Platelet lysate 3D scaffold supports mesenchymal stem cell chondrogenesis: An improved approach in cartilage tissue engineering

Articular lesions are still a major challenge in orthopedics because of cartilage's poor healing properties. A major improvement in therapeutics was the development of autologous chondrocytes implantation (ACI), a biotechnology-derived technique that delivers healthy autologous chondrocytes after in vitro expansion. To obtain cartilage-like tissue, 3D scaffolds are essential to maintain chondrocyte differentiated status. Currently, bioactive 3D scaffolds are promising as they can deliver growth factors, cytokines, and hormones to the cells, giving them a boost to attach, proliferate, induce protein synthesis, and differentiate. Using mesenchymal stem cells (MSCs) differentiated into chondrocytes, one can avoid cartilage harvesting. Thus, we investigated the potential use of a platelet-lysate-based 3D bioactive scaffold to support chondrogenic differentiation and maintenance of MSCs. The MSCs from adult rabbit bone marrow (n=5) were cultivated and characterized using three antibodies by flow cytometry. MSCs (1×105) were than encapsulated inside 60μl of a rabbit platelet-lysate clot scaffold and maintained in Dulbecco's Modified Eagle Medium Nutrient Mixture F-12 supplemented with chondrogenic inductors. After 21 days, the MSCs-seeded scaffolds were processed for histological analysis and stained with toluidine blue. This scaffold was able to maintain round-shaped cells, typical chondrocyte metachromatic extracellular matrix deposition, and isogenous group formation. Cells accumulated inside lacunae and cytoplasm lipid droplets were other observed typical chondrocyte features. In conclusion, the usage of a platelet-lysate bioactive scaffold, associated with a suitable chondrogenic culture medium, supports MSCs chondrogenesis. As such, it offers an alternative tool for cartilage engineering research and ACI. © 2013 Informa UK Ltd.

RADIOGRAFIA E MACROSCOPIA do JOELHO APÓS ESTABILIZAÇÃO EXTRA-ARTICULAR UTILIZANDO FÁSCIA LATA, FIO DE POLIÉSTER TRANÇADO OU FIO DE POLIAMIDA PARA CORREÇÃO da RUPTURA do LIGAMENTO CRUZADO CRANIAL em CÃES

A ruptura do ligamento cruzado cranial é uma das principais doenças ortopédicas que afetam os cães. Muitas técnicas cirúrgicas foram descritas no intuito de aliviar a dor, restaurar a estabilidade biomecânica do joelho e prevenir a progressão da osteoartrite. Fáscia lata, fio de poliéster trançado e fio de poliamida foram empregados na estabilização do joelho após excisão do ligamento cruzado cranial em cães, os quais foram submetidos à avaliação radiográfica e macroscópica da articulação. Neste estudo, foram utilizados 18 cães com massa corporal superior a 15Kg (peso médio - 19,67kg), separados em 3 grupos eqüitativos correspondentes a cada técnica, avaliados durante 30 e 60 dias. Ao exame radiográfico, independentemente de grupo, os cães apresentaram evidência de efusão articular moderada a severa, distensão da cápsula articular e, na maioria dos casos, ausência de sinais de doença articular degenerativa. Ao exame macroscópico da articulação do joelho observou-se espessamento da cápsula articular e tecidos moles periarticulares, erosão da cartilagem articular dos côndilos femorais em todos os grupos e afrouxamento dos fios nos cães submetidos às técnicas de estabilização extra-articular com fio de poliéster trançado e fio de poliamida.

Estudo comparativo do uso de meloxicam por via intra-articular e sistêmica no controle da osteoartrite experimentalmente induzida em joelho de coelhos

Objective: This study aimed to evaluate morphologic changes, as well as chondroprotective and intra-articular effects of meloxicam on joint repair in rabbits induced by experimental trochleoplasty, minimizing possible adverse side effects. Methods: Thirty-five rabbits were divided into four groups: the control group, which did not undergo surgery, and operated groups, which used different ways of administering the anti-inflammatory agent: systemic, 0.2 mg/kg; intra-articular, 0.5 mg/kg; positive group control, without meloxicam. Each operated group was divided according to the periods of 7 or 30 days evaluation after surgery. Results: Regarding macroscopic and histological evaluation of cartilage, after 30 days, most animals showed almost complete joint repair, the presence of few or no inflammatory cells; whereas part of the animals treated with meloxicam presented necrosis in the trochlear ridge and absence of inflammatory cells after 7 days. In positive control group, it was observed moderate inflammation and connective tissue proliferation. None of the animals in the operated groups showed irregularities 30 days after surgery. Conclusion: Either intra-articular or systemic, meloxicam revealed to be favorable to be used for joint repair and control of inflammatory reaction. © 2013 Sociedade Brasileira de Ortopedia e Traumatologia.

Estenose da fossa intercondilar após estabilização articular com retalho de fáscia lata em cães

Determinaram-se os índices de largura da fossa intercondilar (FI), após transecção do ligamento cruzado cranial em nove cães adultos submetidos à estabilização articular com retalho de fáscia lata. Os joelhos foram alocados em dois grupos, sendo o joelho direito (GI) submetido à incisuroplastia troclear (ITR) e posterior estabilização articular, e o joelho esquerdo submetido somente à substituição ligamentar (GC). Cada grupo foi dividido em três subgrupos correspondentes aos momentos de eutanásia aos 30, 90 e 180 dias de pós-operatório. Os índices de largura da FI foram determinados, macroscópica e radiograficamente, pela mensuração da abertura cranial da FI nos terços cranial, médio e caudal, e indexados em relação à largura epicondilar. Observou-se aumento significativo dos índices macroscópicos e radiográficos nas articulações do GI, sendo estes estatisticamente diferentes daqueles das articulações de GC. Não foi observada estenose intercondilar nos joelhos de GC após a estabilização articular. Conclui-se que a estabilização articular com retalho de fáscia lata preveniu a estenose da fossa intercondilar, e que a ITR promoveu o alargamento permanente dessa estrutura.

Macro- and microstructural organization of the rabbit's celiac-mesenteric ganglion complex (Oryctolagus cuniculus)

The macro- and microstructures of the rabbit celiac-mesenteric ganglion complex are described in 20 young animals. We found ten celiac ganglia, twenty-seven cranial mesenteric ganglia and eleven celiac-mesenteric ganglia. The celiac ganglia had a rectangular shape in nine cases (90%) and a circular one in one case (10%). The cranial mesenteric ganglia presented triangular (66.7%), rectangular (11.1%), L-shape (18.5%) and semilunar (3.7%) arrangements. The celiac-mesenteric ganglia were organized in three patterns: a single left celiac-mesenteric ganglion having a caudal portion (72.7%); celiac-mesenteric ganglia without a caudal portion (18.2%) and a single celiac-mesenteric ganglion with two portions: left and right (9.1%).The microstructure was investigated in nine celiac-mesenteric ganglia. The results showed that the celiac-mesenteric ganglion is actually a ganglion complex constituted of an agglomerate of ganglionic units separated by nerve fibers, capillaries and septa of connective tissue. Using the semi-thin section method we described the cellular organization of the celiac-mesenteric ganglion complex. Inside of each ganglionic unit, there were various cell types: principal ganglion neurons (PGN), glial cells (satellite cells) and SIF cells (small intensely fluorescent cells or small granular cells), which are the cytologic basis for each ganglionic unit of the rabbit's celiac-mesenteric ganglion complex.

Comparison of synthetic hydroxyapatite and porous polyethylene implants in eviscerated rabbit eyes

Purpose: Synthetic hydroxyapatite and porous polyethylene (Polipore) spheres were placed in rabbits' eviscerated cavities to evaluate tissue reaction and volume maintenance.Methods. Fifty-six Norfolk albino rabbits underwent unilateral evisceration and implantation of synthetic hydroxyapatite (H group, 28 animals) or porous polyethylene spheres (P group, 28 animals). Postoperative reactions, animal behavior, and socket conditions were monitored. Light microscopy and morphometric evaluation with statistical analysis of the exenterated orbits were performed at 7, 15, 30, 60, 90, 120, and 180 days. Scanning electron microscopy was appraised 7, 60, and 180 days after surgery.Results: Two animals from the H group and 1 from the P group had extrusion 7 days after surgery. Throughout the experimental period, the synthetic hydroxyapatite caused more inflammation than the porous polyethylene material. Ingrowth in the sphere occurred 7 to 15 days after the surgery in both groups, and the tissue reaction became denser at approximate to60 to 90 days, when bony metaplasia began in the H group. Volume maintenance was better in the P group and with a smaller pseudocapsule surrounding the implanted sphere than in the H group.Conclusions: Clinical findings demonstrated mild inflammation inside the sphere and in the pseudocapsule surrounding it and better cavity volume maintenance in the P group animals. The authors consider porous polyethylene a more suitable material than synthetic hydroxyapatite for use in anophthalmic cavity reconstruction.

Effects of Inhaled Nitric Oxide on Oxidative Stress and Histopathological and Inflammatory Lung Injury in a Saline-Lavaged Rabbit Model of Acute Lung Injury

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

High-frequency oscillatory ventilation attenuates oxidative lung injury in a rabbit model of acute lung injury

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Progression of articular and extraarticular damage in oligoarticular juvenile idiopathic arthritis

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Development of a rabbit's urethral sphincter deficiency animal model for anatomical-functional evaluation

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)