RESUMEN
OBJECTIVE: Dura mater healing is crucial to prevent cerebrospinal fluid (CSF) leaks after neurosurgical procedures. Biological mechanisms leading to dural closure are only partially understood and have been studied in animals exclusively. We studied an in vitro model of dural closure which uses human cells. MATERIALS AND METHODS: We used human dura intended for disposal after surgery. Explant primary cultures were performed. Cells were characterized through common staining and immunohistochemistry. A cell growth curve was elaborated and the effect of dexamethasone on cell count was assessed. Spongostan®, oxidized regenerated cellulose and autologous plastic materials were also evaluated for their effect on cellular growth. RESULTS: All specimens showed growth in fusiform cells, which project pseudopods and fuse into spindles. Cells showed desmin and vimentin positivity, and were negative for all the other stains, behaving phenotypically like fibroblasts. No collagen base was necessary for cell growth. Dexamethasone decreased cell count in the primary culture as well as in the explant, and reduced the cell proliferation marker Ki-67. Spongostan® was successfully used as a graft, and fibroblast cultures were additionally developed with muscle, pericranium, galea, and fascia. Oxidized cellulose induced cell death by lowering the pH of the solution. DISCUSSION: According to the findings, unlike mini-pigs and rabbits, in humans, dural fibroblast sensitivity to collagen seems to be lower. Dexamethasone inhibits fibroblast invasion, which is the biological base of wound dehiscence in cranial surgery. Although Spongostan is useful, Surgicel® can lower the media pH, thereby inhibiting cellular growth.
Asunto(s)
Técnicas de Cultivo de Célula/métodos , Duramadre/citología , Fibroblastos/citología , Modelos Biológicos , Cicatrización de Heridas , Antiinflamatorios/farmacología , Proliferación Celular/efectos de los fármacos , Celulosa Oxidada/farmacología , Dexametasona/farmacología , Duramadre/efectos de los fármacos , Espuma de Fibrina/farmacología , Fibroblastos/efectos de los fármacos , Hemostáticos/farmacología , Humanos , Inmunohistoquímica , Cicatrización de Heridas/efectos de los fármacosRESUMEN
OBJECTIVE: To compare the classical neurosurgical technique with a new simplified technique for prenatal repair of a myelomeningocelelike defect in sheep. METHODS: A myelomeningocele-like defect (laminectomy and dural excision) was created in the lumbar region on day 90 of gestation in 9 pregnant sheep. Correction technique was randomized. In Group 1 the defect was corrected using the classic neurosurgical technique of three-layer suture (dura mater, muscle and skin closure) performed by a neurosurgeon. In Group 2, a fetal medicine specialist used a biosynthetic cellulose patch to protect the spinal cord and only the skin was sutured above it. Near term (day 132 of gestation) fetuses were sacrificed for pathological analysis. RESULTS: There were two miscarriages and one maternal death. In total, six cases were available for pathological analysis, three in each group. In Group 1, there were adherence of the spinal cord to the scar (meningo-neural adhesion) and spinal cord architecture loss with posterior funiculus destruction and no visualization of grey matter. In Group 2, we observed in all cases formation of a neo-dura mater, separating the nervous tissue from adjacent muscles, and preserving the posterior funiculus and grey matter. CONCLUSION: The new simplified technique was better than the classic neurosurgical technique. It preserved the nervous tissue and prevented the adherence of the spinal cord to the scar. This suggests the current technique used for the correction of spina bifida in humans may need to be reassessed.
OBJETIVO: Comparar a técnica neurocirúrgica clássica a uma nova técnica simplificada, para correção de mielomeningocele, em fetos de ovelhas. MÉTODOS: Em 9 fetos, foi criado um defeito semelhante à mielomeningocele (laminectomia e excisão de dura-máter) no 90º dia de gestação. O tipo de correção foi randomizado. No Grupo 1, o defeito foi corrigido usando a técnica neurocirúrgica clássica, com a sutura de três camadas (dura-máter, músculo e pele), realizada por um neurocirurgião. No Grupo 2, um especialista em Medicina Fetal utilizou a técnica simplificada, colocando um fragmento de celulose biossintética sobre a medula e suturando apenas da pele sobre a celulose. Próximo ao termo da gestação (132 dias), os fetos foram sacrificados para análise anatomopatológica. RESULTADOS: Ocorreram dois casos de aborto e uma morte materna, restando seis casos para avaliação - três em cada grupo. No Grupo 1, todos os casos mostraram aderência da medula à cicatriz (meningoadesão) e perda da arquitetura medular, com destruição do funículo posterior e perda da visualização da substância cinzenta. No Grupo 2, observou-se, em todos os casos, a formação de uma neodura-máter, separando o tecido nervoso do músculo adjacente, sendo que o funículo posterior e a substância cinzenta estavam preservados. CONCLUSÃO: A técnica simplificada foi superior à neurocirúrgica, com maior preservação da medula e evitando as aderências do tecido nervoso. Os presentes achados sugerem que a técnica utilizada atualmente na correção de mielomeningocele em fetos humanos deva ser reavaliada.
Asunto(s)
Animales , Femenino , Embarazo , Terapias Fetales , Fetoscopía/métodos , Feto/cirugía , Meningomielocele/cirugía , Procedimientos Neuroquirúrgicos/métodos , Modelos Animales de Enfermedad , Duramadre/citología , Duramadre/cirugía , Invenciones , Trabajo de Parto Prematuro , OvinosRESUMEN
OBJECTIVE: To compare the classical neurosurgical technique with a new simplified technique for prenatal repair of a myelomeningocele-like defect in sheep. METHODS: A myelomeningocele-like defect (laminectomy and dural excision) was created in the lumbar region on day 90 of gestation in 9 pregnant sheep. Correction technique was randomized. In Group 1 the defect was corrected using the classic neurosurgical technique of three-layer suture (dura mater, muscle and skin closure) performed by a neurosurgeon. In Group 2, a fetal medicine specialist used a biosynthetic cellulose patch to protect the spinal cord and only the skin was sutured above it. Near term (day 132 of gestation) fetuses were sacrificed for pathological analysis. RESULTS: There were two miscarriages and one maternal death. In total, six cases were available for pathological analysis, three in each group. In Group 1, there were adherence of the spinal cord to the scar (meningo-neural adhesion) and spinal cord architecture loss with posterior funiculus destruction and no visualization of grey matter. In Group 2, we observed in all cases formation of a neo-dura mater, separating the nervous tissue from adjacent muscles, and preserving the posterior funiculus and grey matter. CONCLUSION: The new simplified technique was better than the classic neurosurgical technique. It preserved the nervous tissue and prevented the adherence of the spinal cord to the scar. This suggests the current technique used for the correction of spina bifida in humans may need to be reassessed.