The inability of the avascular region of the meniscus to regenerate has led to the use of tissue engineering to treat meniscal injuries. The aim of this study was to evaluate the ability of fibrochondrocytes preseeded on PLDLA/PCL-T [poly(L-co-D,L-lactic acid)/poly(caprolactone-triol)] scaffolds to stimulate regeneration of the whole meniscus. Porous PLDLA/PCL-T (90/10) scaffolds were obtained by solvent casting and particulate leaching. Compressive modulus of 9.5±1.0 MPa and maximum stress of 4.7±0.9 MPa were evaluated. Fibrochondrocytes from rabbit menisci were isolated, seeded directly on the scaffolds, and cultured for 21 days. New Zealand rabbits underwent total meniscectomy, after which implants consisting of cell-free scaffolds or cell-seeded scaffolds were introduced into the medial knee meniscus; the negative control group consisted of rabbits that received no implant. Macroscopic and histological evaluations of the neomeniscus were performed 12 and 24 weeks after implantation. The polymer scaffold implants adapted well to surrounding tissues, without apparent rejection, infection, or chronic inflammatory response. Fibrocartilaginous tissue with mature collagen fibers was observed predominantly in implants with seeded scaffolds compared to cell-free implants after 24 weeks. Similar results were not observed in the control group. Articular cartilage was preserved in the polymeric implants and showed higher chondrocyte cell number than the control group. These findings show that the PLDLA/PCL-T 90/10 scaffold has potential for orthopedic applications since this material allowed the formation of fibrocartilaginous tissue, a structure of crucial importance for repairing injuries to joints, including replacement of the meniscus and the protection of articular cartilage from degeneration.
Cell adhesion on materials surface is critical because this phenomenon occurs before other events, as cell spreading, cell migration and cell differentiation. It is commonly accepted that the adhesion of cells on solid substrate is influenced by several substratum surface properties, such as wettability, surface charge, roughness and topography. Plasma technique is a convenient method for modifying surface properties of materials without affecting physical properties. In this study, poly(lactide-co-glycolide), PLGA, membranes were modified by oxygen and nitrogen plasma to improve polymer hydrophilicity and verify their effect on Vero cells culture. The PLGA membranes, which were characterized by SEM and contact angle, showed increased surface rugosity and narrower contact angles. Cell adhesion, cytotoxicity assay, SEM and cytochemistry analysis showed that plasma treatment was beneficial to cell growth by improving cell-polymer interaction.
A aplicação de polímeros bioreabsorvíveis como suporte para cultura de células é um método alternativo para o tratamento de lesões e perdas teciduais. A modificação da superfície desses polímeros por plasma é uma técnica efetiva e econômica para torná-los mais hidrofílicos e melhorar a adesão celular. O objetivo deste trabalho foi estudar as interações entre células Vero e suportes de PLGA previamente tratados por plasma de oxigênio, com o objetivo de aumentar a hidrofilicidade da superfície desses materiais. As amostras foram caracterizadas através das análises de ângulo de contato, MEV e citoquímica. O tratamento por plasma melhorou a adesão e a proliferação celular, em relação às membranas sem tratamento.
The physiological repair of osteochondral lesions requires the development of a scaffold that is compatible with the structure of the damaged tissue, cartilage and bone. The aim of this study was to evaluate the biological performance of a PLDLA/PCL-T (90/10) scaffold for repairing osteochondral defects in rabbits. Polymeric scaffolds containing saccharose (75% w/v) were obtained by solvent casting and then implanted in the medial knee condyles of 12 New Zealand rabbits after osteochondral damage with a trephine metallic drill (diameter: 3.3 mm) in both medial femoral condyles. Each rabbit received the same treatment, i.e., the polymeric scaffold was implanted on the right side while no material was implanted on the left side (control). Four and 12 weeks later histological examination revealed bone neoformation in the implant group, with the presence of hyaline cartilage and mesenchymal tissue. In contrast, the control group showed bone neoformation with necrosis, exacerbated superficial fibrosis, inflammation and cracks in the neoformed tissue. These findings indicate that the PLDLA/PCL-T scaffold was biocompatible and protected the condyles by stabilizing the lesion and allowing subchondral bone tissue and hyaline cartilage formation.
Resumo: Buscando estratégias que repercutam na melhoria da interação entre materiais poliméricos biorreabsorvíveis e o crescimento celular, o presente estudo in vitro teve como objetivo estudar a influência de blendas de PPD/PHB na adesão celular e crescimento de fibrocondrócitos obtidos a partir de cultura primária. As blendas de PPD/PHB foram preparadas pelo método de evaporação de solvente nas composições 100/0, 60/40 e 50/50 e caracterizadas por microscopia eletrônica de varredura (MEV). Observações ultra-estruturais mostraram alterações na morfologia celular, sugerindo que os fibrocondrócitos podem responder a alterações no substrato alterando seu perfil fenotípico. As análises com MTT demonstraram que as blendas não apresentaram citotoxicidade e permitiram a adesão e proliferação dos fibrocondrócitos sobre os substratos em todas as suas composições. O ensaio colorimétrico com Sirius Red evidenciou a capacidade de manutenção da síntese de matriz extracelular colágena sobre as amostras, concluindo-se que as blendas de PPD/PHB podem ser indicadas para o cultivo celular. Palavras-chave: Blenda, poli(p-dioxanona), poli(hidroxi butirato), fibrocondrócitos. Cytocompatibility of Poly(p-dioxanone)/ Poly(hydroxybutic) (PPD/PHB) Blends to Cartilage Tissue EngineeringAbstract: In order of seek strategies to improve the interaction between bioreabsorbable polymer materials and cellular growth, this work aimed at evaluating in vitro the influence of PPD/PHB blends on cell adhesion and fibrochondrocytes growth. Fibrochondrocytes cells were obtained by primary extraction from enzymatic digestion methods. The PPD/PHB blends were prepared by casting with 100/0, 60/40 and 50/50 compositions, and were characterized by scanning electron microscopy (SEM). After 6, 48, 120 and 168 hours in culture, ultrastructural observations showed changes in cell morphology, suggesting that the fibrochondrocytes can respond to substrate modifications, changing their phenotypic profile. The MTT analyses showed that the blends did not present cytotoxicity and allowed fibrochondrocytes adhesion and proliferation on the membranes in all compositions. The colorimetric Sirius Red test revealed the capability of extracellular matrix synthesis on the blends, from which one can conclude that the PPD/PHB blends are not cytotoxic and can be indicated for cell culture.
Resumo: Polímeros sintéticos biorreabsorvíveis podem ser utilizados sob a forma de membranas para sustentar e guiar o crescimento celular, através do processo de reparação tecidual. Este trabalho avaliou membranas de poli(ácido lático), PLLA, com adição de 10% de trietil-citrato usadas como curativos de feridas cutâneas agudas em ratos Wistar. Inicialmente uma ferida de 2cm 2 foi provocada na região dorsal de 24 animais. Estes foram divididos em 2 grupos: tratamento, nos quais as feridas foram recobertas pela membrana polimérica e controle, com feridas permanecendo cruentas. Os resultados obtidos em 1, 3, 7 e 15 dias mostraram uma resposta inflamatória mais satisfatória nas feridas protegidas pelas membranas, com reparação precoce e colágeno mais organizado quando comparadas com as áreas incialmente mantidas sem proteção. Além do que, as áreas protegidas pelas membranas não mostraram alterações inflamatórias irritativas que pudessem ser imputadas ao uso da membrana polimérica. Diante disso, conclui-se que a membrana de PLLA/Trietil-citrato protegeu efetivamente as feridas, permitindo o processo de reparação e mostrando-se promissora como curativo cutâneo. Palavras-chaves: PLLA/Trietil-citrato, reparação tecidual, curativo cutâneo. PLLA/Triethyl Citrate Membrane as an Alternative for the Treatment of Skin WoundsAbstract: Bioresorbable polymers can be applied as membranes to sustain and guide cell growth through the regeneration process. This study evaluated poly(acid lactide), PLLA, membranes with addition of 10% triethyl citrate as skin wound healing in Wistar rats. Initially a 2cm 2 skin wound was exercised of the back of 24 animals. The animals were divided into two groups: treated, whose the polymer membrane was implanted, and control, in which the wound was kept exposed. The results obtained after 1, 3, 7 and 15 days showed an inflammatory response more satisfactory in the implanted wounds, with early repair and collagen more organized when compared to exposed wounds. In addition to, the protected areas showed no irritant inflammatory response which could be attributed to the membrane. Thus, we conclude that the PLLA/Triethyl citrate membrane has effectively protected the wounds, allowing the repair and presenting itself as a promising skin dressing. Keywords: PLLA/triethyl citrate, tissue repair, skin wound. IntroduçãoUma ferida é representada pela interrupção da continuidade de um tecido corpóreo, em maior ou em menor extensão, causada por qualquer tipo de trauma físico, químico, mecânico ou desencadeada por uma afecção clínica, que aciona as frentes de defesa orgânica [1] . Uma vez estabelecida a lesão, a pele possui dois mecanismos para reparação tissular: a regeneração e a cicatrização. A regeneração envolve a substituição do tecido lesado por tecido novo, equivalente ao lesado, através de um processo inflamatório agudo. Isso ocorre quando a camada basal da epiderme mantém-se intacta, e assim as bordas epiteliais vão se aproximando em direção ao centro da ferida, ocluindo rapidamente sua superfície. A c...
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