RESUMO
Pediatric traumatic vertebral injuries usually present as stable A (AOspine classification) fractures, whereas B and C injuries are relatively uncommon. In contrast to adults the appropriate treatment strategy in children remains an issue of debate.The data from two pediatric patients admitted with B and C type spinal injuries in 2007 and 2008 were retrospectively analyzed. The initial diagnostics were performed via computed tomography (CT) and an additional magnetic resonance imaging (MRI) was carried out in one case.The clinical and radiological follow-up controls were carried out after 77 and 66 months as well as 123 and 112 months, respectively. In both cases thoracolumbar MRI scans revealed degenerative alterations of the ventral half of the L1/L2 disc with a regular disc signal in the dorsal segment at the first follow-up and a progressive disc degeneration in one patient at the second follow-up.Surgical treatment of pediatric B and C type injuries via open reduction and temporary monosegmental posterior screw and rod instrumentation results in satisfactory clinical and radiological outcomes. In the absence of vertebral burst fractures, the function and stability of discoligamentous injuries in children can be restored without any additional osseous fusion.
Assuntos
Fraturas da Coluna Vertebral , Fusão Vertebral , Traumatismos da Coluna Vertebral , Criança , Humanos , Vértebras Lombares , Estudos Retrospectivos , Fraturas da Coluna Vertebral/cirurgia , Traumatismos da Coluna Vertebral/cirurgia , Vértebras TorácicasRESUMO
Cartilage-specific extracellular matrix synthesis is the prerequisite for chondrocyte survival and cartilage function, but is affected by the pro-inflammatory cytokine TNF-alpha in arthritis. The aim of the present study was to characterize whether the immunoregulatory cytokine IL-10 might modulate cartilage matrix and cytokine expression in response to TNF-alpha. Primary human articular chondrocytes were treated with either recombinant IL-10, TNF-alpha or a combination of both (at 10ng/mL each) or transduced with an adenoviral vector overexpressing human IL-10 and subsequently stimulated with 10ng/ml TNF-alpha for 6 or 24h. The effects of IL-10 on the cartilage-specific matrix proteins collagen type II, aggrecan, matrix-metalloproteinases (MMP)-3, -13 and pro-inflammatory cytokines were evaluated by real-time RT-PCR and immunohistochemistry. Transduced chondrocytes overexpressed high levels of IL-10 which significantly up-regulated collagen type II expression. TNF-alpha suppressed collagen type II and aggrecan, but increased MMP and cytokine expression in chondrocytes compared to the non-stimulated controls. The TNF-alpha mediated down-regulation of aggrecan expression was significantly antagonized by IL-10 overexpression, whereas the suppression of collagen type II was barely affected. The MMP-13 and IL-1beta expression by TNF-alpha was slightly reduced by IL-10. These results suggest that IL-10 overexpression modulates some catabolic features of TNF-alpha in chondrocytes.
Assuntos
Condrócitos/efeitos dos fármacos , Condrócitos/metabolismo , Proteínas da Matriz Extracelular/metabolismo , Regulação da Expressão Gênica/efeitos dos fármacos , Regulação da Expressão Gênica/genética , Glicoproteínas/metabolismo , Interleucina-10/metabolismo , Adenoviridae/genética , Adenoviridae/metabolismo , Idoso , Idoso de 80 Anos ou mais , Artérias/metabolismo , Proteína de Matriz Oligomérica de Cartilagem , Células Cultivadas , Condrócitos/ultraestrutura , Colágeno Tipo II/genética , Colágeno Tipo II/metabolismo , Proteínas da Matriz Extracelular/genética , Vetores Genéticos/genética , Glicoproteínas/genética , Humanos , Interleucina-10/genética , Proteínas Matrilinas , Metaloproteinase 13 da Matriz/genética , Metaloproteinase 13 da Matriz/metabolismo , Metaloproteinase 3 da Matriz/genética , Metaloproteinase 3 da Matriz/metabolismo , Microscopia Eletrônica de Transmissão , Pessoa de Meia-Idade , RNA Mensageiro/genética , Fator de Necrose Tumoral alfaRESUMO
Recent studies have shown that increased lymphocyte apoptosis contributes to sepsis-induced mortality. Furthermore, studies have demonstrated that IL-10 can suppress lymphocyte apoptosis, in part, by upregulating Bcl-2 expression and interfering with activation induced cell death. We have previously shown that intrathymic delivery of IL-10 with an adenoviral vector in wild-type mice significantly improves outcome to sepsis. Presently, we investigated the role of endogenous IL-10 expression on thymocyte apoptosis and outcome in IL-10 null mice subject to induction of generalized polymicrobial peritonitis via cecal ligation and puncture. Compared to wild-type C57BL/6 mice, IL-10 null mice demonstrated increased mortality and enhanced lymphocyte apoptosis. Intrathymic injection with an adenoviral vector expressing human IL-10 prior to cecal ligation and puncture in IL-10 null mice significantly improved outcome and decreased thymic caspase-3 activity. Furthermore, plasma concentrations of IL-6 were also significantly reduced in IL-10 null mice treated with the IL-10 expressing adenovirus. In contrast, injection of a control adenovirus did not improve outcome in IL-10 null mice, nor was caspase-3 activity reduced. Thus, local thymic expression of IL-10 not only improves outcome but also reduces local tissue apoptosis and caspase-3 activity, and appears to attenuate the systemic proinflammatory cytokine response.
Assuntos
Apoptose/imunologia , Interleucina-10/imunologia , Sepse/terapia , Timo/imunologia , Adenoviridae/genética , Animais , Caspase 3/imunologia , Caspase 3/metabolismo , Modelos Animais de Doenças , Feminino , Expressão Gênica , Terapia Genética , Humanos , Interleucina-10/genética , Interleucina-10/metabolismo , Interleucina-6/sangue , Linfócitos/imunologia , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Transgênicos , Sepse/imunologiaRESUMO
The aim of this study is to determine if there is an antagonistic effect between tumour necrosis factor (TNF)-alpha and the immunoregulatory interleukin (IL)-10 on chondrocytes survival. Serum-starved primary human articular chondrocytes were stimulated with either 10 ng/ml recombinant TNF-alpha, IL-10 or a combination of both (at 10 ng/ml each). Chondrocyte apoptosis was determined by measuring caspase-3/7, -8 and -9 activities using caspase assays. Mitochondrial apoptotic inducer bax, and the suppressor bcl-2 were evaluated using western blotting at 48 h. Results indicated that TNF-alpha increased caspase activities and resulted in a significant (p = 0.001) increase in bax/bcl-2 ratio. Stimulation with IL-10 did not alter caspase activities, while co-treatment with IL-10 and TNF-alpha inhibited TNF-alpha induced caspase activities and significantly (p > 0.004) impaired bax/bcl-2 ratio. At 24 h, mRNA levels for collagen type II, TNF-alpha and IL-10 were determined using real-time RT-PCR. Stimulation with TNF-alpha or TNF-alpha and IL-10 significantly inhibited collagen type II and increased IL-10 and TNF-alpha mRNA expression. IL-10 modulated the pro-apoptotic capacity of TNF-alpha in chondrocytes as shown by the decrease in caspase activities and bax/bcl-2 ratio compared to TNF-alpha stimulated chondrocytes, suggesting a mostly antagonistic interplay of IL-10 and TNF-alpha on mitochondrial apoptotic pathways.
Assuntos
Apoptose/efeitos dos fármacos , Cartilagem Articular/imunologia , Condrócitos/imunologia , Interleucina-10/farmacologia , Fator de Necrose Tumoral alfa/farmacologia , Idoso , Apoptose/imunologia , Cartilagem Articular/citologia , Cartilagem Articular/metabolismo , Caspases/imunologia , Caspases/metabolismo , Sobrevivência Celular/efeitos dos fármacos , Sobrevivência Celular/imunologia , Células Cultivadas , Condrócitos/citologia , Condrócitos/metabolismo , Colágeno Tipo II/biossíntese , Colágeno Tipo II/imunologia , Antagonismo de Drogas , Feminino , Regulação da Expressão Gênica/efeitos dos fármacos , Regulação da Expressão Gênica/imunologia , Humanos , Interleucina-10/antagonistas & inibidores , Interleucina-10/imunologia , Interleucina-10/metabolismo , Masculino , Pessoa de Meia-Idade , Mitocôndrias/imunologia , Mitocôndrias/metabolismo , Proteínas Recombinantes/antagonistas & inibidores , Proteínas Recombinantes/imunologia , Proteínas Recombinantes/metabolismo , Proteínas Recombinantes/farmacologia , Fatores de Tempo , Fator de Necrose Tumoral alfa/antagonistas & inibidores , Fator de Necrose Tumoral alfa/biossíntese , Fator de Necrose Tumoral alfa/imunologia , Proteína X Associada a bcl-2/imunologia , Proteína X Associada a bcl-2/metabolismoRESUMO
Dendritic cells (DC) represent a potential target for gene therapy. In their ability to process antigens and present them to T cells, DC have been allocated a unique role as initiators of the immune response in both the innate and acquired immunity. Recent in vitro studies have showed the feasibility of DC transduction with adenoviral recombinants. In cancer therapy, targeting of DC with adenovirus has been proved to be effective in inhibiting tumour growth, as well as in reducing the number of tumour metastases. The aim of our study is to evaluate the feasibility of in vivo transduction of DC in a murine lymphocyte-rich compartment (thymus) as a potential treatment for acute inflammatory diseases. Nearly 50% of the total thymic DC were transduced with a first-generation adenoviral construct following intrathymic injection, and post-transductional inflammation was neglectable. Transduction of thymic cells with adenoviral recombinants was able to induce the expression of an intracellular protein (beta-galactosidase, green fluorescent protein), as well as the secretion of human interleukin-10, within the local compartment. Furthermore, this induction of the latter significantly decreased thymic apoptosis in the applied model of acute bacterial peritonitis (cecal ligation and puncture).