Human adipose-derived stem cells (ADSCs) can release exosomes; however, their specific functions remain elusive. In this study, we verified that exosomes derived from osteogenically differentiated ADSCs can promote osteogenic differentiation of ADSCs. Furthermore, in order to investigate the importance of exosomal microRNAs (miRNAs) in osteogenic differentiation of ADSCs, we used microarray assays to analyze the expression profiles of exosomal miRNAs derived from undifferentiated as well as osteogenically differentiated ADSCs; 201 miRNAs were upregulated and 33 miRNAs were downregulated between the two types of exosomes. Additionally, bioinformatic analyses, which included gene ontology analyses, pathway analysis, and miRNA-mRNA-network investigations, were performed. The results of these analyses revealed that the differentially expressed exosomal miRNAs participate in multiple biological processes, such as gene expression, synthesis of biomolecules, cell development, differentiation, and signal transduction, among others. Moreover, we found that these differentially expressed exosomal miRNAs connect osteogenic differentiation to processes such as axon guidance, MAPK signaling, and Wnt signaling. To the best of our knowledge, this is the first study to identify and characterize exosomal miRNAs derived from osteogenically differentiated ADSCs. This study confirms that alterations in the expression of exosomal miRNAs can promote osteogenic differentiation of ADSCs, which also provides the foundation for further research on the regulatory functions of exosomal miRNAs in the context of ADSC osteogenesis.

Regeneration and repair of bone tissue are pluripotent stem cells through a series of complex process completed. It mainly includes proliferation and differentiation, recognition of extracellular matrix and signal molecules, expression of related factors and targeting. Adiposederived stem cells (ADSCs) are a population of non-hematopoietic adult stem cells with self-renewal ability and multi-lineage potential isolated

Background Human adipose-derived stem cells (hADSCs) are stem cells with the potential to differentiate in multiple directions. miR-204-5p is expressed at low levels during the osteogenic differentiation of hADSCs, and its specific regulatory mechanism remains unclear. Here, we aimed to explore the function and possible molecular mechanism of miR-204-5p in the osteogenic differentiation of hADSCs. Methods The expression patterns of miR-204-5p, Runx2, alkaline phosphatase (ALP), osteocalcin (OCN), forkhead box C1 (FOXC1) and growth differentiation factor 7 (GDF7) in hADSCs during osteogenesis were detected by qRT-PCR. Then, ALP and alizarin red staining (ARS) were used to detect osteoblast activities and mineral deposition. Western blotting was conducted to confirm the protein levels. The regulatory relationship among miR-204-5p, FOXC1 and GDF7 was verified by dual-luciferase activity and chromatin immunoprecipitation (ChIP) assays. Results miR-204-5p expression was downregulated in hADSC osteogenesis, and overexpression of miR-204-5p suppressed osteogenic differentiation. Furthermore, the levels of FOXC1 and GDF7 were decreased in the miR-204-5p mimics group, which indicates that miR-204-5p overexpression suppresses the expression of FOXC1 and GDF7 by binding to their 3′-untranslated regions (UTRs). Overexpression of FOXC1 or GDF7 improved the inhibition of osteogenic differentiation of hADSCs induced by the miR-204-5p mimics. Moreover, FOXC1 was found to bind to the promoter of miR-204-5p and GDF7, promote the deacetylation of miR-204-5p and reduce the expression of miR-204-5p, thus promoting the expression of GDF7 during osteogenic differentiation. GDF7 induced hADSC osteogenesis differentiation by activating the AKT and P38 signalling pathways. Conclusions Our results demonstrated that the miR-204-5p/FOXC1/GDF7 axis regulates the osteogenic differentiation of hADSCs via the AKT and p38 signalling pathways. This study further revealed the regulatory mechanism of hADSC differentiation from the perspective of miRNA regulation.

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3D-printed titanium (Ti) materials have attracted much attention in the field of bone tissue repair. However, the combination strength of traditional alloy materials with bone tissue is lower, and the elastic modulus is higher than that of natural bone tissue, which makes the titanium alloy susceptible to stress shielding phenomena after implantation. Therefore, it is urgent to find better surface modification technology. In this study, the physical and chemical properties, toxicity, and proliferation of adipose stem cells of composite graphene-coated titanium alloy (Gr–Ti) were investigated using 3D-printed titanium alloy as a material model. Physical and chemical property tests confirmed that 3D printing could produce porous titanium alloy materials; the compressive strength and elastic modulus of the titanium alloy scaffolds were 91 ± 3 MPa and 3.1 ± 0.4 GPa, matching the elastic modulus of normal bone tissue. The surface characterization shows that graphene can be coated on titanium alloy by a micro-arc oxidation process, which significantly improves the surface roughness of titanium alloy. The roughness factor (Ra) of the Ti stent was 4.95 ± 1.12 μm, while the Ra of the Gr–Ti stent was 6.37 ± 0.72 μm. After the adipose stem cells were co-cultured with the scaffold for 4 h and 24 h, it was found that the Gr–Ti scaffold could better promote the early cell adhesion. CCK-8 tests showed that the number of ADSCs on the G–Ti scaffold was significantly higher than that on the Ti scaffold (p < 0.01). The relative growth rate (RGR) of ADSCs in Gr–Ti was grade 0–1 (non-toxic). In the in vivo experiment of repairing a critical bone defect of a rabbit mandible, the bone volume fraction in the Gr–Ti group increased to 49.42 ± 3.28%, which was much higher than that in the Ti group (39.76 ± 3.62%) (p < 0.05). In conclusion, the porous graphene–titanium alloy promotes the proliferation and adhesion of adipose stem cells with multidirectional differentiation potential, which has great potential for the application of bone tissue engineering in repairing bone defects in the future.

Distraction osteogenesis (DO) is widely used for bone tissue engineering technology. Immune regulations play important roles in the process of DO like other bone regeneration mechanisms. Compared with others, the immune regulation processes of DO have their distinct features. In this review, we summarized the immune-related events including changes in and effects of immune cells, immune-related cytokines, and signaling pathways at different periods in the process of DO. We aim to elucidated our understanding and unknowns about the immunomodulatory role of DO. The goal of this is to use the known knowledge to further modify existing methods of DO, and to develop novel DO strategies in our unknown areas through more detailed studies of the work we have done.

Titanium and titanium alloys (Ti6Al4V and Ti) have been widely used in bone tissue engineering to repair maxillofacial bone defects caused by traumas and tumors. However, such materials are also bio-inert, which does not match the elastic modulus of bone. Therefore, different surface modifications have been proposed for clinical application. Based on the use of traditional titanium alloy in the field of bone repair defects, we prepared a compound Gr-Ti scaffold with ADSC-derived Exos. The results showed that Gr-Ti scaffolds have low toxicity and good biocompatibility, which can promote the adhesion and osteogenic differentiation of ADSCs. Exos played a role in promoting osteogenic differentiation of ADSCs: the mRNA levels of RUNX2, ALP, and Osterix in the Gr-Ti/Exos group were significantly higher than those in the Gr-Ti group, which process related to the Wnt signaling pathway. Gr-Ti scaffolds with ADSCs and ADSC-derived Exos successfully repaired rabbit mandibular defects. The bone mineral density and the bending strength of the Gr-Ti/Exos group was significantly higher than that of the Gr-Ti group. This study provides a theoretical basis for the research and development of new clinical bone repair materials.

Melatonin is a hormone, synthesized in the pineal gland, which primarily controls the circadian rhythm of the body. In recent years, melatonin has also been shown to regulate metabolism, provide neuroprotection, and act as an anti-inflammatory, free radical scavenger. There has also been a recent research interest in the role of melatonin in regulating mesenchymal stromal cells (MSCs). MSCs are pivotal for their ability to differentiate into a variety of different tissues. There is also increasing evidence for the therapeutic prospects of MSCs via paracrine signaling. In addition to secreting cytokines and chemokines, MSCs can secrete extracellular vesicles (EVs), allowing them to respond to injury and promote tissue regeneration. While there has been a major research interest in the use of MSCs for regenerative medicine, the clinical application is limited by many risks, including tumorigenicity, senescence, and sensitivity to toxic environments. The use of MSC-derived EVs for cell-free therapy can potentially avoid the disadvantages of MSCs, which makes this an exciting prospect for regenerative medicine. Prior research has shown that MSCs, via paracrine mechanisms, can identify receptor-independent responses to melatonin and then activate a series of downstream pathways, which exert a variety of effects, including anti-tumor and anti-inflammatory effects. Here we review the synthesis of melatonin, its mechanisms of action, and the effect of melatonin on MSCs via paracrine signaling. Furthermore, we summarize the current clinical applications of melatonin and discuss future prospects.