Arnaldo Rodrigues Santos

Use of blends of bioabsorbable poly(L-lactic acid)/poly(hydroxybutyrate- co-hydroxyvalerate) as surfaces for Vero cell culture

Vero cells, a cell line established from the kidney of the African green monkey (Cercopithecus aethiops), were cultured in F-10 Ham medium supplemented with 10% fetal calf serum at 37 degrees C on membranes of poly(L-lactic acid) (PLLA), poly(hydroxybutyrate-co-hydroxyvalerate) (PHBV) and their blends in different proportions (100/0, 60/40, 50/50, 40/60, and 0/100). The present study evaluated morphology of cells grown on different polymeric substrates after 24 h of culture by scanning electron microscopy. Cell adhesion was also analyzed after 2 h of inoculation. For cell growth evaluation, the cells were maintained in culture for 48, 120, 240, and 360 h. For cytochemical study, the cells were cultured for 120 or 240 h, fixed, processed for histological analysis, and stained with Toluidine blue, pH 4.0, and Xylidine ponceau, pH 2.5. Our results showed that cell adhesion was better when 60/40 and 50/50 blends were used although cells were able to grow and proliferate on all blends tested. When using PLLA/PHBV (50/50) slightly flattened cells were observed on porous and smooth areas. PLLA/PHBV (40/60) blends presented flattened cells on smooth areas. PLLA/PHBV (0/100), which presented no pores, also supported spreading cells interconnected by thin filaments. Histological sections showed that cells grew as a confluent monolayer on different substrates. Cytochemical analysis showed basophilic cells, indicating a large amount of RNA and proteins. Hence, we detected changes in cell morphology induced by alterations in blend proportions. This suggests that the cells changed their differentiation pattern when on various PLLA/PHBV blend surfaces.

Analysis of the growth pattern of Vero cells cultured on dense and porous poly (L-Lactic Acid) scaffolds

Poly (L-lactic acid) (PLLA) polymers are the most frequently used substrates for cell culture, tissue regeneration and orthopedic prostheses, mainly because of their atoxic characteristics and good biocompatibility. The objective of this study was to evaluate whether a higher density or different pore diameters (less than 45, 180-250, and 250-350 µm) would change the growth pattern of cultured cells. The cells were found to adhere to and spread over all PLLA scaffolds studied. The cells also showed similar proliferation on dense and porous PLLA scaffolds, except for PLLA scaffolds with a smaller pore diameter. The cytochemical data showed high metabolic cellular activity on the various substrates. Overall, the results indicated satisfactory cell growth and proliferation on the different PLLA scaffolds studied, especially for those with pore diameters of 180-250 µm and 250-350 µm.

Cartilage reconstruction using self-anchoring implant with functional gradient

This study presents an innovative and original biomaterial designed to substitute for articular cartilage and mimic its mechanical behavior, including elastic cushioning and the characteristics of fiber-reinforced gel. The material was composed of polyurethane and bioglass microfiber 45S5. It was designed to present a tribological surface to the cartilage of the tibial plateau, and to convert over a functional gradient to an osteointegrable region for self-anchorage to the subchondral bone. The biomaterial samples showed no toxicity and promoted cell spreading. Subsequent in vivo studies in rabbits demonstrated the formation of a rigid structure similar to bone trabeculae in the distal region of the tribological surface of the implant. The tribological surface of the proximal region showed a fibrocartilaginous tissue with highly vascularized chondrocytes, thus validating the proposed concept for the design of the implant incorporating a functional gradient and auto-stability.

Titanium Oxide (TiO2) Coatings Produced on Titanium by Oxygen-Plasma Immersion and Cell Behaviour on TiO2

Plasma immersion process was investigated as a method for producing bioceramics coatings on metallic implants due to its advantages, which include the production of coatings on three-dimensional workpieces, with high density and superior adhesion. In this process, the oxygen plasma was utilized to form titanium oxide on titanium substrate. The structure, composition and surface morphology were studied using scanning electron microscopy (SEM) and X-ray diffraction. In addition a preliminary study has also been carried out, on TiO2-coated and uncoated titanium substrates, to analyse the in vitro biocompatibility (cytotoxicity evaluation and cell morphology).

Technologies Applied to Stimulate Bone Regeneration

Arnaldo Rodrigues Santos Jr.1, Christiane Bertachini Lombello2 and Selma Candelaria Genari3 1Centro de Ciencias Naturais e Humanas (CCNH), Universidade Federal do ABC, Santo Andre, SP; 2Centro de Engenharia e Ciencias Sociais Aplicadas (CECS), Universidade Federal do ABC, Santo Andre, SP; 3Centro Estadual de Educacao Tecnologica Paula Souza, Faculdade de Tecnologia de Bauru (FATEC), Bauru, SP; Brazil

Morphology of Fibroblastic Cells Cultured on Diamond-Like Carbon Coatings Produced by Plasma Immersion Using AFM and SEM

For the potential use of diamond-like carbon (DLC) coating for biomedical applications, it would be important to evaluate the biological effects of these coatings. In this study, DLC coatings were deposited on glass coverslips using the plasma immersion process, which produces films with adhesion properties superior to those prepared with conventional techniques. Scanning electron microscopic and atomic force microscopic observations were used to study the morphology of fibroblasts growth on DLC coatings.