Mesenchymal stem cells (MSCs) have been described to induce angiogenesis in various tissues and have been used for the development of novel cell‐based therapies. Increasing evidence suggests that MSCs execute their paracrine function via the secretion of exosomes, especially under hypoxic conditions. However, the mechanisms by which MSC‐derived exosomes secreted under hypoxia enhance angiogenesis still remain unclear. To study exosome physiology under hypoxic or normoxic conditions, we isolated exosomes from bone marrow mesenchymal stem cells (BMSCs). Furthermore, we detected the uptake of exosomes by human umbilical vein endothelial cells (HUVECs) by immunofluorescence staining. In addition, we determined the effects of exosomes on cell viability, migration and tube formation in HUVECs by Cell Counting Kit‐8, migration and tube formation assays, respectively. We examined the expression of key proteins related to exosome‐induced angiogenesis by BMSCs cultured under hypoxic conditions by western blot. Exosomes released by BMSCs cultured under hypoxic conditions enhanced cell proliferation, migration and angiogenesis of HUVECs. Hypoxia induced the expression of high mobility group box 1 protein (HMGB1) in BMSC‐derived exosomes, and silencing of HMGB1 abolished the angiogenic effect in HUVECs. Furthermore, exosomal HMGB1 activated the JNK signaling pathway and induced hypoxia‐inducible factor‐1α/vascular endothelial growth factor expression, consequently enhancing angiogenesis in HUVECs. Our data reveal that exosomal HMGB1 promotes angiogenesis via JNK/hypoxia‐inducible factor‐1α signaling. Therefore, BMSC exosomes derived under hypoxia may have potential for development of novel treatment strategies for angiogenesis‐related diseases.

Angiogenesis is an essential step in tissue engineering. MSC exosomes play an important role in angiogenesis. Functional biomolecules in exosomes vested by the culture microenvironment can be transferred to recipient cells and affects their effect. 3D culture can improve the proliferation and activity of MSCs. However, whether exosomes derived from 3D culture of MSCs have an enhanced effect on angiogenesis is unclear. Herein, we compared the bioactivity of exosomes produced by conventional 2D culture (2D-exos) and 3D culture (3D-exos) of bone marrow stem cells (BMSCs) in angiogenesis. A series of in vitro and in vivo experiments indicated that 3D-exos exhibited stronger effects on HUVEC cell proliferation, migration, tube formation, and in vivo angiogenesis compared with 2D-exos. Moreover, the superiority of 3D-exos might be attributed to the activation of HMGB1/AKT signaling. These results indicate that exosomes from 3D culture of MSCs may serve as a potential therapeutic approach for pro-angiogenesis.

In the field of bone tissue engineering, the development of effective strategies for bone defect repair remains a major challenge. Herein, we report a novel approach involving the integration of high-yield 3D bioprinted adipose-derived stem cell small extracellular vesicles (3D-sEVs) into an injectable HAMA-CPC hydrogel (HAMA-CPC@3D-sEVs). In vitro, HAMA-CPC@3D-sEVs significantly promoted the proliferation, migration, and osteogenic differentiation of bone marrow stromal cells (BMSCs) and promoted angiogenesis in human umbilical vein endothelial cells (HUVECs). In vivo, in a rat tibial defect model, this treatment strongly promoted bone regeneration, increasing the bone volume fraction (BV/TV), bone volume (BV), and trabecular thickness (Tb.Th) at 4 weeks post-surgery. Mechanistically, proteomic analysis revealed that NAMPT in 3D-sEVs upregulated S1PR1 in HUVECs, leading to increased VEGF expression and angiogenesis. These findings suggest that 3D-sEVs-functionalized HAMA-CPC hydrogels have good potential for bone defect repair. Graphical abstract

Accurate measurement of the minimum distance between bony structures of the humeral head and the acromion or coracoid helps advance a better understanding of the shoulder anatomical features. Our goal was to precisely determine the minimum acromiohumeral distance (AHD), coracohumeral distance (CHD), and glenohumeral distance (GHD) in a sample of the Chinese population as an in vivo anatomical analysis. We retrospectively included 146 patients who underwent supine computed tomography (CT) examination of the shoulder joint. The minimum AHD, CHD, and GHD values were quantitatively measured using three-dimensional (3D) CT reconstruction techniques. The correlation between minimum AHD, CHD, and GHD value and age with different sexes was evaluated using Pearson Correlation Coefficient. The mean value of minimum AHD in males was greater than that in females (male 7.62 ± 0.98 mm versus female 7.27 ± 0.86 mm, p = 0.046). The CHD among different sexes differed significantly (male 10.75 ± 2.40 mm versus female 8.76 ± 1.38 mm, p < 0.001). However, we found no statistical differences in GHD with different sexes (male 2.00 ± 0.31 mm versus female 1.96 ± 0.36 mm, p > 0.05). In terms of age correlation, a negative curve correlation existed between age and AHD among the different sexes (male R² = 0.124, p = 0.030, female R² = 0.112, p = 0.005). A negative linear correlation was found in CHD among the different sexes (male R² = 0.164, p < 0.001, female R² = 0.122, p = 0.005). There were no differences between age and minimum GHD in both sexes. The 3D CT reconstruction model can accurately measure the minimum AHD, CHD, and GHD value in vivo and is worthy of further investigation for standard clinical anatomical assessment. Aging may correlate with AHD and CHD narrowing for both sexes.