RESUMEN En este proyecto se investigan los cambios que acontecen en el desarrollo y crecimiento de conejos hembras de la línea New Zealand (CoNZ) en sus huesos femorales. Los animales fueron mantenidas en jaulas individuales desde las dos semanas de edad, con comida y agua ad libitum y se sacrificaron en tiempos mensualmente consecutivos: 1, 2, 3, 4, 5, y 6 meses. Tras la obtención de las piezas femorales, y a partir de estudios imagenológicos se determinaron los ángulos del cuello femoral (Af), la longitud total (L), la densidad mineral ósea total, del centro óseo y de la metáfisis femoral (DMOt, DMOco y DMOmf respectivamente), analizándose las variaciones intergrupales por el test Wilcoxon, y corrección de Bonferroni. Se realizaron estudios histológicos de los cortes descalcificados de las piezas femorales. Los análisis sobre los Af mostraron un incremento significativo durante el primer mes mientras que L se estabilizó a partir del 4to mes. Los valores de DMOt mostraron un plateau a partir del cuarto mes, si bien las DMOco y DMOmf ya a partir del tercer mes no mostraron incrementos significativos. Histológicamente se observó para el cuarto mes ausencia de las diferentes zonas características del cartílago de crecimiento metafisiario, con presencia únicamente de un pequeño remanente de células condrales. Desde el quinto mes se observa ausencia total de cartílago, con presencia únicamente de tejido osteoide (TO). La interpretación integrada de los resultados nos permite afirmar, que a partir del cuarto mes de desarrollo, el fémur de CoNZ adquiere características compatibles con un periodo de adultez.

ABSTRACT In this project we investigated the changes of femoral development and growth of female New Zealand rabbits (NZr). Animals were maintained in individual cages since they were two weeks old with food and water ad libitum, and were sacrificed monthly consecutively: 1, 2, 3, 4, 5 and 6. Radiological studies were made with femoral pieces to determine femoral neck angle (fnA), total length (L), total bone mineral density (tBMD), bone center mineral density (bcBMD) and femoral metaphysis bone mineral density (fmBMD). We analyzed intergroup variations with Wilcoxon test and Bonferroni correction. We also performed histologic studies with femoral pieces. The fnA analyzes showed a significant increase in the first month while L stabilized since the fourth month. tBMD showed a plateu since the fourth month, even though bcBMD and fmBMD did not show any significant changes since the third month. In histology it was observed the absence of all typical growth cartilage zones since the fourth month, with the only presence of small remaining cartilage cells. In the fifth month we observed complete absence of cartilage, and presence of osteoid tissue only. The integrated interpretation of the results allows us to affirm that since the fourth month of development the femur of NZr acquires characteristics compatible with the adulthood.

Resumo Este estudo avaliou o reparo ósseo em defeitos cirúrgicos de ratos tratados com ácido hialurônico (AH) associado ou não à fração proteica de Hevea brasiliensis (F-1). Foram criados defeitos ósseos em 15 ratos albinos Wistar divididos em 3 grupos (n = 5): Grupo controle (1) - coágulo sanguíneo; Grupo HA (2) - ácido hialurônico 0,5%; Grupo HAF1 (3) - fração proteica F-1 0,1% dissolvida em ácido hialurônico a 0,5%. Após 4 semanas, os animais foram submetidos à eutanásia e o reparo ósseo avaliado por meio de análise histomorfométrica, zimografia e imunohistoquímica. A área óssea neoformada não apresentou diferença significativa (p = 0,757), mas houve tendência de aumento da trabeculação óssea nos grupos HA e HAF1. Para a análise imunoistoquímica, houve diferença na marcação do fator de crescimento endotelial vascular (VEGF) (p = 0,023), sendo maior nos grupos HA e HAF1 do que no grupo controle. Nenhuma diferença significativa na sialoproteína óssea (BSP) (p = 0,681), osteocalcina (p = 0,954), no entanto, diferenças significativas foram encontradas para a molécula de adesão de células endoteliais plaquetárias-1 (CD-31) (p = 0,040), com o grupo HAF1 sendo significativamente inferior ao controle. Para a análise zimográfica, não houve diferença significativa para metaloproteinase-2 (MMP-2) (p = 0,068), mas houve tendência de aumento da MMP-2 no grupo HA. Apesar da influência sobre os fatores angiogênicos e da aparente tendência de maior trabeculação nos grupos HA e HAF1, não houve diferença significativa na área de tecido ósseo neoformado no período analisado.

Abstract This study evaluated the bone repair in surgical defects of rats treated with hyaluronic acid (HA) associated or not with Hevea brasiliensis fraction protein (F-1). Bone defect were created in 15 albino Wistar rats divided into 3 groups (n=5): Control group (1) - blood clot; HA group (2) - 0.5% hyaluronic acid; HAF1 group (3) - 0.1% F-1 protein fraction dissolved in 0.5% hyaluronic acid. After 4 weeks, the animals were euthanized and the bone repair was evaluated through histomorphometric analysis, zymography and immunohistochemistry. The neoformed bone area did not show a significant difference (p = 0.757), but there was a tendency for bone trabeculation to increase in the groups HA and HAF1. For immunohistochemically analysis, there was a difference in vascular endothelial growth factor (VEGF) labeling (p = 0.023), being higher in the groups HA and HAF1 than the control group. No significant difference in bone sialoprotein (BSP) (p = 0.681), osteocalcin (p = 0.954), however, significant difference in platelet endothelial cell adhesion molecule-1 (CD-31) (p = 0.040), with HAF1 group being significantly lower than the control. For zymographic analysis, there was no significant difference for metalloproteinase-2 (MMP-2) (p = 0.068), but there was a tendency to increase MMP-2 in the HA group. Despite the influence on angiogenic factors and the apparent tendency for greater trabeculation in the HA and HAF1 groups, there was no significant difference in the area of newly formed bone tissue in the analyzed period.

ABSTRACT One of the important research topics in tissue engineering is the development of optimum three-dimensional scaffolds for regeneration and growth of bone tissue. The scaffold developed should promote an initial biomechanical support, provide the formation, deposition and organization of the new organic matrix generated, and degrade proportionally to the growth of the new tissue. In this study hybrid scaffolds based on the blend Chitosan (CHI)/Poly (vinyl alcohol) (PVA), with two different CHI:PVA molar ratios (1:1, and 3:1), and Bioactive Glass as the inorganic phase, were developed by a sol-gel route, followed by lyophilization. The materials were cross-linked with Glutaraldehyde. The obtained porous scaffolds were characterized by SEM, FTIR, porosity measurements by Archimedes method, and compression test. The in vitro degradation was studied by immersion in simulated body fluid for several time periods and evaluation of mass loss. Citotoxicity analysis was carried out on samples as prepared and after immersion in PBS solution for 24hrs, using human fibroblast cells and MTT method to evaluate cell viability. The matrices obtained showed promising results, presenting about 96% porosity, pore size varying in the range 20-300 µm, and interconnected pores. The mass loss presented by the matrices with CHI/PVA ratios 3:1 and 1:1 during the degradation test in vitro, was around 10% after a week of testing, with macroscopic preservation of their physical structure. Cytotoxicity tests showed that the samples were toxic as produced and not toxic after treatment with PBS, showing this approach was suitable as a final preparation step of these samples.