RESUMO
An ideal novel treatment for bone defects should provide regeneration without autologous or allogenous grafting, exogenous cells, growth factors, or biomaterials while ensuring spatial and temporal control as well as safety. Therefore, a novel osteoinductive nonviral in vivo gene therapy approach using sonoporation was investigated in ectopic and orthotopic models. Constitutive or regulated, doxycycline-inducible, bone morphogenetic protein 2 and 7 coexpression plasmids were repeatedly applied for 5 days. Ectopic and orthotopic gene transfer efficacy was monitored by coapplication of a luciferase plasmid and bioluminescence imaging. Orthotopic plasmid DNA distribution was investigated using a novel plasmid-labeling method. Luciferase imaging demonstrated an increased trend (61% vs. 100%) of gene transfer efficacy, and micro-computed tomography evaluation showed significantly enhanced frequency of ectopic bone formation for sonoporation compared with passive gene delivery (46% vs. 100%) dependent on applied ultrasound power. Bone formation by the inducible system (83%) was stringently controlled by doxycycline in vivo, and no ectopic bone formation was observed without induction or with passive gene transfer without sonoporation. Orthotopic evaluation in a rat femur segmental defect model demonstrated an increased trend of gene transfer efficacy using sonoporation. Investigation of DNA distribution demonstrated extensive binding of plasmid DNA to bone tissue. Sonoporated animals displayed a potentially increased union rate (33%) without extensive callus formation or heterotopic ossification. We conclude that sonoporation of BMP2/7 coexpression plasmids is a feasible, minimally invasive method for osteoinduction and that improvement of bone regeneration by sonoporative gene delivery is superior to passive gene delivery.
Assuntos
Proteína Morfogenética Óssea 2/genética , Proteína Morfogenética Óssea 7/genética , Técnicas de Transferência de Genes , Vetores Genéticos/metabolismo , Animais , Proteína Morfogenética Óssea 2/metabolismo , Proteína Morfogenética Óssea 7/metabolismo , Regeneração Óssea , Osso e Ossos/diagnóstico por imagem , Doxiciclina/farmacologia , Feminino , Fraturas Ósseas/terapia , Expressão Gênica/efeitos dos fármacos , Terapia Genética , Vetores Genéticos/genética , Vetores Genéticos/uso terapêutico , Masculino , Camundongos , Camundongos Nus , Músculo Esquelético/patologia , Osteogênese , Ratos , Ratos Sprague-Dawley , Sonicação , Nicho de Células-Tronco , Tomografia Computadorizada por Raios XRESUMO
Arthroscopic shoulder stabilization using Bankart's repair technique has become a standard intervention in the treatment of traumatic shoulder instability. Nevertheless, knowledge about sex-related differences postoperatively is scarce. The goal of our study was to compare the outcome of men and women after arthroscopic shoulder stabilization. All patients underwent arthroscopic shoulder stabilization due to anterior shoulder instability after traumatic shoulder dislocation. Patients received a standardized questionnaire for evaluating subjective parameters. Subsequently, patients were clinically examined by orthopedic physicians where their Constant-Murley score was raised, and apprehension and sulcus sign tests were conducted. The study included 24 men (mean age, 29.9 years; mean follow-up, 55 months) and 12 women (mean age, 31.8 years; mean follow-up, 62.3 months). Women showed a significantly decreased Constant-Murley score compared to men (P=.045). A positive apprehension test indicating decreased shoulder stability was found more often in women (P=.018). A positive sulcus sign test indicating increased joint laxity was found in 25% of women and only 12.5% of men without reaching statistical significance (P=.378). Compared to men, shoulder function as well as shoulder stability was significantly decreased in women after arthroscopic shoulder stabilization.
