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1.
Int J Pediatr Otorhinolaryngol ; 77(6): 936-42, 2013 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-23561635

RESUMO

OBJECTIVE: There is no treatment established for congenital sensorineural hearing loss because the majority of the cases are hereditary. Although congenital sensorineural hearing loss is thought to be hereditary, this hearing loss occur postnatally. We hypothesized that the transplantation of MSCs (mesenchymal stem cells) to the cochlea would be an effective therapy for stopping or delaying the progression of sensorineural hearing loss in childhood. METHODS: Cultured mouse MSCs were labeled with EGFP (enhanced green fluorescence protein) using retroviruses. EGFP-MSCs were transplanted into the posterior semicircular canal of mice at 2-3 weeks (young group) and 24-26 weeks (adult group) of age by a novel perilymphatic perfusion technique. Engraftment of MSCs was evaluated immunohistologically at 1 week and 2 weeks after transplantation. RESULTS: In young mice, migrated MSCs were detected in the cochlea tissue by immunofluorescence for EGFP and by immunohistochemistry for fibronectin. The differentiation of migrated MSCs into fibrocyte-like cells was demonstrated by immunofluorescence for connexin 26. There were no adverse effects on auditory function by MSC transplantation, and the auditory brain stem responses threshold did not significantly shift after surgery. In contrast, neither MSC migration nor differentiation was detected in the adult mice canal after MSC transplantation. CONCLUSION: The bone marrow derived MSCs were successfully transplanted into the cochlea of young mice by the perilymphatic perfusion technique and were further differentiated into fibrocyte-like cells without any adverse effects on auditory function.


Assuntos
Cóclea/cirurgia , Potenciais Evocados Auditivos do Tronco Encefálico , Perda Auditiva Neurossensorial/cirurgia , Transplante de Células-Tronco Mesenquimais/métodos , Fatores Etários , Animais , Criança , Cóclea/citologia , Cóclea/patologia , Modelos Animais de Doenças , Rejeição de Enxerto , Sobrevivência de Enxerto , Humanos , Imuno-Histoquímica , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Medição de Risco , Resultado do Tratamento
2.
Otol Neurotol ; 33(4): 655-9, 2012 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-22525215

RESUMO

HYPOTHESIS: The present study assessed how to inject a gene into the mouse vestibule and which is the optimum gene to the mouse vestibule adenovirus (AdV) vector or adeno-associated virus (AAV) vector. BACKGROUND: Loss of vestibular hair cell is seen in various balance disorder diseases. There have been some reports concerning gene delivery to the mouse vestibule in recent years. To effectively induce transgene expression at the vestibule, we assessed the efficiency of inoculating the mouse inner ear using various methods. METHODS: We employed an AdV- and AAV-carrying green fluorescent protein using a semicircular canal approach (via a canalostomy) and round window approach. RESULTS: AAV injection via canalostomy induced gene expression at the hair cells, supporting cells, and fibrocytes at the vestibular organs without auditory or balance dysfunction, suggesting it was the most suitable transfection method. This method is thus considered to be a promising strategy to prevent balance dysfunction. CONCLUSION: AAV injection via canalostomy to the vestibule is the noninvasive and highly efficient transfection method, and this study may have the potential to repair balance disorders in human in the future.


Assuntos
Dependovirus/genética , Técnicas de Transferência de Genes , Vetores Genéticos , Células Ciliadas Auditivas/citologia , Vestíbulo do Labirinto , Animais , Terapia Genética/métodos , Vetores Genéticos/administração & dosagem , Células Ciliadas Auditivas/metabolismo , Humanos , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Vestíbulo do Labirinto/metabolismo , Vestíbulo do Labirinto/patologia , Vestíbulo do Labirinto/fisiologia
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