MSC Administration Research Articles

Mesenchymal Stem Cell-Derived Exosomes Attenuate Murine Cytomegalovirus-Infected Pneumonia via NF-κB/NLRP3 Signaling Pathway.

Reactivation and infection with cytomegalovirus (CMV) are frequently observed in recipients of solid organ transplants, bone marrow transplants, and individuals with HIV infection. This presents an increasing risk of allograft rejection, opportunistic infection, graft failure, and patient mortality. Among immunocompromised hosts, interstitial pneumonia is the most critical clinical manifestation of CMV infection. Recent studies have demonstrated the potential therapeutic benefits of exosomes derived from mesenchymal stem cells (MSC-exos) in preclinical models of acute lung injury, including pneumonia, ARDS, and sepsis. However, the role of MSC-exos in the pathogenesis of infectious viral diseases, such as CMV pneumonia, remains unclear. In a mouse model of murine CMV-induced pneumonia, we observed that intravenous administration of mouse MSC (mMSC)-exos reduced lung damage, decreased the hyperinflammatory response, and shifted macrophage polarization from the M1 to the M2 phenotype. Treatment with mMSC-exos also significantly reduced the infiltration of inflammatory cells and pulmonary fibrosis. Furthermore, in vitro studies revealed that mMSC-exos reversed the hyperinflammatory phenotype of bone marrow-derived macrophages infected with murine CMV. Mechanistically, mMSC-exos treatment decreased activation of the NF-κB/NLRP3 signaling pathway both in vivo and in vitro. In summary, our findings indicate that mMSC-exo treatment is effective in severe CMV pneumonia by reducing lung inflammation and fibrosis through the NF-κB/NLRP3 signaling pathway, thus providing promising therapeutic potential for clinical CMV infection.

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).

Mesenchymal Stem Cell-Derived Exosomal Noncoding RNAs as Alternative Treatments for Myocardial Ischemia-Reperfusion Injury: Current Status and Future Perspectives.

Ischemic cardiomyopathy is treated mainly with thrombolytic drugs, percutaneous coronary intervention, and coronary artery bypass grafting to recanalize blocked vessels. Myocardial ischemia-reperfusion injury (MIRI) is an unavoidable complication of obstructive revascularization. Compared with those of myocardial ischemic injury, few effective therapeutic options are available for MIRI treatment. The pathophysiological mechanisms of MIRI involve the inflammatory response, the immune response, oxidative stress, apoptosis, intracellular Ca2+ overload, and cardiomyocyte energy metabolism. These mechanisms exacerbate MIRI. Mesenchymal stem cell-derived exosomes (MSC-EXOs) can alleviate MIRI through these mechanisms and, to some extent, prevent the limitations caused by direct MSC administration. Therefore, using MSC-EXOs instead of MSCs to treat MIRI is a potentially beneficial cell-free treatment strategy. In this review, we describe the mechanism of action of MSC-EXO-derived noncoding RNAs in the treatment of MIRI and discuss the advantages and limitations of this strategy, as well as possible future research directions.

Local, systemic and immunologic safety comparison between xenogeneic equine umbilical cord mesenchymal stem cells, allogeneic canine adipose mesenchymal stem cells and placebo: a randomized controlled trial.

Mesenchymal stem cells are multipotent cells with a wide range of therapeutic applications, including, among others, tissue regeneration. This work aims to test the safety (EUC-MSC) of intra-articular administration of equine umbilical cord mesenchymal stem cells in young healthy dogs under field conditions following single and repeated administration. This was compared with the safety profile of allogenic canine adipose derived mesenchymal stem cells (CAD-MSC) and placebo in order to define the safety of xenogeneic use of mesenchymal stem cells when administered intra-articular. Twenty-four police working dogs were randomized in three groups in a proportion 1:1:1. EUC-MSCs and CAD-MSCs were obtained from healthy donors and were manufactured following company SOPs and under GMP and GMP-like conditions, respectively, and compliant all necessary controls to ensure the quality of the treatment. The safety of the treatment was evaluated locally, systemically and immunologically. For this purpose, an orthopedic examination and Glasgow test for the assessment of pain in the infiltrated joint, blood tests, clinical examination and analysis of the humoral and cellular response to treatment were performed. No adverse events were detected following single and repeated MSC administration despite both equine and canine MSC generate antibody titres in the dogs. The intra-articular administration of equine umbilical cord mesenchymal stem cells in dogs has demonstrated to be safe.

The Effects of Intranasal Implantation of Mesenchymal Stem Cells on Nitric Monoxide Levels in the Hippocampus, Control of Cognitive Functions, and Motor Activity in a Model of Cerebral Ischemia in Rats

Hypoxia occurs in situations of disbalance between metabolic needs and the supply of oxygen to organs and tissues of the body. In this regard, tissue hypoxia and ischemia are essential components of the pathogenesis of many diseases. One of the promising areas of research into the mechanisms of ischemia is attempting to weaken the negative effect of hypoxia and ischemia in the brain by using a variety of techniques that activate neuroprotective mechanisms. Here, we aimed to assess the dynamics of restoration of motor activity control in an experimental model of ischemic stroke in rats (cerebral ischemia, CI) after intranasal perineural implantation of mesenchymal stem cells into the receptive field of the olfactory nerve. It was found that the perineural administration of MSCs to rats in the acute period of cerebral ischemia was accompanied by clear signs of recovery of cognitive and motor functions within 1 and 3 days after the operation. On the seventh day after ischemia modeling, rats with the introduction of MSCs had no distinctive features in the control of motor activity compared to the period before the operation in the same rats. In the hippocampus of rats after modeling ischemia, a significant decrease in the content of NO by about 50% relative to the initial level is observed after 1 day. In the hippocampus of rats in which ischemia was modeled with simultaneous intranasal administration of MSC, a significant decrease in NO content by 39% relative to the initial level was also observed after 1 day. The content of NO increases slightly, but the difference in the level of NO relative to ischemic rats was not significant. The copper content in the hippocampus in the rats of these two groups did not change. There was a tendency to increase the efficiency of the antioxidant system 1 day after ischemia in both studied groups, and this effect was more pronounced with intranasal administration of MSC.

Involvement of T-bet and GATA3 transcription factors in Mesenchymal stem cells and royal jelly combination treatment in brain stroke

IntroductionMesenchymal stem cells (MSC) therapy is a promising therapeutic strategy to overcome the brain stroke side effects. However, it may be associated with long-term complications, including induction of inflammation. This project was designed to examine the effects of MSC administration and its combination with royal jelly (RJ) on the differentiation of T helper subsets. Material and methodsIn this project, the mice were divided to the six groups, including control (healthy without stroke), stroke (mice model of middle cerebral artery occlusion (MCAO)), treated with mouse MSC (mMSC), royal jelly (RJ), combination of mMSC and RJ (mMSC + RJ) and MSC conditioned medium (SUP). Thereafter, sticky test, brain mRNA levels of T-bet (transcription factor for Th1 subset), GATA3 (transcription factor for Th2 subset), and ROR-γ (transcription factor for Th17 subset) and percentage of myeloperoxidase (MPO) activities were explored in the groups. ResultsAdministration of mMSC and mMSC + RJ improved the sticky test times and decreased the MPO activities. Using mMSCs and RJ was associated with increased expression of T-bet and GATA3 transcription factors. Transplantation of mMSCs in combination with RJ reduced expression of T-bet in the infarcted tissue. ConclusionUsing mMSC may be associated with Th1-related inflammation in the long term. RJ co-administration significantly reduced the risks, hence, to decrease the plausible side effects of MSCs, it can be proposed to use RJ in combination with MSC to reduce stroke complications.

Potential use of mesenchymal stem cells in the treatment of inflammatory bowel disease - a systematic review of the literature.

The incidence of inflammatory bowel disease is increasing worldwide and, despite advances in conservative and surgical treatment, a significant proportion of patients do not achieve complete remission and have to undergo repeated surgical procedures. Stem cell therapy is a new alternative to immunomodulatory therapy. We performed a systematic review of the literature dealing with the use of mesenchymal stem cells for the treatment of IBD. The PubMed database was searched and original studies published between 2015 and 2021 were included. A total of 80 citations were found, of which 21 clinical studies and 47 experimental studies were included in the review. The efficacy of MSCs in the treatment of perianal Crohns disease is at least comparable to current therapies. A positive effect of MSC administration has also been demonstrated in animal models of luminal disease or enterocutaneous fistula. The results after systemic administration in IBD patients are still ambiguous. inflammatory bowel disease.

Comparison of the treatment efficacy of umbilical mesenchymal stem cell transplantation via renal subcapsular and parenchymal routes in AKI-CKD mice

BackgroundMesenchymal stem cells (MSCs) have emerged as a promising cell-based therapy for acute kidney injury (AKI). However, the optimal route of MSC transplantation remains controversial, and there have been no comparisons of the therapeutic benefits of MSC administration through different delivery routes.MethodsIn this study, we encapsulated MSCs into a collagen matrix to help achieve local MSC retention in the kidney and assessed the survival of MSCs in vitro and in vivo. After transplanting collagen matrix-encapsulated-MSCs (Col-MSCs) under the renal capsule or into the parenchyma using the same cell dose and suspension volume in an ischemia/reperfusion injury model, we evaluated the treatment efficacy of two local transplantation routes at different stages of AKI.ResultsWe found that Col-MSCs could be retained in the kidney for at least 14 days. Both local MSC therapies could reduce tubular injury, promote the proliferation of renal tubular epithelial cells on Day 3 and alleviate renal fibrosis on Day 14 and 28. MSC transplantation via the subcapsular route exerts better therapeutic effects for renal functional and structural recovery after AKI than MSC administration via the parenchymal route.ConclusionsSubcapsular MSC transplantation may be an ideal route of MSC delivery for AKI treatment, and collagen I can provide a superior microenvironment for cell–cell and cell–matrix interactions to stabilize the retention rate of MSCs in the kidney.

Comparison of Single and Repeated Dosing of Anti-Inflammatory Human Umbilical Cord Mesenchymal Stromal Cells in a Mouse Model of Polymicrobial Sepsis

SummaryMesenchymal stromal cells (MSCs) ameliorate pre-clinical sepsis and sepsis-associated acute kidney injury (SA-AKI) but clinical trials of single-dose MSCs have not indicated robust efficacy. This study investigated immunomodulatory effects of a novel MSC product (CD362-selected human umbilical cord-derived MSCs [hUC-MSCs]) in mouse endotoxemia and polymicrobial sepsis models. Initially, mice received intra-peritoneal (i.p.) lipopolysaccharide (LPS) followed by single i.p. doses of hUC-MSCs or vehicle. Next, mice underwent cecal ligation and puncture (CLP) followed by intravenous (i.v.) doses of hUC-MSCs at 4 h or 4 and 28 h. Analyses included serum/plasma assays of biochemical indices, inflammatory mediators and the AKI biomarker NGAL; multi-color flow cytometry of peritoneal macrophages (LPS) and intra-renal immune cell subpopulations (CLP) and histology/immunohistochemistry of kidney (CLP). At 72 h post-LPS injections, hUC-MSCs reduced serum inflammatory mediators and peritoneal macrophage M1/M2 ratio. Repeated, but not single, hUC-MSC doses administered at 48 h post-CLP resulted in lower serum concentrations of inflammatory mediators, lower plasma NGAL and reversal of sepsis-associated depletion of intra-renal T cell and myeloid cell subpopulations. Hierarchical clustering analysis of all 48-h serum/plasma analytes demonstrated partial co-clustering of repeated-dose hUC-MSC CLP animals with a Sham group but did not reveal a distinct signature of response to therapy. It was concluded that repeated doses of CD362-selected hUC-MSCs are required to modulate systemic and local immune/inflammatory events in polymicrobial sepsis and SA-AKI. Inter-individual variability and lack of effect of single dose MSC administration in the CLP model are consistent with observations to date from early-phase clinical trials.Graphical

UM171A-induced ROS promote antigen cross-presentation of immunogenic peptides by bone marrow-derived mesenchymal stromal cells

BackgroundMesenchymal stromal cells (MSCs) have been extensively used in the clinic due to their exquisite tissue repair capacity. However, they also hold promise in the field of cellular vaccination as they can behave as conditional antigen presenting cells in response to interferon (IFN)-gamma treatment under a specific treatment regimen. This suggests that the immune function of MSCs can be pharmacologically modulated. Given the capacity of the agonist pyrimido-indole derivative UM171a to trigger the expression of various antigen presentation-related genes in human hematopoietic progenitor cells, we explored the potential use of UM171a as a means to pharmacologically instill and/or promote antigen presentation by MSCs.MethodsBesides completing a series of flow-cytometry-based phenotypic analyses, several functional antigen presentation assays were conducted using the SIINFEKL-specific T-cell clone B3Z. Anti-oxidants and electron transport chain inhibitors were also used to decipher UM171a’s mode of action in MSCs. Finally, the potency of UM171a-treated MSCs was evaluated in the context of therapeutic vaccination using immunocompetent C57BL/6 mice with pre-established syngeneic EG.7T-cell lymphoma.ResultsTreatment of MSCs with UM171a triggered potent increase in H2-Kb cell surface levels along with the acquisition of antigen cross-presentation abilities. Mechanistically, such effects occurred in response to UM171a-mediated production of mitochondrial-derived reactive oxygen species as their neutralization using anti-oxidants or Antimycin-A mitigated MSCs’ ability to cross-present antigens. Processing and presentation of the immunogenic ovalbumin-derived SIINFEKL peptide was caused by de novo expression of the Psmb8 gene in response to UM171a-triggered oxidative stress. When evaluated for their anti-tumoral properties in the context of therapeutic vaccination, UM171a-treated MSC administration to immunocompetent mice with pre-established T-cell lymphoma controlled tumor growth resulting in 40% survival without the need of additional supportive therapy and/or standard-of-care.ConclusionsAltogether, our findings reveal a new immune-related function for UM171a and clearly allude to a direct link between UM171a-mediated ROS induction and antigen cross-presentation by MSCs. The fact that UM171a treatment modulates MSCs to become antigen-presenting cells without the use of IFN-gamma opens-up a new line of investigation to search for additional agents capable of converting immune-suppressive MSCs to a cellular tool easily adaptable to vaccination.Graphical abstract

P–802 The fate and regenerative efficiency of differently administered BMSCs in thin-endometrium rat models

Abstract Study question This study aims to compare the engraftment, retaining time and therapeutic efficiency of differently administered BMSCs and help to select an optimal therapeutic route in clinical settings. Summary answer Compared with intrauterine infusion, BMSCs could better promote angiogenesis by upregulating related cytokines, such as VEGF, when administered through the ipsilateral iliac artery. What is known already MSC-based therapy has become a promising method for endometrial disease(thin endomtrium or Ashernmen’s syndrowe). Therapeutic effects could always be observed even though different MSC administration routes or MSCs of different tissue sources were used in these studies. Only a few studies compared efficacy of different transplantation routes. However, the results seem to be controversial. Comparable therapeutic effects were reported in some studies, while others stated that systematic administration gave a better outcome than local administration. Study design, size, duration Experimental animal study. Forty-eight female Sprague-Dawley (SD) rats were used in this study. They were randomly assigned to 4 groups: normal, injured, intra-arterial and intra-uterine group. For all rats except for normal group, the thin endometrium models were established by infusing 95% ethanol into the uterine horns and BMSCs were transplanted either locally or intra-arterially after modeling. The therapeutic efficacy were evaluated in the following month. Participants/materials, setting, methods The thin endometrium models induced by ethanol in SD rats, GFP/Luciferin labeled BMSCs were injected either locally or intra-arterially. The retaining time and quantitative distribution were assessed by in vivo bioluminescence imaging and immune-histological analysis. The precise location and differentiation of differently administered BMSCs were determined by immunofluorescence methods. The endometrial fibrosis, angiogenesis were detected by immunohistochemistry and western blotting at a consecutive time after treatment to compare the therapeutic efficiency of two administration methods. Main results and the role of chance The engraftment and differentiation abiility were comparable in 2 groups. The luminescent signal both remained distinct and strong in the abdomen in the first 4 days post-treatment(7.98 × 105 and 6.02 × 105p/s for IU and IA group), indicating the precise and concentrated distribution of BMSCs administered both locally or intra-arterially. The luminescent signals disappeared under bioluminescence imaging over time. We further evaluated the precise distribution, differentiation ability and retaining time of the BMSCs delivered in two strategies by immunofluorescence analysis. All the GFP positive cell localized in stroma, but not in the epithelium or myometrium. Furthermore, there are significantly more positive staining in basal layer of the endometrium close to the glands and vessels than the outer layer of the endometrium in the intra-arterial group. At the 28th days post treatment, we could capture a few GFP staining in the basalis layel of endometrium in intra-arterial group and there were no GFP fluorescence signals detected in intra-uterine group(P < 0.05), suggesting a better survival of BMSCs administered intra-arterially. Differentiation ability of differently administered BMSCs were similar. A few BMSCs began to differentiate into stromal cell 12 days after therapy. Limitations, reasons for caution No pregnancy tests were carried out in these rats to further confirm the regeneration of thin endometrium and compare the therapeutic efficacy. Wider implications of the findings: Our study unveiled that the location of MSCs might determined their regenerarive ability and retaining time, and provided an optimal therapeutic route in clinical settings. Trial registration number Not applicable

Anti-fibrotic effect of adipose-derived mesenchymal stem cell conditioned medium in muscle fibrosis.

Recent investigations have demonstrated that the administration of MSC in mouse model of diseases provided beneficial effects. On the other hand, human adipose-derived MSC condition medium (ADSC-CM) is reported as containing beneficial secreted factors, but its role in muscle fibrosis has not been identified. The aim of this study was to investigate the inhibitory effects of MSC-CM in muscle fibrosis in vitro using the C2C12 murine muscle, myoblast cell line. C2C12 cells were cultured overnight in 0.1% albumin-Dulbecco's Modifies Eagle's Medium (DMEM). The cells were then pre-incubated in ADSC-CM for 20 min, treated with 2.5-10 ng/mL human TGFβ1 for 8-72 hours and analyzed using RT-qPCR, Western blot and immunofluorescent staining. Treatment with 20% ADSC-CM for 3 days suppressed αSMA protein expression in TGFβ1 treated C2C12 cells. ADSC-CM stimulated the proliferation of C2C12 cells in a dose-dependent manner. Furthermore, TGFβ1 induced Acta2/αSMA mRNA expression which was inhibited by ADSC-CM treatment for 8 hours. Decorin, one of the dermatan sulfate proteoglycans and an endogenous inhibitor of TGFβ1, was expressed in ADSC-CM, but not in TGFβ1 pre-incubated ADSC-CM. Our studies provide useful information for establishing anti-fibrotic mechanism(s) of ADSC-CM, thus facilitating potential application to prevent muscle fibrosis.

Mesenchymal Stem Cells Can Both Enhance and Inhibit the Cellular Response to DNA Immunization by Genes of Nonstructural Proteins of the Hepatitis C Virus.

Despite extensive research, there is still no vaccine against the hepatitis C virus (HCV). The aim of this study was to investigate whether MSCs can exhibit adjuvant properties during DNA vaccination against hepatitis C. We used the pcNS3-NS5B plasmid encoding five nonstructural HCV proteins and MSCs derived from mice bone marrow. Five groups of DBA mice were immunized with the plasmid and/or MSCs in a different order. Group 1 was injected with the plasmid twice at intervals of 3 weeks; Group 2 with the plasmid, and after 24 h with MSCs; Group 3 with MSCs followed by the plasmid the next day; Group 4 with only MSCs; and Group 5 with saline. When the MSCs were injected prior to DNA immunization, the cell immune response to HCV proteins assessed by the level of IFN-γ synthesis was markedly increased compared to DNA alone. In contrast, MSCs injected after DNA suppressed the immune response. Apparently, the high level of proinflammatory cytokines detected after DNA injection promotes the conversion of MSCs introduced later into the immunosuppressive MSC2. The low level of cytokines in mice before MSC administration promotes the high immunostimulatory activity of MSC1 in response to a DNA vaccine. Thus, when administered before DNA, MSCs are capable of exhibiting promising adjuvant properties.

Human Mesenchymal Stem Cells Treatment for Severe COVID-19: 1-Year Follow-Up Results of a Randomized, Double-Blind, Placebo-Controlled Trial

Background The long-term consequences of human umbilical cord-derived mesenchymal stem cell (UC-MSC) treatment for COVID-19 patients are yet to be reported. This study assessed the 1-year outcomes in patients with severe COVID-19, who were recruited in our previous UC-MSC clinical trial.Methods: In this prospective, longitudinal, cohort study, 100 patients enrolled in our phase 2 trial were prospectively followed up at 3-month intervals for 1 year to evaluate the long-term safety and effectiveness of UC-MSC treatment. The primary endpoint was an altered proportion of whole-lung lesion volumes measured by high-resolution CT. Other imaging outcomes, 6-minute walking distance (6-MWD), lung function, plasma biomarkers, and adverse events were also recorded and analyzed. This trial was registered with ClinicalTrials.gov (NCT04288102).Findings: Within 3 months, MSC administration exerted numerical improvement in whole-lung lesion volume compared with the placebo, leading to a significant difference of −10.82% (95% CI: −20.69%, −1.46%, P=0.030) on day 10. MSC also reduced the proportion of solid component lesion volume compared with the placebo at each follow-up point, with a significant difference of − 9.02% (95%CI: − 17.44%, − 0.10%, P=0.045) at month 9. More interestingly, 17.86% (10/56) of patients in the MSC group had normal CT images at month 12 ( P= 0.013), but none in the placebo group. The incidence of symptoms was lower in the MSC group than in the placebo group at each follow-up time, particularly sleep difficulties at month 3 (OR 0.19, 95% CI 0.07,0.50; P=0.001), and usual activity at month 12 (OR 0.15, 95% CI 0.03,0.79; P=0.018). Neutralizing antibodies were all positive, with a similar median inhibition rate (61.6% vs. 67.55%) in both groups at month 12. No difference in adverse events at the 1-year follow-up and tumor markers at month 12 were observed between the two groups.Interpretation: UC-MSC administration achieves a long-term benefit in the recovery of lung lesions and symptoms in COVID-19 patients.Trial Registration: This trial was registered with ClinicalTrials.gov (NCT04288102).Funding The National Key R&D Program of China, the Innovation Groups of the National Natural Science Foundation of China, and the National Science and Technology Major Project.Declaration of Interest: None to declare. Ethical Approval: This study was approved by the Ethics Committee of the Fifth Medical Center, Chinese PLA General Hospital (2020-013-D).

Mesenchymal stromal (stem) cell therapy modulates miR-193b-5p expression to attenuate sepsis-induced acute lung injury.

Although mesenchymal stromal (stem) cell (MSC) administration attenuates sepsis-induced lung injury in pre-clinical models, the mechanism(s) of action and host immune system contributions to its therapeutic effects remain elusive. We show that treatment with MSCs decreased expression of host-derived microRNA (miR)-193b-5p and increased expression of its target gene, the tight junctional protein occludin (Ocln), in lungs from septic mice. Mutating the Ocln 3' untranslated region miR-193b-5p binding sequence impaired binding to Ocln mRNA. Inhibition of miR-193b-5p in human primary pulmonary microvascular endothelial cells prevents tumour necrosis factor (TNF)-induced decrease in Ocln gene and protein expression and loss of barrier function. MSC-conditioned media mitigated TNF-induced miR-193b-5p upregulation and Ocln downregulation in vitro When administered in vivo, MSC-conditioned media recapitulated the effects of MSC administration on pulmonary miR-193b-5p and Ocln expression. MiR-193b-deficient mice were resistant to pulmonary inflammation and injury induced by lipopolysaccharide (LPS) instillation. Silencing of Ocln in miR-193b-deficient mice partially recovered the susceptibility to LPS-induced lung injury. In vivo inhibition of miR-193b-5p protected mice from endotoxin-induced lung injury. Finally, the clinical significance of these results was supported by the finding of increased miR-193b-5p expression levels in lung autopsy samples from acute respiratory distress syndrome patients who died with diffuse alveolar damage.

The Role of Mesenchymal Stem Cells in the Treatment of a Chronic Rhinosinusitis-An In Vivo Mouse Model.

Objectives/Hypothesis: It is acknowledged that the treatment of chronic rhinosinusitis (CRS) represents an important challenge for rhinology and for social and economic life. At present, one of the most common treatments for CRS is represented by local corticosteroids followed by endoscopic sinus surgery (ESS). Starting from the example of the mesenchymal stem cell’s (MSC) capacity to migrate and to modulate a real response in the nasal mucosa of an allergic rhinitis mouse model, we try to obtain a response in a CRS mouse model, using MSC derived by adipose tissue. The aim of this study is to demonstrate that the MSC can be used in CRS treatment and could change its priorities. Methods: Seventy female mice (6 MSC donor mice) were randomized in two stages of study, 32 Aspergillus fumigatus (Af) exposure mice (20 for histological comparison to 1st control mice and 12 for MSC administration, to CRS/MCS model) and 32 control mice (20 for histological comparison to CRS model and 12 for MSC administration and histological control to MSC model); in the first stage, the Aspergillus fumigatus (Af) CRS mouse model was targeted, in this section were included 64 (n = 32) mice (treated and control group). In order to assess the inflammation level (histological analysis), the animals were euthanized; in the second stage MSCs (1 × 106/animal) were administered intravenously to a total of 24 (n = 24) mice (12 mice from the exposed group and 12 mice from the second control group). Results: After 12 weeks of Af intranasal instillation, the inflammation parameters evaluated indicated a severe diffuse chronic inflammation, associated with diffuse severe hyperplasia and mature diffuse squamous metaplasia. The MSCs’ injection via the ophthalmic vein induced important histopathological changes in the CRS experimental group, starting with the presence of MSCs in all samples and continuing with the important degenerative character of inflammation. Conclusions: MSC administration demonstrated a real improvement of CRS evolution on the CRS mouse model.

Effect of mesenchymal stromal cell infusions on lung function in COPD patients with high CRP levels

BackgroundWe previously reported a Phase 1/2 randomized placebo-controlled trial of systemic administration of bone marrow-derived allogeneic MSCs (remestemcel-L) in COPD. While safety profile was good, no functional efficacy was observed. However, in view of growing recognition of effects of inflammatory environments on MSC actions we conducted a post-hoc analysis with stratification by baseline levels of a circulating inflammatory marker, C-reactive protein (CRP) to determine the effects of MSC administration in COPD patients with varying circulating CRP levels.MethodsTime course of lung function, exercise performance, patient reported responses, and exacerbation frequency following four monthly infusions of remestemcel-L vs. placebo were re-assessed in subgroups based on baseline circulating CRP levels.ResultsIn COPD patients with baseline CRP ≥ 4 mg/L, compared to COPD patients receiving placebo (N = 17), those treated with remestemcel-L (N = 12), demonstrated significant improvements from baseline in forced expiratory volume in one second, forced vital capacity, and six minute walk distance at 120 days with treatment differences evident as early as 10 days after the first infusion. Significant although smaller benefits were also detected in those with CRP levels ≥ 2 or ≥ 3 mg/L. These improvements persisted variably over the 2-year observational period. No significant benefits were observed in patient reported responses or number of COPD exacerbations between treatment groups.ConclusionIn an inflammatory environment, defined by elevated circulating CRP, remestemcel-L administration yielded at least transient meaningful pulmonary and functional improvements. These findings warrant further investigation of potential MSC-based therapies in COPD and other inflammatory pulmonary diseases.Trial registration: Clinicaltrials.gov NCT00683722.

The Role of Mesenchymal Stem Cell in Controlling Liver Function on Liver Fibrosis

ABSTRACT
 Background: Liver fibrosis (LF) is the end-stage of liver damage characterized by the increase concentration of SGOT and SGPT. A previous study, revealed that liver transplantation as the most effective therapy for LF has limited availability and remains a major challenge. Mesenchymal stem may control inflammation and regenerate damaged tissue leading to accelerated liver function repair.
 Aim: The purpose of this study was to determine the optimum doze for MSC administration to recover LF. 
 Method: The research used variations doze of MSCs administration of 1x106 and 2x106. Liver firbosis was induced by twice a week intraperitoneal injection of carbon tetrachloride (CCl4) (Sigma-Aldrich) dissolved with olive oil in 1:1 ratio for eight weeks. The concentration of SGPT SGOT was measured at day 9 post MSC administration. One Way Anova was performed to analyze statictical data.
 Result: This study showed that there was a significant decrease in SGOT concentration (P < 0,05). P2 group treatment showed the lowest concentration of SGOT among the treatment (Figure B). In the same time, the administration of UC-MSCs reduce SGPT concentration insignificantly (P > 0,05).
 Conclusion: The transplantation of MSCs ameliorated LF by reducing SGPT and SGOT concentration

Intravenous administration of bone marrow mesenchymal stem cells alleviates renal failure in diabetic mice.

To study the protective effect and mechanism of intravenously administered bone marrow mesenchymal stem cells (BMSCs) on renal failure in diabetic mice. BMSCs were obtained from the bone marrow of mice, identified by flow cytometry and tri-line differentiation test. Diabetic model (DM) mice were established using STZ and infused with mice BMSCs intravenously, followed by the analysis of fasting blood glucose and proteinuria levels, renal tissue damage by optical microscope and electron microscope, and PI3K/AKT signaling protein expression in kidney by Western blot and endocrine function by immunofluorescence staining. DM mice exhibited gradually increased albumin excretion rate with time, and the MSC-treated diabetic mice presented significantly reduced proteinuria levels. HE staining indicated that MSC administration mitigated renal damage as proved by smaller tubular dilatation, reduced glomerulosclerosis and trace protein cylinders, and recovered kidney ultrastructure as shown by improved mesangial dilatation and podocyte loss. Further, BMSCs treatment activated PI3K/AKT signaling, which was downregulated in diabetic mice. However, MSC administration failed to improve pancreatic endocrine function in DM mice. Intravenous administration of MSCs can effectively prevent renal failure in diabetic mice by activating PI3K/AKT signaling pathway.

Short-term and long-term results of intracoronary introduction of autologous mesenchymal stem cells in exertional angina pectoris

Purpose is to study the effectiveness of intracoronary introduction application of mesenchymal stem cells (MSC’s) in refractory angina pectoris.Material and methods. The study was based on the results founded by the investigation, treatment and dynamic follow-up results of 30 patients with refractory angina pectoris who underwent the stationary therapy in the department of Urgent and Recovery Cardiac surgery of SI «Institute of Urgent and Recovery Surgery named after V.K. Gusak of NAMS of Ukraine» from 2007 till 2014. We formed 2 groups; each of them included 15 patients: in the first group (a control one), the patients received only medicamental therapy; the second group received intracoronary administration of autologous MSC’s. According to NYHA (New York Heart Association) classification, all the patients had II-IV functional class of chronic heart failure (CHF). All the patients earlier went through the different revascularization surgeries. Mesenchymal autologous bone marrow stem cells were received according to the standard method in the laboratory of cellular and tissue cultivation of SI «Institute of Urgent and Recovery Surgery named after V.K. Gusak of NAMS of Ukraine». Using the local anesthesia with catheter application in the coronary vessel mouth was implemented a transplant in the dose of 10 million cells.Results. During the control examination, a clinical improvement was observed in all the patients; it was expressed in the quality of life increase according to the Minnesota Questionnaire (MLHFQ); in a decrease of heart failure grade according to NYHA, and tolerance growth to physical exercise during the treadmill test. The maximal effect, by the way, was developed in month and had been holding on during the 9-12 months. Thus if the quality of life before applying the transplantation of autologous MSC was 58+1,8 points; after the 3 months this rate was 32±2,2 points (by p<0,05); but after the I year, it was 37±2,1 points and was progressively decreased in the future. During the ultrasound imaging in the first group was mentioned decrease of left ventricle (LV) ejection fraction (EF) till 23,2±5,2% during the whole study; in the second group, it was observed an EF increase from 31,2±7,1 to 37,4±5,2% by p<0,05 and to the deadline of 6 months it began to decrease. In this period of time the patients reduced the doses of taken medicines; it was mentioned tolerance intensification to the physical exercises that we approved with the help of treadmill test. During the cardiac mapping with the help of navigation system CARTO XP we observed that in 6 months the amplitude of unipolar signal increased from 7,8-8,7 MB to 9,0-9,4 MB in the front-lateral zone; and from 5,3-12,0 to 7,9-14,1 MB in the b posteroseptal correspondingly; it tells about the improvement of electromechanical association and improvement of working function as well as myocardium perfusion. In three years during the control investigation, it was mentioned a negative dynamics in comparison with the baseline results. Conclusions. As we see, the first experience of intracoronary introduction of autologous MSC in patients with refractory angina pectoris showed the raise of myocardium functional reserve that was reflected in the improvement of life quality; reduction of antianginal agent doses; raise of LV EF; as well as in the amplitude of unipolar myocardium signal increase in the MSC introduction zone.