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Improving Motor Neuron-Like Cell Differentiation of Henscs by the Combination of Epothilone B Loaded Pcl Microspheres in Optimized 3D Collagen Hydrogel Publisher Pubmed



Mahmoodi N1 ; Ai J2 ; Hassannejad Z3 ; Ebrahimibarough S2 ; Hasanzadeh E4 ; Nekounam H5 ; Vaccaro AR6 ; Rahimimovaghar V1
Authors
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Authors Affiliations
  1. 1. Sina Trauma and Surgery Research Center, Tehran University of Medical Sciences, Tehran, Iran
  2. 2. Department of Tissue Engineering and Applied Cell Sciences, School of Advanced Technologies in Medicine, Tehran University of Medical Sciences, Tehran, Iran
  3. 3. Pediatric Urology and Regenerative Medicine Research Center, Tissue, Cell and Gene Research Institute, Tehran University of Medical Sciences, Tehran, Iran
  4. 4. Immunogenetics Research Center, Department of Tissue Engineering and Applied Cell Sciences, School of Advanced Technologies in Medicine, Mazandaran University of Medical Sciences, Sari, Iran
  5. 5. Department of Medical Nanotechnology, School of Advanced Technologies in Medicine, Tehran University of Medical Sciences, Tehran, Iran
  6. 6. Department of Orthopedic Surgery, Rothman Institute, Thomas Jefferson University, Philadelphia, PA, United States

Source: Scientific Reports Published:2021


Abstract

Spinal cord regeneration is limited due to various obstacles and complex pathophysiological events after injury. Combination therapy is one approach that recently garnered attention for spinal cord injury (SCI) recovery. A composite of three-dimensional (3D) collagen hydrogel containing epothilone B (EpoB)-loaded polycaprolactone (PCL) microspheres (2.5 ng/mg, 10 ng/mg, and 40 ng/mg EpoB/PCL) were fabricated and optimized to improve motor neuron (MN) differentiation efficacy of human endometrial stem cells (hEnSCs). The microspheres were characterized using liquid chromatography-mass/mass spectrometry (LC-mas/mas) to assess the drug release and scanning electron microscope (SEM) for morphological assessment. hEnSCs were isolated, then characterized by flow cytometry, and seeded on the optimized 3D composite. Based on cell morphology and proliferation, cross-linked collagen hydrogels with and without 2.5 ng/mg EpoB loaded PCL microspheres were selected as the optimized formulations to compare the effect of EpoB release on MN differentiation. After differentiation, the expression of MN markers was estimated by real-time PCR and immunofluorescence (IF). The collagen hydrogel containing the EpoB group had the highest HB9 and ISL-1 expression and the longest neurite elongation. Providing a 3D permissive environment with EpoB, significantly improves MN-like cell differentiation and maturation of hEnSCs and is a promising approach to replace lost neurons after SCI. © 2021, The Author(s).
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