Journal: Journal of Orthopaedic Translation

Article Title: Elevated calcitonin gene-related peptide expression through prostaglandin E2-EP4 receptor pathway in promoting proliferative periosteum formation in primary hypertrophic osteoarthropathy

doi: 10.1016/j.jot.2026.101096

Figure Lengend Snippet: PGE2 on PDSCs osteogenesis in vivo. (A) Immunofluorescent staining for nuclei staining in the femur in PGE2-treated and vehicle-treated rats (scale bars = 100 μm, 1 image from each of 3 rats per PGE2-treated or vehicle-treated group). (B) Representative FACS profile showed the analysis of CD44 and CD105 expressions. Q1: CD44 positive and CD105 negative; Q2: CD44 positive and CD105 positive; Q3: CD44 negative and CD105 negative; Q4: CD44 negative and CD105 positive. (C) At day 7 of osteogenic induction, mRNA expression of osteogenic-associated genes ( Sp7, Alp , Runx2 , and Bglap ) in PDSCs was isolated from the PGE2-treated rat and vehicle-treated rat femora tissues (n = 3 rats per group). (D) ALP staining in PDSCs isolated from PGE2-treated and vehicle-treated rat femora tissues (n = 5 images, scale bars = 100 μm). (E) At day 7 of osteogenic induction, mRNA expression of Cgrp and Ptger4 in PDSCs was isolated from PGE2-treated and vehicle-treated rat femora tissues (n = 3 rats per group). (F) Immunofluorescent staining and immunohistochemistry staining for CGRP, SP7, RUNX2 and BGLAP in the femur in PGE2 and ADV-shCGRP treated rats (1 image from each of 4 rats per group, scale bars = 200 μm). BM=Bone marrow. Outer cortex = peripheral cortex of the cortical bone. All values were expressed as mean ± SD and were replicated at least two times. p value less than 0.05 were considered statistically significant. ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001.

Article Snippet: The recombinant adenovirus vectors ADV-shCGRP (hU6-shCGRP-CMV-EGFP) and ADV-shNC (hU6-CMV-EGFP) were constructed and synthesized by GeneChem (Shanghai, China).

Techniques: In Vivo, Staining, Expressing, Isolation, Immunohistochemistry

Journal: Journal of Orthopaedic Translation

Article Title: Elevated calcitonin gene-related peptide expression through prostaglandin E2-EP4 receptor pathway in promoting proliferative periosteum formation in primary hypertrophic osteoarthropathy

doi: 10.1016/j.jot.2026.101096

Figure Lengend Snippet: PGE2 mediates PDSCs osteogenesis through EP4 receptor and CGRP. (A) Effect of PGE2 (0 nM and 100 nM) on the mRNA expression of Sp7 , Alp , and Runx2 in PDSCs at early time-point (12 h, day 1, day 3) and late time-point (day 7 and day 14) of osteogenesis was measured using Real-time PCR (n = 3 per group). (B) The effect of PGE2 (0 nM and 100 nM) on the mRNA expression of BGLAP in PDSCs at early time-point (day 3) and late time-point (day 7 and day 14) of osteogenesis were measured using Real-time PCR (n = 3 per group). (C) Effect of PGE2 (0 nM and 100 nM) on the mRNA expression of Cgrp and Ptger4 in PDSCs at early time-point (12 h, day 1, day 3) and late time-point (day 7 and day 14) of osteogenesis were measured using Real-time PCR (n = 3 per group). (D) The effect of PGE2 (0 nM and 100 nM) and ADV- shCGRP on the protein expression of SP7, RUNX2, BGLAP, EP4 and CALCRL in PDSCs at day 3, day 7 and day 14 of osteogenesis were measured using western blotting. (E) ALP staining was used to evaluate the effect of PGE2 on the activity of osteoblast in PDSCs culture at day 1, day 3, and day 7 of osteogenesis induction. All values were expressed as mean ± SD using Welch′s t-test and were replicated three times. p value less than 0.05 were considered statistically significant. ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001.

Article Snippet: The recombinant adenovirus vectors ADV-shCGRP (hU6-shCGRP-CMV-EGFP) and ADV-shNC (hU6-CMV-EGFP) were constructed and synthesized by GeneChem (Shanghai, China).

Techniques: Expressing, Real-time Polymerase Chain Reaction, Western Blot, Staining, Activity Assay

Journal: Journal of Orthopaedic Translation

Article Title: Elevated calcitonin gene-related peptide expression through prostaglandin E2-EP4 receptor pathway in promoting proliferative periosteum formation in primary hypertrophic osteoarthropathy

doi: 10.1016/j.jot.2026.101096

Figure Lengend Snippet: Effect of PGE2 on BMSCs osteogenesis in vitro. (A) Effect of PGE2 (0 nM and 100 nM) on the mRNA expression of Sp7 , Alp , and Runx2 in BMSCs at early time-point (12 h, day 1, day 3) and late time-point (day 7 and day 14) of osteogenesis was measured using Real-time PCR (n = 3 per group). (B) The effect of PGE2 (0 nM and 100 nM) on the mRNA expression of Bglap in BMSCs at late time-point (day 7 and day 14) of osteogenesis were measured using Real-time PCR (n = 3 per group). (C) Effect of PGE2 (0 nM and 100 nM) on the mRNA expression of Cgrp and Ptger4 in BMSCs at early time-point (12 h, day 1, day 3) and late time-point (day 7 and day 14) of osteogenesis were measured using Real-time PCR (n = 3 per group). (D) The effect of PGE2 (0 nM and 100 nM) and ADV- shCGRP on the protein expression of SP7, RUNX2, BGLAP, EP4 and CALCRL in BMSCs at day 3 and day 7 of osteogenesis were measured using western blotting. (E) ALP staining was used to evaluate the effect of PGE2 on the activity of osteoblast in BMSCs culture at day 7 and day 14 of osteogenesis induction. All values were expressed as mean ± SD using Welch′s t-test and were replicated three times. p value less than 0.05 were considered statistically significant. ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001.

Article Snippet: The recombinant adenovirus vectors ADV-shCGRP (hU6-shCGRP-CMV-EGFP) and ADV-shNC (hU6-CMV-EGFP) were constructed and synthesized by GeneChem (Shanghai, China).

Techniques: In Vitro, Expressing, Real-time Polymerase Chain Reaction, Western Blot, Staining, Activity Assay