Living organisms are extremely complex functional systems. At present, there are many in vivo models of spinal cord injury (SCI) that allow the modeling of any type of central nervous system (CNS) injury, however, with some disadvantages. The production of injury models can be a highly invasive and time-consuming process and requires high technical requirements, and costly financial issues should also be taken into account. Of course, a large number of animals have been used to obtain the relevant data of statistical significance. All of these aspects can be reduced by carrying out experiments in in vitro conditions. The primary advantage of in vitro method is that it simplifies the system under study. There are two major groups of in vitro model in use: cell culture and organotypic slice (OTS) culture. OTS is an intermediate system of the screening of in vitro cell culture and animal models and represents the in vitro system preserving the basic tissue architecture that able to closely mimic the cellular and physiological characteristics in vivo. In vitro models are the preferred methods for the study of acute or subacute pathophysiology after a trauma stimulus, enabling precise control on the extracellular environment, easy and repeatable access to the cells.

Despite strong efforts in the field, spinal cord trauma still belongs among the untreatable neurological conditions at present.Given the complexity of the nervous system, an effective therapy leading to complete recovery has still not been found. One of the potential tools for supporting tissue regeneration may be found in mesenchymal stem cells, which possess anti-inflammatory and trophic factor-producing properties. In the context of transplantations, application of degradable biomaterials which could form a supportive environment and scaffold to bridge the lesion area represents another attractive strategy. In the present study, through a combination of these two approaches we applied both alginate hydrogel biomaterial alone or allogenic transplants of MSCs isolated from bone marrow seeded in alginate biomaterial into injured rat spinal cord at three weeks after spinal cord compression performed at Th8-9 level. Following three-week survival, using immunohistochemistry we studied axonal growth (GAP-43 expression) and both microglia (Iba-1) and astrocyte (GFAP) reactions at the lesion site and in the segments below and above the lesion. To detect functional improvement, during whole survival period we performed behavioral analyses of locomotor abilities using a classical open field test (BBB score) and a Catwalk automated gait analyzing device (Noldus). We found that despite the absence of locomotor improvement, application of both alginate and MSCs caused significant increase in the number of GAP-43 positive axons.

Objectives In this study, a new approach was used with an in vitro model in which neural cells were exposed to conditioned media from the injured spinal cord (SCI-CM) mimicking a local inflammatory microenvironment . Subsequently, the neuroprotective effect of rat adipose tissue-derived msesenchymal stem cell-conditioned media (ATMSC-CM) was investigated through a cell-free based therapy, which was used to treat cortical neurons and astrocytes under inflammation. Methods Primary cell cultures isolated from postnatal day (P6) Wistar rat brain cortex were exposed to SCI-CM derived from the central lesion, rostral and caudal segments of injured spinal cord. After 48 h incubation, the SCI-CM was replaced and primary cultures were cultivated either in DMEM media alone or in ATMSC-CM for 72 h. The impact of ATMSC-CM on the viability of neurons and astrocytes was assessed using a CyQUANT® Direct Cell Proliferation Assay Kit as well as immunocytochemistry analysis. Results Immunocytochemical analysis revealed significant decrease in the number of MAP2 positive neurons exposed to SCI-CM compared to Control. Protection by ATMSC-CM was associated with increased survival of neurons compared to primary culture cultivated in DMEM media alone. The ATMSC-CM effect on astrocytes was more variable and without any significant impact. Conclusion The results demonstrate that SCI-CM mimicking inflammation can reduce cortical neuron survival, and subsequent exposure to ATMSC-CM can stabilize the neuronal population most likely via released neuroprotective and trophic factors. In addition, astrogliosis was not affected by ATMSC-CM.

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Ефективність лікування захворювань стовбуровими клітинами, найвірогідніше, пов'язана з їх паракринною активністю, ніж зі здатністю диференціюватися в різні типи клітин. У цьому дослідженні вперше було in vitro піддано нервові клітини впливу середовища, кондиційованого клітинами травмованого спинного мозку (ТСМ-КС), що моделювало місцеве запальне мікросередовище. На кортикальних нейронах та астроцитах оцінено при запаленні нейропротекторний ефект середовищ, кондиційованих замороженими-відтаяними мезенхімальними стовбуровими клітинами, отриманими з жирової тканини (ЖТМСК-КС).Первинні культури клітин, виділені з кори головного мозку 6-денних щурів лінії Wistar, культивували протягом 48 годин у ТСМ-КС, потім замінювали середовище на DMEM (нелікована група) або на ЖТМСК-КС (лікована група) і культивували подальші 72 години. Вплив ТМСК-КС на життєздатність нейронів і астроцитів оцінювали за допомогою тестів для проліферації клітин CyQUANT ® та імуноцитохімічного аналізу. Аналогічні процедури виконували з первинними культурами кори головного мозку, що піддавалися впливу середовища, кондиційованого тканиною неушкодженого спинного мозку (контроль).Імуноцитохімічний аналіз показав суттєве зменшення числа MAP2-позитивних нейронів після культивування у ТСМ-КС відносно контролю. Захисний ефект ЖТМСК-КС був пов'язаний зі збільшенням виживання нейронів у порівнянні з первинною культурою, культивованою лише в середовищах DMEM. Вплив ЖТМСК-КС на астроцити не мав значущого впливу.Встановлено, що ТСМ-КС через 48 годин зменшує кількість нейронів, але не астроцитів. Дія ЖТМСК-КС підвищувала виживання MAP2-позитивних нейронів. Популяція GFAP-позитивних астроцитів, присутня у первинній клітинній культурі, не змінювалася після впливу ТСМ-КС, проте після інкубування з ЖТМСК-КС незначно зменшувалася кількості астроцитів. Загальна відповідь астроцитів показала високу варіативність без значних змін. Можливо, що ЖТМСК-КС має нейропротекторний ефект за рахунок паракринних трофічних факторів, але наявна варіабельність впливу вимагає подальшого вивчення.Таким чином, моделювання запалення за допомогою ТСМ-КС може призвести до зменшення виживання кортикальних нейронів, а наступний вплив ЖТМСК-КС сприяє стабілізації популяції нейронів, вірогідно, через вивільнення нейропротекторних та трофічних факторів. Крім того, ЖТМСК-КС не впливала на астрогліоз.Робота була підтримана в рамках проекту VEGA 2/ 0145/16. The importance of mesenchymal stem cells in the treatment of CNS diseases, described in recent studies, is attributed to their paracrine activity rather than their ability to differentiate into different cell types. In this study we demonstrate a new approach with an in vitro model in which neural cells were exposed to conditioned media from the injured spinal cord (SCI-CM) mimicking a local inflammatory microenvironment. Subsequently, the neuroprotective effect of frozen rat adipose tissue-derived mesenchymal stem cell-conditioned media (ATMSC-CM) was investigated through a cell-free based therapy, which was used to treat cortical neurons and as...

Chemotherapy-induced peripheral neuropathy is one of the most frequent dose-limiting side effects, observed in patients receiving antineoplastic agents, persisting for up to two years after completing treatment, greatly affecting both the course of chemotherapy and patients' quality of life. Approximately 20 to 85% of patients treated with neurotoxic chemotherapy will develop peripheral neuropathy and there is considerable variability in its severity among patients. The main symptoms are numbness, paresthesia, and burning pain in a "glove and stocking" distribution. The prevalence of chemotherapy-induced peripheral neuropathy will likely increase as cancer survival rates continue to improve. Currently, there are only a few therapeutic options available for the prevention or successful therapy because the mechanisms of chemotherapy-induced peripheral neuropathy remain unclear. A better understanding of the risk factors and underlying mechanisms of chemotherapy-induced peripheral neuropathy is needed to develop effective preventive and therapeutic strategies.