MSC Groups Research Articles

The Effect of Intrahepatic and Intrasplenic Administration of Mesenchymal Stem Cell to Liver Function and Degree of Liver Fibrosis in Common Bile Duct Ligation Model in Rabbit

BackgroundMesenchymal stem cells (MSC) is a promising alternative method in liver cirrhosis management. Several administration routes of MSC have been studied, but few studies compared one to another. The purpose of this study is to compare the intrahepatic and intrasplenic route of MSC administration in terms of liver function and degree of liver fibrosis in the bile duct ligation model in rabbits. MethodExperimental study was conducted using rabbits (Oryctolagus cuniculus) model undergoing bile duct ligation (BDL). The subjects were randomized into 4 groups: sham surgery; bile duct ligation; bile duct ligation followed by intrahepatic route of MSC (BDL + IH MSC), and bile duct ligation followed by intrasplenic route of MSC (BDL + IS MSC). Umbilical cord mesenchymal stem cell (UC MSC) was administered on the fifth day after bile duct ligation, and the subjects were observed until the fourteenth day after bile duct ligation. The liver function was evaluated with alanine aminotransferase (ALT), aspartate aminotransferase (AST), and total and direct bilirubin. The degree of fibrosis was evaluated with Laennec score, fibrosis area fraction, the number of viable and necrosis hepatocytes, and the number of hepatic progenitor cells. ResultThe subjects were randomized into 4 groups: 2 in sham surgery group, and 7 in each of the following groups: BDL, BDL + IH MSC and BDL + IS MSC groups. The mortality rate in BDL group was 57.1 %, while mortality in BDL + IH MSC and BDL + IS MSC groups were 14.3 % and 28.6 % respectively. No significant difference was found regarding liver function in each group, such as AST, ALT, total, and direct bilirubin. Histopathology examination in almost every subject undergone bile duct ligation (regardless of MSC administration) showed degree of fibrosis of Laennec 4B. Fibrosis area fraction, the number of viable and necrotic hepatocytes, and progenitor cells were analyzed; no significant difference was found between BDL + IH MSC and BDL + IS MSC groups, but the groups administered with MSC showed a larger number of viable hepatocytes compared to BDL group. No difference was found between BDL + IH MSC and BDL + IS MSC groups in terms of liver function and histologic parameters. ConclusionAdministration of MSC increases the number of viable hepatocytes, but no difference was found in terms of liver function and degree of liver fibrosis between the intrahepatic route and intrasplenic route of administration. Type of StudyAnimal Research, Randomized Controlled Study. Level of EvidenceLevel I? (animal research is not indicated in the levels of evidence table in the journal website).

A112Vital conversations for student podiatrists: a ‘real’ simulated placement

It is recognized that clinical placements are sometimes limited for allied health professionals (AHPs) [1]. This, coupled with the ‘People Plan’ [2], which highlights the need for increasing numbers of AHPs in the workforce, has led to considerations of alternatives to traditional clinical placements. We were approached by a podiatry undergraduate programme to assist with delivering a podiatry simulated placement, for BSc and separately for MSc podiatry students, each for three days, for the academic year 2022/2023. Development of the simulated placement took five weeks; it was collaborative, with input from HEI academics, subject specialists, and simulation-based educationalists. Six scenarios were created which aligned with the HCPC Standards of Proficiency for Podiatrists [3]. These reflected the breadth of experiences students might have experienced in a clinical placement; they focused on communication and behaviours. Actors were involved in playing the roles of simulated colleagues, patients, and relatives, with experienced facilitators setting up a safe, non-threatening, immersive learning environment, covering triggers and time outs in the pre-brief. The following were areas covered: Scope of practice and autonomous practice Professional judgement Culture, equality, diversity and non-discriminatory practice Confidentiality and professionalism Team working Communication skills, face to face and telephone Safe practice environments The learning was underpinned by a communication skills framework enabling students to structure their conversations and behaviours appropriately. Reflection was used post debrief and explored the subsequent day. All scenarios were presented as either forum theatre or fishbowl simulation, with all students being present in the same space as the simulation. Evaluation was positive from both BSc and MSc groups. Students highly rated the structured approach provided by the communication framework stating it helped them converse effectively and build rapport with patients, relatives and colleagues. The inclusion of actors, although daunting for some initially, added hugely to their learning experience. Students commented on how they felt more prepared for real-world situations and how they hadn’t appreciated the breadth and impact of their practice. Simulated placements can offer a safe and controlled environment for podiatry students to develop their skills and engage in vital conversations with patients, relatives and colleagues. Facilitators should adapt to different confidence levels and learning styles of the students and actors fully briefed and in line with these adaptations. These simulated real-life placements are replicable and can help in preparing a workforce fit for purpose. Authors confirm that all relevant ethical standards for research conduct and dissemination have been met. The submitting author confirms that relevant ethical approval was granted, if applicable.

Ectopic expression of insulin in a type 1 diabetic rat model by injection of manipulated mesenchymal stem cells with an insulin construct driven by a glucose-sensitive promoter in the port vein.

The treatment of type 1 diabetes through islet cell transplantation is a complex process, facing challenges such as allograft rejections and a limited supply of donors. One potential solution is to utilize the liver as an alternative for natural insulin production, as hepatocytes can secrete proteins and respond to glucose levels. Recent research has shown promising results in using mesenchymal stem cells as a potential cure for diabetes. The study utilized a diabetic rat model, confirmed through blood sugar measurement. A plasmid vector was designed with specific genetic components, synthesized by biotech company, and then Inserted vector into a plasmid with resistance genes and bacterial origin. Bone marrow-derived mesenchymal stem cells (BM-MSCs) were cultured and transfected with the plasmid using Lipofectamine 3000. Polymerase chain reaction wasemployed to confirm successful transfection using specific primers. For the animal study, 30 male Wistar rats were divided into six groups, each comprising five rats. The control group did not receive any treatment, while the second group received MSCs via Portal Vein Injection. The third group received MSCs transfected with a specific construct via Portal Vein Injection. The fourth group was induced to develop diabetes through streptozotocin(STZ) injection, the fifth group developed diabetes and received untransfected MSCs via Portal Vein Injection, and the sixth group received MSCs transfected with the specific construct via Portal Vein Injection. To manage Pain, appropriate pain control was administered to the rats for 3 days after the surgery. Fixed liver tissues obtained from the euthanized rats were utilized for immunohistochemistry. In this study, immunohistochemical techniques were used to examine insulin expression in different groups of rats. The control groups showed high levels of insulin expression, while the diabetic groups exhibited lower expression. However, there was a significant difference between the diabetic groups treated with MSC and transgenic MSC cells. All groups had similar baseline glucose levels, but the diabetic groups showed a significant increase after STZ injection, whereas the control and MSC groups did not. Postintervention, both the control and MSC groups had similar glucose levels to the post-STZ levels. However, diabetes-induced groups experienced a significant decrease in glucose levels, with the transfected MSCs showing a greater decrease than the untransfected MSCs. The study suggested that treatment with MSCs, especially transfected ones, can effectively reduce glucose levels in rats with diabetes. In this research, rat BM-MSCs were utilized to create insulin-producing mesenchymal cells with glucose-sensitive insulin expression. The cells were transferred to the liver of diabetic rats via portal vein injection, leading to an increase in insulin expression. This study proposes a novel approach for cell therapy and delivery in the treatment of type 1 diabetes.

Treatment of Acute Respiratory Distress Syndrome Caused by COVID-19 with Human Umbilical Cord Mesenchymal Stem Cells

This study aimed to identify the impact of mesenchymal stem cell transplantation on the safety and clinical outcomes of patients with severe COVID-19. This research focused on how lung functional status, miRNA, and cytokine levels changed following mesenchymal stem cell transplantation in patients with severe COVID-19 pneumonia and their correlation with fibrotic changes in the lung. This study involved 15 patients following conventional anti-viral treatment (Control group) and 13 patients after three consecutive doses of combined treatment with MSC transplantation (MCS group). ELISA was used to measure cytokine levels, real-time qPCR for miRNA expression, and lung computed tomography (CT) imaging to grade fibrosis. Data were collected on the day of patient admission (day 0) and on the 7th, 14th, and 28th days of follow-up. A lung CT assay was performed on weeks 2, 8, 24, and 48 after the beginning of hospitalization. The relationship between levels of biomarkers in peripheral blood and lung function parameters was investigated using correlation analysis. We confirmed that triple MSC transplantation in individuals with severe COVID-19 was safe and did not cause severe adverse reactions. The total score of lung CT between patients from the Control and MSC groups did not differ significantly on weeks 2, 8, and 24 after the beginning of hospitalization. However, on week 48, the CT total score was 12 times lower in patients in the MSC group (p ≤ 0.05) compared to the Control group. In the MSC group, this parameter gradually decreased from week 2 to week 48 of observation, whereas in the Control group, a significant drop was observed up to week 24 and remained unchanged afterward. In our study, MSC therapy improved lymphocyte recovery. The percentage of banded neutrophils in the MSC group was significantly lower in comparison with control patients on day 14. Inflammatory markers such as ESR and CRP decreased more rapidly in the MSC group in comparison to the Control group. The plasma levels of surfactant D, a marker of alveocyte type II damage, decreased after MSC transplantation for four weeks in contrast to patients in the Control group, in whom slight elevations were observed. We first showed that MSC transplantation in severe COVID-19 patients led to the elevation of the plasma levels of IP-10, MIP-1α, G-CSF, and IL-10. However, the plasma levels of inflammatory markers such as IL-6, MCP-1, and RAGE did not differ between groups. MSC transplantation had no impact on the relative expression levels of miR-146a, miR-27a, miR-126, miR-221, miR-21, miR-133, miR-92a-3p, miR-124, and miR-424. In vitro, UC-MSC exhibited an immunomodulatory impact on PBMC, increasing neutrophil activation, phagocytosis, and leukocyte movement, activating early T cell markers, and decreasing effector and senescent effector T cell maturation.

Effect of High Efficiency Digestion and Utilization of Organic Iron Made by Saccharomyces cerevisiae on Antioxidation and Caecum Microflora in Weaned Piglets.

Organic iron is expected to replace inorganic iron used in diets as an iron source. Organic iron possesses high absorption efficiency and low fecal iron excretion. This study aims to study the effect of organic iron produced by Saccharomyces cerevisiae (yeast iron) on digestion, utilization, antioxidation and caecum microflora in weaned piglets. In total, 20 piglets that had been weaned after 28 days were divided into 4 groups, each of which followed a different basal diet. The basal diet of each of these 4 groups contained, respectively, 104 mg/kg iron (ferrous sulfate, CON), 84 mg/kg iron (yeast iron, LSC), 104 mg/kg iron (yeast iron, MSC) or 124 mg/kg iron (yeast iron, HSC). This experiment lasted 35 d. The apparent digestibility of iron in LSC, MSC and HMS was higher than that in CON (p < 0.01) and the fecal iron content in LSC, MSC and HMS was lower than that in CON (p < 0.01). Serum iron contents in LSC, MSC and HMS were higher than that in CON (p < 0.01). The iron contents of the heart, lungs, liver, kidney and left gluteus muscle in the MSC and HMS groups were higher than that in CON and LSC (p < 0.05). Serum catalase, glutathione peroxidase, superoxide dismutase activity, superoxide anion, glutathione, hydroxyl free radical scavenging rate, total antioxidant capacity, and liver superoxide anion clearance rate and peroxidase in MSC and HMS were higher than that in CON and LSC (p < 0.05). The contents of nitric oxide and peroxide of the weaned piglets in MSC and HMS were lower than that in CON and LSC (p < 0.05). The abundance of Firmicutes, Blautia and Peptococcus in LSC, HSC and MSC was higher than that in CON (p < 0.01). The abundance of Lactobacillus in CON and LSC was higher than that in MSC and HSC (p < 0.01). The abundance of Acinetobacter, Streptococcus and Prevotella in LSC, MSC and HSC was lower than that in CON (p < 0.01). The results suggested that a diet containing 84 mg/kg iron of yeast iron has the same effect as a diet containing 104 mg/kg iron of ferric sulfate, and that a diet containing 104 or 124 mg/kg iron of yeast iron is superior to a diet containing 104 mg/kg iron of ferric sulfate.

Лечение остеоартроза коленных суставов на основе применения клеточных технологий

Osteoarthritis (OA) is the most common chronic disease of joint and a major cause of joint pain and disability. Traditional therapies for knee osteoarthritis show little clinical benefit, and the terminal treatment option is joint replacement. Currently, cell therapy is prospective for the treatment of knee osteoarthritis and has shown significant results in animal studies and human clinical trials. The aim: to investigate the treatment results in patients with knee osteoarthritis using adipose tissue stromal-vascular fraction and synovial mesenchymal stem cells.Methods. 60 patients with knee osteoarthritis were included in the study. The stage of knee osteoarthritis was assessed using the Kellgren-Lawrence classification, and patients with stage II-III of osteoarthritis were included in this study. After arthroscopic debridement, patients were divided into 2 groups. The first group was injected with the stromal-vascular fraction (SVF) of synovial membrane, and the second group was injected with adipose tissue mesenchymal stem cells (SVF). The minimum clinical and radiologic follow-up period was 12 months. Results. The assessment of the joints condition after 12 months using different scales showed that in both experimental groups there was a significant improvement of the functional status of the knee of the patients after treatment with MSC and SVF. Functional indicators of knee joints on the WOMAC scale improved by 14.7 and 13.4, on the KOOS scale by 23.6 and 26.4 in the SVF and MSC groups, respectively. Pain syndrome regressed by 1.8 and 2.1 on the VAS scale in the SVF and MSC groups, respectively.Conclusion. The results of both study groups showed that there is a significant difference between WOMAC, VAS and KOOS values before and after surgery (8, 12 weeks and 12 months) in the compared groups towards improvement

Human Umbilical Cord Wharton Jelly Cells Treatment Prevents Osteoporosis Induced by D-Galactose.

Methods Sixteen male mice were randomly divided into 4 groups: control (ordinary feeding), D-gal (D-galactose) group, D-gal + MSC (human umbilical cord Wharton jelly cells), and D-gal + MSC-TNFα groups. Except for the control group (fed with same amount of saline solution), other mice received gastric feeding of 250 mg/kg D-galactose every day for 8 weeks. TNFα (10 ng/mL for 24 h) cocultured or noncocultured HUCWJCs (5 × 105) were suspended in 0.1 ml of sterile PBS and injected into tail veins every other week in D-gal + MSC-TNFα and D-gal + MSC groups, respectively, and only 0.1 ml of sterile PBS for control and D-gal groups. The bone mass was detected by qPCR, ELISA, microcomputed tomography (μCT), and hematoxylin-eosin staining. Proliferation, apoptosis, and differentiation of periosteal-derived osteoblasts (POB) were assessed. Transwell assay and scratch healing were performed to detect POB migration and invasion ability. The effect of HUCWJCs on POB signaling pathway expression was evaluated by immunoblotting. Results The malondialdehyde (MDA) in serum was higher and superoxide dismutase (SOD) was lower in the D-gal group compared to the other groups (p < 0.05). Mice in D-gal group showed significantly decreased bone mass when compared to the control group, while HUCWJCs treatment partially rescued the phenotype, as demonstrated by μCT and histology (p < 0.05). Mechanically, HUCWJCs treatment partially promoted proliferation and migration and decreased apoptosis of POB induced by oxidative stress via activating the mitogen-activated protein kinase (MAPK) signaling pathway. Conclusion HUCWJCs can prevent the progression of osteoporosis by inhibiting oxidative stress, which may act by regulating osteoblasts fate through the MAPK signaling pathway.

Combination of Haploidentical Hematopoietic Stem Cell Transplantation with Umbilical Cord-Derived Mesenchymal Stem Cells in Patients with Severe Aplastic Anemia: A Retrospective Controlled Study

Objective:We retrospectively compared the outcomes of patients with severe aplastic anemia (SAA) who received haploidentical hematopoietic stem cell transplantation (haplo-HSCT) combined or not combined with umbilical cord-derived mesenchymal stem cells (UC-MSCs).Materials and Methods:A total of 101 patients with SAA were enrolled in this study and treated with haplo-HSCT plus UC-MSC infusion (MSC group, n=47) or haplo-HSCT alone (non-MSC group, n=54).Results:The median time to neutrophil engraftment in the MSC and non-MSC group was 11 (range: 8-19) and 12 (range: 8-23) days, respectively (p=0.049), with a respective cumulative incidence (CI) of 97.82% and 97.96% (p=0.101). Compared to the non-MSC group, the MSC group had a lower CI of chronic graft-versus-host disease (GVHD) (8.60±0.25% vs. 24.57±0.48%, p=0.048), but similar rates of grades II-IV acute GVHD (23.40±0.39% vs. 24.49±0.39%, p=0.849), grades III-IV acute GVHD (8.51±0.17% vs. 10.20±0.19%, p=0.765), and moderate-severe chronic GVHD (2.38±0.06% vs. 7.45±0.18%, p=0.352) were observed. The estimated 5-year overall survival (OS) rates were 78.3±6.1% and 70.1±6.3% (p=0.292) while the estimated 5-year GVHD-free, failure-free survival (GFFS) rates were 76.6±6.2% and 56.7±6.9% (p=0.045) in the MSC and non-MSC groups, respectively.Conclusion:In multivariate analysis, graft failure was the only adverse predictor for OS. Meanwhile, graft failure, grades III-IV acute GVHD, and moderate-severe chronic GVHD could predict worse GFFS. Our results indicated that haplo-HSCT combined with UC-MSCs infusion was an effective and safe option for SAA patients.

Gingival-Derived Mesenchymal Stem Cells Protect Against Sepsis and Its Complications.

ObjectiveIn the present study, we separated and characterized mouse gingival-derived mesenchymal stem cells (GMSCs) and investigated whether GMSCs can improve lipopolysaccharide (LPS)-induced sepsis and its complications.MethodsNinety-six ICR mice were randomly divided into the following groups: the control (Sham), LPS, and LPS + MSC groups. Mice received 5 mg/kg LPS intraperitoneally to induce sepsis. Histopathological micrographs illustrated organ injury. We detected systemic inflammation, blood glucose levels, and serum levels of high-mobility group box 1 (HMGB1) and lactate. In addition, pulmonary inflammation, lung permeability, and oxidative stress-related indicators in lung tissue were measured.ResultsWe successfully separated a novel population of MSCs from mouse gingiva. These cells had MSC-associated properties, such as a typical fibroblast-like morphology, multiple differentiation potential, and certain phenotypes. Cell-based therapy using GMSCs significantly improved the survival rate, systemic inflammation, hypoglycemia, multiple organ dysfunction syndrome (MODS), and aortic injury during sepsis. GMSCs administration reduced pulmonary inflammation, lung permeability, and oxidative stress injury. GMSCs administration reduced neutrophil infiltration partly because GMSCs inhibited neutrophil chemoattractants tumor necrosis factor (TNF-α), C-X-C motif chemokine ligand (CXCL-1), and Interleukin (IL-8). GMSCs impaired LPS-induced HMGB1 and lactate release during sepsis.ConclusionGMSCs administration is a novel therapeutic strategy targeting aerobic glycolysis for the treatment of sepsis because GMSCs impair LPS-induced HMGB1 and lactate release. GMSCs alleviate lung injury partly because GMSCs exert immune effects, inhibit neutrophilic inflammation, and reduce oxidative stress injury.

Manipulating Osteogenic/Adipogenic Balance in Mesenchymal Stromal Cells (MSCs) for Hematopoietic Applications

Aims: Bone marrow-derived mesenchymal stromal cells (BM-MSCs) have the potential to improve hematopoietic transplantation by supporting hematopoietic stem and progenitor cell (HSPC) expansion as well as by promoting HSPC engraftment to the BM. However, culture expanded MSCs becomes a mixture of progenitor cells committed to different cell lineages, each with different potencies for any one desired application. Clinically, the accumulation of adipogenic MSCs in the BM is believed to contribute towards age-related impairment of regeneration and hematopoiesis; this observation, together with our understanding of the inverse relationship between MSC osteogenic and adipogenic differentiation prompted a more in depth study of the ability of these different MSC lineages to support hematopoietic clinical applications. Clinically relevant methods for engineering a pure cell population from heterogeneous MSC culture will also be discussed.Methods: MSCs were derived from the healthy BM of patients aged 30-50 years. At various stages of osteogenic and adipogenic differentiation in vitro , cultures were evaluated for their ability to support cord blood (CB) CD34+ HSC expansion and stimulate regenerative processes such as angiogenesis and cell proliferation. qPCR, coupled with secretome analysis was performed to discover gene expression markers for MSCs with potent hematopoietic supportive capabilities. Finally, due to the lack of unique surface markers for identification of MSC subpopulations, we utilized multivariate biophysical analysis (microfluidic sorting, atomic force microscopy, etc.), correlating these quantitative measures with biomolecular markers as well as in vitro and in vivo functionality. Novel label-free cell separation strategies that take advantage of unique biophysical traits in a potent MSC subpopulation for HSPC transplantation was also developed to enable clinical translation.Results: Differentiated osteoblasts (OB) as well as pre-OB demonstrated a higher ability to support CB CD34+ HSC expansion compared to undifferentiated MSCs or MSCs in the adipogenic state. Over a period of 10 days, the total HSPC fold expansion was 44.4 ± 4.83, 57.8 ± 10.67, 40.2 ± 4.65, 34.2 ± 6.18 and 13.8 ± 5.50 for OB, pre-OB, MSCs, pre-AD and AD, respectively (n = 5). Absolute expanded CD34+ HSCs was highest with pre-OB cultures (~1.7 fold, n = 5) compared to undifferentiated MSCs. Adipogenic MSCs had much reduced capabilities for supporting HSPC expansion. In vitro angiogenic and proliferation assays showed the greatest degree of HUVEC sprouting and proliferation (~ 2.6 fold greater than undifferentiated MSCs) when exposed to a pre-OB secretome. Pre-OBs are distinct in their transcriptome (up-regulated osteopontin, runx2, angiopoietin-1, FGF1, etc) but no surface marker reliably distinguishes them from other MSCs. We subsequently found a minimal set of three biophysical markers: cell diameter, membrane stiffness and nuclear membrane fluctuations, which collectively are predictive of MSC subpopulations (multipotent vs committed progenitors). We hypothesized and experimentally verified that a biophysically derived pre-OB subpopulation, defined biophysically as MSCs with >20 μm diameter, >375Pa stiffness and <1.2 % nuclear fluctuation, exhibits significantly greater efficacy over other MSC subpopulations for promoting bone marrow repair (~2-3 fold improvements in medial survival following BM injury) and expansion of HSPCs (~1.5-2 fold increase in HSPC numbers). Following co-transplantation with HSPCs, biophysically derived pre-OB promoted faster engraftment of donor cells in the peripheral blood by week 4 (~ 9% vs < 5% in other MSC groups, P<0.05).Conclusion: These results demonstrate a correlation between MSCs with high osteogenic activity and the capabilities for BM repair as well as hematopoietic support, making such cells ideal for augmentation of HSC transplantations. In the absence of reliable surface markers for isolation of these pre-OBs, we developed a clinically relevant biophysical isolation strategy for derivation of pre-OBs that can markedly improve the efficacy of MSC-based approaches for augmentation of hematopoietic transplants. DisclosuresNo relevant conflicts of interest to declare.

Se-Methylselenocysteine Alleviates Liver Injury in Diethylnitrosamine (DEN)-Induced Hepatocellular Carcinoma Rat Model by Reducing Liver Enzymes, Inhibiting Angiogenesis, and Suppressing Nitric Oxide (NO)/Nitric Oxide Synthase (NOS) Signaling Pathway.

BackgroundHepatocellular carcinoma is the third leading cause of cancer-associated mortality. This study aimed to investigate the effects of se-methylselenocysteine (MSC) on oncogenesis of diethylnitrosamine (DEN)-induced hepatocellular carcinoma.Material/MethodsA hepatocellular carcinoma rat model was established by administering DEN. Rat models were divided into Model (0.1 mg/kg MSC), Model+0.3 mg/kg MSC, Model+1 mg/kg MSC, and Model+3 mg/kg MSC groups. A Normal control group consisted of mice not administered MSC. Hematoxylin and eosin staining was used to determine liver injury. Immunohistochemical analysis was conducted to identify CD34 and vascular endothelial growth factor (VEGF) expression. VEGF gene transcription was detected with RT-PCR. Biochemical analyses were performed to determine alanine aminotransferase, aspartate aminotransferase, total bilirubin, γ-glutamyl transpeptidase, alkaline phosphatase, and albumin levels in serum, and nitric oxide (NO)/nitric oxide synthase (NOS) levels in liver tissues. Transmission electron microscopy was used to observe the ultra-microstructures of hepatocytes.ResultsMSC treatment markedly alleviated liver injury and nuclear lesions in the treatment groups compared to the Model group. MSC treatment significantly improved liver functions in the treatment groups compared to the Model group (P<0.05). MSC treatment significantly decreased CD34 expression and NO and NOS levels in liver tissues and suppressed VEGF expression compared to the Model group (all P<0.05).ConclusionsMSC administration alleviated liver injury in a DEN-induced hepatocellular carcinoma rat model through reducing liver enzymes, inhibiting angiogenesis, and suppressing the NO/NOS signaling pathway.

Mindful Self-Compassion program for chronic pain patients: A randomized controlled trial.

Although evidence-based psychological treatments for chronic pain (CP) have been demonstrated to be effective for a variety of outcomes, modest effects observed in recent reviews indicate scope for improvement. Self-compassion promotes a proactive attitude towards self-care and actively seeking relief from suffering. Consequently, more compassionate people experience better physical, psychological and interpersonal well-being. We conducted a single-blind, randomized, controlled trial to examine the effects of a Mindful Self-Compassion program (MSC) on relevant clinical outcomes in patients with CP. Patients were randomly assigned to one of the two intervention arms: MSC or cognitive-behavioural therapy (CBT). The protocols of both intervention arms were standardized and consisted of a 150-min session once a week during 8weeks formatted to groups of no more than 20 participants. The primary outcome was self-compassion, measured with the Self-Compassion Scale (SCS). The secondary outcomes were other pain-related scores, quality-of-life measures, and anxiety and depression scores. In all, 62 and 61 patients were assigned to the MSC and CBT groups, respectively. The MSC intervention was more effective than CBT for self-compassion (average treatment effect [ATE]=0.126, p<0.05). The secondary outcomes, pain acceptance (ATE=5.214, p<0.01), pain interference (ATE=-0.393, p<0.05), catastrophizing (ATE=-2.139, p<0.10) and anxiety (ATE=-0.902, p<0.05), were also favoured in the experimental arm (MSC). No serious adverse events were observed. Mindful Self-Compassion is an appropriate therapeutic approach for CP patients and may result in greater benefits on self-compassion and emotional well-being than CBT. This randomized controlled trial compares the novel intervention (MSC program) with the gold standard psychological intervention for CP (CBT). MSC improves the levels of self-compassion, a therapeutic target that is receiving attention since the last two decades, and it also improves anxiety symptoms, pain interference and pain acceptance more than what CBT does. These results provide empirical support to guide clinical work towards the promotion of self-compassion in psychotherapeutic interventions for people with CP.

Efficacy and safety of intra-articular injection of mesenchymal stem cells in the treatment of knee osteoarthritis: A systematic review and meta-analysis.

Objective:To evaluate the effects and safety of intra-articular injection of mesenchymal stem cells on patients with knee osteoarthritis by a systematic review and meta-analysis.Methods:PubMed, EMBASE, and Cochrane Library were retrieved. An assessment of the risk of bias was done through the Cochrane Collaborative Bias Risk Tool, publication bias was assessed by plotting funnel plots and Egger tests. Pain and functional improvements in patients with knee osteoarthritis were determined by changes in VAS scores and WOMAC scores at baseline and follow-up endpoints. For the evaluation of MRI, the WORMS score and changes in cartilage volume were used. In addition, the number of adverse events in the intervention group and the control group were counted to explore the safety.Results:A total of 10 randomized controlled trials involving 335 patients were included. In the pooled analysis, compared with the control groups, the VAS scores of MSC groups decreased significantly (MD,−19.24; 95% CI: −26.31 to −12.18, P < .00001. All of the WOMAC scores also improved significantly: the total scores (SMD, − 0.66; 95% CI: − 1.09 to −0.23, P = .003), pain scores (SMD, − 0.46; 95% CI: − 0.75 to −0.17, P = .002), stiffness scores (SMD, −0.32; 95% CI: −0.64 to 0.00 P = 0.05), and functional scores (SMD, −0.36; 95% CI: −0.69 to −0.04, P = .03). Two studies with non-double-blind designs were the main source of heterogeneity. In terms of cartilage repair, there was no significant difference in the WORMS score, but there was a significant increase in cartilage volume in the MSC group (SMD, 0.69; 95% CI: 0.25 to 1.13, P = .002). The proportion of patients with adverse events in the MSCs treatment group was significantly higher than that in the control group (OR, 3.20; 95% CI: 1.50 to 6.83, P = .003).Conclusions:Intra-articular injection of mesenchymal stem cells is effective and safety to relieve pain and improve motor function of patients with knee osteoarthritis in a short term which is different to conclusions of previous study.

Recovery Dynamics of Intestinal Bacterial Communities of CCl4-Treated Mice with or without Mesenchymal Stem Cell Transplantation over Different Time Points.

Liver injury has caused significant illness in humans worldwide. The dynamics of intestinal bacterial communities associated with natural recovery and therapy for CCl4-treated liver injury remain poorly understood. This study was designed to determine the recovery dynamics of intestinal bacterial communities in CCl4-treated mice with or without mesenchymal stem cell transplantation (i.e., MSC and CCl4 groups) at 48 h, 1 week (w), and 2 w. MSCs significantly improved the histopathology, survival rate, and intestinal structural integrity in the treated mice. The gut bacterial communities were determined with significant changes in both the MSC and CCl4 groups over time, with the greatest difference between the MSC and CCl4 groups at 48 h. The liver injury dysbiosis ratio experienced a decrease in the MSC groups and a rise in the CCl4 groups over time, suggesting the mice in the MSC group at 48 h and the CCl4 group at two weeks were at the least gut microbial dysbiosis status among the corresponding cohorts. Multiple OTUs and functional categories were associated with each of the bacterial communities in the MSC and CCl4 groups over time. Among these gut phylotypes, OTU1352_S24-7 was determined as the vital member in MSC-treated mice at 48 h, while OTU453_S24-7, OTU1213_Ruminococcaceae, and OTU841_Ruminococcus were determined as the vital phylotypes in CCl4-treated mice at two weeks. The relevant findings could assist the diagnosis of the microbial dysbiosis status of intestinal bacterial communities in the CCl4-treated cohorts with or without MSC transplantation.

Effect of Combining Low Temperature Plasma, Negative Pressure Wound Therapy, and Bone Marrow Mesenchymal Stem Cells on an Acute Skin Wound Healing Mouse Model.

Low-temperature plasma (LTP; 3 min/day), negative pressure wound therapy (NPWT; 4 h/day), and bone marrow mesenchymal stem cells (MSCs; 1 × 106 cells/day) were used as mono- and combination therapy in an acute excisional skin wound-healing ICR mouse model. These therapies have been beneficial in treating wounds. We investigated the effectiveness of monotherapy with LTP, NPWT, and MSC and combination therapy with LTP + MSC, LTP + NPWT, NPWT + MSC, and LTP + NPWT + MSC on skin wounds in mice for seven consecutive days. Gene expression, protein expression, and epithelial thickness were analyzed using real time polymerase chain reaction (RT-qPCR), western blotting, and hematoxylin and eosin staining (H&E), respectively. Wound closure was also evaluated. Wound closure was significantly accelerated in monotherapy groups, whereas more accelerated in combination therapy groups. Tumor necrosis factor-α (TNF-α) expression was increased in the LTP monotherapy group but decreased in the NPWT, MSC, and combination therapy groups. Expressions of vascular endothelial growth factor (VEGF), α-smooth muscle actin (α-SMA), and type I collagen were increased in the combination therapy groups. Re-epithelialization was also considerably accelerated in combination therapy groups. Our findings suggest that combination therapy with LPT, NPWT, and MSC exert a synergistic effect on wound healing, representing a promising strategy for the treatment of acute wounds.

Inflammatory licensing alters MSC metabolism of exogenous fatty acids and preserves immunomodulatory potency in lipid-rich environments

Background & Aim MSC immunomodulation has been central to the expanded use of MSCs within clinical trials. Inflammatory cytokine licensing of an anti-inflammatory MSC phenotype has emerged as a promising driver of enhanced MSC immunosuppressive capacity, however, the mechanism(s) that facilitate this enhancement, as well as critical modifiers of this licensing process are only beginning to emerge. Recent research has demonstrated that metabolic adaptations displayed by MSCs in response to inflammatory licensing are critical to the enhanced immunomodulatory profile of MSCs after licensing. In our previous work, we found that naive, non-licensed MSCs became pro-inflammatory within lipid-rich environments. However, inflammatory licensing allowed MSCs to remain immunosuppressive and non-inflammatory within these same environments. In our current study, we hypothesize that alterations in the underlying metabolic programs elicited by inflammatory licensing are critical to the preservation of MSC immunomodulatory potency within lipid-rich environments. Methods, Results & Conclusion To test our central hypothesis, we performed a metabolomic screen using gas chromatography-mass spectroscopy to identify metabolic programs through relative abundance analyses within four MSC groups: naive MSCs, naive MSCs exposed to the fatty acid palmitate, licensed MSCs, and licensed MSCs exposed to palmitate. Palmitate alone was used as a representative fatty acid because it is the fatty acid in highest abundance within human serum and elevated serum levels are associated with the development of metabolic pathologies including type 2 diabetes. Six umbilical cord MSC donors were used and a total of 80 metabolites covering several critical nodes of central metabolism were assayed. Each treatment condition elicited a unique metabolic profile, with at least 5 or more metabolites showing significant differences in between-group comparisons. Interestingly, in response to palmitate exposure, naive MSCs show less adaptation than their paired licensed counterparts, suggesting that licensing may enhance metabolic adaptability (A). Additionally, metabolites within the distal aspect of the TCA cycle [Aspartate (B), Malate (C), Fumarate (D), and Succinate (E)] showed differential regulation between naive and licensed groups, indicating that this may be a critical pathway elicited by licensing that aids in the preservation of MSC immunomodulation within lipid-rich environments.

Genetically modified il-4 over-expressing mscs enhance bone healing

Background & Aim Healing of large critical size bone defects is challenging in cases of fracture non-union, wear particle disease, osteonecrosis etc. Traditionally, substantial amounts of autologous bone are harvested from the patient's iliac crest for grafting. However, this procedure has substantial potential morbidity; furthermore, bone graft may be insufficient in quantity or quality due to local or systemic factors (e.g. chronic diseases, antagonistic medications, advanced age etc.). New approaches have been devised including the use of autologous concentrated cells harvested from the iliac crest, together with an osteoinductive or osteoconductive carrier. We developed lentivirus-transduced Interleukin-4 (IL-4) over-expressing MSCs (IL-4 MSCs) that continuously produce the anti-inflammatory, pro-regenerative cytokine IL-4, for healing of long bone defects. We report the potential of IL-4 MSCs for healing of large bone defects. Methods, Results & Conclusion Methods: MSCs encapsulated in a unique microribbon (μRB) hydrogel scaffold (Sc) were tested in an established murine femoral critical sized defect model. Ten-week-old male BALB/c mice were fitted with MouseExFix devices (RISystem AG, Switzerland) and a 2 mm bone defect was made in the femoral diaphysis. The three treatments included Sc only, Sc + (unmodified) MSCs, and Sc + IL-4 MSCs. In the Sc only and the Sc + MSC groups, μRB-Sc were placed immediately after the bone defects were made; in the Sc + IL-4 MSCs group, μRB-Sc with IL-4 MSCs were placed 3 days after the defect was created in a second surgery, to allow for the initial mandatory inflammatory response for optimal bone healing to occur. 1.0 × 107 MSCs/mL were used in the scaffold. MicroCT, immune and histomorphometric analyses of the defect were performed 6 weeks after surgery. Results The Sc + IL-4 MSC group had significantly decreased defect size at 6 weeks, compared to the other two groups (Figs. 1 and 2). Histological analysis showed more trabecular bone and M2 macrophages in the defect area in Sc + IL-4 MSCs group, compared to the other 2 groups. Conclusion IL-4 has been shown to promote osteogenesis and M2 macrophage polarization, after an initial mandatory period of acute inflammation lasting 48-72 hours. IL-4 overexpressing MSCs in a unique microribbon hydrogel scaffold, when delivered at the appropriate time and dosage, appears to be a promising strategy to treat critical size long bone defects.

Wound healing improvement by curcumin‐loaded electrospun nanofibers and BFP‐MSCs as a bioactive dressing

AbstractWound healing, one of the most complex processes of the body involving the cooperation of several important biomolecules and pathways, is one of the major therapeutic and economic issues in regenerative medicine. The present study aimed to introduce a novel electrospun curcumin (Cur)‐incorporated chitosan/polyvinyl alcohol/carbopol/polycaprolactone nanofibrous composite for concurrent delivery of the buccal fat pad‐derived mesenchymal stem cells (BFP‐MSCs) and Cur to a full‐thickness wound on the mouse model. Scaffolds were characterized structurally using scanning electron microscopy (SEM), fluorescence microscopy imaging and Fourier‐transform infrared spectroscopy, and toxicity of the scaffolds was also evaluated after BFP‐MSC seeding by SEM imaging and 3‐(4,5 dimethyiazol‐2‐1)‐2‐5‐diphenyl tetrazolium bromide (MTT) assay. Then, its influence on the wound‐healing process was investigated as a wound dressing for a full‐thickness skin defect in mouse model. Results demonstrated that the designed composite scaffolds have the capability for cell seeding and support their growth and proliferation. Macroscopic and histopathological characteristics were evaluated at the end of the 7 and 14 days after surgery, and their results showed that our designed scaffold groups accelerated the wound‐healing process compared with the control group. Among those, scaffold/Cur, scaffold/Cur/BFP‐MSC and scaffold/BFP‐MSC groups demonstrated more wound repair efficacy. These results indicated that the combined grafts can be used to improve the wound‐healing process, and therefore, the electrospun nanofibers presented in this study, Cur and BFP‐MSC together, were demonstrated to have promising potential for wound‐dressing applications.

Therapeutic Potential of Allogenic Bone Marrow-Derived Mesenchymal Stem Cells for Aortic Aneurysm

Introduction - We have previously reported effectiveness of autologous bone marrow derived mesenchymal stem cells (BM-MSCs) for regression of aortic aneurysm (AA). However, they have various disadvantage in terms of cell quality and quantity. Meanwhile, it is known that the stem cell has ability of immunological tolerance. The purpose of this study were to investigate whether treatment of allogeneic MSCs is effective for angiotensin II (AT II)-induced apolipoprotein E-deficient (apoE−/−) already-formed AA mouse. Methods - To identify characterization of each MSCs, the surface markers were checked by flow cytometry. AA was induced in apoE−/− mice by Ang II-infusion for 28 days through subcutaneously implanted osmotic mini-pump. The mice were randomly divided into 3 groups: (ⅰ) 0.2ml saline was intra-veinously injected (n=10, group S), (ⅱ) 1 × 106 autologous BM-MSCs(isolated from C57BL/6) in 0.2 ml saline (n=10, group Au), (ⅲ) 1 × 106 allogeneic BM-MSCs (isolated from BALB/C) (n=10, group Al) via tail vein. Mice were sacrificed at 2weeks after injection. Aortic diameter, matrix metalloproteinase (MMP)-2 and -9 enzymatic activity and cytokine concentrations were measured in obtained aortic tissue. Results - Flow cytometric analysis demonstrated that the both MSCs were similar in surface markers .The Aortic diameters of group Au and Al were significantly decreased compared to group S (group S vs Au vs Al; 2.29 vs 1.40 vs 1.36 (mm), p<0.01). The enzymatic activities of MMP-2 and -9 were significantly reduced in both MSCs groups (active-MMP2: group S vs Au vs Al; 0.45 vs 0.28 vs 0.24 (unit/ml), p<0.05 amd active-MMP9: group S vs Au vs Al; 0.34 vs 0.16 vs 0.14 (unit/ml), p<0.05). Inflammatory cytokines were down-regulated in the both MSC groups (interleukin-6: group S vs Au vs Al; 3399.5 vs 1475.6 vs 937.5 (pg/ml), p<0.01 and monocyte chemotactic protein-1: group S vs Au vs Al; 352.7 vs 208.0 vs 165.1 (pg/m), p<0.05).Insulin-like growth factor (IGF)-1 and tissue inhibitor of metalloproteinase (TIMP)-2 were up-regulated in the MSC groups (IGF-1: group S vs Au vs Al; 2.0 vs 4.7 vs 5.7 (pg/ml), p<0.01; TIMP-2: group S vs Au vs Al; 4.0 vs 9.5 vs 11.1 (pg/ml), p<0.01). Both MSCs injection inhibited infiltration of M1 macrophages in the aortic tissue. Conclusion - Our results suggest that the therapeutic effect of allogeneic MSCs might be similar to that of autologous MSCs for treatment of AA. Allogeneic MSCs injection could provide an alternative therapeutic strategy for the treatment of AA.

Comparing the Osteogenic Potentials and Bone Regeneration Capacities of Bone Marrow and Dental Pulp Mesenchymal Stem Cells in a Rabbit Calvarial Bone Defect Model.

The bone regeneration efficiency of bone marrow mesenchymal stem cells (BMSCs) and dental pulp mesenchymal stem cells (DPSCs) combined with xenografts in the craniofacial region remains unclear. Accordingly, this study commenced by comparing the cell morphology, cell proliferation, trilineage differentiation, mineral synthesis, and osteogenic gene expression of BMSCs and DPSCs in vitro. Four experimental groups (empty control, Bio-Oss only, Bio-Oss+BMSCs, and Bio-Oss+DPSCs) were then designed and implanted in rabbit calvarial defects. The BMSCs and DPSCs showed a similar morphology, proliferative ability, surface marker profile, and trilineage-differentiation potential in vitro. However, the BMSCs exhibited a higher mineral deposition and expression levels of osteogenic marker genes, including alkaline phosphatase (ALP), runt related transcription factor 2 (RUNX2), and osteocalcin (OCN). In the in vivo studies, the bone volume density in both MSC groups was significantly greater than that in the empty control or Bio-Oss only group. Moreover, the new bone formation and Collagen I / osteoprotegerin protein expressions of the scaffold+MSC groups were higher than those of the Bio-Oss only group. Finally, the Bio-Oss+BMSC and Bio-Oss+DPSC groups had a similar bone mineral density, new bone formation, and osteogenesis-related protein expression. Overall, the DPSCs seeded on Bio-Oss matched the bone regeneration efficacy of BMSCs in vivo and hence appear to be a promising strategy for craniofacial defect repair in future clinical applications.