MicroRNA‐7 promotes neural differentiation of trabecular meshwork mesenchymal stem cell on nanofibrous scaffold

Abstract

The purpose of this study was to investigate miR‐7 overexpression effects on neural differentiation of mesenchymal stem cells (MSCs) on both two‐dimensional (2D) and three‐dimensional (3D) culture systems. We upregulated miR‐7 through lentiviral vector in trabecular meshwork MSCs (TMMSCs) and polymers of poly l‐lactic acid/polycaprolactone fibrous scaffold were fabricated by electrospinning and characterized using scanning electron microscopy (SEM) and Fourier transform infrared (FTIR). Neural markers expression was evaluated through quantitative‐polymerase chain reaction (q‐PCR) and immunostaining. The results showed that the high percentage of cell transduction (84.9%) and miR‐7 expression (folds: 677.93 and 556.4) was detected in TMMSCs‐miR‐7(+). SEM and FTIR established the fabrication of the hybrid scaffold. q‐PCR analysis showed that on days 14 and 21 of transduction, the expression level of Nestin and glial fibrillary acidic protein (GFAP) genes were significantly higher in the scaffold (3D) compared with tissue culture polystyrene (2D) culture. The expression of microtubule‐associated protein‐2 (MAP‐2) and GFAP genes in TMMSCs‐miR‐7(+) cells were significantly higher than those miR‐7(−) cells after 21 days of cell culture. Also, MAP‐2 and Nestin proteins were detected in TMMSCs‐miR‐7(+) cells. Our results demonstrate that miR‐7 is involved in neural differentiation of TMMSCs and scaffold can improve differentiate into glial and neural progenitor cells. These findings provided some information for future use of microRNAs and scaffold in tissue engineering and cell therapy for neurological diseases.

miR‑217‑5p regulates myogenesis in skeletal muscle stem cells by targeting FGFR2

“…Previous studies have reported that mirna-217-5p regulates pluripotent stem cell proliferation and differentiation (17) and is involved in metabolic processes in various cells, such as endothelial (18) and colorectal cancer (19) cells. These findings provide insights into the application of miRNAs in regenerative and cell therapy for muscle diseases (14). However, a limited number of studies have explored the potential roles of mir-217-5p in SkMScs.…”

“…accumulating evidence suggests that a number of micrornas (mirnas), such as mir-99a-5p (10), mir-9-5p (11), mir-208b (12), serve important roles in skeletal muscle myogenesis by regulating gene expression, and that their abnormal expression is associated with a number of muscle diseases including, muscle atrophy and ischemic injury (6,13,14). regarding the molecular mechanism of mirnas in myogenesis, studies have reported a role for these molecules in the differentiation of SkMScs (13,15).…”

“…a recent study has reported that mir-483-3p is involved in the osteogenic differentiation of bone marrow-derived mesenchymal stem cells (BMScs) by targeting STaT1 and may serve as a potential therapeutic target for bone loss due to aging (16). in addition, studies have revealed that mir-7 regulates the neural differentiation of trabecular meshwork mesenchymal stem cells (TMMScs), and that the poly (l-lactate) (Plla)/poly (e-caprolactone) (Pcl) scaffold, termed a three-dimensional (3d) culture system, can promote their differentiation towards glial and neural progenitor cells (14). Previous studies have reported that mirna-217-5p regulates pluripotent stem cell proliferation and differentiation (17) and is involved in metabolic processes in various cells, such as endothelial (18) and colorectal cancer (19) cells.…”

Fundamental Biomaterial Considerations in the Development of a 3D Model Representative of Primary Open Angle Glaucoma

“…The effect of nanoscale to submicrometre topographic features on influencing TM cell alignment, migration and gene expression has been investigated in several studies [ 53 , 95 , 96 , 97 ]. Similar observations were noted: a loss of in vivo characteristic cell properties when cultured on tissue culture plastic or planar surfaces, such as myocillin [ 98 ], and TM/stem cell expression [ 53 ] and stabilisation of these necessary in vivo markers in the presence of randomised nanopatterns.…”

“…It was illustrated that presence of these specific patterns had retained myocillin and verscian protein expression in comparison to TM cultures grown on tissue culture plastic or aligned surfaces [ 95 , 96 ]. This has led other studies to look deeper into the manipulation of TM cells through topographical features, with electrospun, non-aligned, nano-fibrous scaffolds containing poly L-lactic acid/polycaprolactone improving the differentiation of TM cells into glial and neural progenitor cells [ 97 ]. In addition to the embedding of TM cells into Matrigel ® , a BM mixture that is rich in nanotopographical features [ 101 ] was deemed favourable for the expansion and manipulation of human TM cells into adipocytes, chondrocytes and endothelial cells [ 52 , 53 ].…”