Mantas não tecidas de nanofibras de três polímeros biodegradáveis poli(ácido láctico), PDLLA, poli(Ε-caprolactona), PCL, e poli(butileno adipato-co-tereftalato), PBAT e seus nanocompósitos com uma nanoargila montmorilonita (MMT) foram produzidas por eletrofiação. A morfologia, o comportamento térmico e a estrutura interna das nanofibras foram analisados por microscopia eletrônica de varredura e transmissão, calorimetria diferencial de varredura e difração de raios X, respectivamente. Observou-se que as nanofibras dos nanocompósitos possuíam diâmetros menores do que os correspondentes polímeros puros e que as nanofibras de PBAT puro e de PBAT/MMT apresentavam a menor cristalinidade de todas as mantas. A viabilidade celular de todas as nanofibras foi analisada pela técnica de redução do sal de tetrazolium pelo complexo enzimático piruvato desidrogenase presente na matriz de mitocôndrias (teste MTT). Os resultados mostraram que nenhuma manta nanofibrílica apresentou toxicidade às células e que as nanofibras de PBAT puro e seu nanocompósito propiciaram ainda um ambiente mais favorável ao desenvolvimento celular de fibroblastos de cardiomiócitos do que as condições oferecidas pelo controles, provavelmente por apresentarem menores diâmetros e baixa cristalinidade em relação às demais nanofibras. Estes resultados mostram o potencial de uso destas mantas nanofibrílicas como suportes de crescimento celular.

Non-woven mats of nanofibers of three biodegradable polymers, viz. poly(lactic acid), PDLLA, poly(Ε-caprolactone), PCL, and poly(butylene adipate-co-terephthalate), PBAT, and their nanocomposites with montmorillonite nanoclay (MMT) were produced by electrospinning. The morphology, thermal behavior and internal structure of the nanofibers were analyzed by scanning and transmission electron microscopy, differential scanning calorimetry and wide angle X-ray diffraction, respectively. The nanofibers of the nanocomposites had lower diameters than the nanofibers of the corresponding neat polymers, while the nanofibers from PBAT and PBAT/MMT were the least crystalline. The cell viability of all the nanofibers was analyzed by reduction of the tetrazolium salt by the pyruvate dehydrogenase enzymatic complex present in the mitochondria (MTT test). None of the nanofibers was toxic to the cells and the PBAT and PBAT/MMT nanofibers exhibited a more favorable environment for developing fibroblasts from cardiomyocytes than the control, probably due to their low crystallinity. These results demonstrated the potential use of nanofibers´ mats as scaffolds for cell growth.

OBJETIVO: O objetivo deste estudo foi avaliar o efeito da T3 na expressão da osteocalcina, osteopontina e colágeno I durante a diferenciação osteogênica das células-tronco mesenquimais (CTM). MATERIAIS E MÉTODOS: As células da medula óssea de ratas Wistar jovens foram extraídas, cultivadas e separadas em cinco grupos: controle (indiferenciado), diferenciado (estímulo osteogênico) e diferenciado com T3 (10-3 nM, 10-2 nM e 100 nM). Para cada grupo, foram cultivadas quatro amostras que foram analisadas por RT-PCR tempo real aos 7, 14 e 21 dias, para quantificação dos transcritos gênicos para osteocalcina, osteopontina e colágeno I. RESULTADOS: Todos os grupos diferenciados sem T3 ou com T3 independentemente da concentração apresentaram expressão de colágeno I significativamente menor e expressão de osteocalcina e osteopontina significativamente maior em comparação a das CTM indiferenciadas. Mas o grupo T3 100 nM apresentou concentração de osteocalcina mais elevada e semelhante à da cultura de osteoblastos. CONCLUSÃO: Conclui-se que a triiodotironina não altera a expressão de osteopontina e de colágeno pelas CTM, mas aumenta a expressão da osteocalcina durante a diferenciação osteogênica in vitro, sendo esse efeito dose-dependente.

OBJECTIVE: The aim of this study was to evaluate the effect of T3 on the expression of osteocalcin, osteopontin and collagen I during osteogenic differentiation of mesenchymal stem cells (MSC). MATERIALS AND METHODS: The bone marrow cells of Wistar rats with 30 days of age were extracted, cultured and separated into five groups: control (undifferentiated), differentiated (osteogenic stimulus) and differentiated with T3 (10-3 nM, 10-2 nM and 100 nM). For each group, four samples were cultured and were analyzed by real time RT-PCR at 7, 14 and 21 days for quantification of gene transcripts for osteocalcin, osteopontin and collagen I. RESULTS: All the different groups without T3 or with T3 regardless of the concentration, showed the collagen I expression significantly lower expression, and osteocalcin and osteopontin expression significantly greater than that of undifferentiated MSC. Nevertheless, the group T3 100 nM showed higher expression of osteocalcin and a similar expression of the osteoblast culture. CONCLUSION: In conclusion, the triiodothyronine does not affect the expression of osteopontin and collagen I, but increases ostecalcin expression during osteogenic differentiation in vitro of the MSC, and this effect is dose-dependent.

Polyurethanes (PUs) have been considered good candidates to be used in biomedical temporary devices that require mechanical properties comparable to soft tissues. However, toxicity of some PUs is still a concern, since these polyurethanes can contain potential toxic components and residual organic solvents derived from their synthesis. In this work, in vitro tests to measure viability and proliferation of human mesenchymal stem cells (hMSCs) in contact with PUs with tunable biodegradability were performed by employing MTT, alkaline phosphatase and collagen secretion assays. PUs were produced in an aqueous environment by employing isophorone diisocyanate/hydrazine (hard segment) and poly(caprolactone diol)/2,2-bis (hydroxymethyl) propionic acid (soft segment) as the main reagents. Three series of PUs having different soft segment contents were synthesized. These PUs had their chemical structure, morphology and hydrolytic degradation investigated. The rate of hydrolysis of the obtained PUs was tailored by modifying the soft segment content of the polymers. In vitro results showed that PUs can provide a satisfactory environment for the adhesion and proliferation of hMSCs.

A citocompatibilidade de compósitos sintéticos hidroxiapatita/colágeno dopados ou não com Zn+2 foi avaliada, usando cultura primária de osteoblastos. A hidroxiapatita (HAP) foi sintetizada tendo como precursores o hidróxido de cálcio e o ácido ortofosfórico. Um novo compósito de HAP foi desenvolvido adicionando 1,05% em peso de Zn(NO3)2.6H2O, obtendo-se HAPZn. Colágeno tipo I puro (COL) foi obtido de pericárdio bovino pelo método da digestão enzimática. Os compósitos HAP/COL e HAPZn/COL foram desenvolvidos e caracterizados através de MEV/EDS. A viabilidade celular e a produção de fosfatase alcalina na presença dos compósitos foram avaliadas pelos ensaios de MTT e NBT-BCIP, respectivamente, e comparadas às células osteoblásticas do controle. Três experimentos individuais foram realizados em triplicatas e submetidos à análise de variância e pós-teste de Bonferroni com significância estatística p<0,05. Os compósitos HAPZn/COL não estimularam a proliferação e o aumento da produção de fosfatase alcalina das células osteoblásticas. Os compósitos testados não alteraram a viabilidade celular nem causaram alterações na morfologia celular em 72 h, mostrando propriedades adequadas para aplicações biológicas.

The cytocompatibility of synthetic hydroxyapatite/collagen composites alone or doped with Zn+2 was tested by using primary culture of osteoblasts. The hydroxyapatite (HAP) was synthesized having calcium hydroxide and orthophosphoric acid as precursors. A new HAP composite was developed adding 1.05 w% of Zn(NO3)2.6H2O forming HAPZn. The pure type I collagen (COL) was obtained from bovine pericardium by enzymatic digestion method. The HAP/COL and HAPZn/COL composites were developed and characterized by SEM/EDS. The cell viability and alkaline phosphatase activity in the presence of composites were evaluated by MTT assay and NBT-BCIP assay, respectively, and compared to osteoblastic cells of the control. Three individual experiments were accomplished in triplicates and submitted to the variance analysis and Bonferroni’s post-test with statistically significant at p<0.05. The HAPZn/COL composite did not stimulate the proliferation and increasing of alkaline phosphatase activity of the osteoblastic cells. The tested composites did not alter the cellular viability neither caused alterations in the cellular morphology in 72 h showing adequate properties for biological applications.

We have developed an in vitro model of granuloma formation for the purpose of studying the immunological components of delayed type hypersensitivity granuloma formation in patients infected with Schistosoma mansoni. Our data show that 1) granulomatous hypersensitivity can be studied by examining the cellular reactivity manifested as multiple cell layers surrounding the antigen conjugated beads; 2) this reactivity is a CD4 cell dependent, macrophage dependent, B cell independent response and 3) the in vitro granuloma response is antigenically specific for parasite egg antigens. Studies designed to investigate the immune regulation of granulomatous hypersensitivity using purified populations of either CD4 or CD8 T cells have demonstrated the complexity of cellular interactions in the suppression of granulomatous hypersensitivity. The anti-S. mansoni egg immune responses of individual patients with chronic intestinal schistosomiasis can be classified either as soluble egg antigen (SEA) hypersensitive with maximal granulomatous hypersensitivity or SEA suppressive with activation of the T cell suppressor pathway with effective SEA granuloma modulation. Our data suggest that T cell network interactions are active in the generation of effective granuloma modulation in chronic intestinal schistosomiasis patients.