Regenerative Potential Of Stem Cells Research Articles

Tissue mosaicism following stem cell aging: blood as an exemplar.

Loss of stem cell regenerative potential underlies aging of all tissues. Somatic mosaicism, the emergence of cellular patchworks within tissues, increases with age and has been observed in every organ yet examined. In the hematopoietic system, as in most tissues, stem cell aging through a variety of mechanisms occurs in lockstep with the emergence of somatic mosaicism. Here, we draw on insights from aging hematopoiesis to illustrate fundamental principles of stem cell aging and somatic mosaicism. We describe the generalizable changes intrinsic to aged stem cells and their milieu that provide the backdrop for somatic mosaicism to emerge. We discuss genetic and nongenetic mechanisms that can result in tissue somatic mosaicism and existing methodologies to detect such clonal outgrowths. Finally, we propose potential avenues to modify mosaicism during aging, with the ultimate aim of increasing tissue resiliency.

Exploring the Regenerative Potential of Stem Cells for Treating Eye Diseases: A Review of the New Findings

The escalating prevalence of vision loss due to eye diseases has instigated a quest for innovative therapies, given that conventional approaches often fall short in repairing and regenerating damaged eye tissues, particularly the retina. Stem cell-based interventions have emerged as a promising avenue, with numerous studies in animal models and human trials exploring their potential to enhance visual acuity. Beyond addressing conditions like age-related macular degeneration (AMD) and diabetic retinopathy (DR), stem cell therapies demonstrate efficacy in treating genetic disorders such as retinitis pigmentosa (RP). In severe eye damage necessitating regeneration, stem cells play a pivotal role, leveraging their regenerative capabilities. Noteworthy is the transplantation of retinal pigment epithelial (RPE) cells derived from embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs), showcasing promising results in preclinical models and clinical studies, leading to improved retinal function without severe side effects. Mesenchymal stem cells (MSCs) have successfully treated optic neuropathy, RP, DR, and glaucoma, yielding positive clinical outcomes. The safety of adult stem cells, particularly MSCs derived from adipose tissue or bone marrow, has been firmly established. This review highlights significant advancements in utilizing human ESC-derived retinal pigmented epithelium and iPSCs for treating eye injuries. While cell-based therapy is relatively nascent, with numerous clinical trials pending review, stem cells' regenerative potential and clinical applications in addressing eye diseases offer substantial promise. This study aims to comprehensively examine the applications of stem cells in the context of eye diseases and their potential role in regenerative medicine.

Comparing Older Matched Related to Younger Matched Unrelated Donors in Acute Myeloid Leukemia in Remission Using Post-Transplant Cyclophosphamide

Backgroud. Over the past 25 years, the landscape of allogeneic stem cell transplantation (allo-SCT) changed from being rarely performed in patients 50 years or older to accounting for a little less than half of the transplantations reported in USA and Europe. Older patients will on average have older matched related siblings (MRD) being considered as donors with issues such as comorbidities, risk of clonal haematopoiesis of indeterminate potential, and impaired immune and regenerative potential of stem cells. Although younger HLA-matched unrelated donors (MUD) are an important alternative in this context, it remains unclear whether the use of MUD is associated with better outcomes. This is especially true in the context of post-transplant cyclophosphamide (PTCy) as graft-versus-host disease (GvHD) prophylaxis. Methods. The aim of this study was to compare the graft-relapse-free survival (GRFS) for patients diagnosed with acute myeloid leukemia (AML) in first complete remission (CR1), aged 50y or more, and having received a first allo-SCT from a 10/10 MUD younger than 40y or a MRD older than 50y using PTCy for in-vivo T-cell depletion. All transplants were performed between 2015-2021 at the EBMT centres. Results. A total of 410 consecutive patients were included, 172pts receiving a MRD, 238pts transplanted from a MUD. All patients with detailed conditioning regimen dosages were classified according to the transplant conditioning intensity (TCI) score in low (1-2), intermediate (2.5-3.5), high (4-6). TCI-score was available for 345 patients and was as follows: low in 53pts (36.8%), intermediate in 79pts (54.9%) and high in 12pts (8.3%) for MRD, low in 91pts (45.3%), intermediate in 93pts (46.3%) and high in 17pts (8.5%) for MUD (p=0.26). Only patients with available TCI-score were analysed. In univariate analysis the 2-year GRFS was 58% (49.8-65.3%) for TCI intermediate-high and 43.1% [34.3-51.5] for TCI low (p=0.005). In TCI-score intermediate-high non-relapse mortality (NRM) wasn't higher compared to TCI-score low (12.7% [12.2-24] vs 13.7% [8.4-20.3], p=0.53) but relapse incidence (RI) was lower (17.7% [12.2-24] vs 33.2% [25.1-41.5], p=0.003). As a strong interaction between TCI-score and type of donor was found, we compared transplant outcomes between MRD and MUD separately for TCI-score low and TCI-score intermediate-high. In TCI-score low, 2-year GRFS was 44.4% [33.5-54.8] for MUD and 40.9% [26.4-54.9] for MRD (p=0.83) (Figure 1A). In multivariable analysis (MVA) type of donor was not a risk factors for neither GRFS (HR 0.98, CI: 0.57-1.69; p=0.95) nor for any other transplant outcome. In TCI-score intermediate-high, 2-year GRFS was 67.2% [55.8-76.2] for MUD and 46.1% [34.4-57.1] for MRD (p=0.001) (Figure 1B). In MVA MUD were associated to better GRFS (HR 0.41, CI: 0.24-0.7; p=0.001), lower NRM (HR 0.19, CI: 0.07-0.55; p=0.002) and lower RI (HR 0.32, CI: 0.14-0.73; p=0.007) without advantages in terms of neither aGvHD nor cGvHD. Conclusions. In patients with AML in CR1, aged 50y or more and receiving a conditioning regimen with TCI-score ≥ 2.5 in the PTCy setting, a MUD younger than 40y is associated to longer GRFS, lower NRM and lower RI compared to a MRD older than 50y. For patients receiving regimens with TCI-score low transplant outcomes are independent on the type of HLA-matched donor.

Effect of Honokiol on culture time and survival of Alzheimer's disease iPSC-derived neurons.

Patient-derived induced pluripotent stem cells (iPSCs) have been widely used as disease models to test new therapeutic strategies. Moreover, the regenerative potential of stem cells can be improved with the use of biologically active compounds. Our study was designed to explore the effect of honokiol, a small polyphenol molecule extracted from Magnolia officinalis, on the survival and culture time of iPSC-derived neurons from a sporadic Alzheimer's disease (AD) patient. This study aimed to generate iPSCs from peripheral blood mononuclear cells (PBMCs) of an AD patient using episomal plasmids with a nucleofector system and differentiate them into neurons. These iPSC-derived neurons were used to investigate the effect of honokiol extracted from M. officinalis on their survival and long-term cultures. IPSCs were generated from PBMCs of an AD patient by introducing Oct-3/4, Sox2, Klf4, L-Myc, and Lin28 using NucleofectorTM Technology. Differentiation of neurons derived from iPSCs was carried out using inducers and recognized by biomarkers. The viability of iPSC-derived neurons with the addition of honokiol extracted from the bark of M. officinalis was determined by the MTT analytical kit. IPSCs were generated by reprogramming AD patient-derived PBMCs and subsequently converted into neurons. The survival and growth of iPSC-derived neurons were significantly enhanced by adding honokiol in the experiment conditions. AD iPSC-derived neurons had a high viability rate when cultured in the presence of honokiol. These results have shown that AD iPSC-derived neurons can be an excellent model for screening neurotrophic agents and improving the conditions for long-term cultures of human iPSC-derived neurons. Honokiol proves to be a potential candidate for cellular therapeutics against neurodegenerative disorders.

HSF-1 enhances cardioprotective potential of stem cells via exosome biogenesis and their miRNA cargo enrichment.

Stem cell therapy provides a hope to no option heart disease patient group. Stem cells work via different mechanisms of which paracrine mechanism is reported to justify most of the effects. Therefore, identifying the control arms for paracrine cocktail production is necessary to tailor stem cell functions in disease contextual manner. In this study, we describe a novel paracrine cocktail regulatory axis, in stem cells, to enhance their cardioprotective abilities. We identified that HSF1 knockout resulted in reduced cardiac regenerative abilities of mesenchymal stem cells (MSCs) while its overexpression had opposite effects. Altered exosome biognesis and their miRNA cargo enrichment were found to be underlying these altered regenerative abilities. Decreased production of exosomes by MSCs accompanied their loss of HSF1 and vice versa. Moreover, the exosomes derived from HSF1 depleted MSCs showed significantly reduced candidate miRNA expression (miR-145, miR-146, 199-3p, 199b and miR-590) compared to those obtained from HSF1 overexpressing MSCs. We further discovered that HSF1 mediates miRNAs' enrichment into exosomes via Y binding protein 1 (YBX1) and showed, by loss and gain of function strategies, that miRNAs' enrichment in mesenchymal stem cell derived exosomes is deregulated with altered YBX1 expression. It was finally demonstrated that absence of YBX1 in MSCs, with normal HSF1 expression, resulted in significant accumulation of candidate miRNAs into the cells. Together, our data shows that HSF1 plays a critical role in determining the regenerative potential of stem cells. HSF1 does that by affecting exosome biogenesis and miRNA cargo sorting via regulation of YBX1 gene expression.

MTOR signalling pathway in stem cell bioactivities and angiogenesis potential.

The mammalian target of rapamycin (mTOR) is a protein kinase that responds to different stimuli such as stresses, starvation and hypoxic conditions. The modulation of this effector can lead to the alteration of cell dynamic growth, proliferation, basal metabolism and other bioactivities. Considering this fact, the mTOR pathway is believed to regulate the diverse functions in several cell lineages. Due to the pleiotropic effects of the mTOR, we here, hypothesize that this effector can also regulate the bioactivity of stem cells in response to external stimuli pathways under physiological and pathological conditions. As a correlation, we aimed to highlight the close relationship between the mTOR signalling axis and the regenerative potential of stem cells in a different milieu. The relevant publications were included in this study using electronic searches of the PubMed database from inception to February 2023. We noted that the mTOR signalling cascade can affect different stem cell bioactivities, especially angiogenesis under physiological and pathological conditions. Modulation of mTOR signalling pathways is thought of as an effective strategy to modulate the angiogenic properties of stem cells.

Abstract 620: Hsf-1 Enhances Cardioprotective Potential Of Stem Cells Via Exosome Biogenesis And Their Mirna Cargo Enrichment.

Introduction: Stem cells work via different mechanisms of which paracrine mechanism is able to justify most of the effects. Therefore, identifying the control arms for paracrine cocktail production is necessary to tailor stem cell functions. Objective: This study shows how HSF1 determines stem cell potential to repair ischemic heart via affecting production of paracrine factors including exosome and their miRNA cargo. Methods and Results: CRISPR-Cas9 gene editing tool was used to knock out HSF1 gene in nMSCs while lentiviral hsRNA constructs for HSF1 were used to overexpress it in aMSCs. We identified that HSF1 knockout resulted in reduced cardiac regeneration abilities in nMSCs while its overexpression in aMSCs had opposite effects as depicted by functional characteristics including LVEF, LVEDV, LVESV and LVFS in our rat ischemic model. Further, we found that exosome number in nMSCs decreased significantly after HSF1 knockout while it went up in aMSCs after HSF1’s overexpression as determined by NanoSite particle tracking analysis. The exosomes from nMSCs (nEXOs), aMSCs (aEXOs) and their genetically modified versions (nKO-EXOs) and aOE-EXOs) respectively) were analysed for miRNA cargo using RT-PCR. nKO-EXOs showed significantly reduced candidate miRNA expression (miR-145, miR-146, 199-3p, 199b and miR-590) than nEXOs while aOE-EXOs had increased candidate miRNA expression compared to aEXOs. Since HSF1 does not bind to RNA molecules, we hypothesised that YBX1, another protein highly expressed by nMSCs than aMSCs and has RNA binding nature, under the influence of HSF1 sorts miRNAs into exosomes. YBX1 expression was found to be controlled by HSF1 as shown by RT-PCR and western results. HSF1 controls YBX1 expression, at transcriptional level, is strongly supported by our finding that YBX1 promoter possesses heat shock elements (HSEs) as HSF1 binding sites. We further went on to see whether YBX1 alteration affects miRNA loading of exosomes and it was demonstrated that absence of YBX1 resulted in accumulation of candidate miRNAs into the cells. Conclusion: Together, our data shows that HSF1 plays a critical role in determining the regenerative potential of stem cells. HSF1 does that by affecting exosome biogenesis and miRNA cargo sorting via YBX1 protein.

Recent clinical trials with stem cells to slow or reverse normal aging processes.

Aging is associated with a decline in the regenerative potential of stem cells. In recent years, several clinical trials have been launched in order to evaluate the efficacy of mesenchymal stem cell interventions to slow or reverse normal aging processes (aging conditions). Information concerning those clinical trials was extracted from national and international databases (United States, EU, China, Japan, and World Health Organization). Mesenchymal stem cell preparations were in development for two main aging conditions: physical frailty and facial skin aging. With regard to physical frailty, positive results have been obtained in phase II studies with intravenous Lomecel-B (an allogeneic bone marrow stem cell preparation), and a phase I/II study with an allogeneic preparation of umbilical cord-derived stem cells was recently completed. With regard to facial skin aging, positive results have been obtained with an autologous preparation of adipose-derived stem cells. A further sixteen clinical trials for physical frailty and facial skin aging are currently underway. Reducing physical frailty with intravenous mesenchymal stem cell administration can increase healthy life expectancy and decrease costs to the public health system. However, intravenous administration runs the risk of entrapment of the stem cells in the lungs (and could raise safety concerns). In addition to aesthetic purposes, clinical research on facial skin aging allows direct evaluation of tissue regeneration using sophisticated and precise methods. Therefore, research on both conditions is complementary, which facilitates a global vision.

Electrically Triggered Quercetin Release from Polycaprolactone/Bismuth Ferrite Microfibrous Scaffold for Skeletal Muscle Tissue.

Skeletal muscle tissue engineering presents a promising avenue to address the limitations pertaining to the regenerative potential of stem cells in case of injury or damage. The objective of this research was to evaluate the effects of utilizing novel microfibrous scaffolds, containing the compound quercetin (Q), on skeletal muscle regeneration. Morphological test results showed us that the combination of bismuth ferrite (BFO), polycaprolactone (PCL), and Q were bonded and well-ordered with each other, and a uniform microfibrous structure was obtained. Antimicrobial susceptibility testing of PCL/BFO/Q was conducted, and microbial reduction was found to be over 90% in the highest concentration of Q-loaded microfibrous scaffolds with the most inhibitory effect on S. aureus strains. Further, biocompatibility was investigated by performing MTT testing, fluorescence testing, and SEM imaging on mesenchymal stem cells (MSCs) to determine whether they could act as suitable microfibrous scaffolds for skeletal muscle tissue engineering. Incremental changes in the concentration of Q led to increased strength and strain, allowing muscles to withstand stretching during the healing process. In addition, electrically conductive microfibrous scaffolds enhanced the drug release capability by revealing that Q can be released significantly more quickly by applying the appropriate electric field, compared with conventional drug-release techniques. These findings suggest a possible use for PCL/BFO/Q microfibrous scaffolds in skeletal muscle regeneration by demonstrating that the combined action of both guidance biomaterials was more successful than Q itself acting alone.

Thyroid-stimulating hormone receptor signaling restores skeletal muscle stem cell regeneration in rats with muscular dystrophy.

Duchenne muscular dystrophy (DMD) is a severe and progressive myopathy leading to motor and cardiorespiratory impairment. We analyzed samples from patients with DMD and a preclinical rat model of severe DMD and determined that compromised repair capacity of muscle stem cells in DMD is associated with early and progressive muscle stem cell senescence. We also found that extraocular muscles (EOMs), which are spared by the disease in patients, contain muscle stem cells with long-lasting regenerative potential. Using single-cell transcriptomics analysis of muscles from a rat model of DMD, we identified the gene encoding thyroid-stimulating hormone receptor (Tshr) as highly expressed in EOM stem cells. Further, TSHR activity was involved in preventing senescence. Forskolin, which activates signaling downstream of TSHR, was found to reduce senescence of skeletal muscle stem cells, increase stem cell regenerative potential, and promote myogenesis, thereby improving muscle function in DMD rats. These findings indicate that stimulation of adenylyl cyclase leads to muscle repair in DMD, potentially providing a therapeutic approach for patients with the disease.

ADIPOSE-DERIVED STEM CELLS (ADSCS) PRETREATED WITH VASCULAR ENDOTHELIAL GROWTH FACTOR (VEGF) PROMOTED WOUND HEALING IN RAT SKIN BURN MODEL

Stem cells are extensively used for regenerative purposes as they are unspecialized cells capable of renewal and differentiation. Growth factors like vascular endothelial growth factor (VEGF) play a crucial role in enhancing the regenerative potential of stem cells. The present study was designed to elucidate the effects of VEGF preconditioning in accelerating the regenerative potential of adipose-derived stem cells (ADSCs) for wound healing. In vivo study was carried out using female Sprague Dawley (SD) rats randomly divided into three groups, i.e., VEGF preconditioned ADSCs transplanted group (Pre-Tx), normal ADSCs transplanted group (N-Tx) and control group. ADCS were isolated from female SD rats and treated with VEGF for the Pre-Tx group. At 21st day post-transplantation, the wound in the Pre-Tx group was completely closed. However, the wound was not fully healed in the N-Tx group and the Control group. For further analysis, the experimental area of skin tissues was taken from all groups and examined histologically. The cellularity and granulation were thicker in the pre-Tx group and thicker in the pre-Tx group and thicker, indicating rapid wound recovery. Furthermore, the polymerase chain reaction results also confirmed the down-regulation of apoptotic marker caspase3, which indicates less cell death at the injury site and up-regulation of certain growth factors like IGF, sdf1α and some other markers like E cadherin and vimentin. RT-PCR analysis revealed significant up-regulation of all these factors in the pre-Tx group compared to other groups. These results suggest that VEGF pre-conditioning improves the reparative potency of ADSCs by increasing their survival rate and stimulating the secretion of various growth factors having crucial involvement in angiogenesis and recovery of damaged tissue.

Prospects of cell therapy for hematopoietic syndrome of acute radiation sickness

Cellular therapy of hematopoietic acute radiation syndrome is currently at the forefront of scientific interest due to the high regenerative potential of stem cells, their anti-inflammatory and antifibrotic effects. Stem cells have a high ability to stimulate the proliferation and differentiation of hematopoietic cells. It has been shown that after administration stem cells are able to migrate to lesions under the influence of chemotactic factors. At the same time, they retain their functions and can differentiate into cells of affected tissues and contribute to their recovery.
 The article analyzes the current trends in the use of cellular products for the treatment of hematopoietic acute radiation syndrome. The processes of cell proliferation and differentiation in the blood system are regulated by feedback mechanisms. This occurs in accordance with signals coming from more differentiated pools, as well as signals from other organs and tissues. They come along the nervous and humoral pathways. The hematopoietic microenvironment created by fibroblasts, macrophages, endotheliocytes, lymphocytes and other cells is play an important role in these processes. The influence of this microenvironment can be carried out both through direct contacts between cells and through the production of regulators (cytokines).
 At present cellular technologies are one of the most promising areas for the treatment of acute radiation syndrome along with the use of cytokines. Cell therapy of hematopoietic acute radiation syndrome has advantages over the use of cytokines and growth factors. It does not require a basic level of stem and progenitor cells, which are the main target for the cytokine effect.
 The presented data testify that cell therapy has been a promising approach in treatment of radiation injury of bone marrow caused by high doses of radiation exposure.

Evaluation of Hypothalamic Neural Stem Cell Niche – Implications on Obesity‐Induced Sympathoexcitation

The newly discovered hypothalamic neural stem cell niche is emerging as a key player in the regulation of systemic homeostasis. Previous studies have shown that inflammation‐associated degenerative processes in the hypothalamic neural stem cell niche impairs central regulation of metabolism and contributes to the development of metabolic syndrome in obesity. In addition to energy homeostasis, hypothalamic nuclei specifically paraventricular nucleus, is a well‐known player in the regulation of sympathetic nerve activity and in turn cardiovascular function. How obesity‐induced degeneration of neural stem cell niche affects sympathetic regulation of blood pressure is still not understood. In the present study, we aim to characterize the neural stem cell population from the hypothalamic niche. In this pilot study, 3‐month‐old C57BL/6J mice fed with standard diet were sacrificed and brains were collected for further processing. Hypothalami were freshly isolated and minced. After series of enzymatic dissociation, washing and filtration, NSCs were cultured. On 14th day of culturing, neurospheres were observed without any changes in their structural integrity. Neurosphere counts were recorded, and cells were collected for RNA extraction to perform RT‐PCR analysis for characterization. Our PCR analysis confirmed the expression of NSC markers like nestin, SOX1 and SOX2 suggesting successful isolation and culturing of NSCs in our lab. Earlier studies in our lab, suggest that HFD‐induced obesity causes cellular senescence in the PVN, a region adjacent to the neurogenic niche of hypothalamus. Senescent cells are known to impair the regenerative potential of stem cells by impairment of tissue microenvironment through their secretome. Having established these techniques, future studies will investigate the link between hypothalamic neurogenesis, senescence (in the PVN) and sympathoexcitation in obesity.

Strategies to improve regenerative potential of mesenchymal stem cells.

In the last few decades, stem cell-based therapies have gained attention worldwide for various diseases and disorders. Adult stem cells, particularly mesenchymal stem cells (MSCs), are preferred due to their significant regenerative potential in cellular therapies and are currently involved in hundreds of clinical trials. Although MSCs have high self-renewal as well as differentiation potential, such abilities are compromised with “advanced age” and “disease status” of the donor. Similarly, cell-based therapies require high cell number for clinical applications that often require in vitro expansion of cells. It is pertinent to note that aged individuals are the main segment of population for stem cell-based therapies, however; autologous use of stem cells for such patients (aged and diseased) does not seem to give optimal results due to their compromised potential. In vitro expansion to obtain large numbers of cells also negatively affects the regenerative potential of MSCs. It is therefore essential to improve the regenerative potential of stem cells compromised due to “in vitro expansion”, “donor age” and “donor disease status” for their successful autologous use. The current review has been organized to address the age and disease depleted function of resident adult stem cells, and the strategies to improve their potential. To combat the problem of decline in the regenerative potential of cells, this review focuses on the strategies that manipulate the cell environment such as hypoxia, heat shock, caloric restriction and preconditioning with different factors.

Extracellular vesicles as novel approaches for the treatment of osteoarthritis: a narrative review on potential mechanisms.

Osteoarthritis (OA) is a progressive degeneration of articular cartilage with involvement of synovial membrane, and subchondral bone. Current treatment approaches have focused on controlling the OA symptoms, pain, and inflammation. Recently, cell-based therapies, including the application of stem cells such as mesenchymal stem cells (MSCs), have been introduced for restoration of the articular cartilage. Despite promising outcomes, there are some limitations in the application of MSCs for OA treatment. It has been demonstrated that the regenerative potential of stem cells is related to the production of paracrine factors. Extracellular vehicles (EVs), the main component of cell secretome, are membrane-bounded structures that deliver biologically active agents. The delivery of molecules (e.g., nucleic acids, proteins, and lipids) leads to cell-to-cell communication and the alteration of cell functions. In this review, general characteristics of EVs, as well as their potential mechanisms in the prevention and treatment of OA were considered. Based on in vitro and in vivo studies, EVs have shown to contribute to cartilage regeneration via suppression of degenerative factors and regulation of chondrocyte function in the synthesis of extracellular matrix components. Also, they inhibit the progression of OA or protect the cartilage from degradation via their impact on inflammatory cytokines. The different signaling pathways of EVs against the pathologic features of OA were summarized in this review. According to the results obtained from several investigations, more investigations should be design to prove the safety and effectiveness of EVs in the treatment and prevention of OA progression.

Human cardiac stem cells rejuvenated by modulating autophagy with MHY-1685 enhance the therapeutic potential for cardiac repair

Stem cell-based therapies with clinical applications require millions of cells. Therefore, repeated subculture is essential for cellular expansion, which is often complicated by replicative senescence. Cellular senescence contributes to reduced stem cell regenerative potential as it inhibits stem cell proliferation and differentiation as well as the activation of the senescence-associated secretory phenotype (SASP). In this study, we employed MHY-1685, a novel mammalian target of rapamycin (mTOR) inhibitor, and examined its long-term priming effect on the activities of senile human cardiac stem cells (hCSCs) and the functional benefits of primed hCSCs after transplantation. In vitro experiments showed that the MHY-1685‒primed hCSCs exhibited higher viability in response to oxidative stress and an enhanced proliferation potential compared to that of the unprimed senile hCSCs. Interestingly, priming MHY-1685 enhanced the expression of stemness-related markers in senile hCSCs and provided the differentiation potential of hCSCs into vascular lineages. In vivo experiment with echocardiography showed that transplantation of MHY-1685‒primed hCSCs improved cardiac function than that of the unprimed senile hCSCs at 4 weeks post-MI. In addition, hearts transplanted with MHY-1685-primed hCSCs exhibited significantly lower cardiac fibrosis and higher capillary density than that of the unprimed senile hCSCs. In confocal fluorescence imaging, MHY-1685‒primed hCSCs survived for longer durations than that of the unprimed senile hCSCs and had a higher potential to differentiate into endothelial cells (ECs) within the infarcted hearts. These findings suggest that MHY-1685 can rejuvenate senile hCSCs by modulating autophagy and that as a senescence inhibitor, MHY-1685 can provide opportunities to improve hCSC-based myocardial regeneration.

Regenerative medicine: potential applications for foot and ankle disorders

Introduction. Regenerative therapies have recently gained popularity in orthopedics and we overview the available scientific evidence on the topic.Methods. A narrative literature review with three relevant case reports.Results. Scientific evidence on regenerative medicine is growing, but some established findings can de underscored. First, a persistent inflammatory response plays a key role in tissue reparation because it inhibits the activity of stem cells: therefore, regenerative therapy is effective if it can reduce local inflammation, thus allowing stem cells to regenerate the damage. Secondly, the regenerative potential of stem cells is regulated by local immunity and, in particular, by macrophages, which are involved in damage response and tissue regeneration. Among others, the concentrate of peripheral blood mononuclear cells (PBMNCs), rich in monocytes, lymphocytes, and CD34+ hematopoietic stem cells, appears an interesting cell-based therapeutic strategy to promote tissue regeneration in several orthopedic disorders.Discussion. Regenerative medicine can offer new valuable therapeutic strategies. In particular, potential applications of PB-MNCs in foot and ankle disorders are discussed with some explanatory cases from clinical practice.

Distinctive cytokine profiles of stem cells from human exfoliated deciduous teeth and dental pulp stem cells

Background/purposeSHED and DPSC have stem cell regenerative potential, but comparative research on their cytokine profile is rare. This study aimed to investigate and compare cytokine profiles secreted from stem cells from human exfoliated deciduous teeth (SHED) and dental pulp stem cells (DPSCs). Materials and methodsSHED-conditioned medium (CM) and DPSC-CM were extracted using seven primary and permanent teeth each. Cytokine membrane array was performed for each CM to quantify and compare the secretomes of 120 cytokines. Enzyme-linked immunosorbent assay, immunocytochemistry, and immunohistochemistry analysis were performed to demonstrate cytokine membrane array analysis. ResultsSignificant differences were observed in the expression levels of 68 cytokines–27 and 41 cytokines were 1.3-fold more strongly expressed in SHED-CM and DPSC-CM, respectively. Cytokines involved in immunomodulation, odontogenesis and osteogenesis were more strongly expressed in SHED-CM. Cytokines involved in angiogenesis were detected more strongly in DPSCs-CM. SHED and DPSCs have distinctive cytokine profiles and characteristics in terms of their stem cell regenerative potential. ConclusionThese observations suggest that SHED may have a better cytokine profile related to inflammatory, proliferative, osteogenic, and odontogenic potential.

N-Cadherin Regulates the Odontogenic Differentiation of Dental Pulp Stem Cells via β-Catenin Activity.

Dental pulp stem cell (DPSC) transplantation has shown new prospects in dental pulp regeneration, and is of great significance in the treatment of pulpitis and pulp necrosis. The fate and regenerative potential of stem cells are dependent, to a great extent, on their microenvironment, which is composed of various tissue components, cell populations, and soluble factors. N-cadherin-mediated cell–cell interaction has been implicated as an important factor in controlling the cell-fate commitment of mesenchymal stem cells. In this study, the effect of N-cadherin on odontogenic differentiation of DPSCs and the potential underlying mechanisms, both in vitro and in vivo, was investigated using a cell culture model and a subcutaneous transplantation mouse model. It was found that the expression of N-cadherin was reversely related to the expression of odontogenic markers (dentin sialophosphoprotein, DSPP, and runt-related transcription factor 2, Runx2) during the differentiation process of DPSCs. Specific shRNA-mediated knockdown of N-cadherin expression in DPSCs significantly increased the expression of DSPP and Runx2, alkaline phosphatase (ALP) activity, and the formation of mineralized nodules. Notably, N-cadherin silencing promoted nucleus translocation and accumulation of β-catenin. Inhibition of β-catenin by a specific inhibitor XAV939, reversed the facilitating effects of N-cadherin downregulation on odontogenic differentiation of DPSCs. In addition, knockdown of N-cadherin promoted the formation of odontoblast-like cells and collagenous matrix in β-tricalcium phosphate/DPSCs composites transplanted into mice. In conclusion, N-cadherin acted as a negative regulator via regulating β-catenin activity during odontogenic differentiation of DPSCs. These data may help to guide DPSC behavior by tuning the N-cadherin-mediated cell–cell interactions, with implications for pulp regeneration.

Reviewing the evidence for shockwave- and cell-based regenerative therapies in the treatment of erectile dysfunction.

Erectile dysfunction (ED) is both a common and complex disease process. Existing ED treatments do not always achieve adequate results. There is clinical interest in employing regenerative therapies, including low-intensity extracorporeal shockwave therapy (Li-ESWT), platelet rich plasma (PRP), and stem cell therapy (SCT), in the treatment of ED as adjunct or alternative treatments. Here, we present evidence for emerging shockwave- and cell-based regenerative therapies for the treatment of ED following a thorough review of the existing PubMed literature pertaining to Li-ESWT, PRP, and SCT in relation to the treatment of ED. Li-ESWT causes microtrauma in tissue that hypothetically upregulates angiogenesis and recruits stem cells. Several large-scale systematic reviews and meta-analyses have reported that Li-ESWT improved ED in humans. Additionally, evidence has commenced to show that Li-ESWT may be effective against two recognized and complex etiologies of ED: diabetic and neurogenic. PRP delivers an autologous sample rich in growth factors to damaged tissue. Animal model studies have demonstrated improved erectile function recovery as well as preservation of cavernous nerve axons. Studies with PRP in humans are limited. SCT utilizes the regenerative potential of stem cells for healing of damaged tissue. In the treatment of ED, SCT has been used in the setting of diabetic and post-prostatectomy ED. Results of human studies are varied, although SCT treatments did result in increased erectile rigidity with some patients recovering the ability to achieve penetration. While these regenerative therapies show potential to augment the current treatment regimen for ED, there is a paucity of evidence to support the safety and efficacy of these treatments. Further research is necessary to define the role of these alternative therapies in the treatment of ED.