como os últimos relatos sobre a atividade biológica desta molécula.

Biflorin is an o-naphthoquinone with proven cytotoxic effects on tumor cells showing antimicrobial, antitumor and antimutagenic activities. Biflorin is an isolated compound taken from the roots of the plantCapraria biflora L. (Schrophulariaceae), indigenous of the West Indies and South America, which is located in temperate or tropical areas. This compound has shown to be strongly active against grampositive and alcohol-acid-resistant bacteria. It has been efficient in inhibiting the proliferation tumor cell lines CEM, HL-60, B16, HCT-8 and MCF-7. Recently, SK-Br3 cell line was treated with biflorin showing important cytotoxic effects. In this article, information related to the first structural characterization studies are presented, as well as the latest reports concerning the biological activity of this molecule.

Revisar o potencial papel das células-tronco para o tratamento de doença degenerativa do disco intervertebral (IVD). Foi realizada uma revisão dos trabalhos da base de dados Medline em relação a células-tronco e doença degenerativa discal (DDD). Para fins de discussão dos dados, os trabalhos foram divididos em: revisão, in vitro, experimentais e clínicos. Os tratamentos disponíveis atualmente focam basicamente na redução dos sintomas, e não na reversão do processo degenerativo do DIV. O uso de células-tronco mesenquimais (CTM) vem sendo proposto como uma opção de tratamento para DDD. Estudos in vitro demonstraram que as CTM são capazes de se diferenciarem em células do NP e que as CTM também diminuem os níveis inflamatórios do DIV degenerado. Além disso, estudos experimentais demonstraram que as CTM permaneciam viáveis quando injetadas no DIV e que eram capazes de regenerar parcialmente do DIV degenerado e sua estrutura. Os poucos estudos clínicos presentes na literatura apresentam resultados divergentes. O uso de CTM esta sendo amplamente estudado e mostra resultados promissores para o tratamento da DDD. Embora muitos avanços venham sendo alcançados em estudos in vitro e experimentais, ainda faltam estudos clínicos para comprovar o papel das CTM no manejo da DDD.

To review the potential role of stem cells in treating degenerative disc disease of the intervertebral disc (IVD). A review was performed of articles from the Medline database concerning stem cells and degenerative disc disease (DDD). To discuss the data, the papers were classified as: review, in vitro, experimental, and clinical. The currently available treatments were basically for symptom reduction, not to revert the IVD degenerative process. The use of mesenchymal stem cells (MSC) is being proposed as an option of treatment for DDD. In vitro studies have shown that the MSC are able to differentiate into NP cells and that the MSC also reduce the inflammatory levels of the degenerated IVD. Besides, experimental studies demonstrated that the MSC remained viable when injected into the IVD, and that they were able to regenerate partially from the degenerated IVD and its structure. The few clinical studies found in the literature presented diverging results. The use of MSC is being widely studied and shows promising results for the treatment of DDD. Although many advances are being achieved in studies in vitro and experimental, there is a lack of clinical studies to prove the role of MSC in DDD management.

Revisar el papel potencial de las células madre en el tratamiento de la enfermedad degenerativa del disco intervertebral (IVD). Se realizó una revisión de los artículos de la base de datos Medline sobre células madre y la enfermedad degenerativa del disco (DDD). Para fines de discusión de los datos, los trabajos se dividieron en: revisión, in vitro, experimentales y clínicos. Los tratamientos disponibles actualmente enfocan básicamente la reducción de los síntomas, y no la reversión del proceso degenerativo del IVD. El uso de células madre mesenquimales (MSC) se propone como una opción de tratamiento para DDD. Estudios in vitro demostraron que las MSC son capaces de diferenciarse en células NP y que las MSC también reducen los niveles inflamatorios de la IVD degenerado. Además, estudios experimentales demostraron que las MSC se mantuvieron viables cuando inyectadas en el IVD y que eran capaces de regenerarse parcialmente del IVD degenerado y su estructura. Los pocos estudios clínicos presentes en la literatura presentan resultados divergentes. El uso de MSC se está estudiando ampliamente y muestra resultados prometedores en el tratamiento de la DDD. Aunque se hayan logrado muchos avances en estudios in vitro y experimentales, aún faltan estudios clínicos para comprobar el papel de las MSC en el manejo de la DDD.