Mesenchymal Stem Cell Transplantation Research Articles

Fibrous Dysplasia: From Historical Approaches to Modern Treatment Perspectives

Objective. This study aims to analyze scientific literature on fibrous dysplasia (FD) and highlight the state of diagnosis and treatment of orthopedic pathology in patients with various forms of FD. Materials and Methods. This article is based on a retrospective review of papers by various authors on the treatment of patients with FD. Articles were selected from scientific databases such as SCOPUS, Web of Science, and Google Scholar. The selected articles were reviewed in chronological order according to the topic of the study. Results and Discussion. The article provides a systematic analysis of treatment approaches for patients with FD at different stages of the development of science. Modern diagnostic and therapeutic approaches are also analyzed, with particular attention given to the molecular-genetic aspects of the disease’s etiology, particularly mutations in the GNAS gene, which underlie bone growth regulation disorders. Current treatment approaches, including the use of bisphosphonates and various types of surgical interventions, are discussed. Conclusions. A review of the scientific literature and analysis of historical aspects related to FD indicate both significant achievements of the orthopedic community in the diagnosis and treatment of FD, as well as significant challenges that will require further resolution. Changing views on the etiology and pathogenesis of the disease, as well as the identification of genetic mutations during early fetal development, have led to the recognition of FD as a stem cell disease. This provides new leverage for doctors and researchers to use stem cells in the treatment of patients with this pathology. Research on FD and the identification of this pathology as a result of stem cell disease has allowed scientists to explore new therapeutic approaches based on the use of mesenchymal stem cells (MSCs). MSCs can be used to correct bone tissue disorders since they can differentiate into osteoblasts and promote bone tissue remodeling. This opens the possibility for MSC transplantation or modification of their functioning to restore bone structures affected by FD. In our opinion, one promising area is the study of the osteogenic activity of MSCs from pathological sites, as this may help develop new regeneration strategies. There is a limited number of studies indicating the potential for bone defect reconstruction through the combination of MSCs and allomaterial.

Mesenchymal stem cells transplantation in the treatment of tendon disorders

Introduction and purpose: Tendon disorders are conditions that lead to pain, reduced mobility, and are often associated with recurrences and complications. A novel therapeutic approach for these conditions is the transplantation of mesenchymal stem cells (MSCs) into the affected area. This paper presents knowledge on the use of MSCs for the treatment of tendon disorders. Additionally, it presents advances in research regarding the application of MSC transplantation in the following conditions: rotator cuff tendon injuries, lateral epicondylitis, Achilles tendon disorders, and patellar tendon injuries. Material and methods: A review of the PubMed and Google Scholar databases was conducted. Results: It is suggested that MSCs support tendon regeneration through the regulation of inflammatory processes, their capacity to differentiate into various cell types, their influence on collagen synthesis, and their effect on the migration and proliferation of tenocytes. Research indicates that transplantation into damaged tendons enhances tendon function and reduces pain. While many studies have not reported the occurrence of side effects, some have noted such cases. Conclusions: Further research is required to develop a deeper understanding of the role that MSCs play in the treatment of tendon disorders. Additionally, there is a need to establish precise treatment protocols and evaluate the safety of this procedure.

Mesenchymal Stem Cells With Polydopamine-Coated NaGdF4 Nanoparticles with Ca2+ Chelation Ability for Ischemic Stroke Therapy.

Mesenchymal stem cells (MSCs) transplantation is a promising therapeutic strategy for ischemic stroke. However, the survival of transplanted MSCs is often compromised by the excessive levels of reactive oxygen species (ROS) and calcium ions (Ca2+) in the ischemic microenvironment following blood flow occlusion. In this study, a protective strategy is developed using functional nanomaterials to escort and shield MSCs. Specifically, NaGdF4@PDA-ALD nanoparticles (NPANs) are synthesized, featuring a NaGdF4 core coated with polydopamine (PDA) for ROS scavenging and further modified with alendronate sodium (ALD) for Ca2+ chelation. The internalization of NPANs by MSCs protected them from oxidative damage and calcium overload, thereby promoting their viability and functionality. Furthermore, NaGdF4 generated T1 signal enhancement, enabling in vivo tracking of MSCs via magnetic resonance imaging. The NPANs-treated MSCs demonstrated improved survival and migration to the ischemic region, promoting blood flow restoration and angiogenesis. These findings confirm the feasibility of employing functional nanoparticles to augment MSCs-based therapies, offering a promising strategy to improve their therapeutic efficacy in ischemic stroke treatment.

Intrathecal transplantation of human umbilical cord mesenchymal stem cells enhances spinal cord injury recovery: Role of miR‑124‑3p as a biomarker.

Spinal cord injury (SCI) is a severe condition that often leads to permanent functional impairments. The current treatment options are limited and there is a need for more effective treatments. Human umbilical cord mesenchymal stem cells (hUCMSCs) have shown promise in promoting neuroregeneration and modulating immune response. In addition, miR-124-3p has been identified as a potential biomarker for monitoring the progress of neural repair, making it a focus of the present study, which used a rat model of SCI to evaluate the effects of intrathecal hUCMSC transplantation. The present study included three groups: A sham-operated group, an SCI model group receiving PBS and an SCI group receiving hUCMSCs. Neurological function was assessed using the Basso, Beattie and Bresnahan locomotor rating scale and Rivlin inclined plane test on days 1, 3, 7, 14 and 21 post-injury. Histological analysis included hematoxylin and eosin staining to assess tissue morphology, Nissl staining to evaluate neuron survival and immunofluorescence to detect bromodeoxyuridine (BrdU)+/neuron-specific enolase (NSE)+ cells, which indicate neurogenesis. Detection of brain-derived neurotrophic factor (BDNF) protein expression at various time points in rats with spinal cord injury using western blotting. miR-124-3p expression was quantified using reverse transcription-quantitative (RT-q)PCR to assess its potential as a biomarker for SCI recovery. The hUCMSC group showed significant improvements in motor function compared with the control group, particularly on days 7 and 14 post-injury. Histological analysis revealed reduced scar tissue formation and increased neuron survival in the hUCMSC group. Immunofluorescence analysis showed a higher number of BrdU+/NSE+ cells in the hUCMSC group, indicating enhanced neurogenesis. The expression of the neurorepair-related protein BDNF was markedly higher in the hUCMSCs group compared with the control group. Furthermore, RT-qPCR analysis demonstrated a marked upregulation of miR-124-3p in the hUCMSC group, which was correlated with improved functional recovery. The present study demonstrated that intrathecal transplantation of hUCMSCs notably enhanced recovery following SCI, probably by promoting neurogenesis and modulating miR-124-3p expression. miR-124-3p upregulation in the hUCMSC group highlighted its potential as a biomarker for tracking the progress of SCI recovery. These findings provided a foundation for the future clinical applications of hUCMSCs in SCI treatment and the use of miR-124-3p as a monitoring tool.

P0792 Remission rates in Complex Perianal Fistulizing Crohn's Disease – Real-life experience of a tertiary reference center

Abstract Background The treatment of complex perianal fistula (cPF) in Crohn’s Disease (CD) requires an integrated and multidisciplinary approach (1) This phenotype affects up to a third of CD patients (2,3) and significantly harms their quality of life (4). This study aims to review the experience of a tertiary reference center and identify prognostic factors related to non-response to treatment. Methods Adult patients with CD who underwent surgery for cPF, in a tertiary reference center, between 2015 and 2023 were retrospectively reviewed. Extracted data included baseline demographic data, clinical recurrence of perianal disease, and resolution of perianal disease. Cases were stratified according to clinical response to treatment of perianal disease after surgery: complete clinical response (clinical remission), partial clinical response, clinical nonresponse or clinical recurrence. Results Seventy-nine patients (50.6% male) were included; the average age was 28 years (Q1=21 and Q3=38). Median follow-up of 85 months (interquartile range, 37-124); the multidisciplinary team discussed all of them. At the time of first surgery, 53.2% of patients had trans-sphincteric cPF, 22.8% had supra-sphincteric cPF, and 15.2% had rectovaginal cPF. In 88.6% of patients, the first procedure was seton placement. All but two patients were treated with 1 to 4 biologic drugs. The two patients treated with mesenchymal stem cells transplantation achieved a complete clinical response at 24 weeks, maintained at 52 weeks of follow-up. Female sex was a protective factor for nonresponse to treatment, and supra-esfincteric fistula was factor associated with the risk of nonresponse or recurrence. At end of follow-up, 78.5% of patients had a complete or partial response, and 21.5% had a nonresponse or a recurrence. Due to perineal destruction 3 patients required an ostomy, and the other 3 a proctectomy. Conclusion This real-life study showed that a multidisciplinary approach, including surgical intervention, can completely heal Crohn's disease perianal fistula in more than seventy-eight percent of patients. Supra-esphincteric fistula was a risk factor for worse disease outcomes, and female sex was a protective factor for nonresponse.

Innovative treatment of age-related hearing loss using MSCs and EVs with Apelin

Utilizing single-cell transcriptome sequencing (scRNA-seq) technology, this study explores the viability of employing mesenchymal stem cells (MSCs) as a therapeutic approach for age-related hearing loss (ARHL). The research demonstrates MSCs' ability to differentiate into inner ear cell subpopulations, particularly hair cells, delivering Apelin via extracellular vesicles (EVs) to promote M2 macrophage polarization. In vitro experiments show reduced inflammation and preservation of hair cell health. In elderly mice, MSCs transplantation leads to hair cell regeneration, restoring auditory function. These findings highlight the regenerative capabilities of MSCs and EV-mediated therapeutic approaches for ARHL.

The Diurnal Variation in Mitochondrial Gene in Human Type 2 Diabetic Mesenchymal Stem Cell Grafts.

The application of regenerative therapy through stem cell transplantation has emerged as a promising avenue for the treatment of diabetes mellitus (DM). Transplanted tissue homeostasis is affected by disturbances in the clock genes of stem cells. The aim of this study is to investigate the diurnal variation in mitochondrial genes and function after transplantation of adipose-derived mesenchymal stem cells (T2DM-ADSCs) from type 2 diabetic patients into immunodeficient mice. Diurnal variation in mitochondrial genes was assessed by next-generation sequencing. As a result, the diurnal variation in mitochondrial genes showing troughs at ZT10 and ZT22 was observed in the group transplanted with adipose-derived mesenchymal stem cells derived from healthy individuals (N-ADSC). On the other hand, in the group transplanted with T2DM-ADSCs, diurnal variation indicative of troughs was observed at ZT18, with a large phase and amplitude deviation between the two groups. To evaluate the diurnal variation in mitochondrial function, we quantified mitochondrial DNA copy number using the Human mtDNA Monitoring Primer Set, measured mitochondrial membrane potential using JC-1, and evaluated mitophagy staining. The results showed a diurnal variation in mitochondrial DNA copy number, mitophagy, mitochondrial membrane potential, and NF-kB signaling in the N-ADSC transplant group. In contrast, no diurnal variation was observed in T2DM-ADSC transplants. The diurnal variation in mitochondrial function revealed in this study may be a new marker for the efficiency of T2DM-ADSC transplantation.

Effect of mesenchymal stem cells transplantation in treatment of amikacin induced- kidney injury in male mice

Effect of mesenchymal stem cells transplantation in treatment of amikacin induced- kidney injury in male mice

Therapeutic effects of intracerebral transplantation of human modified bone marrow-derived stromal cells (SB623) with voluntary and forced exercise in a rat model of ischemic stroke.

Ischemic stroke results in significant long-term disability and mortality worldwide. Although existing therapies, such as recombinant tissue plasminogen activator and mechanical thrombectomy, have shown promise, their application is limited by stringent conditions. Mesenchymal stem cell (MSC) transplantation, especially using SB623 cells (modified human bone marrow-derived MSCs), has emerged as a promising alternative, promoting neurogenesis and recovery. This study evaluated the effects of voluntary and forced exercise, alone and in combination with SB623 cell transplantation, on neurological and psychological outcomes in a rat model of ischemic stroke. Male Wistar rats that had undergone middle cerebral artery occlusion (MCAO) were divided into six groups: control, voluntary exercise (V-Ex), forced exercise (F-Ex), SB623 transplantation, SB623 + V-Ex, and SB623 + F-Ex. Voluntary exercise was facilitated using running wheels, while forced exercise was conducted on treadmills. Neurological recovery was assessed using the modified neurological severity score (mNSS). Psychological symptoms were evaluated through the open field test (OFT) and forced swim test (FST), and neurogenesis was assessed via BrdU labeling. Both exercise groups exhibited significant changes in body weight post-MCAO. Both exercises enhanced the treatment effect of SB623 transplantation. The forced exercise showed a stronger treatment effect on ischemic stroke than voluntary exercise alone, and the sole voluntary exercise improved depression-like behavior. The SB623 + F-Ex group demonstrated the greatest improvements in motor function, infarct area reduction, and neurogenesis. The SB623 + V-Ex group was most effective in alleviating depression-like behavior. Future research should optimize these exercise protocols and elucidate the underlying mechanisms to develop tailored rehabilitation strategies for stroke patients.

Advancements in the treatment of cerebral ischemia-reperfusion injury: Acupuncture combined with mesenchymal stem cells transplantation.

Cerebral ischemia-reperfusion injury (CIRI) constitutes a significant etiology of exacerbated cerebral tissue damage subsequent to intravenous thrombolysis and endovascular mechanical thrombectomy in patients diagnosed with acute ischemic stroke. The treatment of CIRI has been extensively investigated through a multitude of clinical studies. Acupuncture has been demonstrated to be effective in treating CIRI. Recent 5 years studies have identified potential mechanisms of acupuncture, including regulation of autophagy, promotion of angiogenesis, inhibition of inflammation and apoptosis, modulation of cell activation, neuroplasticity regulation, and promotion of nerve regeneration. The transplantation of mesenchymal stem cells (MSCs) can effectively suppress apoptosis, modulate immune responses, and enhance the proliferation and migration of endogenous neural stem cells (NSCs), thereby compensating for the NSCs deficiency following cerebral ischemia/reperfusion injury. The combination of acupuncture and MSCs transplantation demonstrates superiority over individual treatments, significantly enhancing the survival rate of MSCs. Moreover, it facilitates the secretion of various cytokines to promote their homing and differentiation into functional neurons, thereby providing a novel approach for clinical treatment of CIRI.

Synergistic Effects of Hydrogen Peroxide Preconditioning and Valproic Acid on Hepatic Differentiation of Mesenchymal Stem Cells.

Ex vivo preconditioning increases the therapeutic potential of mesenchymal stem cells (MSCs) in terms of antioxidant activity, growth factor production, homing, differentiation, and immunomodulation. Therefore, it is considered an effective strategy to be used before transplantation and therapeutic application of MSCs. Histone deacetylase inhibitor (HDACi), valproic acid (VPA), has been reported to induce hepatic differentiation in MSCs. Although individual studies have shown that preconditioning and epigenetic modification enhance the survival and differentiation of MSCs, the combined effects of these therapies have not been fully explored. This study aims to investigate the combined effect of hydrogen peroxide (H2O2) preconditioning and HDACi (valproic acid) on the differentiation of bone marrow-derived mesenchymal stem cells (BM-MSCs) into hepatic-like cells. MSCs were first preconditioned with H2O2 and then cultured with VPA. The migration and proliferation potential of the treated cells were evaluated using wound healing and colony-- forming unit assays. Furthermore, the expression of hepatic genes (FOXA2, CK8, CK18, TAT) and proteins (AFP, ALB, TAT) was evaluated in all treated groups. The combined therapy group exhibited enhanced cell migration and proliferation, as evidenced by wound healing and colony-forming unit assays. Additionally, the combined treatment group showed higher expression of FOXA2, CK8, and CK18 hepatic genes and TAT protein, suggesting an improved differentiation of stem cells into hepatocytes. In conclusion, the combination of H2O2 and VPA emerges as an important factor in promoting hepatocyte differentiation. However, further studies are required to optimize this protocol for future therapeutics.

PLGA scaffold combined with MSCs transplantation improved neural function and brain tissue structure in rats with traumatic brain injury.

Poly (lactic-co-glycolic acid) (PLGA) is an important biomaterial for tissue defect repair, but its application in replacing missing brain tissue needs improvement. Mesenchymal stem cells (MSCs) have been used to treat various neurological diseases, but they face challenges when filling large tissue defects. The purpose of this study was to investigate the effects of PLGA combined with MSCs transplantation on brain structure and neural function in rats with traumatic brain injury (TBI), and explore its possible mechanism. The results showed that both PLGA transplantation and PLGA+MSCs transplantation could improve the brain structure and promote nerve function recovery in rats with TBI, with PLGA+MSCs transplantation being superior to PLGA transplantation. Furthermore, compared to PLGA transplantation alone, PLGA+MSCs transplantation further reduced brain injury and cell apoptosis, promoted neuron survival, and improved synaptic plasticity. Overall, the adhesion of MSCs to PLGA can enhance the therapeutic efficacy of PLGA in rats following TBI.

Therapeutic Potential of Different Injection Methods for Bone Marrow Mesenchymal Stem Cell Transplantation in Buslfan-Induced Male Rat Infertility.

Background: In recent years, bone marrow derived mesenchymal stem cells (BM-derived MSCs) have emerged as a powerful cell-based therapy for various diseases, including male infertility. Aim: Demonstrating the efficiency of BM-derived MSCs transplantation by different routes of injection to home and repair testis of busulfan-induced azoospermic rats. Material and methods: In the present study, rat BM-derived MSC was isolated and characterized for mesenchymal &hematopoietic markers using flow-cytometry. Induction of infertility was induced by two successive doses of 10 mg/kg of busulfan. Azoospermic rats were treated by BM-derived MSCs which were injected via various routes (IP, IV, and local in testis). After 60 days; sperm analyses were performed beside mainly Biochemical, histopathological, immunohistological, and ultrastructural investigations. Results: BM-derived MSCs were expressed by CD44+ve, CD105+ve, CD106+ve, CD73+ve, CD34-ve, and CD45-ve. Sperm analysis showed a substantial improvement in sperm morphology, motility, and count following treatment with BM-derived MSCs. Caspase-3 and PCNA immunoexp ression accompanied with the levels of FSH, LH, testosterone, SOD, GSH and MDA depicted a considerable restoration of healthy levels after BM-derived MSCs treatment. The seminiferous tubules showed healthy morphology and spermatozoa were detected in their lumen according to the histopathological and ultrastructural analysis of BM-derived MSCs treated rats. Interestingly, BM-derived MSCs intravenous injection revealed the most significant infertility repair outcomes (P<0.05). Conclusion: Transplanted BM-derived MSCs had the potential to home in rat azoospermic testes and restore spermatogenesis. Consequently, the distinctive characteristics of BM-derived MSCs, such as their ability to differentiate and home, make them a promising cell-based therapeutic option for male infertility.

Role of the nuclear factor-kappa B signaling pathway in the repair of white matter injury in neonatal rats through human umbilical cord mesenchymal stem cell transplantation

To observe the reparative effects of human umbilical cord mesenchymal stem cell (hUC-MSC) transplantation on white matter injury (WMI) in neonatal rats and explore its mechanism through the nuclear factor-kappa B (NF-κB) signaling pathway mediated by microglial cells. Sprague-Dawley rats, aged 2 days, were randomly divided into three groups: sham-operation,WMI, and hUC-MSC (n=18 each). Fourteen days after modeling, hematoxylin-eosin staining was used to observe pathological changes in the white matter, and immunofluorescence staining was used to measure the expression level of ionized calcium-binding adapter molecule 1 (Iba1). Western blotting was used to measure the protein expression levels of inhibitory subunit of nuclear factor-kappa B alpha (IκBα), phosphorylated IκBα (p-IκBα), phosphorylated NF-κB p65 (p-NF-κB p65), myelin basic protein (MBP), and neuron-specific nuclear protein (NeuN). Quantitative real-time PCR was used to assess the mRNA expression levels of tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), MBP, and NeuN. Immunohistochemistry was used to measure the protein expression levels of MBP and NeuN. On day 28, the Morris water maze test was used to evaluate spatial cognitive ability. Fourteen days after modeling, the sham-operation group exhibited intact white matter structure with normal cell morphology and orderly nerve fiber arrangement. In the WMI group, large-scale cell degeneration and necrosis were observed, and nerve fiber arrangement was disordered. The hUC-MSC group showed relatively normal cell morphology and more orderly nerve fibers. Compared with the sham-operation group, the WMI group had significantly higher proportions of Iba1-positive cells, increased protein levels of p-IκBα and p-NF-κB p65, and higher mRNA levels of TNF-α and IL-1β. The protein expression of IκBα and the positive expression of MBP and NeuN, as well as their protein and mRNA levels, were significantly reduced in the WMI group (P<0.05). Compared with the WMI group, the hUC-MSC group showed reduced proportions of Iba1-positive cells, decreased protein levels of p-IκBα and p-NF-κB p65, and lower mRNA levels of TNF-α and IL-1β. Furthermore, IκBα protein expression and MBP and NeuN expression (both at the protein and mRNA levels) were significantly increased in the hUC-MSC group (P<0.05). On day 28, the Morris water maze results showed that compared with the sham-operation group, the WMI group had significantly longer escape latency and fewer platform crossings (P<0.05). In contrast, the hUC-MSC group had significantly shorter escape latency and more platform crossings than the WMI group (P<0.05). hUC-MSC transplantation can repair WMI in neonatal rats, promote the maturation of oligodendrocytes, and support neuronal survival, likely by inhibiting activation of the NF-κB signaling pathway mediated by microglial cells.

Engraftment of self-renewing endometrial epithelial organoids promotes endometrial regeneration by differentiating into functional glands in rats.

Extensive trauma frequently disrupts endometrial regeneration by diminishing endometrial stem cells/progenitor cells, affecting female fertility. While bone marrow mesenchymal stem cell (BMSC) transplantation has been suggested as an approach to address endometrial injury, it comes with certain limitations. Recent advancements in endometrial epithelial organoids (EEOs) have displayed encouraging potential for endometrial regeneration. Therefore, this study aims to explore whether EEOs surpass BMSCs in their ability to repair injured endometrium and to examine whether the restoration process involves the integration of EEOs into the endometrial tissue of the recipient. We developed rat EEOs (rEEOs) mimicking the features of the rat endometrium. Subsequently, we created a rat model of endometrial injury to compare the effects of rEEOs and rat BMSCs (rBMSCs) on endometrial regeneration and reproductive recovery. Bulk RNA-sequencing analysis was conducted to further investigate the capacity of rEEOs for endometrial regeneration and to identify discrepancies between rEEOs and rBMSCs. Additionally, to track the fate of the transplanted cells in vivo, we transplanted green fluorescent protein (GFP) -labelled rEEOs or red fluorescent protein (RFP) -labelled rBMSCs. In a rat model of endometrial injury, we observed that fertility recovery in rats transplanted with rEEOs was more comparable to that of normal rats than in those treated with rBMSC. rEEOs possess a high concentration of endometrial epithelial stem/progenitor cells and secrete vascular endothelial growth factor (VEGF)-A to promote endometrial neovascularization. Significantly, we observed that cells from GFP-labelled rEEOs could integrate and differentiate into functional glands within the injured endometrium of recipient rats. EEOs offer a transformative approach to address the challenges of endometrial trauma. Their remarkable regenerative potential holds promise for the restoration of damaged endometrium. As we venture into the future, the concept of utilizing patient-specific EEOs for transplantation emerges as a tantalizing prospect. However, the EEOs in our experiments were mainly cultured in Matrigel, which has barriers to clinical translation as a biomaterial, a new biomaterial to be explored. Secondly, our experiments have been successful only in rat models, and more efforts need to be made before clinical translation.

The role of stem cells in the management of neonatal posthemorrhagic hydrocephalus.

Neonatal intraventricular hemorrhage (IVH) is a common complication of prematurity as it affects 12.4% of preterm infants weighing under 1500g. Posthemorrhagic hydrocephalus (PHH) is an important complication of neonatal IVH and can have serious long-term consequences such as cognitive impairment and cerebral palsy. The purpose of this review is to determine whether stem cell transplantation can play a role in the treatment of neonatal IVH mainly focusing on the prevention of the catastrophic sequelae of neonatal IVH, as well as to the improve outcome of these patients. A literature search was performed using the PubMed/MEDLINE and Scopus databases, and after meticulous screening, eight articles were finally selected. The authors included both animal and human studies in this narrative review. Our review included eight articles, five animal studies and three human studies, including one phase 1 clinical trial, one pilot study, and one case report. Intraventricular transplantation of mesenchymal stem cells (MSCs) early after IVH diagnosis seems to prevent the development of PHH, improve myelination, and reduce periventricular cell death, inflammation, and reactive gliosis. It also seems to be a safe and well-tolerated procedure in preterm infants. Animal and human study findings regarding stem cell transplantation in the treatment of IVH show promising results in reducing the risk of PHH. Further research with larger series is needed to better determine its safety and efficacy. Larger studies such as randomized controlled trials could establish the efficacy and tolerability of the treatment.

Circadian Rhythms of Clock Genes After Transplantation of Mesenchymal Stem Cells with Type 2 Diabetes Mellitus.

Regenerative therapy involving stem cell transplantation has become an option for the radical treatment of diabetes mellitus. Disruption in the clock genes of stem cells affects the homeostasis of transplanted tissues. We examined the circadian rhythm of genes in transplanted adipose-derived mesenchymal stem cells derived from a patient with type 2 diabetes mellitus (T2DM-ADSC). The clock genes (PER2, CLOCK1, CRY1, and ARNTL[BMAL1]) exhibited similar daily fluctuations in phase and amplitude between a group transplanted with adipose-derived mesenchymal stem cells derived from a healthy individual (N-ADSC) and a group transplanted with T2DM-ADSC. The findings demonstrated that clock genes in stem cells are synchronized with those in living organisms. Next-generation sequencing was then employed to categorize genes that exhibited variation in expression between N-ADSC and T2DM-ADSC. MTATP8P1 and NDUFA7_2 gene expression was significantly reduced at two time points (ZT6 and ZT18), and daily fluctuations were lost. The present study reports, for the first time, that the circadian rhythms of MTATP8P1 and NDUFA7_2, genes involved in mitochondrial processes, are altered in T2DM-ADSC.

Atg5-deficient mesenchymal stem cells protect against non-alcoholic fatty liver by accelerating hepatocyte growth factor secretion

Background/aimsMesenchymal stem cells (MSCs) have shown promising therapeutic potential in treating liver diseases, such as non-alcoholic fatty liver disease (NAFLD). Genetic modification has been employed to enhance the characteristics of MSCs for more effective disease treatment. Here, we present findings on human adipose-derived MSCs with Atg5 deficiency, investigating their therapeutic impact and the associated mechanisms in NAFLD.MethodsIn vitro, lentiviral transduction was employed to downregulate Atg5 or HGF in human adipose-derived MSCs using short hairpin RNA (shRNA). Subsequently, experiments were conducted to evaluate cell senescence, proliferation, cell cycle, apoptosis, and other pertinent aspects. In vivo, a non-alcoholic fatty liver mouse model was established by feeding them a high-fat diet (HFD), and the effects of MSCs transplantation were assessed through serological, biochemical, and pathological analyses.ResultsOur research findings indicate that Atg5-deficient MSCs display heightened proliferative activity. Subsequent co-culturing of MSCs with hepatocytes and the transplantation of Atg5-deficient MSCs into NAFLD mouse models demonstrated their ability to effectively reduce lipid accumulation in the NAFLD disease model by modulating the AMPKα/mTOR/S6K/Srebp1 pathway. Furthermore, we observed that Atg5 deficiency enhances the secretion of hepatocyte growth factor (HGF) by promoting recycling endosome (RE) production. Lastly, our study revealed that 3-MA-primed MSCs can improve the characteristics of NAFLD by boosting the secretion of HGF.ConclusionsOur research findings suggest that Atg5-deficient MSCs protect against NAFLD by accelerating HGF secretion. This indicates that Atg5 gene-modified MSCs may represent a promising strategy for treating NAFLD.

Synergistic effects of repeated transcranial magnetic stimulation and mesenchymal stem cells transplantation on alleviating neuroinflammation and PANoptosis in cerebral ischemia

BackgroundNeuronal death is the primary cause of poor outcomes in cerebral ischemia. The inflammatory infiltration in the early phase of ischemic stroke plays a vital role in triggering neuronal death. Either transplantation of mesenchymal stem cells (MSCs) derived from humans or repetitive transcranial magnetic stimulation (rTMS) have respectively proved to be neuroprotective and anti-inflammatory in cerebral ischemia. However, either treatment above has its limitations. Whether these two therapies have synergistic effects on improving neurological function and the underlying mechanisms remains unclear. This investigation aims to elucidate the synergistic effects and underlying mechanisms of MSCs combined with rTMS treatment on the neurological function recovery post-ischemia.MethodsA Sprague-Dawley rat model of cerebral infarction was induced via transient middle cerebral artery occlusion (tMCAO). The rats were divided into five groups (n = 50): sham, tMCAO, rTMS, MSCs, and MSCs + rTMS groups. Transplantation of human umbilical cord MSCs and rTMS intervention were performed 24 h post-stroke. Neurological function was further assessed via several behavioral tests and the 2,3,5-triphenyltetrazolium chloride (TTC) staining companied with Nissl staining were used to assess neuronal survival. TUNEL staining, western blotting, immunofluorescence, immunohistochemistry, ELISA, and flow cytometry were employed to measure the levels of neuroinflammation and PANoptosis. The molecular mechanisms underlying the special role of rTMS in the combined therapy were distinguished with transcriptome sequencing via PC12 cells in oxygen-glucose deprivation/reoxygenation (OGD/R) conditions.ResultsThe combined therapy efficiently reduced lesion volume and improved neuronal survival (P < 0.05), subsequently improving functional recovery after ischemic stroke. MSCs + rTMS treatment ameliorated the PANoptosis in neurons (P < 0.05), accompanied by decreased levels of inflammatory factors in the cerebral tissue and serum during the subacute phase of cerebral infarction. To further explore the roles of either therapy on synergistic effect, we found that the transplanted MSCs primarily localized in the spleen and reduced cerebral inflammatory infiltration after ischemia via suppressed splenic inflammation. Meanwhile, rTMS significantly protects neurons from PANoptosis in MSCs-inhibited inflammatory conditions by downregulating REST unveiled by transcriptome sequencing.ConclusionsOur study elucidates an unidentified mechanism by which the combination of MSCs and rTMS could synergistically promote neuronal survival and suppress neuroinflammation during the subacute phase of cerebral infarction, thus improving neurological outcomes. The downregulating REST induced by rTMS may potentially contribute to the neuroprotective effect against PANoptosis in MSCs-inhibited inflammatory conditions. These results are expected to provide novel insights into the mechanisms of MSCs and rTMS combination therapy in synergistically protecting against cerebral ischemia injury and potential targets underlying neuronal PANoptosis in the early phase of stroke.

The effect of human umbilical cord-derived mesenchymal stem cell transplantation on the reparative properties of muscle-aponeurotic defect reconstruction in the anterior abdominal wall of rats

Postoperative ventral hernias represent a significant challenge in modern surgery, arising as complications following abdominal operations due to weakness or defects in the musculoaponeurotic structure of the anterior abdominal wall. Mesenchymal stem cells (MSCs) possess high reparative potential due to their paracrine ability to stimulate the regeneration of damaged recipient tissues. Objective. To investigate the effect of intraperitoneal transplantation of human umbilical cord-derived MSCs on the regeneration of muscle fibers in the area of musculoaponeurotic defect repair in the anterior abdominal wall of rats. Materials and Methods. A surgical model of a musculoaponeurotic defect (2 cm in diameter) in the anterior abdominal wall was created in 72 white rats. Animals were divided into four groups based on the method of defect correction: 1) repair with autologous tissues; 2) repair with autologous tissues combined with MSC injection; 3) repair using a polypropylene mesh; 4) repair using a polypropylene mesh combined with MSC injection. Human umbilical cord-derived MSCs were isolated using enzymatic methods, validated by flow cytometry for surface marker expression, and administered intraperitoneally at a dose of 1 million cells per kilogram of body weight. On the 10th and 30th days, histological analysis of tissue samples from the repair site was conducted, assessing the presence of granulation tissue, collagen fibers, newly formed connective tissue, and cellular infiltration. Morphometric comparisons included the relative areas of granulation and fibrous reticular tissue, microcirculatory vessels, and counts of neutrophilic leukocytes, lymphohistiocytic elements, and fibroblasts per square millimeter of section. Results. In Group 1, the repair area demonstrated loose connective tissue rich in fibroblasts and histiocytes. In Group 2, a significantly reduced cellular reaction was observed, along with the formation of denser connective tissue bundles; transverse striation of muscle fibers was more clearly visualized at the sites of damage. On day 10, the granulation tissue area was 34.6% smaller, and the microcirculatory vessel area was 15.9% smaller compared to Group 1, and 28.1% and 57.2% smaller, respectively, compared to Group 3. In Group 3, granulation tissue formed at sites of necrosis and muscle fiber destruction. The best results were observed with the combination of polypropylene mesh and MSCs, where complete resolution of the inflammatory response and scar tissue formation was recorded. In Group 4, on day 10, the granulation tissue area was 2.5 times smaller than in Group 1 and 1.3 times smaller than in Group 2. The relative area of microcirculatory vessels was 1.8 times smaller than in Group 1 and 1.3 times smaller than in Group 2, whereas the fibrous reticular tissue area was 3.2 and 1.2 times larger, respectively. By day 30, granulation tissue, microcirculatory vessels, and leukocytes were no longer detected. The fibrous reticular tissue area was the largest among all groups at this time point. Conclusion. Intraperitoneal transplantation of human umbilical cord-derived MSCs in conjunction with polypropylene mesh repair of musculoaponeurotic defects in the anterior abdominal wall of rats improves regeneration at the defect site and promotes faster healing.