RESUMO
In the present work, microporous membranes based on poly(ε-caprolactone) (PCL) and PCL functionalized with amine (PCL-DMAEA) or anhydride groups (PCL-MAGMA) were realized by solvent-non solvent phase inversion and proposed for use in Guided Tissue Regeneration (GTR). Nanowhiskers of hydroxyapatite (HA) were also incorporated in the polymer matrix to realize nanocomposite membranes. Scanning Electron Microscopy (SEM) showed improved interfacial adhesion with HA for functionalized polymers, and highlighted substantial differences in the porosity. A relationship between the developed porous structure of the membrane and the chemical nature of grafted groups was proposed. Compared to virgin PCL, hydrophilicity increases for functionalized PCL, while the addition of HA influences significantly the hydrophilic characteristics only in the case of virgin polymer. A significant increase of in vitro degradation rate was found for PCL-MAGMA based membranes, and at lower extent of PCL-DMAEA membranes. The novel materials were investigated regarding their potential as support for cell growth in bone repair using multipotent mesenchymal stromal cells (MSC) as a model. MSC plated onto the various membranes were analyzed in terms of adhesion, proliferation and osteogenic capacity that resulted to be related to chemical as well as porous structure. In particular, PCL-DMAEA and the relative nanocomposite membranes are the most promising in terms of cell-biomaterial interactions.
Assuntos
Regeneração Óssea , Durapatita/química , Membranas Artificiais , Células-Tronco Mesenquimais/metabolismo , Nanocompostos/química , Poliésteres/química , Adesão Celular , Diferenciação Celular , Proliferação de Células , Células Cultivadas , Feminino , Humanos , Masculino , Células-Tronco Mesenquimais/citologia , Osteogênese , PorosidadeRESUMO
OBJECTIVES: The aim of this study was to analyze the features of an oxidized titanium implant surface and to evaluate its effects on the response of human gingival fibroblasts. METHODS: 10mm×10mm×1mm turned (control) and oxidized (test) titanium samples (P.H.I. s.r.l., Italy) were examined by scanning electron microscopy and atomic force microscopy and characterized by height, spatial and hybrid roughness parameters. Primary cultures of human gingival fibroblasts were seeded on titanium samples, and cell morphology, adhesion, proliferation and extracellular matrix deposition, in terms of type I collagen synthesis, were evaluated. RESULTS: Control and test surfaces appeared considerably different at the microscopic analyses: turned samples were grooved, whereas oxidized surfaces showed a more complex micro- and nano-scaled texture, as evidenced by roughness parameters. Cell adhesion and proliferation rate, as well as collagen synthesis, were greater on oxidized vs turned surfaces. CONCLUSIONS: Although both control and test samples were in the range of average roughness proper of smooth surfaces, they exhibited significantly different topographic properties in terms of height and, mostly, hybrid parameters. Furthermore, oxidized surfaces enhanced human gingival fibroblast adhesion, proliferation and extracellular matrix deposition, and this could be due to the different structure at micro- and nano-scale levels. CLINICAL SIGNIFICANCE: Oxidized nanostructured titanium surfaces could have a significant clinical utilization in virtue of their affinity for soft tissue attachment at the implant neck and/or at the transmucosal portion of the prosthetic abutment.
Assuntos
Materiais Dentários/química , Fibroblastos/fisiologia , Gengiva/citologia , Nanoestruturas/química , Titânio/química , Adulto , Adesão Celular/fisiologia , Técnicas de Cultura de Células , Proliferação de Células , Forma Celular , Células Cultivadas , Colágeno Tipo I/análise , Colágeno Tipo I/ultraestrutura , Matriz Extracelular/química , Matriz Extracelular/ultraestrutura , Feminino , Gengiva/fisiologia , Humanos , Imageamento Tridimensional/métodos , Masculino , Teste de Materiais , Microscopia de Força Atômica , Microscopia Confocal , Microscopia Eletrônica de Varredura , Pessoa de Meia-Idade , Pseudópodes/ultraestrutura , Propriedades de SuperfícieRESUMO
PURPOSE: In the framework of a project aiming to improve the properties of poly(ε-caprolactone) (PCL)-based devices, we prepared novel composites and tested their in vitro biocompatibility and osteogenic capacity on human mesenchymal stromal cells (MSC) from bone marrow. METHODS: We prepared two functionalized derivatives, PCL-g-MAGMA and PCL-g-DMAEA, by insertion of anhydride groups by radical grafting of maleic anhydride (MA) and glycidyl-methacrylate (GMA) molecules, and by insertion of N-(dimethylamino)ethylacrylate (DMAEA) of tertiary amines groups, respectively. In addition, in order to improve the osteoconductive properties of the materials, we also prepared the corresponding composites containing the mineral component of bone, namely hydroxyapatite (HA). Mesenchymal stromal cells (MSC) derived from bone marrow were prepared, plated onto a number of discs obtained from these functionalized derivatives and tested in terms of adhesion and vitality (by MTT test and SEM observation), and the expression of alkaline phosphatase, the early marker of osteoblastic phenotype. RESULTS: The biological in vitro assessment of the functionalized materials, PCL-g-MAGMA and PCL-g-DMAEA, appeared promising only in part, in particular the cells exhibited very poor adhesion to PCL-g-MAGMA. On the contrary, the related composites, PCL-g-MAGMA-HA and PCL-g-DMAEA-HA clearly showed that the addition of HA greatly ameliorated the cell-material interaction. In particular, a surprisingly increased response characterized PCL-g-MAGMA-HA, either in terms of adhesion and vitality or in terms of alkaline phosphatase activity. CONCLUSIONS: Altogether these studies showed that the addition of HA nanowhiskers resulted for all basic materials, in particular PCL-g-MAGMA, in improved cell adhesion and performance.
Assuntos
Biopolímeros/química , Osso e Ossos/fisiologia , Células-Tronco Mesenquimais/citologia , Engenharia Tecidual , Fosfatase Alcalina/metabolismo , Biopolímeros/farmacologia , Células da Medula Óssea/citologia , Regeneração Óssea , Substitutos Ósseos/química , Substitutos Ósseos/metabolismo , Adesão Celular/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Durapatita/química , Compostos de Epóxi/química , Humanos , Anidridos Maleicos/química , Células-Tronco Mesenquimais/metabolismo , Metacrilatos/química , Nanoestruturas/química , Poliésteres/químicaRESUMO
BACKGROUND: The introduction of specific BCR-ABL inhibitors in chronic myelogenous leukemia therapy has entirely mutated the prognosis of this hematologic cancer from being a fatal disorder to becoming a chronic disease. Due to the probable long lasting treatment with tyrosine-kinase inhibitors (TKIs), the knowledge of their effects on normal cells is of pivotal importance. DESIGN AND METHODS: We investigated the effects of dasatinib treatment on human bone marrow-derived mesenchymal stromal cells (MSCs). RESULTS: Our findings demonstrate, for the first time, that dasatinib induces MSCs adipocytic differentiation. Particularly, when the TKI is added to the medium inducing osteogenic differentiation, a high MSCs percentage acquires adipocytic morphology and overexpresses adipocytic specific genes, including PPARγ, CEBPα, LPL and SREBP1c. Dasatinib also inhibits the activity of alkaline phosphatase, an osteogenic marker, and remarkably reduces matrix mineralization. The increase of PPARγ is also confirmed at protein level. The component of osteogenic medium required for dasatinib-induced adipogenesis is dexamethasone. Intriguingly, the increase of adipocytic markers is also observed in MSCs treated with dasatinib alone. The TKI effect is phenotype-specific, since fibroblasts do not undergo adipocytic differentiation or PPARγ increase. CONCLUSIONS: Our data demonstrate that dasatinib treatment affects bone marrow MSCs commitment and suggest that TKIs therapy might modify normal phenotypes with potential significant negative consequences.
Assuntos
Adipócitos/citologia , Antineoplásicos/farmacologia , Células-Tronco Mesenquimais/citologia , Proteínas Tirosina Quinases/antagonistas & inibidores , Pirimidinas/farmacologia , Tiazóis/farmacologia , Adipogenia/efeitos dos fármacos , Células da Medula Óssea/efeitos dos fármacos , Diferenciação Celular/efeitos dos fármacos , Dasatinibe , Humanos , Células K562 , Mesoderma/efeitos dos fármacos , Neoplasias/tratamento farmacológico , Osteoblastos/citologia , Fenótipo , Inibidores de Proteínas Quinases/farmacologia , Células Estromais/efeitos dos fármacosRESUMO
OBJECTIVES: Titanium nitride (TiN) coating has been proposed as an adjunctive surface treatment aimed to increase the physico-mechanical and aesthetic properties of dental implants. In this study we investigated the surface characteristics of TiN-coated titanium plasma sprayed (TiN-TPS) and uncoated titanium plasma sprayed (TPS) surfaces and their biological features towards both primary human bone marrow mesenchymal stem cells (BM-MSC) and bacterial cultures. METHODS: 15 mm×1 mm TPS and TiN-TPS disks (P.H.I. s.r.l., San Vittore Olona, Milano, Italy) were topographically analysed by confocal optical profilometry. Primary human BM-MSC were obtained from healthy donors, isolated and expanded. Cells were seeded on the titanium disks and cell adhesion, proliferation, protein synthesis and osteoblastic differentiation in terms of alkaline phosphatase activity, osteocalcin synthesis and extracellular mineralization, were evaluated. Furthermore, adhesion and proliferation of Streptococcus pyogenes and Streptococcus sanguinis on both surfaces were also analysed. RESULTS: TiN-TPS disks showed a decreased roughness (about 50%, p < 0.05) and a decreased bacterial adhesion and proliferation compared to TPS ones. No difference (p > 0.05) in terms of BM-MSC adhesion, proliferation and osteoblastic differentiation between TPS and TiN-TPS surfaces was found. CONCLUSIONS: TiN coating showed to modify the topographical characteristics of TPS titanium surfaces and to significantly reduce bacterial adhesion and proliferation, although maintaining their biological affinity towards bone cell precursors.
Assuntos
Aderência Bacteriana , Células da Medula Óssea/fisiologia , Materiais Revestidos Biocompatíveis , Implantes Dentários , Células-Tronco Mesenquimais/fisiologia , Titânio , Fosfatase Alcalina/metabolismo , Calcificação Fisiológica , Adesão Celular , Diferenciação Celular , Proliferação de Células , Células Cultivadas , Humanos , Osteoblastos/citologia , Osteoblastos/metabolismo , Osteocalcina/biossíntese , Gases em Plasma , Streptococcus/fisiologia , Propriedades de SuperfícieRESUMO
The effect of the surface charge of different immobilizing hydrogels on biohydrogen production in batch cultures was investigated using a novel isolate associated to the genus Thermoanaerobacterium. Two crosslinked polysaccharide-based hydrogels and two acrylic hydrogels were tested as polymeric carriers for cell adsorption. Immobilization improved both substrate conversion and hydrogen cumulative production compared to the suspended culture, and a yield of 1.9 mol H(2)/mol glucose was observed after 24h for alginate-supported cultures. Cationic carriers dramatically increased cell immobilization, leading to markedly faster kinetics of substrate degradation and hydrogen production in batch operation, with a peak of 3.6 mol H(2)/mol glucose for the acrylic hydrogel HM92. Accumulation of gaseous and acidic metabolites inhibited further H(2) production, shifting the carbon flow to reduced end-products and biomass synthesis. Preliminary tests showed that all the tested hydrogels had good durability and allowed hydrogen production on repeated batch runs.