Growth Inhibition of Osteosarcoma Cell Lines in 3D Cultures: Role of Nitrosative and Oxidative Stress.

BACKGROUND: 3D cell cultures have revolutionized the understanding of cell behavior, allowing culture of cells with the possibility of resembling in vivo intercellular signaling and cell-extracellular matrix interaction. AIM: The effect of limited oxygen penetration into 3D culture of highly metastatic osteosarcoma 143B cells in terms of expression of nitro-oxidative stress markers was investigated and compared to standard 2D cell culture. MATERIALS AND METHODS: Human osteosarcoma (143B cell line) cells were cultured as monolayers, in collagen and Matrigel. Cell viability, gene expression of nitro-oxidative stress markers, and vascular endothelial growth factor were determined using Trypan blue assay, quantitative polymerase chain reaction and enzyme-linked immunosorbent assay, respectively. RESULTS: Three-dimensional environments modify nitro-oxidative stress and influence gene expression and cell proliferation of OS 143B cells. CONCLUSION: Commercial cell lines might not constitute a good model of 3D cultures for bone tissue engineering, as they are highly sensitive to hypoxia, and hypoxic conditions can induce oxidation of the cellular environment.

Spatial and temporal distribution of growth factors receptors in the callus: implications for improvement of distraction osteogenesis.

Management of bone deficits by distraction osteogenesis is an appreciated but lengthy procedure. To accelerate the consolidation of newly formed distraction callus, an administration of growth factors into the distraction gap has been suggested. Changes in expression of growth factors receptors in the distracted callus during consolidation were studied in order to improve our understanding of the underlying molecular mechanisms and to provide a scientific basis for clinical application of growth factors. In a model of rat bone lengthening the expression of receptors for: vascular endothelial growth factor, transforming growth factor beta1, insulin like growth factor and platelet derived growth factor were evaluated semiquantitatively with immunohistochemistry and quantitatively with real time PCR in various callus zones at zero, one and two weeks of consolidation. Overall growth factors receptors' expression was highest at the beginning of consolidation. It was strongest in the trabecular bone and weakest in the fibrous zone. Transforming growth factor beta receptor 1 was most abundant and vascular endothelial growth factor receptor 1, although scarce, showed the most consistent expression. In contrast to the osteogenic zones, the fibrous zone demonstrated a dramatic loss of the growth factors receptors over time. High growth factors receptors expression shortly after termination of the distraction may warrant the maximal callus' response to injected growth factors. Rapid decline of growth factors receptors in the fibrous zone may imply its decreasing sensitivity to growth factors and, as a consequence, a declining osteogenic potential.

Mesomelia-synostoses syndrome results from deletion of SULF1 and SLCO5A1 genes at 8q13.

Mesomelia-synostoses syndrome (MSS) or mesomelic dysplasia with acral synostoses Verloes-David-Pfeiffer type is a rare autosomal-dominant disorder characterized by mesomelic limb shortening, acral synostoses, and multiple congenital malformations. So far, five patients in four unrelated families have been reported worldwide with MMS. By using whole-genome oligonucleotide array CGH, we have identified an interstitial deletion at 8q13 in all patients. The deletions vary from 582 Kb to 738 Kb in size, but invariably encompass only two genes: SULF1, encoding the heparan sulfate 6-O-endosulfatase 1, and SLCO5A1, encoding the solute carrier organic anion transporter family member 5A1. SULF1 acts as a regulator of numerous growth factors in skeletal embryonic development whereas the function of SLCO5A1 is yet unknown. Breakpoint sequence analyses performed in two families showed nonrecurrent deletions. Real-time quantitative RT-PCR analysis showed the highest levels of SULF1 transcripts in human osteoblasts and cartilage whereas SLCO5A1 was highly expressed in human fetal and adult brain and heart. Our results strongly suggest that haploinsufficiency of SULF1 contributes to this mesomelic chondrodysplasia, highlighting the critical role of endosulfatase in human skeletal development. Codeletion of SULF1 and SLCO5A1--which does not result from a low-copy repeats (LCRs)-mediated recombination event in at least two families--was found in all patients, so we suggest that haploinsufficiency of SULF1 combined with haploinsufficiency of SLCO5A1 (or the altered expression of a neighboring gene through position effect) could be necessary in the pathogenesis of MSS.

A case of mesomelic dysplasia Kantaputra type--new findings and a new diagnostic approach.

Mesomelic dysplasia is a heterogeneous group of rare bone diseases characterized by disproportionate shortness of middle segments of limbs and short stature. This study reports a male patient with an early diagnosed mesomelic dysplasia Kantaputra type (MIM*156232), characterized by symmetric, bilateral forearm and lower leg shortening with feet malformations and ankle-tarsal synostoses. As a result of several diagnostic biases in defining the mesomelic syndromes, we attempted to categorize symptoms and to clarify hitherto difficult discrimination between mesomeliae. Given that specific ankle joint alignment distortions seem to be pathognomonic for Kantaputra type (especially a fibulo-calcanear and, to a lesser extent, a tibio-talar synostosis), but not detectable with plane radiography, we encouraged use of MRI evaluation as an indispensable modality for proper diagnosis and further preoperative planning.

[Results of the treatment of the distal femoral epiphysiolysis]. / Wyniki leczenia zluszczen dalszej nasady kosci udowej.

INTRODUCTION: Distal femoral epiphysiolysis (DFE) is rare. In most of the cases it occures as an open epiphysiolysis. In the past DFE had often resulted in a tissue necrosis and required an amputation. Among complications limb shortening due to premature epiphyseal closure and deviation of the limb axis are evidenced. AIM: This study was conduced in order to evaluate the results of the treatment of DFE. MATERIAL: Eight children suffering from DFE who had undergone a treatment between 1990 and 2005 were reviewed. The mean age at the time of injury was 15.3 years. All cases of epiphysiolysis have been classified as Salter-Harris type II. The treatment consisted of reduction and 6 weeks immobilisation in 3 cases. Reduction followed by K-wire stabilization and immobilisation was applied in all other cases. METHODS: The follow-up evaluation covered a clinical and roentgenographic examination. The angular deformmity of tibia was measured in saggital and frontal plane in comparison to intact side. The range of motion and stability of both knee joints were recorded as well. The comparative X-ray pictures of the femur and knee joint were taken in A-P and lateral positions. RESULTS: The study revealed a premature closure of the injured distal femoral epiphysis in four children. There was a limb shortening of over 2 cm in 3 cases and over 3cm in one case. CONCLUSION: The distal femoral epiphysiolysis is a rare type of injury. It often leads to leg length discrepancy.

Late reduction for a chronic trans-scaphoid perilunate dislocation in a skeletally immature 15-year-old boy: a case report.

Fracture dislocation of the wrist is a rare injury in adolescents, and therefore it is easily ignored at the initial treatment. Once ignored, an alternative treatment such as proximal row carpectomy is indicated, but surgical outcome is not as good as that of an early reduction. We have experienced a chronic case of fracture dislocation in a 15-year-old, skeletally immature boy and treated it by scaphoid osteotomy, associated with bone grafting, screw fixation and ligament repair. The patient had no difficulties in daily activities nine years post-operatively, however the X-ray showed slight deformity of the scaphoid. Even for a chronic case, late reduction with ligamentous repair should be considered in adolescents.

Evaluating degenerative changes in the wrist after surgical treatment for scaphoid pseudarthrosis using a corticocancellous graft for anatomical reconstruction.

Background. Pseudoarthrosis of the scaphoid results in the development of carpal instability and severe posttraumatic osteoarthritis. Modern methods of treatment focus on reconstruction of the anatomical shape and length of the scaphoid. The purpose of our study was to determine the occurrence of carpal osteoarthritis after surgical treatment of scaphoid pseudoarthrosis using a cortico-cancellous bone grafting technique. Material and methods. 48 patients who had undergone surgical treatment for scaphoid union failure were examined. The surgical method included the anatomical reconstruction of the scaphoid bone, which resulted in normal ligament tension and carpal stability. The clinical examination included pain evaluation, return to work, range of wrist motion, and grip strength. On comparative x-rays of both wrists we evaluated union, the length and shape of the scaphoid bone, the scapho-lunate angle, the carpal height ratio, and the carpal ulnar shift. Results. Union was obtained in 42 cases. In 26 patients there were no signs of osteoarthritis, while 11 patients had slight degenerative changes in the radio-scaphoid joint. Conclusions. According to our analysis, cortico-cancellous bone grafting is an effective surgical procedure, useful not only in obtaining union, but also in reconstructing the shape and length of the scaphoid and preventing posttraumatic carpal asteoarthritis.

Use of pedicled bone grafts in the treatment of nonunion of the scaphoid.

The purpose of this article is to present some contemporary views regarding the treatment of scaphoid nonunion using pedicled bone grafts. Based on a review of the literature, the authors discuss surgical technique, including the advantages and disadvantages of various methods and their modifications, as well as outcomes. Among the methods discussed are pronator quadratus pedicled bone grafts, grafts pedicled on arteries (radial artery branches), and free vascularized bone grafts. Experimental studies of the use of free vascularised bone-ligament-bone autografts for the reconstruction of the scapholunate ligament are described. The clinical application of scaphoid allograft is also discussed.