Aplicação da membrana de látex natural (NRL) como suporte para cultura das células osteogênicas MC3T3-E1 para regeneração óssea guiada (GBR)

Pós-graduação em Biotecnologia - IQ

Natural latex films as carrier for casearia sylvestris swartz extract associated with ciprofloxacin

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Desenvolvimento inicial de clones IAC de seringueira em São José do Rio Preto, SP

In this study the development of four clones (IAC 35; 40; 300 e 301) of rubber tree [Hevea brasiliensis (Willd. ex A. Juss.) Müell. Arg.] selected by Instituto Agronômico de Campinas were evaluated during the first 24 months of cultivation. The experiment was laid out in randomized block design with three replications and four plants for parcel following the 1.5mx1.0m spacing at São José do Rio Preto, SP. The clone RRIM 600 was used as check. The variables analyzed were the trunk girth, the number of matured whorl, the photosynthesis rate and the concentrations of chlorophyll a, b, total and carotenoids. At the end of the experiment the values of trunk girth at 0.5m above the budgrafting union varied from 5.85 cm (IAC 301) to 10.53 cm (IAC 300) and the mean number of matured whorl varied from 2.58 (IAC 301) to 3.91 (RRIM 600). During the dry season, when the plants were 22 month old, the mean value of photosynthesis rate of IAC 40 and IAC 301 (12mol m-2 s-1) were lower compared to the other clones (15mol m-2 s-1). The chlorophyll a, b and total carotenoids concentrations were equal or superior to the check, never lower. Considering the variables analyzed, excluding the IAC 301, the other clones show performance comparable to RRIM 600.

A new heterologous fibrin sealant as scaffold to recombinant human bone morphogenetic protein-2 (rhBMP-2) and natural latex proteins for the repair of tibial bone defects

Tissue engineering has special interest in bone tissue aiming at future medical applications Studies have focused on recombinant human bone morphogenetic protein-2 (rhBMP-2) and natural latex proteins due to the osteogenic properties of rhBMP-2 and the angiogenic characteristic of fraction 1 protein (P-1) extracted from the rubber tree Hevea brasiliensis. Furthermore, heterologous fibrin sealant (FS) has been shown as a promising alternative in regenerative therapies. The aim of this study was to evaluate these substances for the repair of bone defects in rats. A bone defect measuring 3 mm in diameter was created in the proximal metaphysis of the left tibia of 60 rats and was implanted with rhBMP-2 or P-1 in combination with a new heterologous FS derived from snake venom. The animals were divided into six groups: control (unfilled bone defect), rhBMP-2 (defect filled with 5 mu g rhBMP-2), P-1 (defect filled with 5 mu g P-1), FS (defect filled with 8 mu g FS), FS/rhBMP-2 (defect filled with 8 mu g FS and 5 mu g rhBMP-2), FS/P-1 (defect filled with 8 mu g FS and 5 mu g P-1). The animals were sacrificed 2 and 6 weeks after surgery. The newly formed bone projected from the margins of the original bone and exhibited trabecular morphology and a disorganized arrangement of osteocyte lacunae. Immunohistochemical analysis showed intense expression of osteocalcin in all groups. Histometric analysis revealed a significant difference in all groups after 2 weeks (p < 0.05), except for the rhBMP-2 and FS/rhBMP-2 groups (p > 0.05). A statistically significant difference (p < 0.05) was observed in all groups after 6 weeks in relation to the volume of newly formed bone in the surgical area. In conclusion, the new heterologous fibrin sealant was found to be biocompatible and the combination with rhBMP-2 showed the highest osteogenic and osteoconductive capacity for bone healing. These findings suggest a promising application of this combination in the regeneration surgery.

Genetic diversity strategy for the management and use of rubber genetic resources: more than 1,000 wild and cultivated accessions in a 100-genotype core collection

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Estudo da incorporação e liberação de peptídeos hormonais utilizando membranas de látex natural como carreador

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Natural rubber latex LbL films: Characterization and growth of fibroblasts

The recent biomedical applications of natural rubber (NR) latex, mostly in dry membranes, have motivated research into novel, more noble uses of this low-cost biomaterial. In this article, we provide the first report on the fabrication of layer-by-layer (LbL) films of NR alternated with the polyelectrolytes polyethylenimine (PEI) and polyallylamine hydrochloride (PAH). Stable (PAH/NR)n and (PEI/NR)n LbL films displayed similar physicochemical properties, but differed in terms of film morphology according to atomic force microscopy (AFM) and scanning electron microscopy (SEM) data. Most significantly, (PEI/NR)5 LbL films were made of smaller and flattened particles, which were not efficient for the growth and proliferation of normal human fibroblasts (NHF). In contrast, efficient NHF proliferation could be obtained with (PAH/NR)n LbL films, with the fibroblasts exhibiting the expected elongated morphology. Furthermore, cell growth did not occur for cast films of NR, thus demonstrating the suitability of the LbL method for this biologically related application. The differences between the two polyelectrolytes illustrate the importance of the film architecture and morphology, which open the way for exploiting the molecular control inherent in the LbL technique for further applications of NR-containing films. (c) 2012 Wiley Periodicals, Inc. J Appl Polym Sci, 2012

Oral treatment with Hev b 13 ameliorates experimental colitis in mice

Hev b 13 is an allergenic esterase obtained from the rubber tree Hevea brasiliensis, which has been shown recently to induce human mononuclear cells to release interleukin (IL)-10 in vitro. This immunoregulatory cytokine appears to play an important role in preventing inflammation and mucosal damage in animal models of colitis and in Crohn's disease patients. The aim of this study was to evaluate the therapeutic effect of Hev b 13 in mice with 2,4,6-trinitrobenzene sulphonic acid (TNBS)-induced colitis. Two hours following colonic instillation of the haptenizing agent, and daily thereafter for 5 days, Hev b 13 was administered by oral gavage. In mice treated with daily doses of either 0.5 mg/kg or 5.0 mg/kg of Hev b 13, the clinical signs of diarrhoea, rectal prolapse and body weight loss and also histological damage of the distal colon, were reduced significantly, in comparison with water-treated diseased mice. These findings suggest a potent anti-inflammatory activity of Hev b 13; this activity is speculated to be related to its interaction with cells from the immune system.

Oral treatment with Hev b 13 prevents experimental arthritis in mice

Hev b 13 is an allergenic esterase obtained from the rubber tree Hevea brasiliensis, which has been shown recently to induce human monocytes to release interleukin (IL)-10 in vitro, and to exert a potent anti-inflammatory effect in vivo. Moreover, Hev b 13 has been shown to reduce clinical signs of inflammation and also histological damage to the distal colon of mice with 2,4,6-trinitrobenze sulphonic acid (TNBS)-induced colitis after its oral administration. The aim of this study was to investigate the effect of Hev b 13 on human mononuclear cells, as well as its therapeutic use in the methylated bovine serum albumin (mBSA) model of antigen-induced arthritis. Five days before the intra-articular challenge, and daily thereafter for 8 days, Hev b 13 was administered by oral gavage. In mice treated with a dose of 0.5 mg/kg of Hev b 13, the severity of oedema, leucocyte infiltration, pannus formation and cartilage erosion were reduced significantly. These findings underscore the anti-inflammatory activity suggested previously for Hev b 13, an activity speculated to be related to its interaction with monocytes/macrophages and the consequent stimulation of IL-10 release and reduction of tumour necrosis factor (TNF) release. The study also opens a wide range of possible applications in the field of immune-mediated inflammatory diseases.

Estrutura da vegetação e sua relação com a diversidade, abundância e similaridade de coleópteros bioindicadores em diferentes sistemas vegetacionais, Piracicaba, SP

A Mata Atlântica é considerada um dos biomas mais importantes do mundo devido à sua alta biodiversidade e funções ecossistêmicas. Entretanto, encontra-se fragmentada em porções de pequenas dimensões esparsas em uma matriz predominantemente agrícola, composta principalmente por extensas pastagens e monoculturas. Desse modo, os sistemas agroflorestais por apresentarem uma estrutura diferenciada dos monocultivos e similar às condições naturais, podem ser utilizados como uma alternativa para o manejo e a conservação da biodiversidade nos remanescentes florestais. A fragmentação provoca modificações no ambiente que irão refletir na perda e no deslocamento da biodiversidade, estando os insetos entre os grupos mais afetados. Uma das formas de se avaliar o estado de conservação dos fragmentos e o impacto antrópico nos sistemas vegetacionais, é estudar a presença e distribuição de organismos bioindicadores. Dentre esses, os insetos ocupam posição de destaque. Os insetos da família Scarabaeidae e da subfamília Scolytinae são bons indicadores de distúrbios, pois são muito sensíveis ás mudanças ambientais. Neste trabalho hipotetisou-se que a presença desses insetos está relacionada com a estrutura da vegetação e as condições de vida proporcionadas pelas diferentes formas de uso-da-terra. O objetivo desta pesquisa foi avaliar a diversidade de espécies, o padrão de abundância e a similaridade entre as populações de coleópteros (Scarabaeidae e Scolytinae) em diferentes sistemas vegetacionais de diferentes estruturas: i) Fragmento de floresta estacional semidecidual dividido em três áreas: beira do rio, centro e borda; ii) Sistema Agroflorestal (SAF) (interface entre o fragmento e o pasto); iii) Pasto composto de Brachiaria decumbens (L.); iv) Monocultivo de café (Coffea arábica L.); v) Monocultivo de seringueira (Hevea brasiliensis Müell. Arg.); vi) SAF de café e seringueira - todos situados numa região de domínio anterior de floresta estacional semidecidual em Piracicaba-SP. Os sistemas foram caracterizados quanto à sua estrutura e condições micrometeorológicas. Os insetos foram coletados mensalmente entre agosto/2013 e julho/2014 utilizando-se dois tipos de armadilhas: Pitfall e etanol modelo ESALQ-84. Foram coletados 1.047 espécimes distribuídos em 21 espécies da família Scarabaeidae e 1.833 indivíduos de 38 espécies da subfamília Scolytinae. A maior quantidade de espécies de Scarabaeidae foi encontrada na borda do fragmento florestal, enquanto que a maior abundância ocorreu no fragmento florestal perto do rio. A subfamília Scolytinae apresentou a maior riqueza de espécies no sistema agroflorestal misto (borda) e a maior abundância no sistema agroflorestal café-seringueira. A abundância e riqueza de espécies da família Scarabaeidae foram correlacionadas positivamente com a temperatura do ar, temperatura e umidade do solo e a precipitação. Por outro lado, a abundância e a riqueza de espécies da subfamília Scolytinae apresentaram correlação negativa com a temperatura do ar e a temperatura e umidade do solo. Ambos os grupos de insetos apresentaram a maior abundância e riqueza de espécies nas áreas com estrutura vegetacional mais complexa, sendo influenciadas pelas condições microclimáticas dentro de cada local.

Biological cycle of Tenuipalpus heveae Baker (Acari, Tenuipalpidae) on leaflets of three rubber tree clones

Life cycle of Tenuipalpus heveae Baker (Acari, Tenuipalpidae) on leaflets from three rubber tree clones. The biological cycle of Tenuipalpus heveae Baker, 1945 (Tenuipalpidae), a potential rubber tree pest mite, was studied by the observation of individuals reared on leaflets of the clones GT 1, PB 235 and RRIM 600, in controlled environmental conditions. Three daily observations were done of 60 eggs on leaflets from each clone in order to verify the development of immature stages and the female oviposition. The fertility life table was constructed based in the collected data. Mites reared on PB 235 had faster rate of development, requiring less time in days, to double its population in number (TD), and had the highest values for egg production, female longevity, net reproductive rate (Ro), intrinsic rate of natural increase (r m) and finite rate of increase (λ). Lower reproductive values and the longest time necessary to reach adult stage were recorded for the mites on GT 1. In all studied clones, the deutonymphal phase had the highest viability, while the larval phase had the lowest, highlighted by the survivorship curve that indicated high mortality during this life stage. The clone PB 235 allowed the most suitable conditions for the development of T. heveae, followed by RRIM 600, while GT 1 was the less suitable substratum to rear this mite species.

Divergence and genetic variability among superior rubber tree genotypes

The objective of this work was to estimate the genetic variability and divergence among 22 superior rubber tree (Hevea sp.) genotypes of the IAC 400 series. Univariate and multivariate analyses were performed using eight quantitative traits (descriptors), including yield. In the univariate analyses, the estimated parameters were: genetic and environmental variances; genetic and environmental coefficients of variation; and the variation index. The Mahalanobis generalized distance, the Tocher agglomerative method and canonical variables were used for the multivariate analyses. In the univariate analyses, variability was verified among the genotypes for all the variables evaluated. The Tocher method grouped the genotypes into 11 clusters of dissimilarity. The first four canonical variables explained 87.93% of the cumulative variation. The highest genetic variability was found in rubber yield-related traits, which contributed the most to the genetic divergence. The most divergent pairs of genotypes are suggested for crossbreeding. The genotypes evaluated are suitable for breeding and may be used to continue the IAC rubber tree breeding program.

Assessment of growth and yield performance of rubber tree clones of the IAC 500 series

The objective of this work was to evaluate the performance of 15 clones of the IAC 500 series of Hevea brasiliensis, developed at Instituto Agronômico (IAC), over a 12-year period, in the northwest region of São Paulo State, Brazil. The 15 new clones evaluated are primary clones obtained from selected ortets within half-sib progenies. The clone RRIM 600, of Malaysian origin, was used as the control. Dry rubber yield performance over a four-year period, mean girth at the tenth year, girth increment before and during tapping, thermal properties of the natural rubber produced and other characters of the laticiferous system were evaluated. Forty percent of the clones were superior in comparison to the control for yield. Clone IAC 500 recorded the highest yield (66.81 g per tree per tapping) over four years of tapping, followed by IAC 502 (62.37 g per tree per tapping), whereas the control recorded 48.71 g per tree per tapping. All selected clones were vigorous in growth. The natural rubber from this IAC clones showed thermal stability up to 300ºC. No differences were observed in the thermal behavior of rubber among the IAC series and the RRIM 600 clones. The clones IAC 500, IAC 501, IAC 502, IAC 503 and IAC 506 are the more promising for small-scale plantations, due to growth and yield potential.

Genetic diversity of cultivated accessions and wild species of rubber tree using EST‑SSR markers

The objective of this work was to evaluate the efficiency of EST‑SSR markers in the assessment of the genetic diversity of rubber tree genotypes (Hevea brasiliensis) and to verify the transferability of these markers for wild species of Hevea. Forty‑five rubber tree accessions from the Instituto Agronômico (Campinas, SP, Brazil) and six wild species were used. Information provided by modified Roger's genetic distance were used to analyze EST‑SSR data. UPGMA clustering divided the samples into two major groups with high genetic differentiation, while the software Structure distributed the 51 clones into eight groups. A parallel could be established between both clustering analyses. The 30 polymorphic EST‑SSRs showed from two to ten alleles and were efficient in amplifying the six wild species. Functional EST‑SSR microsatellites are efficient in evaluating the genetic diversity among rubber tree clones and can be used to translate the genetic differences among cultivars and to fingerprint closely related materials. The accessions from the Instituto Agronômico show high genetic diversity. The EST‑SSR markers, developed from Hevea brasiliensis, show transferability and are able to amplify other species of Hevea.

Genetic parameters and estimated genetic gains in young rubber tree progenies

The objective of this work was to assess the genetic parameters and to estimate genetic gains in young rubber tree progenies. The experiments were carried out during three years, in a randomized block design, with six replicates and ten plants per plot, in three representative Hevea crop regions of the state of São Paulo, Brazil. Twenty-two progenies were evaluated, from three to five years old, for rubber yield and annual girth growth. Genetic gain was estimated with the multi-effect index (MEI). Selection by progenies means provided greater estimated genetic gain than selection based on individuals, since heritability values of progeny means were greater than the ones of individual heritability, for both evaluated variables, in all the assessment years. The selection of the three best progenies for rubber yield provided a selection gain of 1.28 g per plant. The genetic gains estimated with MEI using data from early assessments (from 3 to 5-year-old) were generally high for annual girth growth and rubber yield. The high genetic gains for annual girth growth in the first year of assessment indicate that progenies can be selected at the beginning of the breeding program. Population effective size was consistent with the three progenies selected, showing that they were not related and that the population genetic variability is ensured. Early selection with the genetic gains estimated by MEI can be made on rubber tree progenies.