Journal: Progress in Biomaterials

Article Title: Nanohydroxyapatite incorporated photocrosslinked gelatin methacryloyl/poly(ethylene glycol)diacrylate hydrogel for bone tissue engineering

doi: 10.1007/s40204-021-00150-x

Figure Lengend Snippet: Physical and biological properties of GMP/GMPH hydrogels. a Equilibrium swelling ratio (ESR) of GMP and GMPH hydrogels (n = 6, unpaired two-tailed t test, P = 0.0195, *P < 0.05); b Equilibrium water content (EWC) of GMP and GMPH hydrogels (n = 6, unpaired two-tailed t test, P = 0.0190, *P < 0.05); c Gelation ratio of GMP and GMPH hydrogels (n = 6, unpaired two-tailed t test, P = 0.0114, *P < 0.05); d Biodegradation assay showed an accelerated degradation rate (%) of GMP hydrogel in week 2 (n = 6, unpaired two-tailed t test, P = 0.0015, *P < 0.05). GMPH hydrogel showed a slower degradation rate than GMP hydrogel in week 2 (n = 6, unpaired two-tailed t test, P = 0.0004, *P < 0.05); e MTT assay showing the viability of MG63 cells treated with the extracts of hydrogels. All error bars represent mean ± s.e.m. GM (GelMA), GMP (GelMA/PEGDA), GMPH (GelMA/PEGDA-nanohydroxyapatite)

Article Snippet: The degradation rate (DR) of the hydrogel scaffolds was obtained from the following calculation: DR % = W 0 - W f W 0 ∗ 100 5 Cell culture MG63 osteosarcoma cell line was procured from the National Centre for Cell Science, Pune, India.

Techniques: Two Tailed Test, MTT Assay

Journal: Cancer Cell International

Article Title: Tumor-derived exosomal linc00881 induces lung fibroblast activation and promotes osteosarcoma lung migration

doi: 10.1186/s12935-023-03121-3

Figure Lengend Snippet: OS-derived exosomes induce lung fibroblasts activation and promote OS lung migration. A , B . Exosomes from MG63 and HOS cells were measured using electron microscopy and Nanosight particle tracking analysis. Scale bar: 100 nm. C . Western blot analysis was performed to examine the expression of marker proteins in exosomes derived from MG63 and HOS cells. D . Confocal imaging indicates that Dil-labeled exosomes (red) were delivered to DAPI-labeled HFL-1 cells (blue). The green arrow indicates the delivered exosomes and representative images. Scale bar: 25 μm. E. qRT-PCR detection of the expression of α-SMA in the exosomes-stimulated HFL-1 cells. F. Schematic diagram of cell co-culture in-vitro model. G and H. Representative images and quantitative analysis of transwell assay of OS cells migrated to HFL-1 cells when stimulated with OS-derived exosomes, respectively. I. Representative images of lung metastasis of mice stimulated with MG63-derived exosomes. J. qRT-PCR detection of the relative expression of IL-1β, IL-6, and IL-8 in HFL-1 cells stimulated with the two types of exosomes. (* p < 0.05; ** p < 0.01; *** p < 0.0001)

Article Snippet: A , B . Exosomes from MG63 and HOS cells were measured using electron microscopy and Nanosight particle tracking analysis.

Techniques: Derivative Assay, Activation Assay, Migration, Electron Microscopy, Western Blot, Expressing, Marker, Imaging, Labeling, Quantitative RT-PCR, Co-Culture Assay, In Vitro, Transwell Assay

Journal: Cancer Cell International

Article Title: Tumor-derived exosomal linc00881 induces lung fibroblast activation and promotes osteosarcoma lung migration

doi: 10.1186/s12935-023-03121-3

Figure Lengend Snippet: OS-derived exosomal linc00881 promotes OS lung migration and induces lung fibroblasts activation. A . RNA sequencing in a heatmap. B , C . Scatter plots of lncRNA expression in different groups. D . Relative expression of linc00881 in HFL-1 cells simulated by two OS exosomes. E . qRT-PCR detection of the expression linc00881 in exosomes with inhibited or overexpressed linc00881 in MG63 or HOS cells. F . Schematic diagram of the cell co-culture in vitro model. G – J . Representative images of MG63 ( G ) and HOS ( I ) cells migrated to HFL-1 cells with overexpressed or inhibited linc00881 in MG63 or HOS exosomes by transwell assay and the corresponding quantitative analysis ( H , J ). K – N . Representative images of MG63 ( K ) and HOS ( M ) cells migrated to HFL-1 cells with or without an inhibited expression of linc00881 in HFL-1 cells by transwell assay and the corresponding quantitative analysis ( L , N ). O , P . qRT-PCR detection of the relative expression of IL-1β, IL-6, IL-8, and α-SMA in HFL-1 cells with inhibited expression of linc00881 in MG63 or HOS exosomes. (* p < 0.05; ** p < 0.01; *** p < 0.0001)

Article Snippet: A , B . Exosomes from MG63 and HOS cells were measured using electron microscopy and Nanosight particle tracking analysis.

Techniques: Derivative Assay, Migration, Activation Assay, RNA Sequencing, Expressing, Quantitative RT-PCR, Co-Culture Assay, In Vitro, Transwell Assay

Journal: Cancer Cell International

Article Title: Tumor-derived exosomal linc00881 induces lung fibroblast activation and promotes osteosarcoma lung migration

doi: 10.1186/s12935-023-03121-3

Figure Lengend Snippet: Linc00881 sponge miR-29c-3p in lung fibroblasts. A . Distribution of linc00881 in HFL-1 cells assessed by the FISH assay. B . Duplex structure of miR-29c-3p and linc00881. C . Relative expression of miR-29c-3p in HFL-1 cells simulated by two OS exosomes. D . qRT-PCR detection of relative expression of linc00881 using the control or miR-29c-3p probe to perform the pull-down assay. E . qRT-PCR detection of the expression of linc00881 in HFL-1 cells transfected with linc00881 siRNA or vector. F . qRT-PCR detection of the expression of miR-29c-3p in HFL-1 cells transfected as described in E. G – K . Transwell assay of MG63 or HOS cells migrated to the HFL-1 cells transfected with: a. control mimic + control vector; b. miR-29c-3p mimic + control vector; c. control mimic + linc00881 vector; d. miR-29c-3p mimic + linc00881 vector, representative images and quantitative analysis of MG63 and HOS cells are shown in H&I and J&K, respectively. (* p < 0.05; ** p < 0.01; *** p < 0.0001)

Article Snippet: A , B . Exosomes from MG63 and HOS cells were measured using electron microscopy and Nanosight particle tracking analysis.

Techniques: Expressing, Quantitative RT-PCR, Control, Pull Down Assay, Transfection, Plasmid Preparation, Transwell Assay

Journal: Cancer Cell International

Article Title: Tumor-derived exosomal linc00881 induces lung fibroblast activation and promotes osteosarcoma lung migration

doi: 10.1186/s12935-023-03121-3

Figure Lengend Snippet: miR-29c-3p directly regulates the expression of MMP2 in lung fibroblasts cells. A . Duplex structure of MMP2 3′-UTR (Untranslated Region) and miR-29c-3p. B . The relative expression of MMP2 was detected at the mRNA level by qRT-PCR using a control or miR-125a-5p probe to perform the pull-down assay. C . The miR-29c-3p expression was detected via qRT-PCR after the control/miR-29c-3p mimic or control/miR-29c-3p inhibitor was transfected into HFL-1 cells. D – F . Representative images ( D ) and quantitative analysis ( E , F ) of Western blotting analysis and qRT-PCR of MMP2 expression in HFL-1 cells after transfected as described in C . G , H . Representative images ( G ) and quantitative analysis ( H ) of Western blotting analysis of MMP2 expression in HFL-1 cells transfected as follows: a. control mimic and control vector; b. control mimic and the miR-29c-3p-overexpressed plasmid; c. miR-29c-3p mimic and control vector; d. miR-29c-3p mimic and MMP2 vector. I – L . Representative images ( I MG63, K HOS) and quantitative analysis ( J , L ) of transwell assay of MG63 and HOS cells migrated to HFL-1 cells after transfected as described in G . (* p < 0.05; ** p < 0.01; *** p < 0.0001)

Article Snippet: A , B . Exosomes from MG63 and HOS cells were measured using electron microscopy and Nanosight particle tracking analysis.

Techniques: Expressing, Quantitative RT-PCR, Control, Pull Down Assay, Transfection, Western Blot, Plasmid Preparation, Transwell Assay

Journal: Cancer Cell International

Article Title: Tumor-derived exosomal linc00881 induces lung fibroblast activation and promotes osteosarcoma lung migration

doi: 10.1186/s12935-023-03121-3

Figure Lengend Snippet: Exosomal Linc00881 regulates the expression of MMP2 in lung fibroblasts by sponging miR-29c-3p. A , B . Representative images ( A ) of Western blotting analysis of MMP2 expression in HFL-1 cells treated with blank, MG63, or HOS exosomes and the corresponding quantitative analysis ( B ). C . Relative miR-29c-3p concentration was detected in HFL-1 cells simulated by the two types of OS-derived exosomes. D - F . Representative images ( D ) of Western blotting analysis of MMP2 expression in HFL-1 cells transfected with linc00881 siRNA or vector and the corresponding quantitative analysis ( E , F ). G . qRT-PCR detection of the expression of linc00881 in HFL-1 cells transfected with linc00881 siRNA or vector. H . qRT-PCR detection of the expression of miR-29c-3p in HFL-1 cells transfected as described in B . I – K . Representative images ( I ) of Western blotting analysis of MMP2 expression in HFL-1 cells transfected with miR-29c-3p mimic and stimulated with MG63 or HOS cells-derived exosomes, and the corresponding quantitative analysis ( J , K ). L – O . qRT-PCR detection of the relative expression of linc00881 or miR-29c-3p in HFL-1 cells treated in I . (* p < 0.05; ** p < 0.01; *** p < 0.0001)

Article Snippet: A , B . Exosomes from MG63 and HOS cells were measured using electron microscopy and Nanosight particle tracking analysis.

Techniques: Expressing, Western Blot, Concentration Assay, Derivative Assay, Transfection, Plasmid Preparation, Quantitative RT-PCR

Journal: Cancer Cell International

Article Title: Tumor-derived exosomal linc00881 induces lung fibroblast activation and promotes osteosarcoma lung migration

doi: 10.1186/s12935-023-03121-3

Figure Lengend Snippet: Linc00881 activates lung fibroblasts through the NF-κB axis. A , B . Western blotting analysis of the proteins of NF-κB axis in HFL-1 cells treated with exosomes from MG63 and HOS cells. C , D . Western blotting analysis of the expression of MMP2 and N-cadherin in HFL-1 cells treated with MG63 or HOS cell-derived exosomes. E , F . Western blotting analysis of the proteins of NF-κB axis in HFL-1 cells treated with: a. control vector, b. linc00881 vector, c. control siRNA, d. MMP2 siRNA, e. control MMP2, and f. MMP2 vector. G , H . Western blotting analysis of the expression of MMP2 and N-cadherin in HFL-1 cells in different treatments as described in E . (* p < 0.05; ** p < 0.01; *** p < 0.0001)

Article Snippet: A , B . Exosomes from MG63 and HOS cells were measured using electron microscopy and Nanosight particle tracking analysis.

Techniques: Western Blot, Expressing, Derivative Assay, Control, Plasmid Preparation

Journal: Cancer Cell International

Article Title: Tumor-derived exosomal linc00881 induces lung fibroblast activation and promotes osteosarcoma lung migration

doi: 10.1186/s12935-023-03121-3

Figure Lengend Snippet: Activated fibroblasts by exosomal linc00881 accelerate OS progression. A . IL-6 secretion from HFL-1 treated by exosomes from two OS cells was detected via ELISA assay. B . IL-6 secretion from HFL-1 treated by exosomes from two OS cells or exosomes from linc00881 interrupted two OS cells and was detected via ELISA assay. C . Spheroid formation ability of MG63 treated with indicated CM. Representative Images and quantitative analysis were shown.100 × . D – E . Spheroid formation ability of MG63 treated with CM containing: a. IL-6 neutralizing antibody, b. IgG control antibody, c. linc00881 siRNA exo, d. IL-6 neutralizing antibody plus linc00881 siRNA exo. Representative Images and quantitative analysis were shown.100 × . F . Spheroid formation ability of HOS treated with indicated CM. Representative Images and quantitative analysis were shown.100 × . G . Spheroid formation ability of MG63 treated with CM as indicated in D . Representative Images and quantitative analysis were shown.100 × . H . Migration assay of MG63 treated with indicated CM. Representative Images and quantitative analysis were shown.100 × . I . Migration assay of MG63 treated with indicated CM indicate in D . Representative Images and quantitative analysis were shown.100 × . J , K . Migration assay of HOS treated with indicated CM. Representative Images and quantitative analysis were shown.100 × . L . Migration assay of HOS treated with CM indicated in D . Representative Images and quantitative analysis were shown.100 × . M , N . Western blotting analysis of the expression of MMP2 in MG63 and HOS cells treated with indicated CM. (* p < 0.05; ** p < 0.01; *** p < 0.0001)

Article Snippet: A , B . Exosomes from MG63 and HOS cells were measured using electron microscopy and Nanosight particle tracking analysis.

Techniques: Enzyme-linked Immunosorbent Assay, Control, Migration, Western Blot, Expressing