Treatment of knee cartilage by cultured stem cells and three dimensional scaffold: a phase I/IIa clinical trial.

PURPOSE: Damage of the knee cartilage is a common condition manifesting itself mainly by pain and/or swelling that may substantially reduce the quality of life while ultimately leading to osteoarthritis in affected patients. Here, we aimed to evaluate the safety and efficacy of cultured autologous bone marrow mesenchymal stem cells (BM-MSCs) attached to the 3D Chondrotissue® scaffold by autologous blood plasma coagulation (BiCure® ortho MSCp) in the treatment of knee cartilage defects. METHODS: The primary endpoint of this phase I/IIa clinical trial was to evaluate the safety of the treatment. The secondary objective was to determine the short-to-medium-term therapeutic outcomes by standardized scoring questionnaires including Lysholm Knee Scoring Scale (Lysholm score), Knee Injury and Osteoarthritis Outcome Score (KOOS), and pain Visual Analogue Scale (VAS) systems and imaging (X-ray and magnetic resonance imaging, MRI). A total of six patients were included and followed for 12 months after the surgery. RESULTS: BiCure® ortho MSCp was well tolerated with no adverse events associated with the investigational medicinal product. Significant improvements were observed in Lysholm scores and KOOS while X-ray showed no deterioration of the arthritis and MRI revealed a persistent filling of the chondral defects by the implant. CONCLUSION: Overall, our data demonstrate the safety of the tested investigational medicinal product. The function of the treated knee improved within one year after surgery in all enrolled patients. TRIAL REGISTRATION NUMBER AND DATE OF REGISTRATION: EudraCT No.: 2018-004,067-31; October 18 2018.

Small-scale regional engineering geological study of the Czech Republic evaluating the relationship between slope gradients and engineering geological zones.

The aim of the small-scale regional engineering geological study of the Czech Republic was to evaluate the relationship between slope gradient and engineering geological zones. The research motivation was to determine the average slope gradient, 25%, 50% (median) and 75% quantiles related to the different engineering geological zones. This scientific information is critical from the perspectives of engineering geology, geotechnical engineering, and foundation engineering because an increasing slope gradient evokes the need to create a cut respectively foundation excavation or another excavation in the geological structure, which increases the probability of occurrence of the problem in terms of differential settlement and bearing capacity of the structures. The research was carried out in the territory of the Czech Republic in 8 Quaternary zones with soil foundation ground and 10 pre-Quaternary zones with rocks and semi-rocks and their eluvia. A significant difference in the statistical characteristics of slope gradients was found in the group of Quaternary engineering geological zones (evaluated group I) compared to the group of pre-Quaternary zones (evaluated group II). The value range of the average slope gradient was 1.65° (16.9%) to 5.89° (60.3%) for the Quaternary engineering geological zones (soil foundation ground), representing 43.4% difference. Whereas for the over-quaternary engineering geological zones (rocks, semi-rocks and their eluvia), the difference was much higher, 3.59° (36.8%) to 9.76° (100%-value determined as a referential because it was the maximum), which is also reflected in a more significant percentage difference of 63.2%.

A comparative study of workability classes using seven case studies of engineering-geological investigations of sewer systems in Northern Moravia.

While the main focus of numerous engineering-geological investigations is to determine load-bearing capacity and settlement in engineering structures, this article aims to point at the specificity of sewer system engineering-geological investigations, which focuses on workability of soils and rocks. The study deals with workability class assessment of seven different sewer system localities. The significance of this research lies in the mutual comparison of workability class assessment of these seven localities in terms of two different workability standards. Each of the standards represents an independent model of assessment and classification of workability. The first standard (CSN 73 1001) classifies soils and rocks into seven workability classes, while the second (EN ISO 14688) comprises only three workability classes. Each of the approaches has its advantages and disadvantages. In comparison to the first one, the second standard permits faster and easier classification of rocks, but may be less fair to investors or developers when considering the real engineering-geological conditions and costs of implementation. Rocks were newly classified into three (easy, medium and difficult) categories of earthwork realization difficulty. In the study, 33 layers were classified in the category of easy realization of earthworks, 8 layers in the category of a medium degree of earthwork realization difficulty, and two layers in the category of a difficult realization of earthworks.

Co-transplantation of olfactory ensheathing glia and mesenchymal stromal cells does not have synergistic effects after spinal cord injury in the rat.

BACKGROUND AIMS: Olfactory ensheathing glia (OEG) and mesenchymal stromal cells (MSC) are suitable candidates for transplantation therapy of spinal cord injury (SCI). Both facilitate functional improvement after SCI by producing trophic factors and cytokines. In this study, the co-transplantation of both types of cells was studied to clarify their additive and/ or synergistic effects on SCI. METHODS: A balloon-induced compression lesion was used to produce SCI in rats. OEG, MSC or both OEG and MSC (3 x 10(5) cells of each cell type) were implanted by intraspinal injection 1 week after SCI. The effect of transplantation was assessed using behavioral, electrophysiologic and histologic methods. RESULTS: Hindlimb function was examined with Basso, Beattie and Bresnahan (BBB) and Plantar tests. Improvement was found in all three groups of transplanted rats with different time-courses, but there was no significant difference among the groups at the end of the experiment. Motor-evoked potentials after SCI decreased in amplitude from 7 mV to 10 microV. Linear regression analysis showed a modest recovery in amplitude following transplantation, but no change in the control rats. Histologic findings showed that the white and gray matter were significantly spared by transplantation after SCI. CONCLUSIONS: Functional improvement was achieved with transplantation of OEG and/or MSC, but the co-transplantation of OEG and MSC did not show synergistic effects. The poor migration of OEG and MSC might prevent their concerted action. Pre-treatment with a Rho antagonist and a combination of intraspinal and intravenous injection of the cells might be beneficial for SCI therapy.

Properties and growth of human bone marrow mesenchymal stromal cells cultivated in different media.

BACKGROUND AIMS: Human mesenchymal stromal cells (hMSC) are a promising tool for future clinical application, but their use requires rapid cell expansion in media suitable for clinical use. Therefore, we tested the influence of several culture media on colony formation, population doubling (PD) time, cell cycle and surface marker expression. METHODS: hMSC isolated from human bone marrow (BM) obtained from healthy donors were seeded and expanded in different culture media: alpha-minimum essential medium (MEM) supplemented with 2.5%, 5%, 10% or 20% fetal bovine serum (FBS), 5% or 10% human cord blood serum (hCBS), 5% or 10% human blood serum from AB adult donors (hABS), or mesenchymal stem cell growth medium (MSCGM). The number, diameter and total area of the colonies formed and PD time were determined, and the cell cycle and 16 surface markers were analyzed. RESULTS: Colony-forming efficiency was best in alpha-MEM/hCBS and alpha-MEM/hABS, good in MSCGM and worst in alpha-MEM/FBS. The shortest PD time was achieved in media enriched with human sera or MSCGM, while the time was increased in alpha-MEM/FBS. The largest proliferating fraction was seen in MSCGM followed by media enriched with human sera; the fraction was smallest in alpha-MEM/FBS. Staining for CD34, CD45, CD235a and CD271 was negative, while that for CD29, CD44, CD73, CD90, CD105 and human leukocyte antigen (HLA)-A, -B, -C was positive in all media tested. Media with human serum did not adversely affect the differentiation potential of hMSC, and differentiation into osteoblasts was enhanced. CONCLUSIONS: The choice of serum influences hMSC expansion and cell properties; alpha-MEM supplemented with hABS seems to be a promising candidate for clinical use.

Metabolic changes in the rat brain after a photochemical lesion treated by stem cell transplantation assessed by 1H MRS.

OBJECT: Metabolite changes in an experimental lesion in the rat cortex and in the contralateral hemisphere after the intravenous administration of mesenchymal stem cells (MSCs) were assessed by proton MR spectroscopy to verify the impact of the cell treatment on the brain tissue. MATERIALS AND METHODS: Wistar rats with a photochemical cortical lesion and transplanted MSCs or sham transplanted rats were examined. Proton spectra were obtained from the lesion and from the contralateral cortex. RESULTS: Magnetic resonance spectroscopy revealed a gradual recovery of the damaged tissue; however, we found no significant differences in metabolite concentrations in the lesioned hemisphere between treated and untreated animals. Higher concentrations of glutamate and N-acetyl aspartate were found in the contralateral hemisphere in cell-treated animals compared to untreated ones. Lesioned animals showed neurogenesis in the contralateral hemisphere; the number of newly generated cells in stem cell-treated animals was 50% higher than those observed in untreated animals. CONCLUSION: No direct impact of cell transplantation was observed in the lesion. However, changes in the contralateral hemisphere suggest that the transplanted MSCs might stimulate repair processes and plasticity resulting in the generation of newborn cells, which might enable the faster adoption of the damaged tissue's function by healthy tissue.

Transplantation of Mesenchymal Stromal Cells in Patients With Amyotrophic Lateral Sclerosis: Results of Phase I/IIa Clinical Trial.

Amyotrophic lateral sclerosis (ALS) is a progressive untreatable neurodegenerative disorder, leading to the death of the cortical and spinal motoneurons (MNs). Bone marrow-derived mesenchymal stem/stromal cells (BM-MSCs) may represent a new approach to slowing down the progression of ALS by providing neurotrophic support to host MNs and by having an anti-inflammatory effect. We have designed a prospective, nonrandomized, open-label clinical trial (phase I/IIa, EudraCT No. 2011-000362-35) to assess the safety and efficacy of autologous multipotent BM-MSCs in ALS treatment. Autologous BM-MSCs were isolated and expanded under GMP conditions. Patients received 15 ± 4.5 × 106 of BM-MSCs via lumbar puncture into the cerebrospinal fluid. Patients were monitored for 6 months before treatment and then for an 18-month follow-up period. Potential adverse reactions were assessed, and the clinical outcome was evaluated by the ALS functional rating scale (ALSFRS), forced vital capacity (FVC), and weakness scales (WSs) to assess muscle strength on the lower and upper extremities. In total, 26 patients were enrolled in the study and were assessed for safety; 23 patients were suitable for efficacy evaluation. After intrathecal BM-MSC application, about 30% of the patients experienced a mild to moderate headache, resembling the headaches after a standard lumbar puncture. No suspected serious adverse reactions (SUSAR) were observed. We found a reduction in ALSFRS decline at 3 months after application (p < 0.02) that, in some cases, persisted for 6 months ( p < 0.05). In about 80% of the patients, FVC values remained stable or above 70% for a time period of 9 months. Values of WS were stable in 75% of patients at 3 months after application. Our results demonstrate that the intrathecal application of BM-MSCs in ALS patients is a safe procedure and that it can slow down progression of the disease.