Monika Barczewska

MR monitoring of minimally invasive delivery of mesenchymal stem cells into the porcine intervertebral disc.

Purpose Bone marrow stem cell therapy is a new, attractive therapeutic approach for treatment of intervertebral disc (IVD) degeneration; however, leakage and backflow of transplanted cells into the structures surrounding the disc may lead to the formation of undesirable osteophytes. The purpose of this study was to develop a technique for minimally invasive and accurate delivery of stem cells. Methods Porcine mesenchymal stem cells (MSCs) were labeled with superparamagnetic iron oxide nanoparticles (SPIO, Molday ION rhodamine) and first injected into the explanted swine lumbar IVD, followed by ex vivo 3T MRI. After having determined sufficient sensitivity, IVD degeneration was then induced in swine (n=3) by laser-evaporation. 3 x 106 SPIO-labeled cells embedded within hydrogel were injected in 2 doses using a transcutaneous cannula and an epidural anesthesia catheter. T2-weighted MR images were obtained at 3T before and immediately after cell infusion. Two weeks after injection, histological examination was performed for detection of transplanted cells. Results MSCs were efficiently labeled with Molday ION rhodamine. Cells could be readily detected in the injected vertebral tissue explants as distinct hypointensities with sufficient sensitivity. MR monitoring indicated that the MSCs were successfully delivered into the IVD in vivo, which was confirmed by iron-positive Prussian Blue staining of the tissue within the IVD. Conclusion We have developed a technique for non-invasive monitoring of minimally invasive stem delivery into the IVD at 3T. By using a large animal model mimicking the anatomy of IVD in humans, the present results indicate that this procedure may be clinically feasible.

Proliferative Enteropathy (PE)—Induced Changes in the Calbindin-Immunoreactive (CB-IR) Neurons of Inferior Mesenteric Ganglion Supplying the Descending Colon in the Pig

A subpopulation of the pig inferior mesenteric ganglia (IMG) neurons projecting to the colon exhibit calbindin-like immunoreactivity. It is not known if there are any changes in the chemical coding patterns of these neurons during porcine proliferative enteropathy (PE). To answer this question, juvenile Large White Polish pigs with clinically diagnosed Lawsonia intracellularis infection (PE; n = 3) and a group of uninfected controls (C; n = 3) were compared. The retrograde tracer fast blue (FB) was injected into the descending colons of all animals and then tissue comprising IMGs from both groups was processed for double-labeling immunofluorescence with calbindin-D28k (CB) in combination with either tyrosine hydroxylase (TH), neuropeptide Y (NPY), somatostatin (SOM), vasoactive intestinal polypeptide (VIP), nitric oxide synthase, Leu-enkephalin, substance P, vesicular acetylcholine transporter, galanin, or pituitary adenylate cyclase-activating polypeptide. Immunohistochemistry revealed changes in the chemical coding pattern of calbindin-immunoreactive neurons in the inferior mesenteric ganglia of the pig. In control animals, FB/CB-positive neurons were immunoreactive to TH, NPY, SOM, and VIP. In the experimental group, TH-expressing neurons were unaffected, NPY-expressing neurons were increased, whereas the number of neurons immunoreactive to SOM or VIP was reduced. Changes in chemical coding of CB neurons during PE may play an important role in adaptation of these IMG cells under pathological conditions.

Therapeutic Potential of Olfactory Ensheathing Cells and Mesenchymal Stem Cells in Spinal Cord Injuries

Spinal cord injury (SCI) is a devastating neurological condition that affects individuals worldwide, significantly reducing quality of life, for both patients and their families. In recent years there has been a growing interest in cell therapy potential in the context of spinal cord injuries. The present review aims to discuss and compare the restorative approaches based on the current knowledge, available spinal cord restorative cell therapies, and use of selected cell types. However, treatment options for spinal cord injury are limited, but rehabilitation and experimental technologies have been found to help maintain or improve remaining nerve function in some cases. Mesenchymal stem cells as well as olfactory ensheathing cells seem to show therapeutic impact on damaged spinal cord and might be useful in neuroregeneration. Recent research in animal models and first human trials give patients with spinal cord injuries hope for recovery.

Inflammation-Induced Changes in the Chemical Coding Pattern of Colon-Projecting Neurons in the Inferior Mesenteric Ganglia of the Pig

The present study examines the chemical coding of the inferior mesenteric ganglia after chemically induced colitis in the pig animal model. In all animals (n = 6), a median laparotomy was performed under anesthesia, and the Fast Blue retrograde tracer was injected into the descending colon wall. In experimental animals (n = 3), the thick descending colon were injected with formalin solution to induce inflammation. The animals were euthanized and the inferior mesenteric ganglion was harvested and processed for double-labeling immunofluorescence for calbindin-D28k (CB) in combination with either tyrosine hydroxylase (TH), neuropeptide Y (NPY), somatostatin (SOM), vasoactive intestinal polypeptide (VIP), nitric oxide synthase (NOS), Leu-enkephalin (LENK), substance P (SP), vesicular acetylcholine transporter (VAChT), or galanin (GAL). Immunohistochemistry revealed significant changes in the chemical coding pattern of inferior mesenteric ganglion neurons. In control animals, Fast Blue-positive neurons were immunoreactive to TH, NPY, SOM, VIP, LENK, CB, and NOS. In the experimental group, TH, NPY, SOM, VIP, and LENK expressing neurons were reduced, whereas the number of neurons immunoreactive to CB, NOS, and GAL were increased. The increase of so-called neuroprotective neuropeptides suggests that the changes in the chemical coding of inferior mesenteric ganglion neurons reflect adaption under pathological conditions to promote their own survival.

Corrigendum to “Therapeutic Potential of Olfactory Ensheathing Cells and Mesenchymal Stem Cells in Spinal Cord Injuries”

[This corrects the article DOI: 10.1155/2017/3978595.].

Mesenchymal Stem Cell (MSC) Transplantation in Patients with Amyotrophic Lateral Sclerosis (ALS): Is there a âResponder Populationâ?

Objective: To analyze the safety and clinical effect of stem cell therapy in ALS. Methods: In phase I of the trial, ALS subjects have been intrathecally transplanted with autologous bone marrowderived mesenchymal stem cells (MSCs) using a surgical procedure. Results: We present the results of a 6-month interim analysis of the ongoing study. Intrathecal administration of MSCs into ALS patients was feasible and safe. We showed a clinical benefit evident for the entire group of patients (n=25). The mean rate of ALSFRS-R score change (decrease) pre-transplant was 1.76 ± 1.36 points/period whereas the mean post-transplant rate was 1.06 ± 0.9 points/period (p=0.014). The key finding of our study is that there appears to be a group of patients, whom we call “responders” whose reaction to the treatment was different from the reaction of other patients we call “nonresponders”. Conclusion: In our study the “responders” progressed faster prior to the treatment than “non-responders”. Hence, we hypothesize that the pre-treatment progression rate may play a role as a predictive factor and a criterion for selecting ALS patients for cell-based therapies.

The feasibility of the CD271+ and CD271– mesenchymal stromal cell enrichment toward nucleus pulposus-like cells

INTRODUCTION Factors promoting nerve cell ingrowth are considered responsible for chronic back pain resulting from the intervertebral disc degeneration (IDD). One of the recent exploratory IDD treatments is stem cell transplantation therapy. The CD271 (low-affinity nerve growth factor receptor) has been identified as a mark-er of the most homogeneous mesenchymal stem cell (MSC) subset. It is capable of promoting differentiation along adipogenic, osteogenic and chondrogenic lineages and producing significantly higher levels of cytokines as compared to the total population of plastic adherence-mesenchymal stem cells (PA-MSCs). We investigated the ability of CD271+ MSCs to differentiate into chondrocyte-like cells of the nucleus pulposus (NP) of intervertebral disc. We also examined CD271- MSCs, using PA-MSCs as a control cell population. MATERIAL AND METHODS Bone marrow derived PA-MSCs and its two subsets, CD271- MSCs and CD271+ MSCs, were seeded in collagen scaffolds. After two weeks of growth in NP-differentiation medium, RNA was isolated from cells-scaffold constructs and was analyzed by q-PCR for expression of NP markers. Glycosaminoglycans were analyzed biochemically directly in cells-scaffold constructs. RESULTS Expression of NP markers - extracellular matrix components such as aggrecan, collagen type II and glycosaminoglycans on both RNA and the protein levels - was significantly higher in CD271- MSCs compared to the CD271+ MSCs and PA-MSCs cell populations. CONCLUSIONS CD271- MSCs may be superior candidates for NP restorative treatment compared to CD271+ MSCs and PA-MSCs due to their ability of expressing NP-supporting extracellular matrix components at levels higher than the other two studied MSC subsets.

Methodology for Assessing the Degree of Degeneration of the Porcine Intervertebral Discs Based on Magnetic Resonance Imaging

Degeneration of intervertebral discs can lead to various health problems, among which low back pain is the most common. Studies concerning methods of dealing with that problem are being conducted and magnetic resonance imaging is one of the most popular methods of assessing their progress. Many of these studies are based on animal models. The paper links these two facts and so it contains the description of methodology that can be used for assessing the degree of degeneration of the porcine intervertebral discs based on scans obtained in MRI study. Proposed method uses Otsu method of automatic thresholding and next basic operators of morphological image processing. The outcome is binary mask giving direct information about locations of intervertebral discs in MRI scans. The obtained mask can be also used to separate locations of intervertebral discs in following MRI studies of the same animals, in cases where they are much harder to delineate due to their degeneration. The paper shows also exemplary results demonstrating usefulness of proposed methodology for the evaluation of discs treatment using mesenchymal stem cells.