lxw7  (Biosynth Carbosynth)

Journal: Bioactive materials

Article Title: Bioactive extracellular matrix scaffolds engineered with proangiogenic proteoglycan mimetics and loaded with endothelial progenitor cells promote neovascularization and diabetic wound healing.

doi: 10.1016/j.bioactmat.2021.08.017

Figure Lengend Snippet: Fig. 2. Specificity and binding affinity of LXW7 ligand to ZDF-EPCs and expression of αvβ3 integrin on ZDF-EPCs A. On-bead cell binding assay for testing the binding affinity and specificity of ligands to ZDF-EPCs showing high binding of ZDF-EPCs to LXW7 coated beads (right panel) compared to THP-1 monocytes (left panel). B. Quantification of the number of THP-1 monocytes and ZDF-EPCs bound to the LXW7 coated beads. C. Expression of αvβ3 integrin on ZDF-EPCs. D. Flow cytometry analysis of the binding affinity of LXW7 showing the blocking of its binding by anti-αvβ3 antibody. Scale bar = 100 μm. Data are expressed as mean ± SD. ****p < 0.0001, n = 3.

Article Snippet: Before purifying, LXW7 was cyclized by oxidizing cysteine residues using ClearOx resin (Peptides International) according to manufacturer’s protocol.

Techniques: Binding Assay, Expressing, Cell Binding Assay, Flow Cytometry, Blocking Assay

Journal: Bioactive materials

Article Title: Bioactive extracellular matrix scaffolds engineered with proangiogenic proteoglycan mimetics and loaded with endothelial progenitor cells promote neovascularization and diabetic wound healing.

doi: 10.1016/j.bioactmat.2021.08.017

Figure Lengend Snippet: Fig. 3. Effect of LXW7 and LXW7-DS-SILY ligands on the attachment, growth and viability of ZDF-EPCs on ligand modified surfaces. A-B. Representative images of attached ZDF-EPCs on surfaces treated by D-biotin (A; control), LXW7 (B) after 20 min incubation. C. Quantification and the correlative statistical analysis of remaining cells shown in A-B. D-E. Representative images of adhered EPCs on untreated (D; control) or LXW7-DS-SILY modified SIS surface (E) after 20 min incubation. F. Quantification and the correlative statistical analysis of remaining cells shown in D-E. G. Growth and viability of ZDF-EPCs on LXW7 treated tissue culture surfaces and D-biotin treated surfaces (control) was assessed by CCK-8 assay. H. Growth and viability of ZDF-EPCs on LXW7-DS-SILY or DS-SILY treated SIS scaffold surfaces was assessed by CCK-8 assay. Scale bar = 200 μm. Data are expressed as mean ± SD. ****p < 0.0001, **p < 0.01, *p < 0.05, n = 3.

Article Snippet: Before purifying, LXW7 was cyclized by oxidizing cysteine residues using ClearOx resin (Peptides International) according to manufacturer’s protocol.

Techniques: Modification, Control, Incubation, CCK-8 Assay

Journal: Bioactive materials

Article Title: Bioactive extracellular matrix scaffolds engineered with proangiogenic proteoglycan mimetics and loaded with endothelial progenitor cells promote neovascularization and diabetic wound healing.

doi: 10.1016/j.bioactmat.2021.08.017

Figure Lengend Snippet: Fig. 4. SIS/LXW7-DS-SILY scaffolds accelerate wound closure and support ZDF-EPC survival under non-ischemic condition. A. Representative images of healing in wounds treated with different groups at days 0, 3, 7, 11 and 14. B. Quantification of the healing rate of different groups. a) SIS/LXW7-DS-SILY group was significantly better compared to SIS/DS-SILY and SIS groups. b) SIS/LXW7-DS-SILY/EPCs group was significantly better than the SIS/DS-SILY/EPCs and SIS/EPCs groups. c) SIS/LXW7-DS-SILY/EPCs group compared to SIS/LXW7-DS-SILY group showed no significant difference. d) SIS/DS-SILY/EPCs group was signifi cantly better compared to SIS/DS-SILY group. e) There was a trend that the SIS/EPCs group was better than the SIS group but the difference was not statis tically significant. Data are expressed as mean ± SEM. SIS group, SIS/LXW7-DS-SILY group, and SIS/EPCs group: n = 8; SIS/DS-SILY group: n = 7; SIS/DS-SILY/ EPCs group: n = 12; SIS/LXW7-DS-SILY/EPCs group: n = 13. C. Bioluminescence imaging of wounds transplanted with different modified SIS scaffolds loaded with Td-Tomato/luciferin-labeled ZDF-EPCs. D. Quantification of the signal intensity of biolumi nescence (Most of the SIS scaffolds spontaneously fall off the wounds by day 11). Data are expressed as mean ± SEM. n = 5 per group. ***p < 0.001, **p < 0.01, *p < 0.05.

Article Snippet: Before purifying, LXW7 was cyclized by oxidizing cysteine residues using ClearOx resin (Peptides International) according to manufacturer’s protocol.

Techniques: Imaging, Modification, Labeling

Journal: Bioactive materials

Article Title: Bioactive extracellular matrix scaffolds engineered with proangiogenic proteoglycan mimetics and loaded with endothelial progenitor cells promote neovascularization and diabetic wound healing.

doi: 10.1016/j.bioactmat.2021.08.017

Figure Lengend Snippet: Fig. 7. SIS/LXW7-DS-SILY constructs accelerate wound closure and support survival of ZDF-EPCs under ischemic condition. A. Representative images of healing in wounds treated with different groups at days 0, 3, 7, 11 and 14. B. Quantification of the wound closure rates of different groups. a) SIS/LXW7-DS-SILY group was significantly better compared to SIS group. There was a trend that the SIS/DS-SILY group was better than the SIS group but the difference was not statistically significant. b) SIS/LXW7-DS-SILY/EPCs group was significantly better than the SIS/DS-SILY/EPCs and SIS/EPCs groups. c) SIS/LXW7-DS-SILY/EPCs group was significantly better compared to SIS/LXW7-DS- SILY group. d) SIS/DS-SILY/EPCs group compared to SIS/DS-SILY group showed no significant differ ence. e) SIS/EPCs group compared to SIS group showed no significant difference. Data are expressed as mean ± SEM. SIS group, SIS/LXW7-DS-SILY group, and SIS/EPCs group: n = 8; SIS/DS-SILY group: n = 7; SIS/DS-SILY/EPCs group: n = 11; SIS/LXW7-DS- SILY/EPCs group: n = 12. C. Bioluminescence image of wounds transplanted with different modified scaf folds loaded with ZDF-EPCs. D. Quantification of the bioluminescence signals intensity (Most of the SIS scaffolds spontaneously fall off the wounds by day 11). Data are expressed as mean ± SEM. n = 5 per group. **p < 0.01, *p < 0.05.

Article Snippet: Before purifying, LXW7 was cyclized by oxidizing cysteine residues using ClearOx resin (Peptides International) according to manufacturer’s protocol.

Techniques: Construct, Modification

Journal: Acta biomaterialia

Article Title: Rapid endothelialization of small diameter vascular grafts by a bioactive integrin-binding ligand specifically targeting endothelial progenitor cells and endothelial cells

doi: 10.1016/j.actbio.2020.03.005

Figure Lengend Snippet: Binding specificity of LXW7 to ECFCs in blood circulation. (A) Beads displaying LXW7 were incubated with HECFCs, CD34− MNCs, HCASMCs and platelets for 2 h respectively. LXW7 efficiently supported binding of HECFCs (a) but did not support binding of CD34− MNCs (b), HCASMCs (c) and platelets (d). Scale bar =50 μm. (B) The numbers of different types of cells and platelets bound on each bead displaying LXW7 were quantified and statistical analyses were performed. Data were expressed as mean ± standard deviation: ***p < 0.001 (n = 5).

Article Snippet: For platelet-bead binding assay, 2 mL fresh PRP was added to an ultra-low attachment 35-mm Petri dish (Corning Incorporated) followed by LXW7-modified resin beads.

Techniques: Binding Assay, Incubation, Standard Deviation

Journal: Acta biomaterialia

Article Title: Rapid endothelialization of small diameter vascular grafts by a bioactive integrin-binding ligand specifically targeting endothelial progenitor cells and endothelial cells

doi: 10.1016/j.actbio.2020.03.005

Figure Lengend Snippet: ECFC attachment on LXW7-modified electrospun grafts. (A) Fluorescence images of remaining HECFC attachment on untreated grafts (a) and grafts modified with 10 nmol/cm2 LXW7 (b), 25 nmol/cm2 LXW7 (c), 50 nmol/cm2 LXW7 (d), 100 nmol/cm2 LXW7 (e), and 150 nmol/cm2 LXW7 (f) after 30 min incubation. (B) The numbers of HECFCs attached on grafts modified with different concentrations of LXW7 were quantified and statistical analyses were performed. Data were expressed as mean ± standard deviation: *p < 0.05, **p < 0.01, ***p < 0.001 (n = 5).

Article Snippet: For platelet-bead binding assay, 2 mL fresh PRP was added to an ultra-low attachment 35-mm Petri dish (Corning Incorporated) followed by LXW7-modified resin beads.

Techniques: Modification, Fluorescence, Incubation, Standard Deviation

Journal: Acta biomaterialia

Article Title: Rapid endothelialization of small diameter vascular grafts by a bioactive integrin-binding ligand specifically targeting endothelial progenitor cells and endothelial cells

doi: 10.1016/j.actbio.2020.03.005

Figure Lengend Snippet: X-ray photoelectron spectroscopy (XPS) wide spectra of electrospun grafts conjugated with LXW7.

Article Snippet: For platelet-bead binding assay, 2 mL fresh PRP was added to an ultra-low attachment 35-mm Petri dish (Corning Incorporated) followed by LXW7-modified resin beads.

Techniques: Spectroscopy

Journal: Acta biomaterialia

Article Title: Rapid endothelialization of small diameter vascular grafts by a bioactive integrin-binding ligand specifically targeting endothelial progenitor cells and endothelial cells

doi: 10.1016/j.actbio.2020.03.005

Figure Lengend Snippet: Amino acid analysis (AAA) of electrospun grafts conjugated with LXW7.

Article Snippet: For platelet-bead binding assay, 2 mL fresh PRP was added to an ultra-low attachment 35-mm Petri dish (Corning Incorporated) followed by LXW7-modified resin beads.

Techniques:

Journal: Acta biomaterialia

Article Title: Rapid endothelialization of small diameter vascular grafts by a bioactive integrin-binding ligand specifically targeting endothelial progenitor cells and endothelial cells

doi: 10.1016/j.actbio.2020.03.005

Figure Lengend Snippet: ECFC adhesion, proliferation, and differentiation on electrospun grafts conjugated with or without LXW7 treated. (A) Immunofluorescence staining results of remaining HECFCs attached on untreated (a) and LXW7-modified grafts (b) after 2 h incubation at day 1. The proliferation and differentiation of the attached HECFCs on untreated (c) and LXW7-modified grafts (d) at day 7. The numbers of cells attached on grafts at day 1 and day 7 (B), cell proliferation rates (C), and the number of CD31+ cells on grafts at day 1 and day 7 (D) were quantified and statistical analyses were performed. Data were expressed as mean ± standard deviation: **p < 0.01, ***p < 0.001 (n = 5).

Article Snippet: For platelet-bead binding assay, 2 mL fresh PRP was added to an ultra-low attachment 35-mm Petri dish (Corning Incorporated) followed by LXW7-modified resin beads.

Techniques: Immunofluorescence, Staining, Modification, Incubation, Standard Deviation

Journal: Acta biomaterialia

Article Title: Rapid endothelialization of small diameter vascular grafts by a bioactive integrin-binding ligand specifically targeting endothelial progenitor cells and endothelial cells

doi: 10.1016/j.actbio.2020.03.005

Figure Lengend Snippet: ECFC penetration on electrospun grafts conjugated with or without LXW7 treated. (A) HECFCs (green) penetrated into (a, c) untreated grafts and (b, d) LXW7 treated grafts. (c) and (d) are the magnifying images of part of (a) and (b). The area between the white lines were the sections of the grafts. (B) The numbers and (C) distribuation areas of HECFCs penetrated into grafts were quantified and statistical analyses were performed. Data were expressed as mean ± standard deviation: **p < 0.01 (n = 5).

Article Snippet: For platelet-bead binding assay, 2 mL fresh PRP was added to an ultra-low attachment 35-mm Petri dish (Corning Incorporated) followed by LXW7-modified resin beads.

Techniques: Standard Deviation

Journal: Acta biomaterialia

Article Title: Rapid endothelialization of small diameter vascular grafts by a bioactive integrin-binding ligand specifically targeting endothelial progenitor cells and endothelial cells

doi: 10.1016/j.actbio.2020.03.005

Figure Lengend Snippet: Platelet adhesion on electrospun grafts. (A) SEM revealed the morphology and adhesion of platelet on untreated electrospun grafts (a), PEG treated electrospun grafts (b) and LXW7-PEG treated electrospun grafts (c). White arrows showed the platelet aggregation.. (B) The numbers of platelets attached on grafts with different treatments were quantified and statistical analyses were performed. Data were expressed as mean ± standard deviation: **p < 0.01, (n = 5).

Article Snippet: For platelet-bead binding assay, 2 mL fresh PRP was added to an ultra-low attachment 35-mm Petri dish (Corning Incorporated) followed by LXW7-modified resin beads.

Techniques: Standard Deviation

Journal: Acta biomaterialia

Article Title: Rapid endothelialization of small diameter vascular grafts by a bioactive integrin-binding ligand specifically targeting endothelial progenitor cells and endothelial cells

doi: 10.1016/j.actbio.2020.03.005

Figure Lengend Snippet: Examination of explanted grafts. New capillary formation was exhibited on untreated grafts (A) and LXW7-modified grafts (B) at week 6 after implantation. Arrows in (B) indicate new capillary formation was present on the LXW7-modified grafts. Representative H&E staining of the cross sections of the vascular grafts at week 6 after implantation showed that the untreated graft had a significant amount of thrombus formation (C) and LXW7-modified graft exhibited a widely open lumen and no thrombus formation (D). (E) Immunostaining of cross sections of LXW7-modified grafts with mature EC marker CD31. (F) The patency of the grafts was examined at various time points. Each group at each time point included 6 animals.

Article Snippet: For platelet-bead binding assay, 2 mL fresh PRP was added to an ultra-low attachment 35-mm Petri dish (Corning Incorporated) followed by LXW7-modified resin beads.

Techniques: Modification, Staining, Immunostaining, Marker

Journal: Acta biomaterialia

Article Title: Rapid endothelialization of small diameter vascular grafts by a bioactive integrin-binding ligand specifically targeting endothelial progenitor cells and endothelial cells

doi: 10.1016/j.actbio.2020.03.005

Figure Lengend Snippet: Luminal cellularization of vascular grafts. En face DAPI staining was performed for longitudinal sections of the explanted grafts. Untreated grafts (A) and LXW7-modified grafts (B) at week 1 after implantation. Untreated grafts (C) and LXW7-modified grafts (D) at week 2 after implantation. Untreated grafts (E) and LXW7-modified grafts (F) at week 6 after implantation. Higher magnification images of representative areas on the luminal surfaces of the grafts in each group were shown in panels G-L correspondingly. (M) The nucleus coverage on grafts with different treatments and native rat carotid artery were quantified and statistical analyses were performed. Dashed lines indicate the anastomotic sites. Data were expressed as mean ± standard deviation: *p < 0.05, **p < 0.01 (n = 5).

Article Snippet: For platelet-bead binding assay, 2 mL fresh PRP was added to an ultra-low attachment 35-mm Petri dish (Corning Incorporated) followed by LXW7-modified resin beads.

Techniques: Staining, Modification, Standard Deviation

Journal: Acta biomaterialia

Article Title: Rapid endothelialization of small diameter vascular grafts by a bioactive integrin-binding ligand specifically targeting endothelial progenitor cells and endothelial cells

doi: 10.1016/j.actbio.2020.03.005

Figure Lengend Snippet: LXW7 enhanced EPC recruitment and migration and endothelialization on vascular grafts. En face immunostaining for CD31 (green) and CD34 (red) of proximal, middle, and distal portions of grafts. Untreated grafts (A-C) and LXW7-modified grafts (D-F) at week 1 after implantation. Untreated grafts (G-I) and LXW7-modified grafts (J-L) at week 2 after implantation. Untreated grafts (M-O) and LXW7-modified grafts (P-R) at week 6 after implantation. The ration of cells of different phenotypes at week 1 (S), week 2 (T) and week 6 (U) were quantified and statistical analyses were performed. Data were expressed as mean ± standard deviation: *p < 0.05 (n = 5). (V) Shape index of the cell elongation was quantified and statistical analyses were performed. Data were expressed as mean ± standard deviation: *p < 0.05 (n = 10).

Article Snippet: For platelet-bead binding assay, 2 mL fresh PRP was added to an ultra-low attachment 35-mm Petri dish (Corning Incorporated) followed by LXW7-modified resin beads.

Techniques: Migration, Immunostaining, Modification, Standard Deviation