Transfection of arginine decarboxylase gene increases the neuronal differentiation of neural progenitor cells

Growing evidence suggests that the clinical use of neural progenitor cells (NPCs) is hampered by heterogeneity, poor neuronal yield and low survival rate. Recently, we reported that retrovirus-delivered human arginine decarboxylase (hADC) genes improve cell survival against oxidative insult in murine NPCs in vitro. This study investigates whether the induced expression of hADC gene in mNPCs induces any significant change in the cell fate commitment. The evaluation of induced hADC gene function was assessed by knockdown of hADC gene using specific siRNA. The hADC gene delivery triggered higher expression of N-CAM, cell adhesion molecule and MAP-2, neuronal marker. However, the hADC gene knockdown showed downregulation of N-CAM and MAP-2 expression suggesting that hADC gene delivery favors cell fate commitment of mNPCs towards neuronal lineage. Neurite outgrowth was significantly longer in the hADC infected cells. The neurotrophic signal, BDNF aided in the neuronal commitment, differentiation, and maturation of hADC-mNPCs through PI3K and ERK1/2 activation. The induction of neuron-like differentiation is believed to be regulated by the expression of GSK-3β and Wnt/β-catenin signaling pathways. Our findings suggest that hADC gene delivery favors cell fate commitment of mNPCs towards neuronal lineage, bring new advances in the field of neurogenesis and cell therapy

Transfection of arginine decarboxylase gene increases the neuronal differentiation of neural progenitor cells

Growing evidence suggests that the clinical use of neural progenitor cells (NPCs) is hampered by heterogeneity, poor neuronal yield and low survival rate. Recently, we reported that retrovirus-delivered human arginine decarboxylase (hADC) genes improve cell survival against oxidative insult in murine NPCs in vitro. This study investigates whether the induced expression of hADC gene in mNPCs induces any significant change in the cell fate commitment. The evaluation of induced hADC gene function was assessed by knockdown of hADC gene using specific siRNA. The hADC gene delivery triggered higher expression of N-CAM, cell adhesion molecule and MAP-2, neuronal marker. However, the hADC gene knockdown showed downregulation of N-CAM and MAP-2 expression suggesting that hADC gene delivery favors cell fate commitment of mNPCs towards neuronal lineage. Neurite outgrowth was significantly longer in the hADC infected cells. The neurotrophic signal, BDNF aided in the neuronal commitment, differentiation, and maturation of hADC-mNPCs through PI3K and ERK1/2 activation. The induction of neuron-like differentiation is believed to be regulated by the expression of GSK-3β and Wnt/β-catenin signaling pathways. Our findings suggest that hADC gene delivery favors cell fate commitment of mNPCs towards neuronal lineage, bring new advances in the field of neurogenesis and cell therapy. © 2016 The Authors. Published by Elsevier B.V. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).2

Prevention of Corneal Myofibroblastic Differentiation In Vitro Using a Biomimetic ECM Hydrogel for Corneal Tissue Regeneration

Corneal scarring is one of the major causes of blindness, affecting millions worldwide. Despite recent advancements in surgical strategies, there is an unmet need for a clinically feasible material and methods to prevent scarring following corneal injury. In this study, we report the potential utility of a hydrogel derived from cadaveric animal corneas, using a decellularized corneal matrix hydrogel (abbreviated as dCMH), which is prepared by a simple method. This hydrogel is easily injectable, biocompatible, and has the ability to maintain good shape-retention properties at 37 °C, which make it suitable for in vivo applications. Furthermore, our gene expression studies and immunofluorescence studies indicate that dCMH maintains the morphology and function of keratocytes in vitro and prevents their transdifferentiation to myofibroblasts. From the above results, it is evident that dCMH maintains the keratocytes with the ability to regenerate the corneal defect without scar. We thus suggest a simple yet effective approach for corneal tissue decellularization and that dCMH can be a promising material for prophylaxis against blinding scar formation in an injured cornea

Video1_COVID-19 repellent cloth.MP4

In this research work, for the first time, we have developed and demonstrated a COVID-19 repellent coating on cotton cloth that not only repels the virus but also most of the human body fluids (superhemophobic). The coating was tested in the BSL3 lab. The controlled experiments revealed no significant increase in the log viral particles on coated fabric compared to the uncoated surface, evidence that the coated fabric resisted the SARS-CoV-2 inoculum. Further, the coated cloth exhibited excellent dust-free nature and stain resistance against body fluids (blood, urine, bovine serum, water, and saliva aerosol). It also shows sufficient robustness for repetitive usage. The fabrication process for the developed COVID-19 repellent cloth is simple and affordable and can be easily scaled up for mass production. Such coating could be applied on various surfaces, including daily clothes, masks, medical clothes, curtains, etc. The present finding could be a mammoth step towards controlling infection spread, including COVID-19.</p

The Multifaceted Effects of Agmatine on Functional Recovery after Spinal Cord Injury through Modulations of BMP-2/4/7 Expressions in Neurons and Glial Cells

<div><p>Presently, few treatments for spinal cord injury (SCI) are available and none have facilitated neural regeneration and/or significant functional improvement. Agmatine (Agm), a guanidinium compound formed from decarboxylation of L-arginine by arginine decarboxylase, is a neurotransmitter/neuromodulator and been reported to exert neuroprotective effects in central nervous system injury models including SCI. The purpose of this study was to demonstrate the multifaceted effects of Agm on functional recovery and remyelinating events following SCI. Compression SCI in mice was produced by placing a 15 g/mm² weight for 1 min at thoracic vertebra (Th) 9 segment. Mice that received an intraperitoneal (i.p.) injection of Agm (100 mg/kg/day) within 1 hour after SCI until 35 days showed improvement in locomotor recovery and bladder function. Emphasis was made on the analysis of remyelination events, neuronal cell preservation and ablation of glial scar area following SCI. Agm treatment significantly inhibited the demyelination events, neuronal loss and glial scar around the lesion site. In light of recent findings that expressions of bone morphogenetic proteins (BMPs) are modulated in the neuronal and glial cell population after SCI, we hypothesized whether Agm could modulate BMP- 2/4/7 expressions in neurons, astrocytes, oligodendrocytes and play key role in promoting the neuronal and glial cell survival in the injured spinal cord. The results from computer assisted stereological toolbox analysis (CAST) demonstrate that Agm treatment dramatically increased BMP- 2/7 expressions in neurons and oligodendrocytes. On the other hand, BMP- 4 expressions were significantly decreased in astrocytes and oligodendrocytes around the lesion site. Together, our results reveal that Agm treatment improved neurological and histological outcomes, induced oligodendrogenesis, protected neurons, and decreased glial scar formation through modulating the BMP- 2/4/7 expressions following SCI.</p></div

CAST analysis showed that agmatine treatment modulated BMP- 2/7 expressions in neurons & oligodendrocytes and BMP- 4 expressions in astrocytes & oligodendrocytes following SCI.

<p>(A) Stereological counting of BMP- 2⁺/MAP-2⁺and (B) BMP- 2⁺/Olig-2⁺ cells in the injured spinal cord (Th 8–Th 10 segments). The number of BMP- 2⁺/MAP-2⁺ and BMP- 2⁺/Olig-2⁺ cells were significantly increased in the Agm treated group (<i>n</i> = 5) compared with the EC group (<i>n</i> = 5) at 7 & 35 DPI and 7 & 14 DPI (C) BMP- 7⁺/MAP-2⁺ and (D) BMP- 7⁺/Olig-2⁺ cells in the Th 8–Th 10 segments of the injured spinal cord. The number of BMP-7⁺/MAP-2⁺ and BMP-7⁺/Olig-2⁺ cells were significantly increased in the Agm treated group (<i>n</i> = 5) compared with the EC group (<i>n</i> = 5) at 7 & 14 days and 7 days respectively following SCI. (E) BMP- 4⁺/GFAP⁺ and (F) BMP- 4⁺/Olig-2⁺ cells in Th 8–Th 10 segments of the injured spinal cord. The number of BMP- 4⁺/GFAP⁺ and BMP- 4⁺/Olig-2⁺ cells were decreased at all the time periods in the Agm treated group (<i>n</i> = 5) compared with the EC group (<i>n</i> = 5) and the decrease was significant at 7 & 14 days and 14 & 35 days respectively following SCI. †, <i>p</i><0.05 NC group vs EC group; #, <i>p</i><0.05 NC group vs Agm treated group; *, <i>p</i><0.05 EC group vs Agm treated group. Results represent mean ± S.E.M.</p

Agmatine treatment enhanced functional outcome following compression SCI.

<p>The BMS score was used to evaluate hindlimb locomotor function. The BMS scores in the Agm treated mice (Agm treated group, <i>n</i> = 30) were significantly higher than the saline treated mice (EC group, <i>n</i> = 30) at 1, 3, 7, 10, 14, 21, 28 and 35 DPI. The Bladder residual urine volumes were measured in Agm treated group (<i>n</i> = 10) and in the EC group (<i>n</i> = 10). The graph showed that the amount of residual urine was significantly decreased in the Agm treated group compared with the EC group until 14 DPI. *, <i>p</i><0.05 EC group vs Agm treated group. Results represent mean ± S.E.M.</p

Confocal microscopic images of BMP- 2/4/7 expressions in neurons, oligodendrocytes and astrocytes after agmatine treatment following SCI.

<p>(A) Dual immunofluorescence was done to localize the BMP- 2 expression in neurons (MAP-2) and oligodendrocytes (Olig-2) after SCI. The BMP- 2⁺/MAP- 2⁺& BMP- 2⁺/Olig-2⁺ cells around the lesion site were higher in the Agm treated group (<i>n</i> = 4) compared with the EC group (<i>n</i> = 4) at7 DPI. Scale bars: 50 µm. (B) The immunostaining of BMP- 7 (red) with MAP-2 (green) showed increased number of BMP- 7⁺/MAP-2⁺ cells in the Agm treated group (<i>n</i> = 5) when compared with the EC group (<i>n</i> = 5) at 7 days after SCI. The immunostaining of BMP- 7 with Olig-2 showed an increased number of BMP- 7⁺/Olig-2⁺ cells in the Agm treated group (<i>n</i> = 5) compared to EC group (<i>n</i> = 5) at 14 DPI. Scale bars: 10 µm. (C) The immuno co-localization of BMP- 4⁺ cells in astrocytes (GFAP) showed reduced number of BMP- 4⁺/GFAP⁺ cellsaround the lesion site in the Agm treated group (<i>n</i> = 4) compared with the EC group (<i>n</i> = 4) at7 DPI. Whereas, BMP- 4⁺/Olig-2⁺ cells were higher in the Agm treated group compared with EC at 35 DPI. The scale bars: 10 µm.</p