tfllr nh2 peptide  (Tocris)

Journal: Scientific reports

Article Title: Osteogenesis in human periodontal ligament stem cell sheets is enhanced by the protease-activated receptor 1 (PAR 1 ) in vivo.

doi: 10.1038/s41598-022-19520-x

Figure Lengend Snippet: Figure 1. (A) Immunofluorescence, flow cytometry and alizarin red staining experiment flow. (B) Flow cytometry analysis showing the phenotypic characterization of PDLSCs for surface stemness, multipotent embryonic biomarkers (SOX2, OCT4, STRO-1) and PAR1 from three individuals (I1, I2 and I3). Unstained cells were used to set positive cell populations, p < 0.05. (C) PAR1 immunofluorescence expression in PDLSCs isolated cell lines. Images in 60X magnification. (D) Alizarin red staining for PDLSCs sheets (I2) after 14 days. Results are given as the mean ± SD. Figure (A) was created using BioRender.

Article Snippet: Subsequently, cell sheets were assigned for one of the following experimental groups: (1) control medium (CTRL) composed of α-MEM (Thermo Fisher Scientific, Waltham, USA) supplemented with 10% of FBS (Thermo Fisher Scientific, Waltham, USA) and 50 μg/mL vitamin C (Sigma-Aldrich, St Louis, USA); (2) osteogenic medium (OST) (CTRL + 100 nM dexamethasone, 5 mM b-glycerophosphate and 50 μg/ml ascorbic acid (Sigma-Aldrich, St Louis, USA) and (3) osteogenic medium with the addition the PAR1 agonist peptide (100 nM TFLLR-NH2) (Tocris Bioscience Inc., Bristol, UK) (OST + PAR1).

Techniques: Immunofluorescence, Flow Cytometry, Staining, Expressing, Isolation

Journal: Scientific reports

Article Title: Osteogenesis in human periodontal ligament stem cell sheets is enhanced by the protease-activated receptor 1 (PAR 1 ) in vivo.

doi: 10.1038/s41598-022-19520-x

Figure Lengend Snippet: Figure 3. (A) Cell sheets in vivo transplant and ex-vivo analysis workflow. (B) Micro-CT analysis of ectopic bone formation in vivo after 60 days of transplantation (B) and the statistical analysis comparing the BV/TV% between the 3 groups CTRL, OST and OST + PAR1. (*) means a p < 0.05 compared to control. Results are given as the mean ± SD. Figure (A) was created using BioRender.

Article Snippet: Subsequently, cell sheets were assigned for one of the following experimental groups: (1) control medium (CTRL) composed of α-MEM (Thermo Fisher Scientific, Waltham, USA) supplemented with 10% of FBS (Thermo Fisher Scientific, Waltham, USA) and 50 μg/mL vitamin C (Sigma-Aldrich, St Louis, USA); (2) osteogenic medium (OST) (CTRL + 100 nM dexamethasone, 5 mM b-glycerophosphate and 50 μg/ml ascorbic acid (Sigma-Aldrich, St Louis, USA) and (3) osteogenic medium with the addition the PAR1 agonist peptide (100 nM TFLLR-NH2) (Tocris Bioscience Inc., Bristol, UK) (OST + PAR1).

Techniques: In Vivo, Ex Vivo, Micro-CT, Transplantation Assay, Control

Journal: Scientific reports

Article Title: Osteogenesis in human periodontal ligament stem cell sheets is enhanced by the protease-activated receptor 1 (PAR 1 ) in vivo.

doi: 10.1038/s41598-022-19520-x

Figure Lengend Snippet: Figure 4. (A) Ex-vivo histological qualitative analysis from H&E, Van Gieson, Masson’s Trichrome and Von Kossa stains from the 3 groups after a 2-month transplantation period in balb/c nude mice. Images at 10x and 20x. CTRL group displayed connective tissue characteristics with necrotic spots (gray arrows) and no bone formation was detected in this group for the 4 stains. Osteoblast clusters (black solid arrows) and blood vessels (red solid arrows) were detected in higher abundance in the OST group when compared with the other groups. The OST + PAR1 group showed greater osteocyte numbers (black dotted circle) and higher calcium deposits (B) (red dotted line).

Article Snippet: Subsequently, cell sheets were assigned for one of the following experimental groups: (1) control medium (CTRL) composed of α-MEM (Thermo Fisher Scientific, Waltham, USA) supplemented with 10% of FBS (Thermo Fisher Scientific, Waltham, USA) and 50 μg/mL vitamin C (Sigma-Aldrich, St Louis, USA); (2) osteogenic medium (OST) (CTRL + 100 nM dexamethasone, 5 mM b-glycerophosphate and 50 μg/ml ascorbic acid (Sigma-Aldrich, St Louis, USA) and (3) osteogenic medium with the addition the PAR1 agonist peptide (100 nM TFLLR-NH2) (Tocris Bioscience Inc., Bristol, UK) (OST + PAR1).

Techniques: Ex Vivo, Transplantation Assay

Journal: Scientific reports

Article Title: Osteogenesis in human periodontal ligament stem cell sheets is enhanced by the protease-activated receptor 1 (PAR 1 ) in vivo.

doi: 10.1038/s41598-022-19520-x

Figure Lengend Snippet: Figure 5. (A) Histological immunohistochemistry quantification analysis for collagen type I, integrin 1β and bone sialoprotein for the 3 groups ex-vivo samples after a 2-month transplantation period in balb/c nude mice. (B) A semi-quantification analysis was performed for the 3 protein markers and the PAR1 activation group showed greater staining for the 3 osteogenic protein markers after 60 days. Images at 10x. (*) p < 0.05. Results are given as the mean ± SD.

Article Snippet: Subsequently, cell sheets were assigned for one of the following experimental groups: (1) control medium (CTRL) composed of α-MEM (Thermo Fisher Scientific, Waltham, USA) supplemented with 10% of FBS (Thermo Fisher Scientific, Waltham, USA) and 50 μg/mL vitamin C (Sigma-Aldrich, St Louis, USA); (2) osteogenic medium (OST) (CTRL + 100 nM dexamethasone, 5 mM b-glycerophosphate and 50 μg/ml ascorbic acid (Sigma-Aldrich, St Louis, USA) and (3) osteogenic medium with the addition the PAR1 agonist peptide (100 nM TFLLR-NH2) (Tocris Bioscience Inc., Bristol, UK) (OST + PAR1).

Techniques: Immunohistochemistry, Ex Vivo, Transplantation Assay, Activation Assay, Staining