RESUMO
BACKGROUND: The golden retriever muscular dystrophy (GRMD) dogs represent the best available animal model for therapeutic trials aiming at the future treatment of human Duchenne muscular dystrophy (DMD). We have obtained a rare litter of six GRMD dogs (3 males and 3 females) born from an affected male and a carrier female which were submitted to a therapeutic trial with adult human stem cells to investigate their capacity to engraft into dogs muscles by local as compared to systemic injection without any immunosuppression. METHODS: Human Immature Dental Pulp Stem Cells (hIDPSC) were transplanted into 4 littermate dogs aged 28 to 40 days by either arterial or muscular injections. Two non-injected dogs were kept as controls. Clinical translation effects were analyzed since immune reactions by blood exams and physical scores capacity of each dog. Samples from biopsies were checked by immunohistochemistry (dystrophin markers) and FISH for human probes. RESULTS AND DISCUSSION: We analyzed the cells' ability in respect to migrate, engraftment, and myogenic potential, and the expression of human dystrophin in affected muscles. Additionally, the efficiency of single and consecutive early transplantation was compared. Chimeric muscle fibers were detected by immunofluorescence and fluorescent in situ hybridisation (FISH) using human antibodies and X and Y DNA probes. No signs of immune rejection were observed and these results suggested that hIDPSC cell transplantation may be done without immunosuppression. We showed that hIDPSC presented significant engraftment in GRMD dog muscles, although human dystrophin expression was modest and limited to several muscle fibers. Better clinical condition was also observed in the dog, which received monthly arterial injections and is still clinically stable at 25 months of age. CONCLUSION: Our data suggested that systemic multiple deliveries seemed more effective than local injections. These findings open important avenues for further researches.
Assuntos
Diferenciação Celular , Polpa Dentária/citologia , Doenças do Cão/terapia , Distrofia Muscular Animal/terapia , Transplante de Células-Tronco , Dente Decíduo/citologia , Animais , Movimento Celular , Células Cultivadas , Criança , Pré-Escolar , Polpa Dentária/transplante , Doenças do Cão/sangue , Doenças do Cão/genética , Doenças do Cão/fisiopatologia , Cães , Distrofina/metabolismo , Imunofluorescência , Genótipo , Humanos , Camundongos , Desenvolvimento Muscular , Músculo Esquelético/patologia , Distrofia Muscular Animal/sangue , Distrofia Muscular Animal/genética , Distrofia Muscular Animal/fisiopatologia , Dente Decíduo/transplanteRESUMO
Os autores apresentam uma alternativa para restabelecimento anátomo-funcional de molares decíduos com destruiçäo coronária total. No cumprimento desse objetivo, utilizam coroa dentária total de dentes decíduos, previamente tratados e mantidos em um "banco de dentes", através de colagem no remanescente dentário do paciente pediátrico