Studying Diabetic Retinopathy?

Check out How Neuromic's Tools Are Used

The authors used four of Neuromics to complete this study-

Sulodexide reduces glucose induced senescence in human retinal endothelial cells A. Gericke, K. Suminska-Jasińska & A. Bręborowicz Scientific Reports volume 11, Article number: 11532 (2021) Cite this article.

Material and Methods

Experiments were performed on HREC (#HEC09, Neuromics, Edina, MN, USA) in in vitro culture. Cells were seeded in 75 cm² culture flasks coated with AlphaBioCoat Solution (#AC00, Neuromics, Edina, MN, USA) and were grown in Endo Growth Medium (EKG001, Neuromics, Edina, MN, USA) supplemented with fetal bovine serum 0.5% until they formed monolayers. Then the cells were harvested with cell detachment solution (#ADF001, Neuromics, Edina, MN, USA) and seeded in quadruplicates into 25 cm² coated, as described above, culture flasks at a density of 1.5 × 10⁵ cells/flask.

We conclude that Sulodexide may have a beneficial effect in cases of diabetic retinopathy. It slows down hyperglycemia-dependent senescence of endothelial cells, which translates into the lower angiogenic and inflammatory impact of these cells. An important observation was that Sulodexide also has effective antiangiogenic and anti-inflammatory effects in the senescent HREC. That means that Sulodexide may be effective in retinal endothelial cells, which are already senescent. Further studies are required to explain the potential effect of Sulodexide on the endothelial glycocalyx structure and permeability of the retinal endothelial layer composed of the senescent cells to molecules with various size and electrical charge.

We are pleased to see our solutions used in this important  study. It could lead to drugs that slow diabetic retinopathy.

Pain Receptors and Diabetic Neuropathy

We Got the Markers
Our Markers for Pain Research are widely used and frequently published. They are often referenced in publications on Diabetic nerve pain.

Here are some of my past blog postings:

• Cytokines and Neuropathic Pain
• Epidermal Nerve Fibers in Neuropathic Pain Model
• Pain Research and Gene Expression Analysis

Here's a model of the actual nerve receptors behind both intractable pain and loss of sensation.

A: Simplified model of nociception under normal conditions. Free nerve endings transduce a painful stimulus into a neural signal, which propagates to DRG centrally and eventually synapses on a nociceptive neuron within the DH of the spinal cord.B: Proposed model of nociception under conditions of diabetic hyperalgesia and allodynia. A pain signal augmented by upregulated pronociceptive ion channels in sensory neurons is carried toward the DH, where it is further augmented by a hypoactive GABAergic system and subsequently diminished inhibition from an inhibitory interneuron. NMDAR, N-Methyl-D-aspartate receptors. https://doi.org/10.2337/dbi15-0006

We will continue to post findings on the root causes and potential treatment for Neuropathy.

New 3-D Eye Model

More in-vivo like Model
We see our world in 3-D. Diseases of the eye compromise this ability.

Neuromics' is pleased to announce that we have a 3-D model aimed at accelerating drug discovery for these diseases. Sight is a terrible thing to lose and the faster new drugs can be discovered, fewer people will have to suffer the loss of sight.

Our 3D Human Retinal Microvascular Angiogenesis model is constructed using GFP‐Tagged human Retinal Microvascular Endothelial cells. They are co-cultured with RFP-Tagged human supporting cells. GFP positive human retinal capillary-like tubule formation can be monitored in real time under fluorescence microscope throughout the whole process of the experiment.

Microvascular Endothelial Cells

Tested, Characterized and Research Ready
Our Microvascular Endothelial Cells continue to work and work in the hands of our customers.

Check out these pubs:
1. Odunayo O. Mugisho, Colin R. Green, Jie Zhang, Nicolette Binz, Monica L. Acosta, Elizabeth Rakoczy and Ilva D. Rupentha. (2017). Immunohistochemical Characterization of Connexin43 Expression in a Mouse Model of Diabetic Retinopathy and in Human Donor Retinas. Int. J. Mol. Sci. doi: 10.3390/ijms18122567
 2. Michael Anthony Ruiz, Biao Feng, and Subrata Chakrabarti. (2015). Polycomb Repressive Complex 2 Regulates MiR-200b in Retinal Endothelial Cells: Potential Relevance in Diabetic Retinopathy. PLoS One.10(4): e0123987. doi: 10.1371/journal.pone.0123987.

In these, our cells are used as "healthy controls" to study Diabetic Retinopathy.

Figure: Connexin43 (green) and GFAP (red) expression in normal and human DR donor retinas in regions of extensive vascular damage. Large cells (white arrows, left column) represent non-specific auto-fluorescent amacrine cells. Connexin43 expression was markedly higher in the GCL of DR donor tissues compared to age-matched controls, and was strongly expressed throughout all retinal layers. GFAP labeling was also markedly higher in DR compared to normal donor eyes representing hyper-reactive Müller cells. Connexin43 expression was increased in regions identified as blood vessels and correlated with increased GFAP labeling at these sites, indicating glial cell activation (white circle). GCL = ganglion cell layer; IPL = inner plexiform layer; INL = inner nuclear layer; OPL = outer plexiform layer; ONL = outer nuclear layer. Scale bar: 200 µm

We stand ready to serve you. Pete Shuster, CEO and Owner, pshuster@neuromics.com

Ischemic Conditioning Prevents Retinopathy

What is Ischemic Conditioning?
I must confess that I had little knowledge of Ischemic Conditioning and its therapeutic potential before accessing this publication by my friend +Laura A. Pasquini and her team at University of Buenos Aires (users of Neuromics' Neuronal-Glial Markers and Neurotrophins Antibodies.

In the conditioning or pre-conditioning process, blood supply to an organ or a tissue is impaired for a short time (usually less than five minutes) then restored so that blood flow is resumed, and the process repeated two or more times, the cells downstream of the tissue or organ are robustly protected from a final ischemic insult when the blood supply is cut off entirely and permanently.

Here the authors used pressure pulses to induce retinal ischemia. Their results suggest that early vision loss in diabetes could be abated by ischemic conditioning which preserved axonal function and structure: Diego C. Fernandez, Laura A. Pasquini, Damián Dorfman, Hernán J. Aldana Marcos, Ruth E. Rosenstein. Ischemic Conditioning Protects from Axoglial Alterations of the Optic Pathway Induced by Experimental Diabetes in Rats. Research Article | published 20 Dec 2012 | PLOS ONE.

They used our PDGFR-α and an O1 marker to compare conditioned, diabetic with unconditioned, diabetic and controls rats to determine protection on ONs of the eye.

Immature OL (O1+ cells) and OL precursor (PDGFR-α+ cells) were evaluated by immunostaining in transverse ON sections. In the diabetic ON from eyes that received a sham treatment, a significant increase in O1(+) and PDGFR-α (+) area was observed, with the presence of disorganized and hypertrophic cells. In the right panel, the area occupied by glial cells (measured as total optical density (OD)) is shown. Ischemic conditioning significantly prevented these alterations and a clear decrease in O1- and PDGFR-α-immunoreactivity, with cells aligned parallel to axon bundles were found. Data are mean ± SEM (n = 6 nerves/group).

Results suggest that early vision loss in diabetes could be abated by ischemic conditioning which preserved distal axonal function and structure before the neuronal soma loss. Moreover, the present results add new potentialities to the therapeutic effects of ischemic tolerance, which is axon protection. Thus, ischemic tolerance could have promise for application in other neurodegenerative axonal diseases. I will keep you updated on progress.

Early Diagnosis of Diabetic Retinopathy

The earlier the diagnosis the better the outcome. This is especially true with autoimmune diseases like Diabetic Retinopathy (DR). DR is the leading cause of blindness among persons of working age in the industrialized world. Here I feature a publication that shows axoglial alterations at the distal portion of the optic nerve could be the first structural change in the diabetic visual pathway. This could prove good news for discovering better therapies thus preventing blindness: Diego C. Fernandez, Laura A. Pasquini, Damián Dorfman, Hernán J. Aldana Marcos, Ruth E. Rosenstein. Early Distal Axonopathy of the Visual Pathway in Experimental Diabetes. doi:10.1016/j.ajpath.2011.09.018

Oligodendrocytes are responsible for insulating axons. Disruptions in the formation of oligodendrocytes could initiate the domino effect that leads to decreasing and eventual total loss of vision. The authors, for example, discovered that in diabetic rats, oligodendrocyte lineage (OL) cells showed hypertrophic somas and a high number of processes.

Images/Data: OL linage evaluation. Immature OL (O1+ cells) and OL precursor (PDGFR-α+ cells) were evaluated by immunostaining and measured as optical density (OD) per section. In the distal ON from animals that were diabetic for 6 weeks, significantly increased O1 and PDGFR-α immunostaining was observed, with the presence of disorganized and hypertrophic cells. Data are mean ± SEM (n = 5 animals per group); *P < 0.01 versus age-matched controls, by Student′s t-test. Scale bar = 50 μm.

At the ultrastructural level, alterations and loss of larger axons were observed in the distal ON from animals that were diabetic for 6 weeks. In these fibers, myelin was highly disorganized, and frequent lamellar membranous bodies were observed.

I will track new develops in this research and post relevant results here.

Diabetic retinopathy blindness-root causes

Diabetic retinopathy is a leading cause of acquired blindness. This publication from our friends at University of Buenos Aires touches on potential root causes: Diego C. Fernandez, Laura A. Pasquini, Damián Dorfman, Hernán J. Aldana Marcos, Ruth E. Rosenstein. Early Distal Axonopathy of the Visual Pathway in Experimental Diabetes. doi:10.1016/j.ajpath.2011.09.018
" In animals that had been diabetic for 6 weeks, a large increase in astrocyte reactivity occurred in the distal (but not the intraorbital) portion, which coincided with significant axon loss. Moreover, profound myelin alterations and altered morphologic features of oligodendrocyte lineage were observed at the distal (but not the proximal) optic nerve portion. The present results suggest that axoglial alterations at the distal portion of the optic nerve could be the first structural change in the diabetic visual pathway."
The authors used our PDGFR Alpha/CD140A Marker to Study the change in Oligodendrocyte Lineage precursor cells. Expression of the protein was increased in these cells with the presence of disorganized and hypertrophic cells. This could disrupt formation of myelin resulting the pathological alteration at the distal portion.