New in the industry. Exosomes give light-curable 3D printed 45S5 ceramic scaffolds to enhance angiogenesis-bone coupling to accelerate bone regeneration.

Recently, a team led by Xi, Yongming of Spine Surgery, Affiliated Hospital of Qingdao University, published in Composites Part B a study entitled Exosomes endow photocurable 3D printing 45S5 ceramic scaffolds to enhance angiogenesis-osteogenesis coupling for accelerated bone regeneration, using silane coupling agent containing double bonds to modify tetrahedral silicate,Thus, the photocurable 45S5 bioactive glass precursor was prepared.

Original link: https://www.sciencedirect.com/science/article/pii/S135983682400266X?dgcid=rss_sd_all
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Figure 1, schematic diagram showing that endothelial progenitor cell-derived exosomes impart light-curable 3D printed 45S5 ceramic scaffolds to enhance angiogenesis, osteogenesis, and accelerate bone regeneration.

2,(A) Modification and synthesis of light-cured 45S5 precursor PG. (B) Photo-crosslinking properties of PG. (C) FTIR spectral analysis of BG, non-crosslinked PG and crosslinked PG. (D-E) Photosensitive rheology results of PG and PG/TCP gels. (F) Light-cured 3D printed PT stent. (G)PG and PT stents were excellent in printability. (H) SEM images of PG, PT, PT/G. XRD images of PG and PT were (I). (J) Stress-strain curves for PG, PT and PT/G scaffolds. (K) Ultimate compressive strength of PG, PT and PT/G scaffolds. (L) Compressive modulus of PG, PT and PT/G scaffolds. (M) Degradation curves of PG, PT, PT/ G scaffolds.

Figure 3,(A) Schematic diagram of exosome extraction process. (B-D) EPC-exos were identified by TEM, NTA and Western blot analysis of Alix, Flot 1 and CD81. (E-F) BMSCs and HUVECs were EPC-exos by fluorescence internalization. (red: EPC-exos, green: cytoskeleton, blue: nucleus). (G) BMSC viability was assessed on days 1, 3 and 5 using CCK-8 detection methods. (H) mRNA expression levels of osteogenic genes in bone marrow mesenchymal stem cells were detected RT-qPCR (* p < 0.05,**p <0.01).

4, cell viability of bone marrow mesenchymal stem cells in each group was detected by CCK-8 (A). (* p < 0.05, **p < 0.01).(B) Live/dead stained images of bone marrow mesenchymal stem cells in each group. (C) Flow cytometry analysis of cell apoptosis in each group after scaffold-cell co-culture. (D) Transwell experiments showed bone marrow mesenchymal stem cell migration by crystal violet staining. (E) Quantitative analysis of migrating cells. (F) ALP staining of bone marrow mesenchymal stem cells on day 7. (G) ALP activity of cells cultured for 7 days. (H-I) Confocal immunofluorescence imaging and quantitative assessment of the osteogenic marker COL1. (J-N) mRNA expression levels of each constituent bone gene (* p < 0.05,**p <0.01).

Figure 5,(A-B) Scratch test and quantitative analysis of HUVECs. (C-D) Confocal immunofluorescence images and quantitative analysis of CD31. (E-H) Angiogenesis gene mRNA expression (* p < 0.05,**p <0.01).

6,(A) Three-dimensional microct images of reconstructed 6w and 12w bone defect repair. (B-E) Quantitative analysis of in vivo regenerative repair effects after implantation.

7, (A) H & E staining for each implant. Each implant was stained with (B) Masson. (C) Immunofluorescence image of in vivo tissue osteogenesis marker cyanate. (D) Immunofluorescence image of in vivo tissue CD31 immunofluorescence image. (E-F) Image J software (**p < 0.01,*** p < 0.001) was used to quantify the CD31 and cyanate positive regions.

8,(a) Heat map of the DEmirna. Red indicates high relative gene expression and blue indicates low relative gene expression. Each column represents a sample and each row represents a significant miRNA. (B) Pathway analysis of DE miRNAs target genes by Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis. (C) Graphene oxide analysis includes functional pathways, biological processes, and cellular components. (D) Relative expression levels of MAPK pathway-related genes in vitro were detected RT-qPCR. (E-K) Expression of JNK, phosphorylated JNK(P-JNK), ERK1/2, phosphorylated ERK1/2(PERK1/2), P38 and phosphorylated P38(P-P38) proteins in BMSCs (* p < 0.05,**p <0.01).

In this study, the 45S5 precursor was modified by double bond functionalization, and the PT scaffold was fabricated by light curing 3D printing method. The EPC-exos encapsulated in GelMA was fixed on the PT scaffold to form a functionalized PT/G @ Exos composite scaffold to promote angiogenesis-osteogenesis coupling, thereby enhancing bone regeneration. Studies have shown that the composite scaffold promotes bone regeneration through the P38/MAPK pathway and provides new insights for the development of advanced bone tissue engineering scaffolds for clinical applications.

△ The above picture is printed by our company using self-developed light-curing ceramic 3D printing equipment.Bioceramic scaffold sampleTo provide more efficient methods for teachers to do biomaterial and structural research verification