Factors Of Mesenchymal Stem Cells Research Articles

Small Extracellular Vesicles Laden Oxygen-Releasing Thermosensitive Hydrogel for Enhanced Antibacterial Therapy against Anaerobe-Induced Periodontitis Alveolar Bone Defect.

Periodontitis is a bacterially induced chronic destructive inflammatory disease that leads to irreversible destruction of the tooth supporting structure, including connective tissue destruction, bone resorption, and even tooth loss. Until now, there has been no effective treatment to repair inflammatory bone loss in periodontitis. Recently, small extracellular vesicles (sEVs) emerged as the essential paracrine factors of mesenchymal stem cells (MSCs) that mediated tissue regeneration. However, limitations of antimicrobial activity associated with the use of sEVs have led to the urgency of new alternative strategies. Currently, we investigated the potential of a biocompatible oxygen-releasing thermosensitive hydrogel laded with sEVs secreted by bone marrow MSCs (BMMSCs) for the alveolar bone defect in periodontitis. The hydrogel composed of different polymers such as chitosan (CS), poloxamer 407 (P407), and cross-linked hyaluronic acid (c-HA) conglomerating is a kind of nanoporous structure material. Then, the gel matrix further encapsulated sEVs and calcium peroxide nanoparticles to realize the control of sEVs and oxygen release. Furthermore, ascorbic acid was added to achieve the REDOX equilibrium and acid-base equilibrium. The experiments in vivo and in vitro proved its good biocompatibility and effectively inhibited the growth of the periodontal main anaerobe, relieved periodontal pocket anaerobic infections, and promoted the periodontal defect regeneration. Therefore, this finding demonstrated that it was a promising approach for combating anaerobic pathogens with enhanced and selective properties in periodontal diseases, even in other bacteria-induced infections, for future clinical application.

The influence of mesenchymal stem cells and its derivatives on the qualitative indicators of animal sperm before and after cryoponservation (review)

During sperm cryopxation, sperm damage occurs due to osmotic stress, which entails deviations into the DNA and deformation of the tail of a sperm. To minimize destructive changes in cells, antioxidants or cryoprotective media supplemented with nutrient components are used. Numerous studies have established that the use of mesenchymal stem cells and their secret increases the percentage of viable and mobile sperm. Also, many of the soluble secretory factors of mesenchymal stem cells can be in extracellular vesicles and act as mediators of many paracrine actions of mesenchymal stem cells. They play the role of vehicles to displace unwanted cellular proteins, but the data of different authors demonstrated their potential role in many biological processes, including contribution to cell regeneration, survival and change in the phenotype of reici cells. The attachment or internalization of microseusiculi in target cells causes a wide range of epigenetic and phenotypic changes in recipient cells, such as a change in the viability of recipient cells, their adhesive properties and sensitivity or resistance to certain environmental factors. Spermeatozoa using microseusicals can be considered as an effective biological approach to maintaining and improving the quality of sperm, in the future, it can improve their viability and progressive mobility after cryopenservation. The wide therapeutic effect of mesenchymal stem cells and biologically active substances secreted by them requires further research to include sperm freezing in the protocols in order to overcome existing restrictions related to their use, and to clarify the exact mechanisms of their action. Further experiments, in particular in vivo, are necessary to confirm the actual clinical effects of these chemicals on the fertilizing ability of a spermatozoa. Thus, in this review, the issue of conducted studies in the field of use of mesenchymal stem cells and the substances they secrete during cryoponservation of animal sperm was covered.

Protective effects of human amniotic membrane derived mesenchymal stem cells (hAMSCs) secreted factors on mouse spermatogenesis and sperm chromatin condensation following unilateral testicular torsion

Testicular torsion is considered a urological disorder that requires immediate detorsion surgery. Ischemia/reperfusion (I/R) injury after testicular torsion detorsion causes of drastic impairment of spermatogenesis and infertility. Cell-free-based approaches seem to be a promising strategy to prevent I/R injury, they have more stable biological properties, and they contain paracrine factors of mesenchymal stem cells. The purpose of this study was to evaluate the protective effects of human amniotic membrane derived mesenchymal stem cells (hAMSCs) secreted factors on mouse sperm chromatin condensation and spermatogenesis improvement after I/R injury. hAMSCs were isolated and characterized by RT- PCR and flow cytometry, preparation of hAMSCs secreted factors was performed. Forty male mice were randomly divided into 4 groups: sham-operated, torsion detorsion, torsion detorsion+ intratesticular injection of DMEM/F-12, and torsion detorsion+ intratesticular injection of hAMSCs secreted factors. After one cycle of spermatogenesis, the mean number of germ cells, Sertoli, Leydig, myoid as well as tubular parameters, Johnson score, and spermatogenesis indexes were evaluated by H& E and PAS stainings. Sperm chromatin condensation and relative expression of c-kit and prm 1 genes were assessed by aniline blue staining and real-time PCR, respectively. The mean number of spermatogenic cells, Leydig, myoid, Sertoli, spermatogenesis parameters, Johnson score, as well as germinal epithelial height and diameters of seminiferous tubules decreased significantly after I/R injury. The thickness of basement membrane and percentage of sperm with excessive histone significantly increased, while the relative expression of c-kit and prm 1 significantly decreased in torsion detorsion group (p 0.001). hAMSCs secreted factors remarkably restored normal sperm chromatin condensation, spermatogenesis parameters and histomorphometric organization of seminiferous tubules via intratesticular injection (p 0.001). Thus, hAMSCs secreted factors may potentially salvage torsion-detorsion-induced infertility.

Co-regulation of Sox9 and TGFβ1 transcription factors in mesenchymal stem cells regenerated the intervertebral disc degeneration.

Intervertebral disc (IVD) shows aging and degenerative changes earlier than any other body connective tissue. Its repair and regeneration provide a considerable challenge in regenerative medicine due to its high degree of infrastructure and mechanical complexity. Mesenchymal stem cells, due to their tissue resurfacing potential, represent many explanatory pathways to regenerate a tissue breakdown. This study was undertaken to evaluate the co-regulation of Sox9 and TGFβ1 in differentiating human umbilical cord mesenchymal stem cells (hUC-MSC) into chondrocytes. The combinatorial impact of Sox9 and TGFβ1 on hUC-MSCs was examined in vitro by gene expression and immunocytochemical staining. In in vivo, an animal model of IVD degeneration was established under a fluoroscopic guided system through needle puncture of the caudal disc. Normal and transfected MSCs were transplanted. Oxidative stress, pain, and inflammatory markers were evaluated by qPCR. Disc height index (DHI), water content, and gag content were analyzed. Histological examinations were performed to evaluate the degree of regeneration. hUC-MSC transfected with Sox9+TGFβ1 showed a noticeable morphological appearance of a chondrocyte, and highly expressed chondrogenic markers (aggrecan, Sox9, TGFβ1, TGFβ2, and type II collagens) after transfection. Histological observation demonstrated that cartilage regeneration, extracellular matrix synthesis, and collagen remodeling were significant upon staining with H&E, Alcian blue, and Masson's trichrome stain on day 14. Additionally, oxidative stress, pain, and inflammatory markers were positively downregulated in the animals transplanted with Sox9 and TGFβ1 transfected MSCs. These findings indicate that the combinatorial effect of Sox9 and TGFβ1 substantially accelerates the chondrogenesis in hUC-MSCs. Cartilage regeneration and matrix synthesis were significantly enhanced. Therefore, a synergistic effect of Sox9 and TGFβ1 could be an immense therapeutic combination in the tissue engineering of cartilaginous joint bio-prostheses and a novel candidate for cartilage stabilization.

Uninterrupted dynamic stiffening microenvironment enhances the paracrine function of mesenchymal stem cells for vascularization through chromatin remodeling

The stiffness of extracellular matrix is dynamically stiffened during bone formation. Paracrine of angiogenic factors in mesenchymal stem cells (MSCs) plays an essential role during tissue regeneration. Whether the paracrine of angiogenic factors in MSCs is influenced by the mechanical microenvironment of materials, especially dynamic matrix mechanics of materials largely remains unknown. A matrix dynamic stiffening material previous designed was used here to explore whether matrix dynamic stiffening could influence paracrine of angiogenic factors in MSCs. The cytoskeleton polymerization, chromatin remodeling and histone acetylation in MSCs were analyzed using phalloidin staining, chromatin condensation parameter analysis and immunofluorescent staining. The results found that the paracrine of MSCs cultured on dynamic stiffening material was higher than that on static stiffness materials. The cytoskeletal polymerization and nuclear deformation degrees of MSCs seeded on dynamic stiffening material were significantly higher than those seeded on materials with static stiffness. The dynamic stiffening substrate could promote the cytoskeletal polymerization to increase the nuclear deformation of MSCs and further induce the paracrine of MSCs. These findings will help to provide clinical guidelines for designing vascularized bone repair materials with appropriate mechanical properties.

Conditioned medium of bone marrow mesenchymal stem cells improves sperm parameters and reduces histological alteration in rat testicular ischaemia/reperfusion model.

Testis ischaemia-reperfusion (I/R) plays a vital role in male infertility. Recent studies have demonstrated that paracrine factors of mesenchymal stem cells exert the transplanted cells' reparative effects. The present experimental study aimed to investigate the effects of conditioned medium (CM) of bone marrow-derived mesenchymal stem cells (BMMSCs). In this study, 21 rats were separated into three groups of 7 animals: sham, I/R and I/R plus CM. Sperm parameters were measured at the end of this study. Moreover, histological parameters were examined. 2-Deoxyuridine 5-triphosphate nick-end labelling (TUNEL) assay was done to assess the apoptotic cells. The count of adhered neutrophils was measured in subtunical venules. Testicular I/R led to a significant reduction in the viability and concentration of sperm and resulted in a significant elevation in the rate of abnormal sperms in comparison with sham. The CM-treated group demonstrated a significant reduction in the rate of abnormal sperm and a significant elevation in the viability and concentration of sperm compared with the I/R group. Based on the morphometric analysis, in the I/R group, epithelial thickness and seminiferous tubule diameter significantly decreased in comparison with sham. A significant reduction was seen between the I/R and sham groups regarding the mean testicular biopsy score (MTBS) value. However, an improvement was observed in the I/R + CM group MTBS value in comparison with the I/R group. TUNEL assay showed that the apoptotic cells in the seminiferous tubules belonging to the I/R group were significantly higher compared with the control. Nevertheless, apoptotic cells were reduced in the I/R + CM group compared with the I/R group. Results of the present study showed that CM of BMMSCs exerts protective effects on the testicular I/R damages.

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

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

Treatment of inflammatory bone loss in periodontitis by stem cell-derived exosomes

Periodontitis is the primary cause of tooth loss, but there is no effective treatment to repair inflammatory bone loss in periodontitis. Exosomes emerge as essential paracrine factors of mesenchymal stem cells (MSCs) that mediated tissue regeneration. Here, we investigated the potential of exosomes secreted by periodontal ligament stem cells (PDLSCs) as therapeutics for the bone defect in periodontitis. Exosomes secreted from PDLSCs derived from healthy periodontal ligaments (h-PDLSCs) and their function were evaluated on PDLSCs isolated from the inflammatory periodontal ligament of periodontitis patients (i-PDLSCs). Treatment of exosomes of h-PDLSCs led to an increase in the formation of mineralized nodules and the expressions of osteogenic genes and proteins in i-PDLSCs. Mechanistically, h-PDLSCs-exosomes suppressed the over-activation of canonical Wnt signaling to recover the osteogenic differentiation capacity of i-PDLSCs. To evaluate the therapeutic of exosomes on inflammatory bone loss, h-PDLSCs-exosomes loaded with Matrigel or β-TCP were employed to repair bone defects in rat models of periodontitis. Compared to the vehicle-treated control group, h-PDLSCs-exosomes-treated rats resulted in more bone formation in the defect of alveolar bone. In conclusion, these results demonstrated that exosomes derived from healthy PDLSCs could rescue the osteogenesis capacity of endogenous stem cells under an inflammatory environment and promote regeneration of alveolar bone. Our findings suggest that MSCs-derived exosome is an effective and practical cell-free MSC therapeutic for the treatment of periodontitis. Statement of significanceThere is no effective treatment to repair inflammatory bone loss in periodontitis. As essential paracrine factors of PDLSCs, exosomes might mediate tissue regeneration during stem cell therapy. Here, we reported that exosomes secreted from healthy PDLSCs promoted the osteogenic differentiation of PDLSCs derived from periodontitis tissue. Healthy PDLSCs-exosomes treatment resulted in accelerated bone formation in the defect of alveolar bone in rat models of periodontitis. Mechanistically, h-PDLSCs-exosomes suppressed the over-activation of canonical Wnt signaling to recover the osteogenic differentiation capacity of inflammatory PDLSCs. These findings suggest that MSCs-derived exosome is an effective and practical cell-free MSC therapeutic for the treatment of periodontitis.

Expression of reprogramming factors in mesenchymal stem cells isolated from equine umbilical cord Wharton’s jelly and amniotic fluid

Stem cells represent the most promising population for regenerative cell therapy and have gained much attention during the recent past. Mesenchymal stem cells (MSCs) are multipotent stem cells that can differentiate into trilineages. Like haematopoietic cells, mesenchymal cells have been shown to proliferate and form fibroblast-like colonies in vitro. Despite major progress in our general knowledge related to the application of adult stem cells, finding alternative sources for bone marrow MSCs has remained a challenge. A wide diversity of isolation procedures for mesenchymal stromal cells from various tissues of the placenta, umbilical cord and Wharton's jelly have been described for humans and other species. In this study, we isolated established umbilical cord Wharton’s jelly as a primary source for isolation of mesenchymal stem cells since it is a rich source of stem cells and no ethical concerns are involved. Equine umbilical cord Wharton’s jelly segments were collected during foaling time and digested enzymatically and cultured in-vitro in culture medium. In addition to the study of their morphology and colony forming units, the expression of reprogramming factors by the isolated MSCs were also studied. The isolated MSCs were observed to be plastic adherent, clonogenic and their morphology were polygonal, star shaped and fibroblast like. They revealed a strong expression of pluripotent stemness markers OCT-4, SOX-2, Nanog and KLF-4. From the current study, it can be concluded that Wharton's jelly is a rich source of stem cells with stemness properties expressing the reprogramming factors and mesenchymal like morphology and could be used as an alternate for the bone marrow derived mesenchymal stem cells for cell based regenerative therapies.

Treatment Of Inflammatory Bone Loss in Periodontitis By Stem Cell-Derived Exosomes

Periodontitis is the primary cause of tooth loss, but there is no effective treatment to repair inflammatory bone loss in periodontitis. Exosomes emerge as essential paracrine factors of mesenchymal stem cells (MSCs) that mediated tissue regeneration. Here, we investigated the potential of exosomes secreted by periodontal ligament stem cells (PDLSCs) as therapeutics for the bone defect in periodontitis. Exosomes secreted from PDLSCs derived from healthy periodontal ligaments (h-PDLSCs) and their function were evaluated on PDLSCs isolated from the inflammatory periodontal ligament of periodontitis patients (i-PDLSCs). Treatment of exosomes of h-PDLSCs led to an increase in the formation of mineralized nodules and the expressions of osteogenic genes and proteins in i-PDLSCs. Mechanistically, h-PDLSCs-exosomes suppressed the over-activation of canonical Wnt signaling to recover the osteogenic differentiation capacity of i-PDLSCs. To evaluate the therapeutic of exosomes on inflammatory bone loss, h-PDLSCs-exosomes loaded with Matrigel or β-TCP were employed to repair bone defects in rat models of periodontitis. Compared to the vehicle-treated control group, h-PDLSCs-exosomes-treated rats resulted in more bone formation in the defect of alveolar bone. In conclusion, these results demonstrated that exosomes derived from healthy PDLSCs could rescue the osteogenesis capacity of endogenous stem cells under an inflammatory environment and promote regeneration of alveolar bone. Our findings suggest that MSCs-derived exosome is an effective and practical cell-free MSC therapeutic for the treatment of periodontitis.

Study of bilateral elbow joint osteoarthritis treatment using conditioned medium from allogeneic adipose tissue-derived MSCs in Labrador retrievers

Canine elbow dysplasia is a common cause of forelimb lameness in dogs and can lead to development of osteoarthritis (OA). A potential alternative to pain management is the use of a safe cell-free based therapy through trophic and paracrine factors of mesenchymal stem cells (MSCs). The aim of study was to identify the profile of selected mediators of potential clinical relevance in synovial fluid (SF) samples of dogs with elbow OA and analyse the range of motion (ROM) before and after cell-free MSCs-based treatment. In this study, conditioned medium from allogeneic canine adipose tissue - derived MSC (CM-AD-MSC) was prepared and administered into both elbow joints with OA in six Labrador retriever dogs (n = 6) on day 0 and 14 without creating a control group with a placebo. The SF of the elbow joints was analysed for the presence of several biomolecules (IL-6, IL-10, IL-8, IL-2, IL-12, TNF-αIFN-γ, MMP-3TIMP-1) before and after intraarticular applications of CM-AD-MSC. Kinematic analysis was used to assess the clinical effect of CM-AD-MSC. Analyses of SF and ROM were performed on days 0, 14 and 42. Concentration levels of MMP-3, TIMP-1, IL-6 and TNF-α in SF showed significant differences before and after the treatment (P < .05). There was a significant improvement in ROM between day 0 and 42 (P < .001). No severe adverse events were observed during the study. Results support the potential supportive effect of CM-AD-MSC as a noninvasive therapeutic tool for pain management of OA elbow joints in dogs.

Condensation of feline adipose tissue-derived mesenchymal stem cells by electrical stimulation

Background & Aim There are many studies that induce differentiation into cartilage cells by treating the growth factors in mesenchymal stem cells (MSCs) or alter the properties of MSCs by genetic modification. However, studies on cartilage differentiation are still insufficient. Recently, many studies have been reported that stem cells can differentiate into various cells by electrical stimulation (ES). An important step in the process of cartilage differentiation is pre-chondrogenic condensation, which is the stage where cartilage formation begins by condensation of stem cells and cartilage cells in the microenvironment of the joint. Methods, Results & Conclusion ES was applied to feline adipose tissue-derived MSCs (fAD-MSCs) using an electrical stimulator, which is a multi-channel stimulator designed for chronic stimulation of bulk quantities of cells in culture. This instrument emits bipolar pulses to culture media immersed carbon electrodes of a 35 mm dishes. ES was applied to fAD-MSCs cultured under conditions of high-density micro-mass (2 × 105 cells/10 μL) under ES of 1∼20 V/cm, with a duration of 8 ms and a frequency of 5.0 Hz. At indicated time points, we observed size of micro-mass, density and shapes. To induce cell to cell communication, make fAD-MSCs into micromass (2 × 105 cells/10 μL), a mass of cells in a small space, and induce condensation through ES. Condensation behaviors of fAD-MSCs in micro-mass culture were examined using phase contrast images during culture for 3 days under ES. Consistently, micro-mass caused compact condensation by ES at specific condition. Time-course observations of condensation behaviors of fAD-MSCs in micro-mass culture were examined with phase contrast images during culture for 3 days in maintenance medium only (control), and maintenance medium under ES, respectively. As a result, time-dependent observations showed that ES induced gradual aggregation of fAD-MSCs into compact structures within 3 days. This study might contribute to the differentiation of MSCs cells into cartilage cells by inducing pre-chondrogenic condensation under specific ES.

Injectable Therapeutic Organoids Using Sacrificial Hydrogels

SummaryOrganoids are becoming widespread in drug-screening technologies but have been used sparingly for cell therapy as current approaches for producing self-organized cell clusters lack scalability or reproducibility in size and cellular organization. We introduce a method of using hydrogels as sacrificial scaffolds, which allow cells to form self-organized clusters followed by gentle release, resulting in highly reproducible multicellular structures on a large scale. We demonstrated this strategy for endothelial cells and mesenchymal stem cells to self-organize into blood-vessel units, which were injected into mice, and rapidly formed perfusing vasculature. Moreover, in a mouse model of peripheral artery disease, intramuscular injections of blood-vessel units resulted in rapid restoration of vascular perfusion within seven days. As cell therapy transforms into a new class of therapeutic modality, this simple method—by making use of the dynamic nature of hydrogels—could offer high yields of self-organized multicellular aggregates with reproducible sizes and cellular architectures.

3D Printed Hydroxyapatite/Silk Fibroin/Polycaprolactone Artificial Bone Scaffold and Bone Tissue Engineering Materials Constructed with Double-Transfected Bone Morphogenetic Protein-2 and Vascular Endothelial Growth Factor Mesenchymal Stem Cells to Repair Rabbit Radial Bone Defects

To examine the effect of bone tissue engineering material constructed via 3D printed nanohydroxyapatite/silk fibroin/polycaprolactone (nHA/SF/PCL) artificial bone scaffolds and doubletransfected BMP-2/VEGF mesenchymal stem cells in repairing rabbit radial bone defects, 60 New Zealand rabbits were randomly selected, and with the establishment of a 15 mm bone defect model upon the middle of the anterior right radius bone. The rabbits were randomly divided into 4 groups, with each group composed of 15 rabbits: Group A (nHA/SF/PCL BMP-2/VEGF), Group B (simple nHA/SF/PCL), Group C (simple BMP-2/VEGF), Group D (blank control). Our team recorded the eating, movement, redness, exudation, and so forth across each sample group following the operation. The bone protein expression levels were measured via western blotting following 12 weeks post-operation. Histological observation was carried performed via tissue sections. Following the operation procedure, the wound healed healthily across the 4 experimental groups, no serious infections occurred, only a few rabbits exhibited redness, exudation, pus, and other phenomena, and there was no significant difference present in each group (P &gt; 0.05). The A group took the shortest time to recover its autonomous activity ability, followed by the B group, and the D group took the longest time to recover its autonomous activity ability. From western blotting tests, the A group possessed the highest BMP-2 and OCN expression levels, and have significant difference existed with the B, C, and D group (P &lt; 0.05). The order of expression level of BMP-2 and OCN across the other three groups were group C &gt; group B &gt; group D, respectively. The results of a histological section performed at 12 weeks post-operation revealed that there was no immune response present and no adverse inflammatory reaction occurring within each experimental group. Among the rabbits, the A group exhibited an apparent plate layer new bone formation, bone trabeculae were arranged regularly, and there was no obvious space within the new bone tissue; furthermore, the repair of the bone defect was completed. The B group still exhibited some bone scaffold remaining within the implantation area, and there was a small gap between the new bone tissue, and several bone defects had not been repaired. The C group of bone defects presented only a small number of new bone formation, and bone defect repair effect was not obvious. The D group still possessed large bone defects and exhibited poor repair of bone defects. The nHA/SF/PCL composite of artificial bone scaffold and double-transfected BMP-2/VEGF mesenchymal stem cells was effective in repairing radial bone defects within rabbits, and was superior to using nHA/SF/PCL bone scaffolds alone.

Optimization system of mesenchymal stem cells for tissue engineering chondrogenic differentiation

Articular cartilage defect is a common joint damage, however, due to the particularity of cartilage tissue, its repair ability is limited. Thus, articular cartilage damage often changes to more severe osteoarthritis which brings huge pain and economic burden to patients. The development of cartilage tissue engineering brings good news to these patients, but also faces great challenges. Because of the characteristics of mesenchymal stem cells(MSCs) including autologous source, amplification and cartilage differentiation potential, MSCs are often used as the seed cells of cartilage tissue engineering and got widely attention. In the commonly used treatment strategy of articular cartilage differentiated from mesenchymal cells, in vitro pre culture and differentiation showed good therapeutic effects.The mesenchymal stem cells collected from the patient were concentrated, cultured and induced differentiation in vitro, a certain number of differentiated chondrocytes were obtained, and then the cells and culture matrix were transplanted to the patient together. However, the tissue engineering cartilage obtained from mesenchymal stem cells cannot fully meet the requirements of clinical treatment. Moreover, due to the differences of disease type, degree and individuality, there are various optimized factors for the treatment of MSCs derived tissue-engineered cartilage transplantation. For the same type of disease treatment, the optimization system still has no unified recognized standard. In order to obtain better adapt to the human body and meet the clinical requirements of articular cartilage, this review focus onoptimized factors of MSCs in the treatment of orthopedic diseases, summarize and analysis the research status, and discuss the induced factorsfrom three aspects: environmental factors, scaffold selection and seed cells.It provides an idea for using mesenchymal stem cells to obtain better tissue-engineered cartilage and to establish a better optimization system.

Resveratrol promotes secretion of wound healing related growth factors of mesenchymal stem cells originated from adult and fetal tissues

Non-healing wounds have demonstrated aberrant regulation of several growth factors, thus using exogenous growth factors and cytokines in the clinical setting may improve the outcomes of non-healing wounds. Mesenchymal stem cells (MSCs) are the source of growth factors that show beneficial effect in promoting impaired wound healing. Certain culture condition should be developed to stimulate growth factor secretion from stem cell. Resveratrol, a small molecule found to increase MSCs therapeutic effectiveness. This study aims to investigate the effect of RV on secretion of wound healing related growth factors. We isolated and characterised MSCs from wharton’s jelly (WJ), amniotic membrane (AM), and adipose tissue. We treated MSCs with serum deprived medium (SDM) supplemented with RV at 0.1 µM, 0.5 µM, 0.8 µM concentration. Our study revealed that RV at 0.1 µM was more effective to increase cell proliferation rate. Resveratrol at 0.1 µM promoted EGF, HGF, PDGF, and TGF-β1 secretion from MSCs. AD-MSCs showed the greatest response to RV stimulation in the term of cell proliferation and growth factors secretion. As conclusion, RV can facilitate cell proliferation and wound healing related growth factors secretion at dosage dependent manner.

Tendon ECM modified bioactive electrospun fibers promote MSC tenogenic differentiation and tendon regeneration

Proper niche environment is the key factor for mesenchymal stem cells (MSCs) involved tendon-lineage differentiation and regeneration. In the present study, soluble tendon derived extracellular matrix (sTECM) was used to modify aligned electrospun fibers with acid neutralizing effect in order to create an optimal niche environment for tendon regeneration, and its efficacy was evaluated using mouse MSC tenogenic differentiation in vitro and rat Achilles tendon regeneration in vivo. The results showed that sTECM modified ultrafine fibers were more cytocompatible and capable of inducing a predominant tenogenic phenotype of mouse MSCs with the enhanced expression of tendon markers including scleraxis, tenomodulin, collagen III, mohawk homeobox, decorin, fibromodulin and biglycan. By contrast, pure sTECM components led to enhanced multi-lineage differentiation of mouse MSCs including tenogenic lineage. This inductive effect was further enhanced when unilateral mechanical loading was applied in vivo. Moreover, the implantation of sTECM modified scaffold created an ideal niche environment including bioactive sTECM components, topological and mechanical signals, upon in vivo implantation for in situ tendon regeneration. As the results, sTECM modified scaffold in comparison with the non-modified one regenerated higher quality tendon tissue with the features of more mature tissue structure, better tissue organization, cell density and alignment as well as better collagen ultrastructure. Quantitative analysis demonstrated significantly bigger collagen fibril diameter, stronger mechanical property and better tissue grading score and higher expression levels of tenogenic markers when compared to those of control scaffold (p < 0.05). This study demonstrated that sTECM modified pH-neutral ultrafine fibers may represent a novel bioactive scaffold for in situ tendon regeneration, which deserves further investigation in large animals.

[Corrigendum] All-trans retinoic acid restored the osteogenic ability of BMP9 in osteosarcoma through the p38 MAPK pathway.

Subsequently to the publication of the above paper, the authors have realized that the images presented in Fig.1A were selected erroneously (essentially, the images for group 'AdBMP9+++' were chosen to represent the group 'AdGFP'). A corrected version of Fig.1, including the correct data for the experiments depicted in Fig.1A, is shown opposite. Note that this change does not affect the results or the conclusions reported in this paper, and all the authors agree to this correction. The authors apologize to the Editor and to the readership of the Journal for any inconvenience caused. [the original article was published in International Journal of Oncology 50: 1363‑1371, 2017; DOI: 10.3892/ijo.2017.3910].

Isolating, Sequencing and Analyzing Extracellular MicroRNAs from Human Mesenchymal Stem Cells.

Extracellular and circulating RNAs (exRNA) are produced by many cell types of the body and exist in numerous bodily fluids such as saliva, plasma, serum, milk and urine. One subset of these RNAs are the posttranscriptional regulators - microRNAs (miRNAs). To delineate the miRNAs produced by specific cell types, in vitro culture systems can be used to harvest and profile exRNAs derived from one subset of cells. The secreted factors of mesenchymal stem cells are implicated in alleviating numerous diseases and is used as the in vitro model system here. This paper describes the process of collection, purification of small RNA and library generation to sequence extracellular miRNAs. ExRNAs from culture media differ from cellular RNA by being low RNA input samples, which calls for optimized procedures. This protocol provides a comprehensive guide to small exRNA sequencing from culture media, showing quality control checkpoints at each step during exRNA purification and sequencing.

RETRACTED: Inflammatory-Related P62 Triggers Malignant Transformation of Mesenchymal Stem Cells through the Cascade of CUDR-CTCF-IGFII-RAS Signaling

Inflammatory and autophagy-related gene P62 is highly expressed in most human tumor tissues. Herein, we demonstrate that P62 promotes human mesenchymal stem cells’ malignant transformation via the cascade of P62-tumor necrosis factor alpha (TNF-α)-CUDR-CTCF-insulin growth factor II (IGFII)-H-Ras signaling. Mechanistically, we reveal P62 enhances IGFII transcriptional activity through forming IGFII promoter-enhancer chromatin loop and increasing METTL3 occupancy on IGFII 3′ UTR and enhances H-Ras overexpression by harboring inflammation-related factors, e.g., TNFR1, CLYD, EGR1, NFκB, TLR4, and PPARγ. Furthermore, the P62 cooperates with TNF-α to promote malignant transformation of mesenchymal stem cells. These findings, for the first time, provide insight into the positive role that P62 plays in malignant transformation of mesenchymal stem cells and reveal a novel link between P62 and the inflammation factors in mesenchymal stem cells.