tracp5b  (Elabscience Biotechnology)

Journal: Bone & Joint Research

Article Title: Melatonin alleviates senile osteoporosis by regulating autophagy and enhancing fracture healing in aged mice

doi: 10.1302/2046-3758.142.bjr-2024-0112.r2

Figure Lengend Snippet: Fig. 1 Aged bone marrow-derived mesenchymal stem cells (BMMSCs) showed degenerative properties in osteogenesis and melatonin (MEL) promoted osteogenesis of aged BMMSCs. a) Haematoxylin and eosin (H&E) staining (40×) of proximal tibiae and b) micro-CT (μCT) imaging of distal femora of aged and young mice. c) Bone microstructure parameters such as bone mineral density (BMD), trabecular bone volume per total volume (BV/TV), trabecular bone thickness (Tb.Th), trabecular bone number (Tb.N), and trabecular bone separation (Tb.Sp). An independent-samples t-test was used to calculate the p-value in Figure 1c. d) Alkaline phosphatase (ALP) staining (20×) and e) Alizarin Red staining (20×) to analyze the osteogenesis of young and aged BMMSCs. f) Gene enrichment analysis of genes related to senile osteoporosis predicted using the Genecard database, DisGeNET database, and OMIM database, highlighting pathways where MEL metabolism and effects showed the highest enrichment in WikiPathways. Chi-squared test was used to calculate the p-value. g) MEL in serum was analyzed by enzyme-linked immunosorbent assay (ELISA). h) ALP staining (5×), i) ALP level analysis, j) Alizarin Red staining (5×), and k) mineralization level analysis were performed. *p < 0.05, significant differences between each indicated group analyzed using one-way analysis of variance (ANOVA). CTR, control group; IL-18, interleukin-18; MAPK, mitogen-activated protein kinase; ns, not significant; PI3K-AKT, phosphoinositide 3-kinase-protein kinase B; VEGFA-VEGFR2, vascular endothelial growth factor-vascular endothelial growth factor receptor 2.

Article Snippet: The concentration of tartrate-resistant acid phosphatase 5 (ACP5), OCN, MEL, in mouse serum were measured with Mouse Tartrate Resistant ACP5 ELISA Kit, Mouse Osteocalcin ELISA Kit, and Mouse Melatonin ELISA Kit (Elabscience Biotechnology, China), respectively, according to the manufacturer’s instructions.

Techniques: Derivative Assay, Staining, Micro-CT, Imaging, Enzyme-linked Immunosorbent Assay, Control

Journal: Bone & Joint Research

Article Title: Melatonin alleviates senile osteoporosis by regulating autophagy and enhancing fracture healing in aged mice

doi: 10.1302/2046-3758.142.bjr-2024-0112.r2

Figure Lengend Snippet: Fig. 3 Inhibition of autophagy could counteract melatonin (MEL)-induced osteogenic promotion in aged mice. a) Micro-CT (μCT) imaging of distal femora and b) haematoxylin and eosin (H&E) staining (40×) of proximal tibiae of aged mice treated with MEL for six weeks (50 mg kg-1 body weight per day). c) Bone microstructure parameters such as bone mineral density (BMD), trabecular bone volume per total volume (BV/TV), trabecular bone thickness (Tb.Th), trabecular bone number (Tb.N), and trabecular bone separation (Tb.Sp) were measured by μCT scanning. d) to f) Immunohistopathology (200×) showed the osteogenic-related protein presentations of d) matricellular molecular osteocalcin (OCN) and e) osteogenic marker Osterix in proximal tibia bone tissue, and f) integral optical density (IOD) was calculated. g) to i) Concentrations of g) bone resorption marker ACP5, h) osteogenic marker OCN, and i) MEL in serum were analyzed by enzyme-linked immunosorbent assay (ELISA). *p < 0.05, significant differences between each indicated group analyzed using Fisher’s exact test, one-way analysis of variance (ANOVA), or Tukey’s post-hoc test. CTR, control group; ns, not significant.

Article Snippet: The concentration of tartrate-resistant acid phosphatase 5 (ACP5), OCN, MEL, in mouse serum were measured with Mouse Tartrate Resistant ACP5 ELISA Kit, Mouse Osteocalcin ELISA Kit, and Mouse Melatonin ELISA Kit (Elabscience Biotechnology, China), respectively, according to the manufacturer’s instructions.

Techniques: Inhibition, Micro-CT, Imaging, Staining, Marker, Enzyme-linked Immunosorbent Assay, Control

Journal: Bone & Joint Research

Article Title: Melatonin alleviates senile osteoporosis by regulating autophagy and enhancing fracture healing in aged mice

doi: 10.1302/2046-3758.142.bjr-2024-0112.r2

Figure Lengend Snippet: Fig. 5 Administration of melatonin (MEL) has therapeutic effects on bone fracture healing in aged mice. a) Representative radiograph and micro-CT (μCT) of femora in aged mice (n = 6 per group) four weeks after open femoral mid-shaft fracture. b) Nonunion frequency and c) μCT measurement of bone volume per total volume (BV/TV) and bone mineral density (BMD) in callus area of the fractured femora four weeks after open femoral mid-shaft fracture. d) Haematoxylin and eosin (H&E) staining (50×) of fractured femur. e) Ratio of newly formed bone, cartilage, and fibrous tissue area and quantification of the area of new bone formation. f) Immunohistopathology (200×) of fractured femur, showing the osteogenic-related protein matricellular protein osteocalcin (OCN) in the fracture callus. The integral optical density (IOD) of OCN was calculated. g) Osteoblast counts in the fracture callus were assessed by counting the number of osteoblasts on each H&E-stained section, with osteoblast number/bone perimeter (N.Ob/B.Pm) (/mm) determined using Image-Pro Plus software (Media Cybernetics, USA). Blue arrows indicate typical osteoblasts in the sections, highlighting their distribution in the fracture callus. h) Evaluation of osteoclasts in fracture callus. The numbers of osteoclasts were counted based on tartrate-resistant acid phosphatase (TRAP)-stained sections, and osteoblast number/total area (N.OC/T.Ar) (/mm2) was determined by Image-Pro Plus software. i) Protein presentations from bone marrow-derived mesenchymal stem cells (BMMSCs) extracted from tibiae after fracture healing of autophagy markers LC3BII, Beclin, and the osteogenesis marker OCN. *p < 0.05, significant differences between each indicated group analyzed using Fisher's exact test, one-way analysis of variance (ANOVA), or Tukey’s post-hoc test. CTR, control group; GelMA, methacrylated gelatin; ns, not significant.

Article Snippet: The concentration of tartrate-resistant acid phosphatase 5 (ACP5), OCN, MEL, in mouse serum were measured with Mouse Tartrate Resistant ACP5 ELISA Kit, Mouse Osteocalcin ELISA Kit, and Mouse Melatonin ELISA Kit (Elabscience Biotechnology, China), respectively, according to the manufacturer’s instructions.

Techniques: Micro-CT, Staining, Software, Derivative Assay, Marker, Control