James Bardill1, Ahmed Gilani2, Melissa R Laughter3, David Mirsky4, Brent O'Neill5, Daewon Park6, Ahmed I Marwan7. 1. Department of Bioengineering, University of Colorado Denver Anschutz Medical Campus, Aurora, Colorado 80045; Division of Pediatric Surgery, Department of Surgery, University of Colorado Denver Anschutz Medical Campus, Aurora, Colorado 80045. Electronic address: james.bardill@cuanschutz.edu. 2. Department of Pathology, University of Colorado Denver Anschutz Medical Campus, Aurora, Colorado 80045. Electronic address: ahmed.gilani@childrenscolorado.org. 3. Department of Bioengineering, University of Colorado Denver Anschutz Medical Campus, Aurora, Colorado 80045. Electronic address: melissa.laughter@cuanschutz.edu. 4. Department of Radiology, University of Colorado Denver Anschutz Medical Campus, Aurora, Colorado 80045. Electronic address: david.mirsky@cuanschutz.edu. 5. Department of Neurosurgery, University of Colorado Denver Anschutz Medical Campus, Aurora, Colorado 80045. Electronic address: brent.o'neill@childrenscolorado.org. 6. Department of Bioengineering, University of Colorado Denver Anschutz Medical Campus, Aurora, Colorado 80045. Electronic address: daewon.park@cuanschutz.edu. 7. Department of Bioengineering, University of Colorado Denver Anschutz Medical Campus, Aurora, Colorado 80045. Electronic address: rony.marwan@gmail.com.
Abstract
BACKGROUND: Prenatal surgical closure of Myelomeningocele (MMC) is considered part of the current age armamentarium. Clinical data has demonstrated the need for innovative patches to maximize the benefits and decrease the risks of this approach. Our team has developed a minimally invasive reverse thermal gel (RTG) patch with cellular scaffolding properties. Here, we demonstrate the initial gross and microscopic histological effects of this RTG patch in the fetal ovine model of MMC. MATERIALS AND METHODS: A fetal ovine MMC defect was created at 68-75 days gestation, RTG patch application or untreated at 100-103 days, and harvest at 135-140 days. The RTG was applied to the defect and secured in place with an overlay sealant. Defect areas underwent gross and microscopic analysis for inflammation and skin development. Brains were analyzed for hindbrain herniation and hydrocephalus. RESULTS: The untreated fetus (n = 1) demonstrated an open defect lacking tissue coverage, evidence of spinal cord injury, increased caspase-3, Iba1 and GFAP in spinal cord tissues, and hindbrain herniation and ventricular dilation. RTG treated fetuses (n = 3) demonstrated defect healing with well-organized dermal and epidermal layers throughout the entire healed tissue area overlaying the defect with minimal inflammation, reduced caspase-3, Iba1 and GFAP in spinal cord tissues, and no hindbrain herniation or ventricular dilation. CONCLUSION: An RTG patch applied to MMC defects in fetal sheep promoted skin coverage over the defect, was associated with minimal inflammation of the spinal cord tissues and prevented brain abnormalities. The present findings provide exciting results for future comprehensive radiological, functional, and mechanistic evaluation of the RTG.
BACKGROUND: Prenatal surgical closure of Myelomeningocele (MMC) is considered part of the current age armamentarium. Clinical data has demonstrated the need for innovative patches to maximize the benefits and decrease the risks of this approach. Our team has developed a minimally invasive reverse thermal gel (RTG) patch with cellular scaffolding properties. Here, we demonstrate the initial gross and microscopic histological effects of this RTG patch in the fetal ovine model of MMC. MATERIALS AND METHODS: A fetal ovine MMC defect was created at 68-75 days gestation, RTG patch application or untreated at 100-103 days, and harvest at 135-140 days. The RTG was applied to the defect and secured in place with an overlay sealant. Defect areas underwent gross and microscopic analysis for inflammation and skin development. Brains were analyzed for hindbrain herniation and hydrocephalus. RESULTS: The untreated fetus (n = 1) demonstrated an open defect lacking tissue coverage, evidence of spinal cord injury, increased caspase-3, Iba1 and GFAP in spinal cord tissues, and hindbrain herniation and ventricular dilation. RTG treated fetuses (n = 3) demonstrated defect healing with well-organized dermal and epidermal layers throughout the entire healed tissue area overlaying the defect with minimal inflammation, reduced caspase-3, Iba1 and GFAP in spinal cord tissues, and no hindbrain herniation or ventricular dilation. CONCLUSION: An RTG patch applied to MMC defects in fetal sheep promoted skin coverage over the defect, was associated with minimal inflammation of the spinal cord tissues and prevented brain abnormalities. The present findings provide exciting results for future comprehensive radiological, functional, and mechanistic evaluation of the RTG.