Delivery Of Stem Cells Research Articles

Injectable 3D‐Printed Porous Scaffolds for Adipose Stem Cell Delivery and Endometrial Regeneration

AbstractStem‐cell‐based therapeutic strategies are promising in the clinical treatment of intrauterine adhesions (IUAs), while endometrial regeneration still hardly restores the structure and function of the endometrium because of the inadequate microenvironment for the grafted stem cells and subsequent limited therapeutic efficiency. Herein, an injectable porous hydrogel scaffold (PH scaffold) with customizable shapes is presented by using a microfluidic‐based 3D printing technique for adipose‐derived stem cells (ADSCs) delivery to enhance endometrial regeneration. These scaffolds display a controllable interconnected porous structure, which not only facilitates the encapsulation of ADSCs within the scaffold but also supports the recovery to their original shapes after injection. Furthermore, the cell viability of the laden ADSCs is well‐maintained post‐injection, exhibiting promotive effects on cell migration, proliferation, and tube formation. Based on these features, an ADSCs‐laden PH scaffold with a hollow endometrium‐mimicking morphology is designed and in situ injected into the damaged endometrium in rats of IUAs. These results show that the ADSCs‐laden PH scaffolds can enhance functional endometrial regeneration by suppressing the inflammatory response, promoting cell proliferation, and improving vascularization. Thus, it is believed that such unique 3D‐printed porous scaffolds are promising candidates for cell delivery, which also provides a minimally‐invasive and effective strategy for endometrial regeneration.

Efficacy of mesenchymal stem cell-delivery using perpendicular multi-needle injections to the skin: Evaluation of post-ejection cellular health and dermal delivery.

Mesenchymal stem cell (MSC)-therapy is increasingly being evaluated in clinical trials. Dermal delivery is not only time consuming but also unreliable, potentially hampering the therapeutic result. Therefore, qualification of cell delivery protocols is essential. This study evaluated a clinically relevant automated multi-needle injection method for cutaneous MSC-therapy, allowing the skin to be readily and timely treated, by assessing both the cellular health post-ejection and dermal delivery. Following dispensation through the injector (31G needles: 9- or 5-pin) the cellular health and potency (perceived- and long-term (12h) viability, recovery, metabolism, adherence, proliferation and IDO1-expression) of adipose-derived stem cells (10-20-50×106 cells/ml) were assessed in vitro in addition to dermal delivery of solution in human skin. No significant detrimental effect on the perceived cell viability, recovery, metabolism, adherence or IDO1-expression of either cell concentration was observed. However, the overall long-term viability and proliferation decreased significantly regardless of cell concentration, nonetheless marginally. An injection depth above 1.0mm resulted in all needles piercing the skin with dermal delivery from up to 89% needles and minimal reflux to the skin surface, and the results were confirmed by ultrasound and histology. The automated injector is capable of delivering dermal cell-doses with an acceptable cell quality.

Magnetic Microsphere Scaffold-Based Soft Microbots for Targeted Mesenchymal Stem Cell Delivery.

A soft microbot assembled from individual magnetic microsphere scaffold (MMS) beads carrying mesenchymal stem cells (MSC) is navigated under magnetic actuation, where an oscillating field induces mechanical flexion to propel the microbot toward the target site. A seven-bead microbot attained a top translational speed of 205.6µms-1 (0.068 body lengths-1 ) under 10mT and 2Hz field oscillation. The shallow flexion angle (10-24.5°) allows precision movements required to navigate narrow spaces. Upon arrival at the target site, the MMS beads unload their MSC cargo following exposure to a phosphate-buffered saline (PBS) solution, mimicking the extracellular fluid's sodium concentration. The released stem cells have excellent viability and vitality, promoting rapid healing (i.e., 83.2% vs 49%) in a scratch-wound assay. When paired with minimally invasive surgical methods, such as laparoscopy and endoscopic surgery, the microbot can provide precise stem cell delivery to hard-to-reach injury sites in the body to promote healing. Moreover, the microbot is designed to be highly versatile, with individual MMS beads customizable for cargoes of live cells, biomolecules, bionanomaterials, and pharmaceutical compounds for various therapeutic requirements.

Small Extracellular Vesicles' miRNAs: Biomarkers and Therapeutics for Neurodegenerative Diseases.

Neurodegenerative diseases are critical in the healthcare system as patients suffer from progressive diseases despite currently available drug management. Indeed, the growing ageing population will burden the country's healthcare system and the caretakers. Thus, there is a need for new management that could stop or reverse the progression of neurodegenerative diseases. Stem cells possess a remarkable regenerative potential that has long been investigated to resolve these issues. Some breakthroughs have been achieved thus far to replace the damaged brain cells; however, the procedure's invasiveness has prompted scientists to investigate using stem-cell small extracellular vesicles (sEVs) as a non-invasive cell-free therapy to address the limitations of cell therapy. With the advancement of technology to understand the molecular changes of neurodegenerative diseases, efforts have been made to enrich stem cells' sEVs with miRNAs to increase the therapeutic efficacy of the sEVs. In this article, the pathophysiology of various neurodegenerative diseases is highlighted. The role of miRNAs from sEVs as biomarkers and treatments is also discussed. Lastly, the applications and delivery of stem cells and their miRNA-enriched sEVs for treating neurodegenerative diseases are emphasised and reviewed.

A composite hydrogel improves the survival and differentiation of human iPSC-derived neural stem cells after ischemic stroke

Stem cell transplantation can replace lost and damaged neurons after ischemic stroke and has been proposed as a potential treatment for stroke. However, stem cell therapy is challenged by poor cell retention, survival, and differentiation following transplantation. In this study, we developed a composite hydrogel incorporating brain decellularized extracellular matrix (dECM) and a matrix metallopeptidase (MMP)-responsive host–guest (HG) hydrogel that acts as a platform for human induced pluripotent stem cell-derived neural stem cells (iPSC-NSCs) delivery to improve transplant stem cell survival and differentiation. The optimized composite hydrogel ratio was screened by rheology analysis and neurite outgrowth assessment. We then examined the effects of HG/dECM composite hydrogels on iPSC-NSCs survival and differentiation on two-dimensional surfaces modified with hydrogel and in a three-dimensional hydrogel network. Moreover, iPSC-NSCs-laden composite hydrogels were injected into the infarct cavity in an ischemic stroke model to replace lost and damaged nerve cells. As expected, the HG/dECM composite hydrogels better facilitated the survival and differentiation of iPSC-NSC in vivo than the control. Likewise, the iPSC-NSCs loaded in HG/dECM composite hydrogels showed superior retention and survival in the cortical infarct cavity, and the composite hydrogels promoted the differentiation of iPSC-NSCs into neurons. Overall, HG/dECM composite hydrogels are a promising platform to regulate stem cell survival and differentiation.

Local Delivery of Adipose Stem Cells Promotes Allograft Survival in a Rat Hind-Limb Model of Vascularized Composite Allotransplantation.

Adipose stem cells (ASCs) are a promising cell-based immunotherapy because of their minimally invasive harvest, high yield, and immunomodulatory capacity. In this study, the authors investigated the effects of local versus systemic ASC delivery on vascularized composite allotransplant survival and alloimmune regulation. Lewis rats received hind-limb transplants from Brown Norway rats and were administered donor-derived ASCs (passage 3 or 4, 1 × 10 6 cells/rat) locally in the allograft, or contralateral limb, or systemically at postoperative day 1. Recipients were treated intraperitoneally with rabbit anti-rat lymphocyte serum on postoperative days 1 and 4 and daily tacrolimus for 21 days. Limb allografts were monitored for clinical signs of rejection. Donor cell chimerism, immune cell differentiation, and cytokine expression in recipient lymphoid organs were measured by flow cytometric analysis. The immunomodulation function of ASCs was tested by mixed lymphocyte reaction assay and ASC stimulation studies. Local-ASC-treated recipients achieved significant prolonged allograft survival (85.7% survived >130 days; n = 6) compared with systemic-ASC and contralateral-ASC groups. Secondary donor skin allografts transplanted to the local-ASC long-term surviving recipients accepted permanently without additional immunosuppression. The increases in donor cell chimerism and regulatory T-cells were evident in blood and draining lymph nodes of the local-ASC group. Moreover, mixed lymphocyte reaction showed that ASCs inhibited donor-specific T-cell proliferation independent of direct ASC-T-cell contact. ASCs up-regulated antiinflammatory molecules in response to cytokine stimulation in vitro. Local delivery of ASCs promoted long-term survival and modulated alloimmune responses in a full major histocompatibility complex-mismatched vascularized composite allotransplantation model and was more effective than systemic administration. ASCs are a readily available and abundant source of therapeutic cells that could decrease the amount of systemic immunosuppression required to maintain limb and face allografts.

Abstract 1816: Locoregional delivery with gene edited and engineered stem cells modulates unique immune profiling in leptomeningeal metastasis

Abstract Patients with advanced melanoma have a high propensity to metastasize to the brain and numerous investigational treatments with targeted agents have shown modest response in melanoma brain metastatic patients. Leptomeningeal metastasis (LM) is one of the severe disease conditions in brain metastasis. Recently, phase I/II clinical studies with immune-checkpoint inhibitors (ICIs) are promising however the clinical benefit of patients with LM is limited mainly due to poor penetration of anti-PD-1 antibodies into the brain and cerebrospinal fluid. Therefore, there is an urgent need to develop innovative therapies for such tumors. Oncolytic herpes simplex virus (oHSV) that selectively replicate in tumor cells and illicit an antitumor effect via oncolysis and production of neoantigens are among the recently approved promising therapies for primary melanoma patients. However, virus neutralization, and inefficient extravasation are the major barriers to the effective systemic delivery of oHSV to target metastatic tumor lesions in the brain. We first established and characterized PTEN deficient metastatic melanoma tumor model and show that brain metastatic tumors are more immunosuppressive compared to primary melanoma and therefore mimics the clinical settings. Next, we developed a multimodal approach by creating tumor-tropic allogeneic twin (T) stem cells (SC): one for producing oncolytic herpes simplex virus (oHSV) and the other one engineered with CRISPR/Cas9 mediated knockout of Nectin 1 (N1) receptor (N1KO) to acquire resistance to oHSV and releasing immunomodulators. Utilizing mouse models of brain metastatic PTEN deficient melanomas, we show that locoregional delivery of twin stem cells releasing oHSV and cytokine, GM-CSF (TSC-G) activated dendritic cell and T cell-mediated immune responses in both syngeneic and patient derived-humanized mouse models. Importantly, our strategy exhibited greater therapeutic efficacy when compared with the existing oncolytic viral therapeutic approaches. Moreover, the use of SC-oHSV with SCN1KO releasing both GM-CSF and single-chain variable fragment (scFv) anti-PD-1 (TSC-G/P) effectively suppressed immunosuppressive PTEN deficient LM. Our findings provide a novel allogeneic SC based immunotherapeutic strategy against melanomas and a roadmap towards clinical application particularly for leptomeningeal disease. Citation Format: Nobuhiko Kanaya, Kok Siong Chen, Yoshinori Kajiwara, Waleed Seddiq, Thijs A. van Schaik, Touraj Aligholipour Farzani, Paulo Borges, Shah Khalid. Locoregional delivery with gene edited and engineered stem cells modulates unique immune profiling in leptomeningeal metastasis [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 1816.

PD19-03 THE DELIVERY OF THE RECOMBINANT PROTEIN COCKTAIL IDENTIFIED BY STEM CELL-DERIVED SECRETOME ANALYSIS ACCELERATES KIDNEY REPAIR AFTER RENAL ISCHEMIA-REPERFUSION INJURY

You have accessJournal of UrologyCME1 Apr 2023PD19-03 THE DELIVERY OF THE RECOMBINANT PROTEIN COCKTAIL IDENTIFIED BY STEM CELL-DERIVED SECRETOME ANALYSIS ACCELERATES KIDNEY REPAIR AFTER RENAL ISCHEMIA-REPERFUSION INJURY Ji Hyun Kim, Heejo Yang, Michael Kim, Kang Su Cho, Doo Sang Kim, Hyung Eun Yim, Zachary Atala, John Jackson, In Kap Ko, and James Yoo Ji Hyun KimJi Hyun Kim More articles by this author , Heejo YangHeejo Yang More articles by this author , Michael KimMichael Kim More articles by this author , Kang Su ChoKang Su Cho More articles by this author , Doo Sang KimDoo Sang Kim More articles by this author , Hyung Eun YimHyung Eun Yim More articles by this author , Zachary AtalaZachary Atala More articles by this author , John JacksonJohn Jackson More articles by this author , In Kap KoIn Kap Ko More articles by this author , and James YooJames Yoo More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000003285.03AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: Renal disease is a worldwide health issue. Stem cell therapies have been proposed to restore damaged kidneys as an alternative to renal replacement therapies. In addition to stem cell therapies, recent studies have shown that stem cell-derived secretomes or conditioned media (CM) can enhance tissue regeneration. Our previous study demonstrated that the controlled delivery of human placental stem cells (hPSC)-derived CM using platelet-rich plasma (PRP) improved histological and functional recovery in an acute kidney injury (AKI) model in rats. The protein analysis of hPSC-CM showed the five highly expressed proteins that promote kidney repair. Based on our results, we propose that these proteins could be used as a recombinant protein cocktail to treat kidney diseases as an alternative to using CM. This study investigated the feasibility of delivering the recombinant protein cocktail for kidney repair after AKI. METHODS: The effects of the recombinant protein cocktail on human renal cell survival, proliferation, and apoptosis were compared with the CM in vitro. The feasibility of delivering the recombinant protein cocktail with a PRP system to achieve structural and functional recovery after AKI was investigated. RESULTS: The pro-proliferative and anti-apoptotic effects of the protein cocktail on renal cells were demonstrated in vitro and in vivo. The intrarenal delivery of these proteins with PRP ameliorates the renal tubular damage and improved renal function in the AKI-induced rats, yielding similar therapeutic effects compared to the CM delivery. CONCLUSIONS: The delivery of RPC can support renal cell survival and proliferation in vitro and in vivo, resulting in the attenuation of renal tubular damages and amelioration of renal function in the AKI model in rats. Our strategy may provide a therapeutic solution to many challenges associated with kidney repair resulting from the lack of suitable off-the-shelf regenerative medicine products. Source of Funding: This work was supported by the State of North Carolina © 2023 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 209Issue Supplement 4April 2023Page: e577 Advertisement Copyright & Permissions© 2023 by American Urological Association Education and Research, Inc.MetricsAuthor Information Ji Hyun Kim More articles by this author Heejo Yang More articles by this author Michael Kim More articles by this author Kang Su Cho More articles by this author Doo Sang Kim More articles by this author Hyung Eun Yim More articles by this author Zachary Atala More articles by this author John Jackson More articles by this author In Kap Ko More articles by this author James Yoo More articles by this author Expand All Advertisement PDF downloadLoading ...

Construction, Characterization, and Regenerative Application of Self-Assembled Human Mesenchymal Stem Cell Aggregates.

Mesenchymal stem cells (MSCs), characterized by their self-renewal ability and multilineage differentiation potential, can be derived from various sources and are emerging as promising candidates for regenerative medicine, especially for regeneration of the tooth, bone, cartilage, and skin. The self-assembled approach of MSC aggregation, which notably constructs cell clusters mimicking the developing mesenchymal condensation, allows high-density stem cell delivery along with preserved cell-cell interactions and extracellular matrix (ECM) as the microenvironment niche. This method has been shown to enable efficient cell engraftment and survival, thus promoting the optimized application of exogenous MSCs in tissue engineering and safeguarding clinical organ regeneration. This paper provides a detailed protocol for the construction and characterization of self-assembled aggregates based on umbilical cord mesenchymal stem cells (UCMSCs), as well as an example of the cranial bone regenerative application. The implementation of this procedure will help guide the establishment of an efficient MSC transplantation strategy for tissue engineering and regenerative medicine.

Transamniotic stem cell therapy (TRASCET): An emerging minimally invasive strategy for intrauterine stem cell delivery

Transamniotic stem cell therapy (TRASCET) is an emerging strategy for prenatal stem cell therapy involving the least invasive method described to date of delivering select stem cells to virtually any anatomical site in the fetus, including the blood and bone marrow, as well as to fetal annexes, including the placenta. Such broad therapeutic potential derives, to a large extent, from unique routing patterns following stem cell delivery into the amniotic fluid, which have commonalities with naturally occurring fetal cell kinetics. First reported experimentally only less than a decade ago, TRASCET has yet to be attempted clinically, though a first clinical trial appears imminent. Despite significant experimental advances, much promise and perhaps excessive publicity, most cell-based therapies have yet to deliver meaningful large-scale impact to patient care. The few exceptions typically consist of therapies based on the amplification of the normal biological role played by the given cells in their natural environment. Therein lays much of the appeal of TRASCET, in that it, too, is in essence a magnification of naturally occurring processes in the distinctive environment of the maternal-fetal unit. As much as fetal stem cells possess unique characteristics compared with other stem cells, so does the fetus when compared with any other age group, converging into a scenario that enables therapeutic paradigms exclusive to prenatal life. This review summarizes the diversity of applications and biological responses associated with the TRASCET principle.

ROS filter coating scaffold protects 3D mesenchymal stem cell spheroids for dual-phase treatment of spinal cord injury

Implanting mesenchymal stem cells (MSCs) has offered a great promise in spinal cord injury (SCI) repair. However, the harsh environment accompanied with excessive ROS following SCI seriously threatens MSC survival and compromises its efficacy. Additionally, the intricate pathology distinguished as the acute phase and chronic phase makes MSC therapeutics alone difficult to alleviate severe inflammation and promote functional recovery simultaneously. Despite scaffold systems for local stem cell delivery being extensively explored, dual-phase management for SCI using one protective scaffold system has rarely been reported. Herein, a composite scaffold system (RF-pGel-MS), which consisted of an outer ROS filter (RF) and an inner porous GelMA hydrogel (pGel) harbored with MSC spheroids (MSs), was designed for SCI repair. Specifically, RF rapidly scavenged ROS, protecting the MS from peroxidation damage in the acute phase. MS constructed via 3D culture of MSCs presented enhanced paracrine effects, which remarkably facilitated neuron repair and functional recovery in the chronic phase. The dual-phase therapeutic efficacy of RF-pGel-MS led to significant locomotor and electrophysiology recovery in the SCI rats. Hence, the currently proposed strategy using RF-pGel-MS is efficient for ROS scavenging and boosting the stem cell therapeutics, which is insightful for the SCI treatment.

Microvascular obstruction identifies a subgroup of patients who benefit from stem cell therapy following ST-elevation myocardial infarction

Microvascular obstruction (MVO) is associated with greater infarct size, adverse left-ventricular (LV) remodeling and reduced ejection fraction following ST-elevation myocardial infarction (STEMI). We hypothesized that patients with MVO may constitute a subgroup of patients that would benefit from intracoronary stem cell delivery with bone marrow mononuclear cells (BMCs) given previous findings that BMCs tended to improve LV function only in patients with significant LV dysfunction. We analyzed the cardiac MRIs of 356 patients (303 M, 53 F) with anterior STEMIs who received autologous BMCs or placebo / control as part of 4 randomized clinical trials that included the Cardiovascular Cell Therapy Research Network (CCTRN) TIME trial and its pilot, the multicenter French BONAMI trial and SWISS-AMI trials. A total of 327 patients had paired imaging data at 1 year. All patients received 100 to 150 million intracoronary autologous BMCs or placebo / control 3 to 7 days following primary PCI and stenting. LV function, volumes, infarct size and MVO were assessed prior to infusion of BMCs and 1 year later. Patients with MVO (n = 210) had reduced LVEF and much greater infarct size and LV volumes compared to patients without MVO (n=146) (P < .01). At 12 months, patients with MVO who received BMCs had significantly greater recovery of LVEF compared to those patients with MVO who received placebo (absolute difference=2.7%; P < .05). Similarly, left-ventricular end-diastolic (LVEDVI) and end-systolic volume indices (LVESVI) demonstrated significantly less adverse remodeling in patients with MVO who received BMCs compared to placebo. In contrast, no improvement in LVEF or LV volumes was observed in those patients without MVO who received BMCs compared to placebo. The presence of MVO on cardiac MRI following STEMI identifies a subgroup of patients who benefit from intracoronary stem cell therapy.

Application of stem cell therapy for osteoarthritis pain management.

Osteoarthritis (OA) is a degenerative bone disease that results in inflammation of the joint cartilage. Approximately 58 million adults in the world are affected by this disease, which is characterized by pain, stiffness, and decrease in bone density. The current medication for the treatment of OA is non-steroidal anti-inflammatory drugs (NSAID), opioids, and cyclooxygenase (COX)-2-specific drugs which reduce the pain but do not improve the cartilage disorder. The marketed available drugs for the treatment of OA have gastrointestinal, cardiovascular, and addiction problems. To overcome the limitations of knee OA drug treatment, stem-cell-based therapy is being utilized to recover the knee from the pain and regenerate the cartilage from its recent damaged state. The objective of this study is to effectively manage osteoarthritis by Human Mesenchymal Stem Cell (hMSC) delivery to bone cartilage. In our study, OA in rat knees was produced by intra-articular papain enzyme injections and was observed by pain behavior assay. Latencies in rats treated by papain enzyme were significantly decreased compared to saline. We observed that female rats produced more arthritis by papain enzyme than male rats. The results of the hot plate test showed that the paw-withdrawal threshold was significantly decreased by stem cell administration. We found 52% response by von Frey test in enzyme treated with stem cell delivery osteoarthritis rats, whereas above 80% response was observed in only enzyme treated rats. In this study, we observed that stem cell delivery contributes to the suppression of cartilage degradation in rat knees. Our research demonstrates that stem cell therapy could be a promising treatment in terms of combating tissue regeneration of rat knees to improve the pain state in the target area.

Basic Science of Autologous Orthobiologics: Part 2. Mesenchymal Stem Cells.

In recent years, autologous biological preparations have emerged as a growing area of medical innovation in interventional orthopedical procedures and surgical interventions. These cellular therapies are often referred to as orthobiologics and are derived from patient's own tissues, like blood, bone marrow, and adipose tissue to prepare platelet-rich plasma (PRP), bone marrow concentrate (BMC), and adipose tissue concentrate (ATC), respectively. In this article, we emphasize and discuss the physiologic variability of autologous prepared BMC and ATC for the delivery of mesenchymal stem cells to support tissue repair processes.

Abstract TP243: Efficacy Of Intra-arterial Stem Cell Therapy In A Canine Model Of Ischemic Stroke: Role Of Anti-inflammatory Action In The Brain

Introduction: Stroke is the 2 nd leading cause of death globally. Despite the current standard of care for stroke, more than 50% of stroke survivors are chronically disabled, leading to more than $46 billion in annual economic burden in the US alone. It is therefore imperative to develop novel therapies for stroke. Preclinical studies in the past two decades have shown promise in cell therapies for stroke. Intra-arterial (IA) delivery of Mesenchymal Stem Cells (MSCs) is minimally invasive making it attractive for clinical translation. Recent studies from our lab have shown the safety and efficacy of IA-MSC treatment in rodent and canine ischemic stroke models. These studies from our group have established the precise timing and effective dosage of IA-MSC therapy that leads to improved stroke outcomes. It is believed that IA-MSCs exert their protective effects by mediating anti-inflammatory actions. However, this mechanism is not very well studied, specifically in the large animal brain. Methods: In this study we determined the expression pattern of microglia, astrocytes, the Nod-like receptor 3 (NLRP3) inflammasome pathway markers, inflammatory cytokines and the activity of Nuclear factor kappa B subunit 1 (NF-κB) in the large animal canine brain following IA MSC therapy after ischemic stroke. Immunohistochemistry staining and immunofluorescence imaging of the canine brain cortex and hippocampus were performed. Results: Numerical quantification showed significant reduction in astrocyte and microglia activation, reduced expression of NLRP3 and Interluekin-1beta at different doses of IA-MSCs (10, 20, and 40 million) at 30 days post-stroke. Results from this study showed a significant dose-dependent reduction in inflammation in the canine hippocampus and the cortex. There was a simultaneous increase in surviving neurons. Conclusions: These novel findings help us understand inflammation in the canine brain after stroke. Our data further reveal how IA-MSC therapy mediates its action via inflammatory changes. Overall, this approach contributes to developing more robust and engineered IA-MSCs therapies for stroke patients, leading to more effective translation.

Abstract TP241: Therapeutic Effects Of Magnet-induced Stem Cells Using Ferrimagnetic Iron Oxide Nanoparticles In Ischemic Stroke Model

Background: The efficiency of targeted delivery of stem cells via transplantation by intravenous injection is limited because of rapid clearance. Thus, more effective, newer methods are required. We hypothesized that combining the use of ferrimagnetic iron oxide nanoparticles (FION) labeling and magnetic fields could enhance the targeted delivery of stem cells after ischemic stroke. Methods: In a typical synthesis of FION nanocubes, iron(III) acetylacetonate was added to a mixture of oleic acid, 4-biphenylcarboxylic acid, and benzyl ether. Therapeutic effects of neural stem cells (NSC) using the magnetic nanoparticles in the cerebral ischemic stroke. Transient middle cerebral artery occlusion (MCAO) was induced in SD-rats. Iron oxide nanoparticle-labeled NSCs were injected into the tail vein. Magnetic resonance tracking and confirming labeled stem cells with Prussian blue staining. Polypeptides derived from receptors for advanced glycation end products and pharmaceutical composition for preventing and treating cerebrovascular disease comprise the same. Results: In the F3-VEGF NSC cell, FION uptake was shown as immunofluorescence. After overexpression of VEGF in F3 cells, it was confirmed by Western blotting. The measurements of cytotoxicity in FION's NSC showed toxicity over 80g but 40g was treated for 7 days and was not toxic. Six hours after MCAO induction, 4x10 6 NSCs were injected, and a magnet hat so that the stem cells implanted in the pathogen could move well. After one day, a behavioral test was conducted every week for up to 35 days. After MCAO induction, the group transplanted with FION-NSCs significantly improved in the behavioral test on Day 35 compared to the control group. In order to measure the distribution of FION in tissue, MRI was performed showing penetrating into the lesions of the white circles. As a result of ICP-MS, the magnet-applioed group was concentrated with FION 7 times more compared with the FION-only group. The distribution in brain tissue was observed through FION's Fluorescence images. Conclusions: Our study suggests that this use of a magnetic field may be useful for improving the efficacy of targeted migration of stem cells in stem-based cell therapy in ischemic brain injury.

Synergistic effects of 3D chitosan-based hybrid scaffolds and mesenchymal stem cells in orthopaedic tissue engineering.

Restoration of damaged bone and cartilage tissue with biomaterial scaffolds is an area of interest in orthopaedics. Chitosan is among the low-cost biomaterials used as scaffolds with considerable biocompability to almost every human tissue. Considerable osteoconductivity, porosity, and appropriate pore size distribution have made chitosan an appropriate scaffold for loading of stem cells and a good homing place for differentiation of stem cells to bone tissue. Moreover, the similarity of chitosan to glycosaminoglycans and its potential to be used as soft gels, which could be lasting more than 1week in mobile chondral defects, has made chitosan a polymer of interest in repairing bone and cartilage defects. Different types of scaffolds using chitosan in combination with mesenchymal stem cells (MSCs) are discussed. MSCs are widely used in regenerative medicine because of their regenerative ability, and recent line evidence reviewed demonstrated that the combination of MSCs with a combination of chitosan with different materials, including collagen type 1, hyaluronic acid, Poly(L-lacticacid)/gelatin/β-tricalcium phosphate, gamma-poly[glutamic acid] polyelectrolyte/titanium alloy, modified Poly(L-Lactide-co-Epsilon-Caprolactone), calcium phosphate, β-glycerophosphate hydrogel/calcium phosphate cement (CPC), and CPC-Chitosan-RGD, can increase the efficacy of using MSCs, and chitosan-based stem cell delivery can be a promising method in restoration of damaged bone and cartilage tissue.

Combination therapy with human amniotic epithelial cells and hyaluronic acid promotes immune balance recovery in type 1 diabetic rats through local engraftment.

Stem cell engraftment is currently a promising approach for type 1 diabetes mellitus (T1DM) treatment. In our previous study, engraftment of a combination of human amniotic epithelial cells (hAECs) and hyaluronic acid (HA) showed potent anti-diabetic effect in streptozotocin (STZ)-induced T1DM mice via tail vein injection. Here, we adopted a different route of stem cell delivery, that is via pancreatic subcapsular transplantation. This combined local engraftment of hAECs and HA in STZ-induced T1DM rats showed potent anti-diabetic activity, leading to stronger hypoglycaemia, more intact islet structure and increased number of insulin-positive cells compared with those with hAECs or insulin treatments. Engraftment of hAECs alone increased the proportion of Th1 and T-reg cells and decreased the proportion of Th2 and Th17 cells to protect islet β cells in STZ-induced T1DM rats, whereas the combined engraftment of hAECs and HA showed more potent regulatory capacity, considerably decreased the level of TNF-α and IL-17 and increased the level of TGF-β1 compared with those by other treatments. The potent synergistic effect of HA contributed to the recovery of immune balance in the diabetic rat model, thereby suggesting a new strategy for effective treatment of T1DM.

Isolation, Expansion, and Endoscopic Delivery of Autologous Enteric Neuronal Stem Cells in Swine

The enteric nervous system (ENS) is an extensive network of neurons and glia within the wall of the gastrointestinal (GI) tract that regulates many essential GI functions. Consequently, disorders of the ENS due to developmental defects, inflammation, infection, or age-associated neurodegeneration lead to serious neurointestinal diseases. Despite the prevalence and severity of these diseases, effective treatments are lacking as they fail to directly address the underlying pathology. Neuronal stem cell therapy represents a promising approach to treating diseases of the ENS by replacing the absent or injured neurons, and an autologous source of stem cells would be optimal by obviating the need for immunosuppression. We utilized the swine model to address key questions concerning cell isolation, delivery, engraftment, and fate in a large animal relevant to human therapy. We successfully isolated neural stem cells from a segment of small intestine resected from 1-month-old swine. Enteric neuronal stem cells (ENSCs) were expanded as neurospheres that grew optimally in low-oxygen (5%) culture conditions. Enteric neuronal stem cells were labeled by lentiviral green fluorescent protein (GFP) transduction, then transplanted into the same swine from which they had been harvested. Endoscopic ultrasound was then utilized to deliver the ENSCs (10,000–30,000 neurospheres per animal) into the rectal wall. At 10 and 28 days following injection, autologously derived ENSCs were found to have engrafted within rectal wall, with neuroglial differentiation and no evidence of ectopic spreading. These findings strongly support the feasibility of autologous cell isolation and delivery using a clinically useful and minimally invasive technique, bringing us closer to first-in-human ENSC therapy for neurointestinal diseases.

Systemic delivery of menstrual blood stem cells is more effective in preventing remote organ injuries following myocardial infarction in comparison with bone marrow stem cells in rat.

Remote organ injury is a phenomenon that could happen following myocardial infarction (MI). We evaluated the potency of menstrual blood stromal (stem) cells (MenSCs) and bone marrow stem cells (BMSCs) to alleviate remote organ injuries following MI in rats. 2 × 106 MenSCs or BMSCs were administrated seven days after MI induction via the tail vein. Four weeks after cell therapy, activities of aspartate aminotransferase (AST), urea, creatinine, and Blood Urea Nitrogen (BUN) were evaluated. The level of tumor necrosis factor-α (TNF-α), interleukin (IL)-1β, and IL-6 were determined by ELISA assay. The expression of Nuclear Factor-κB (NF-κB) was evaluated by immunohistochemical staining. Apoptosis activity and tissue damage were also determined by TUNEL and H&E staining, respectively. MenSCs and BMSCs administration caused a significant reduction in AST, urea, and BUN levels compared with the MI group. In addition, systemic injection of MenSCs significantly decreased the IL-1β level compared with BMSCs and MI groups (P<0.05 and P<0.01 respectively). Apoptosis in injured kidneys was noticeably diminished in MenSCs-treated rats compared with BMSCs administrated and MI groups (P<0.05 and P<0.05, respectively). In hepatic tissue, limited numbers of TUNEL-positive cells were detected in all groups. Interestingly, MenSCs therapy evoked inhibition of NF-κB in the kidney strikingly. Although, no significant NF-κB expression was observed in hepatic tissue in any group (P>0.05). MenSCs are probably more protective than BMSCs on remote organ injuries following MI via decreasing cell death and immunoregulatory properties.