• bone
• bone marrow mesenchymal stem cells
• bone marrow mesenchymal stromal cells
• cartilage
• extracellular matrix
• human
• regenerative medicine
• stem cells
• tissue engineering

• Humans
• Mesenchymal Stem Cells/metabolism
• Extracellular Matrix/metabolism
• CRISPR-Cas Systems
• Gene Editing/methods
• Animals
• Rats
• Tissue Engineering/methods
• Core Binding Factor Alpha 1 Subunit/metabolism
• Core Binding Factor Alpha 1 Subunit/genetics
• Cell Line
• Vascular Endothelial Growth Factor A/metabolism
• Vascular Endothelial Growth Factor A/genetics

• 2019-01864_3 Vetenskapsrådet
• 948588 European Research Council
• IngaBritt och Arne Lundbergs Forskningsstiftelse
• Knut and Alice Wallenberg Foundation
• Lund University Medical Faculty Foundation
• Region Skåne

• Exosomes
• Genomic engineering
• IVD
• Proteomics
• Regeneration
• Transcriptomic modulation

• National Institute of Arthritis and Musculoskeletal and Skin Diseases
• Paracelsus Medical University

• cancer progression
• cancer stem cells
• cell differentiation
• epithelial–mesenchymal transition (EMT)
• human stem cells
• lipid nanoparticles
• microRNA-181 (miR-181)
• nanotechnology
• therapeutic strategies

• Humans
• MicroRNAs/genetics
• MicroRNAs/metabolism
• Neoplastic Stem Cells/metabolism
• Neoplastic Stem Cells/pathology
• Cell Differentiation/genetics
• Epithelial-Mesenchymal Transition/genetics
• Cell Plasticity/genetics
• Animals
• Neoplasms/genetics
• Neoplasms/pathology
• Neoplasms/metabolism
• Signal Transduction

• 426236 CIHR

• ADSC-exo
• Exosomes
• MSCs
• Oral diseases
• Oral submucous fibrosis

• Cartilage regeneration
• Chondrocytes
• Induced pluripotent stem cells
• Mesenchymal stem cells
• Osteoarthritis
• Personalized regenerative medicine

• Humans
• Induced Pluripotent Stem Cells/cytology
• Induced Pluripotent Stem Cells/metabolism
• Regenerative Medicine/methods
• Cell Differentiation
• Chondrogenesis
• Precision Medicine/methods
• Chondrocytes/cytology
• Chondrocytes/metabolism
• Animals

• Heterogeneity
• Immunosuppression
• Mesenchymal stromal cells
• Pooling
• Secretome

• Mesenchymal Stem Cells/cytology
• Mesenchymal Stem Cells/metabolism
• Humans
• Bone Marrow Cells/cytology
• Bone Marrow Cells/metabolism
• Tissue Donors
• Cell Differentiation
• Cryopreservation/methods
• Cell Proliferation
• Cells, Cultured
• Adult
• Cell Culture Techniques/methods

• concentration
• heterogeneity
• mesenchymal stem cells
• phenotype
• tissue source

• L3-3176 Slovenian Research Agency

• mesenchymal stromal cells
• osteogenesis
• porosity
• regenerative scaffold
• solvent-based printing

• RNA-seq
• cell therapy
• iMSCs
• miR-arrays
• osteoarthritis

• Humans
• Transcriptome/genetics
• MicroRNAs/metabolism
• Cell Differentiation/genetics
• Cell- and Tissue-Based Therapy
• Osteoarthritis/genetics
• Osteoarthritis/therapy

• T1EDK-000128 Operational Program Competitiveness, Entrepreneurship and Innovation, under the call RE-SEARCH - CREATE - INNOVATE

• CRISPLD2
• hASCs
• hBMSCs
• hDPSCs
• osteogenic differentiation

• Humans
• Osteogenesis/genetics
• Mesenchymal Stem Cells/metabolism
• Cell Differentiation/genetics
• RNA, Small Interfering/metabolism
• Bone Diseases/metabolism
• Cells, Cultured
• Forkhead Transcription Factors/metabolism
• Cell Adhesion Molecules/genetics
• Cell Adhesion Molecules/metabolism
• Interferon Regulatory Factors/metabolism

• R21 AR076583 NIAMS NIH HHS

• Rho GTPase signaling
• actomyosin contraction
• bioreactor
• bone tissue engineering
• fluid shear stress
• osteogenic differentiation

• BB/E01335X/1 Biotechnology and Biological Sciences Research Council
• BB/L002965/1 Biotechnology and Biological Sciences Research Council
• L. Meltzers Høyskolefond
• Trond Mohn stiftelse

• Cell therapy
• Mesenchymal stem cells
• Osteogenesis imperfecta
• Transplantation

• Biomarker correlation
• Bone formation
• Bone marrow
• Donor heterogeneity
• Functional attribute
• Global microRNA profiling
• Osteogenic differentiation
• Osteogenic potency assay
• Proteomic
• Telomerised MSC

• animal component-free media
• foetal bovine serum
• macromolecular crowding
• mesenchymal stromal cell differentiation
• mesenchymal stromal cell expansion
• xeno-free media

• Science Foundation Ireland
• European Research Council
• H2020 Marie Skłodowska-Curie Actions

Chronic wound healing has long been an unmet medical need in the field of wound repair, with diabetes being one of the major etiologies. Diabetic chronic wounds (DCWs), especially diabetic foot ulcers, are one of the most threatening chronic complications of diabetes. Although the treatment strategies, drugs, and dressings for DCWs have made great progress, they remain ineffective in some patients with refractory wounds. Stem cell-based therapies have achieved specific efficacy in various fields, with mesenchymal stem cells (MSCs) being the most widely used. Although MSCs have achieved good feedback in preclinical studies and clinical trials in the treatment of cutaneous wounds or other situations, the potential safety concerns associated with allogeneic/autologous stem cells and unknown long-term health effects need further attention and supervision. Recent studies have reported that stem cells mainly exert their trauma repair effects through paracrine secretion, and exosomes play an important role in intercellular communication as their main bioactive component. MSC-derived exosomes (MSC-Exos) inherit the powerful inflammation and immune modulation, angiogenesis, cell proliferation and migration promotion, oxidative stress alleviation, collagen remodeling imbalances regulation of their parental cells, and can avoid the potential risks of direct stem cell transplantation to a large extent, thus demonstrating promising performance as novel "cell-free" therapies in chronic wounds. This review aimed to elucidate the potential mechanism and update the progress of MSC-Exos in DCW healing, thereby providing new therapeutic directions for DCWs that are difficult to be cured using conventional therapy.

• Diabetic wounds
• Wound and injuries
• Mesenchymal stem cells
• Exosomes
• Pre-conditioning
• Preclinical translation

• stem-cell biotechnology
• stem-cell research
• predictive medicine
• machine learning

• atrophic alveolar bone
• bone marrow
• bone tissue engineering
• clinical study
• mesenchymal stem cells
• mesenchymal stromal cells

• adipose stem cell
• catalase
• dedifferentiated fat cell
• osteogenesis
• oxidative stress
• reactive oxygen species

• Exosome
• mesenchymal stem cell
• optimization in bioengineering
• temporomandibular joint osteoarthritis
• therapeutic strategy

• Bioactive glass nanoparticles
• Collagen coatings
• Electric field
• Human mesenchymal stem cells
• Osteogenic differentiation
• Stimulation regimes
• Transformer-like coupling

• Cell Differentiation
• Cells, Cultured
• Collagen/pharmacology
• Dexamethasone/pharmacology
• Electric Stimulation
• Humans
• Mechanotransduction, Cellular
• Mesenchymal Stem Cells
• Nanoparticles
• Osteogenesis

• Biomarkers
• Cartilage matrix
• Cytokines
• Dexamethasone
• Mass spectrometry
• Post-traumatic osteoarthritis
• Proteomics

• Adult
• Aged
• Bayes Theorem
• Biomarkers/metabolism
• Cartilage, Articular/metabolism
• Cytokines/metabolism
• Dexamethasone/pharmacology
• Female
• Humans
• Male
• Middle Aged
• Osteoarthritis/drug therapy
• Osteoarthritis/metabolism
• Proteomics
• Young Adult

• UG3 TR002186 NCATS NIH HHS
• UH3 TR002186 NCATS NIH HHS

• stem cells
• mesenchymal stem cells
• pluripotent stem cells

• Biomarkers/metabolism
• Cell Differentiation
• Humans
• Hydrogels/metabolism
• Induced Pluripotent Stem Cells
• Mesenchymal Stem Cells

• New York Stem Cell Foundation
• Ralph and Ricky Lauren Family Foundation
• National Science Foundation

Mesenchymal stem cells (MSCs) are effective therapeutic agents that contribute to tissue repair and regeneration by secreting various factors. However, donor-dependent variations in MSC proliferation and therapeutic potentials result in variable production yields and clinical outcomes, thereby impeding MSC-based therapies. Hence, selection of MSCs with high proliferation and therapeutic potentials would be important for effective clinical application of MSCs. This study is aimed at identifying the upregulated genes in human Wharton's jelly-derived MSCs (WJ-MSCs) with high proliferation potential using mRNA sequencing. Aurora kinase A (AURKA) and dedicator of cytokinesis 2 (DOCK2) were selected as the upregulated genes, and their effects on proliferation, migration, and colony formation of the WJ-MSCs were verified using small interfering RNA (siRNA) techniques. mRNA expression levels of both the genes were positively correlated with the proliferation capacity of WJ-MSCs. Moreover, AURKA from human WJ-MSCs regulated the antiapoptotic effect of skeletal muscle cells by upregulating the chemokine (C motif) ligand (XCL1); this was further confirmed in the mdx mouse model. Taken together, the results indicated that AURKA and DOCK2 can be used as potential biomarkers for proliferation and migration of human WJ-MSCs. In particular, human WJ-MSCs with high expression of AURKA might have therapeutic efficacy against muscle diseases, such as Duchenne muscular dystrophy (DMD).

Impaired wound healing can lead to local hypoxia or tissue necrosis and ultimately result in amputation or even death. Various factors can influence the wound healing environment, including bacterial or fungal infections, different disease states, desiccation, edema, and even systemic viral infections such as COVID-19. Silk fibroin, the fibrous structural-protein component in silk, has emerged as a promising treatment for these impaired processes by promoting functional tissue regeneration. Silk fibroin’s dynamic properties allow for customizable nanoarchitectures, which can be tailored for effectively treating several wound healing impairments. Different forms of silk fibroin include nanoparticles, biosensors, tissue scaffolds, wound dressings, and novel drug-delivery systems. Silk fibroin can be combined with other biomaterials, such as chitosan or microRNA-bound cerium oxide nanoparticles (CNP), to have a synergistic effect on improving impaired wound healing. This review focuses on the different applications of silk-fibroin-based nanotechnology in improving the wound healing process; here we discuss silk fibroin as a tissue scaffold, topical solution, biosensor, and nanoparticle.

Mesenchymal stem cells (MSCs) can differentiate into endoderm lineages, especially parathyroid-hormone (PTH)-releasing cells. We have previously reported that tonsil-derived MSC (T-MSC) can differentiate into PTH-releasing cells (T-MSC-PTHCs), which restored the parathyroid functions in parathyroidectomy (PTX) rats. In this study, we demonstrate quality optimization by standardizing the differentiation rate for a better clinical application of T-MSC-PTHCs to overcome donor-dependent variation of T-MSCs. Quantitation results of PTH mRNA copy number in the differentiated cells and the PTH concentration in the conditioned medium confirmed that the differentiation efficiency largely varied depending on the cells from each donor. In addition, the differentiation rate of the cells from all the donors greatly improved when differentiation was started at a high cell density (100% confluence). The large-scale expression profiling of T-MSC-PTHCs by RNA sequencing indicated that those genes involved in exiting the differentiation and the cell cycle were the major pathways for the differentiation of T-MSC-PTHCs. Furthermore, the implantation of the T-MSC-PTHCs, which were differentiated at a high cell density embedded in hyaluronic acid, resulted in a higher serum PTH in the PTX model. This standardized efficiency of differentiation into PTHC was achieved by initiating differentiation at a high cell density. Our findings provide a potential solution to overcome the limitations due to donor-dependent variation by establishing a standardized differentiation protocol for the clinical application of T-MSC therapy in treating hypoparathyroidism.

• contact inhibition
• differentiation
• donor-dependent variation
• parathyroid-hormone-releasing cells
• tonsil-derived mesenchymal stem cells

• Biomarkers
• Calcium/metabolism
• Cell Culture Techniques
• Cell Differentiation
• Cells, Cultured
• Contact Inhibition
• Extracellular Space/metabolism
• Gene Expression Profiling
• Gene Regulatory Networks
• High-Throughput Nucleotide Sequencing
• Humans
• Mesenchymal Stem Cells/cytology
• Mesenchymal Stem Cells/metabolism
• Palatine Tonsil/cytology
• Parathyroid Hormone/biosynthesis

We investigated whether irradiated brain-derived neurotropic factor (BDNF)-overexpressing engineered human mesenchymal stem cells (BDNF-eMSCs) improve paracrine efficiency and, thus, the beneficial potency of naïve MSCs against severe hypoxic ischemic (HI) brain injury in newborn rats. Irradiated BDNF-eMSCs hyper-secreted BDNF > 10 fold and were >5 fold more effective than naïve MSCs in attenuating the oxygen-glucose deprivation-induced increase in cytotoxicity, oxidative stress, and cell death in vitro. Only the irradiated BDNF-eMSCs, but not naïve MSCs, showed significant attenuating effects on severe neonatal HI-induced short-term brain injury scores, long-term progress of brain infarct, increased apoptotic cell death, astrogliosis and inflammatory responses, and impaired negative geotaxis and rotarod tests in vivo. Our data, showing better paracrine potency and the resultant better therapeutic efficacy of the irradiated BDNF-eMSCs, compared to naïve MSCs, suggest that MSCs transfected with the BDNF gene might represent a better, new therapeutic strategy against severe neonatal HI brain injury.

• brain
• brain derived neurotropic factor
• cell transplantation
• hypoxia-ischemia
• infant
• mesenchymal stem cell transplantation
• newborn
• stem cells

• Animals
• Animals, Newborn
• Apoptosis/physiology
• Brain/metabolism
• Brain Injuries/metabolism
• Brain-Derived Neurotrophic Factor/administration & dosage
• Brain-Derived Neurotrophic Factor/biosynthesis
• Brain-Derived Neurotrophic Factor/genetics
• Cell Death/physiology
• Gene Expression
• Humans
• Hypoxia-Ischemia, Brain/metabolism
• Hypoxia-Ischemia, Brain/therapy
• Male
• Mesenchymal Stem Cell Transplantation/methods
• Mesenchymal Stem Cells/metabolism
• Rats
• Rats, Sprague-Dawley

• HI14C3484, HI18C2433 Korea Health Industry Development Institute

• Clonal selection
• Critical quality attributes
• Potency
• Priming
• Product release criteria
• Quality control
• Therapeutic efficacy

• Cartilage
• Digital volume correlation
• Strain

Osteoarthritis (OA) is a common chronic disease and a significant health concern that needs to be urgently solved. OA affects the cartilage and entire joint tissues, including the subchondral bone, synovium, and infrapatellar fat pads. The physiological and pathological changes in these tissues affect the occurrence and development of OA. Understanding complex crosstalk among different joint tissues and their roles in OA initiation and progression is critical in elucidating the pathogenic mechanism of OA. In this review, we begin with an overview of the role of chondrocytes, synovial cells (synovial fibroblasts and macrophages), mast cells, osteoblasts, osteoclasts, various stem cells, and engineered cells (induced pluripotent stem cells) in OA pathogenesis. Then, we discuss the various mechanisms by which these cells communicate, including paracrine signaling, local microenvironment, co-culture, extracellular vesicles (exosomes), and cell tissue engineering. We particularly focus on the therapeutic potential and clinical applications of stem cell-derived extracellular vesicles, which serve as modulators of cell-to-cell communication, in the field of regenerative medicine, such as cartilage repair. Finally, the challenges and limitations related to exosome-based treatment for OA are discussed. This article provides a comprehensive summary of key cells that might be targets of future therapies for OA.

• cartilage
• exosome
• infrapatellar fat pad
• osteoarthritis
• stem cell
• subchondral bone
• synovium

• cartilage lesions
• cartilage repair
• degenerative joint disease
• exosomes
• extracellular vesicle

Mesenchymal stem cells (MSCs) have to be expanded in vitro to reach a sufficient cell dose for the treatment of various diseases. During the process of expansion, some obstacles remain to be overcome. The purpose of this study was to investigate the effects of storage solutions and heterogeneity on the behavior of MSCs in vitro and in vivo.

Umbilical cord MSCs (UC-MSCs) of similar sizes within normal ranges were suspended in three different storage solutions, phosphate buffer solution, normal saline, and Dulbecco’s modified Eagle medium. Then, the ultrastructure, viability, and safety of these cells were compared. Other two UC-MSC populations of different sizes were categorized based on their mean diameters. The ultrastructure, proliferation, immunosuppression, hepatic differentiation potential, and number of senescent cells were investigated and compared. The survival rates of mice after the infusion of UC-MSCs of different sizes were compared.

For UC-MSCs suspended in different storage solutions, the cell apoptosis rates, ultrastructure, and survival rates of mice were similar, and no differences were observed. Cells with a diameter of 19.14 ± 4.89 μm were categorized as the larger UC-MSC population, and cells with a diameter of 15.58 ± 3.81 μm were categorized as the smaller population. The mean diameter of the larger UC-MSC population was significantly larger than that of the smaller UC-MSC population (p < 0.01). Smaller UC-MSCs had more powerful proliferation and immunosuppressive potential and a higher nucleus-cytoplasm ratio than those of large UC-MSCs. The number of cells positive for β-galactosidase staining was higher in the larger UC-MSC population than in the smaller UC-MSC population. The survival rates of mice receiving 1 × 10⁶ or 2 × 10⁶ smaller UC-MSCs were 100%, both of which were higher than those of mice receiving the same amounts of larger UC-MSCs (p < 0.01). The cause of mouse death was explored and it was found that some larger UC-MSCs accumulated in the pulmonary capillary in dead mice.

Different storage solutions showed no significant effects on cell behavior, whereas heterogeneity was quite prevalent in MSC populations and might limit cells application. Hence, it is necessary to establish a more precise standardization for culture-expanded MSCs.

The online version contains supplementary material available at 10.1186/s13287-021-02450-2.

• Function
• Heterogeneity
• Mesenchymal stem cells
• Morphology
• Storage solution

In the field of regenerative medicine, considerable advances have been made from the technological and biological point of view. However, there are still large gaps to be filled regarding translation and application of mesenchymal stromal cell (MSC)-based therapies into clinical practice. Indeed, variables such as cell type, unpredictable donor variation, and expansion/differentiation methods lead to inconsistencies. Most protocols use bovine serum (FBS) derivatives during MSC expansion. However, the xenogeneic risks associated with FBS limits the use of MSC-based products in clinical practice. Herein we compare a chemically defined, xenogeneic-free commercial growth medium with a conventional medium containing 10% FBS and 5 ng/ml FGF2. Furthermore, the effect of a fibronectin-coated growth surface was investigated. The effect of the different culture conditions on chondrogenic commitment was assessed by analyzing matrix deposition and gene expression of common chondrogenic markers. Chondrogenic differentiation potential was similar between the FBS-containing αMEM and the chemically defined medium with fibronectin coating. On the contrary, the use of fibronectin coating with FBS-containing medium appeared to reduce the differentiation potential of MSCs. Moreover, cells that were poorly responsive to in vitro chondrogenic stimuli were shown to improve their differentiation potential after expansion in a TGF-β1 containing medium. In conclusion, the use of a xenogeneic-free medium provides a suitable alternative for human bone marrow MSC expansion, due the capability to maintain cell characteristic and potency. To further improve chondrogenic potential of BMSCs, priming the cells with TGF-β1 during expansion is a promising strategy.

• CD markers
• Cell phenotype
• Donor characteristics
• Human bone marrow stromal stem cells
• Osteoblastic and adipocytic differentiation

• extracellular vesicle
• mesenchymal stem cell
• mesenchymal stem cell-derived exosome
• osteoarthritis

• the Bio & Medical Technology Development Program (2019M3E5D1A02068105) National Research Foundation of Korea

• TGF-βIII
• bone marrow stem cells
• bone marrow stimulation
• cartilage repair

The treatment measures of medication-related osteonecrosis of the jaw (MRONJ) is a worldwide challenge in oral and maxillofacial surgery because of its unclear pathogenesis. Previous studies suggested that mesenchymal stem cells played important roles in promoting MRONJ lesion healing, but the detailed mechanisms were unknown. Increasing numbers of studies have demonstrated that exosomes derived from mesenchymal stem cells, especially adipose-derived stem cells, have key roles in stem cell-based therapies by accelerating bone remodeling, facilitating angiogenesis, and promoting wound healing. We hypothesized that exosomes derived from adipose-derived stem cells can prevent MRONJ by accelerating gingival healing and enhancing bone remodeling processes. Our results may provide a promising therapeutic option for MRONJ clinical therapy.

In vitro and in vivo analyses are closely connected, and the reciprocal relationship between the two comprises a key assumption with concern to the conducting of meaningful research. The primary purpose of in vitro analysis is to provide a solid background for in vivo and clinical study purposes. The fields of cell therapy, tissue engineering, and regenerative medicine depend upon the high quality and appropriate degree of the expansion of mesenchymal stromal cells (MSCs) under low-risk and well-defined conditions. Hence, it is necessary to determine suitable alternatives to fetal bovine serum (FBS—the laboratory gold standard) that comply with all the relevant clinical requirements and that provide the appropriate quantity of high-quality cells while preserving the required properties. Human serum (autologous and allogeneic) and blood platelet lysates and releasates are currently considered to offer promising and relatively well-accessible MSC cultivation alternatives. Our study compared the effect of heat-inactivated FBS on MSC metabolism as compared to its native form (both are used as the standard in laboratory practice) and to potential alternatives with concern to clinical application—human serum (allogeneic and autologous) or platelet releasate (PR-SRGF). The influence of the origin of the serum (fetal versus adult) was also determined. The results revealed the key impact of the heat inactivation of FBS on MSCs and the effectiveness of human sera and platelet releasates with respect to MSC behaviour (metabolic activity, proliferation, morphology, and cytokine production).

Bone marrow stromal cells (BMSC) show promise in cartilage repair, and sheep are the most common large animal pre-clinical model.

The objective of this study was to characterise ovine BMSC (oBMSC) in vitro, and to evaluate the capacity of chondrogenic micro-pellets manufactured from oBMSC or ovine articular chondrocytes (oACh) to repair osteochondral defects in sheep.

oBMSC were characterised for surface marker expression using flow cytometry and evaluated for tri-lineage differentiation capacity. oBMSC micro-pellets were manufactured in a microwell platform, and chondrogenesis was compared at 2%, 5%, and 20% O₂. The capacity of cartilage micro-pellets manufactured from oBMSC or oACh to repair osteochondral defects in adult sheep was evaluated in an 8-week pilot study.

Expanded oBMSC were positive for CD44 and CD146 and negative for CD45. The common adipogenic induction ingredient, 3-Isobutyl-1-methylxanthine (IBMX), was toxic to oBMSC, but adipogenesis could be restored by excluding IBMX from the medium. BMSC chondrogenesis was optimal in a 2% O₂ atmosphere. Micro-pellets formed from oBMSC or oACh appeared morphologically similar, but hypertrophic genes were elevated in oBMSC micro-pellets. While oACh micro-pellets formed cartilage-like repair tissue in sheep, oBMSC micro-pellets did not.

The sensitivity of oBMSC, compared to human BMSC, to IBMX in standard adipogenic assays highlights species-associated differences. Micro-pellets manufactured from oACh were more effective than micro-pellets manufactured from oBMSC in the repair of osteochondral defects in sheep. While oBMSC can be driven to form cartilage-like tissue in vitro, the effective use of these cells in cartilage repair will depend on the successful mitigation of hypertrophy and tissue integration.

The online version contains supplementary material available at 10.1186/s13287-020-02045-3.

• Bone marrow stromal cells
• Cartilage
• Chondrogenesis
• Differentiation
• Hypertrophy
• Mesenchymal stem cells
• Micro-pellet
• Osteoarthritis
• Oxygen

Mesenchymal stromal cell (MSC)-based cell therapy has received great interest in regenerative medicine. Priming the cells during the culture phase can improve their efficacy and/or survival after injection. The literature suggests that MSC extracellular vesicles (EV) can recapitulate a substantial part of the beneficial effects of the cells they originate from, and that micro-RNAs (miRNAs) are important players in EV biological action. Here, our aim was to determine if two classical priming methods of MSC, interferon-gamma (IFNγ) and hypoxia (HYP), could modify their EV miRNA content. Human bone marrow MSCs (BM-MSCs) from five healthy donors were cultured with IFNγ or in HYP or in control (CONT) conditions. The conditioned media were collected after 48 h in serum-free condition and EV were isolated by ultracentrifugation. Total RNA was isolated, pools of CONT, IFN, and HYP cDNA were prepared, and a miRNA profiling was performed using RT-qPCR. Then, miRNAs were selected based on their detectability and measured on each individual EV sample. Priming had no effect on EV amount or size distribution. A set of 81 miRNAs was detected in at least one of the pools of EVs. They were measured on each individual sample; 41 miRNAs were detected in all samples. The principal component analysis (PCA) failed to discriminate the groups. HYP induced a significant decrease in EV hsa-miR-34a-3p content and IFN induced a significant increase in five miRNAs (hsa-miR-25-3p, hsa-miR-106a-5p, hsa-miR-126-3p, hsa-miR-451a, and hsa-miR-665). Taken together, we found only limited alterations in the miRNA landscape of MSC EV with a high inter-individual variability.

• exosome (vesicle)
• extracellular vesicles
• mesenchymal stem cells
• mesenchymal stromal cells
• miRNA
• priming

• Cell therapy
• Cryopreservation
• Freezing
• Human bone marrow-derived mesenchymal stem cells
• Quantitative
• Thawing

• biomarkers
• extracellular vesicle
• in vitro studies and in vivo studies
• mesenchymal stem cell
• osteoarthritis

• TGF-β2
• TGFB2
• adipose tissue
• bone
• bone marrow
• decorin
• gene expression
• multipotent stromal cells
• osteogensys
• potency assay
• stem cells

• Biomarkers/metabolism
• Bone and Bones/metabolism
• Cells, Cultured
• Extracellular Matrix Proteins/metabolism
• Humans
• Multipotent Stem Cells/metabolism
• Osteogenesis/physiology
• RNA, Messenger/metabolism
• Reproducibility of Results
• Stromal Cells/metabolism
• Transforming Growth Factor beta2/metabolism

Adipose-derived stem cells (ASC) have been used as an alternative to bone marrow mesenchymal stem cells (BMSC) for bone tissue engineering. However, the efficacy of ASC in bone regeneration in comparison with BMSC remains debatable, since inconsistent results have been reported. Comparing ASC with BMSC obtained from different individuals might contribute to this inconsistency in results. Therefore, this study aimed to compare the bone regenerative capacity of donor-matched human ASC and BMSC seeded onto poly(l-lactide-co-ε-caprolactone) scaffolds using calvarial bone defects in nude rats. First, donor-matched ASC and BMSC were seeded onto the co-polymer scaffolds to evaluate their in vitro osteogenic differentiation. Seeded scaffolds and scaffolds without cells (control) were then implanted in calvarial defects in nude rats. The expression of osteogenesis-related genes was examined after 4 weeks. Cellular activity was investigated after 4 and 12 weeks. Bone formation was evaluated radiographically and histologically after 4, 12, and 24 weeks. In vitro, ASC and BMSC demonstrated mineralization. However, BMSC showed higher alkaline phosphatase activity than ASC. In vivo, human osteogenesis–related genes Runx2 and collagen type I were expressed in defects with scaffold/cells. Defects with scaffold/BMSC had higher cellular activity than defects with scaffold/ASC. Moreover, bone formation in defects with scaffold/BMSC was greater than in defects with scaffold/ASC, especially at the early time-point. These results suggest that although ASC have the potential to regenerate bone, the rate of bone regeneration with ASC may be slower than with BMSC. Accordingly, BMSC are more suitable for bone regenerative applications.

• Adipose-derived stem cell
• Bone marrow mesenchymal stem cell
• Bone regeneration
• Calvarial defect
• Osteogenic differentiation