Journal: Journal of Dental Sciences

Article Title: Human umbilical cord mesenchymal stem cells secretome and nanoemulsion propolis combination ameliorate osteoclastogenesis in lipopolysaccharide-induced osteolysis in hyperglycemia rats

doi: 10.1016/j.jds.2025.07.003

Figure Lengend Snippet: Effect of NEP and HUCMSCS combination on LPS-induced osteolysis in hyperglycemic conditions in TRAPase and Ctsk serum levels. (A) Concentration of TRAP in serum was measured by ELISA. (B) Concentration of Ctsk in serum was measured by ELISA. A comparison of (A) TRAPase and (B) Ctsk serum levels in all experimental groups. Results are expressed as the mean ± SD. The statistical significance of differences was determined by t-tests and one-way analysis of variance (ANOVA) by Tukey Honest Significant Different (HSD) with significance set at P ≤ 0.05. n = 4 for each group. CTSK, Cathepsin K; HUCMSCS, human umbilical cord mesenchymal stem cells secretome; LPS, lipopolysaccharide; NEP, nanoemulsion propolis extract; PBS, phosphate-buffered saline; SD, standart deviation; TRAP, tartrate-resistant acid phosphatase.

Article Snippet: The steps of the sandwich ELISA method begin with preparation, sample incubation, washing, target antibody incubation, sample washing, analysis, and interpretation according to the instructions of the rat TRAP and Ctsk ELISA kit (Shanghai Korain Biotech Co., Ltd., Shanghai, China).

Techniques: Concentration Assay, Enzyme-linked Immunosorbent Assay, Comparison, Saline

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: 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: 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: 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) (/mm 2 ) 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

Journal: International journal of cancer

Article Title: Transcriptional profiling on chromosome 19p indicated frequent downregulation of ACP5 expression in hepatocellular carcinoma.

doi: 10.1002/ijc.20684

Figure Lengend Snippet: FIGURE 2 – Transcriptional mapping on chromosome 19p by cDNA microarray. (a) Normalized expression levels of parental tumors (PHC-2, 4, 5 and C1) examined were plotted against the cytoband locations of cDNA clones in sequential order of megabases. Relative expression levels depicted as inverted curves indicated a consistent reduced fold change by 5- to 24-fold of a cDNA clone R05288 that represented the ACP5 gene. (b) An example of hybridized image on the cDNA microarray is shown. Parental tumor was labeled with Cy3 (green color) and normal liver pool with Cy5 (red color). The bracket highlights the duplicate spots of ACP5; a strong red signal found suggested a lower level of mRNA transcripts compared to normal control.

Article Snippet: Plasmid transfection and effect on in vitro cell growth A plasmid clone encoding a full-length human ACP5 cDNA (TC119151) was acquired from OriGene (OriGene Technologies, Rockville, MD, USA).

Techniques: Microarray, Expressing, Clone Assay, Labeling, Control

Journal: International journal of cancer

Article Title: Transcriptional profiling on chromosome 19p indicated frequent downregulation of ACP5 expression in hepatocellular carcinoma.

doi: 10.1002/ijc.20684

Figure Lengend Snippet: FIGURE 3 – ACP5 expression in HCC. (a) The bar chart indicates the log2 ratio of the normalized ACP5 ex- pression in 74 primary HCC tumors relative to the level in paired adjacent nonmalignant liver tissues. (b) Repre- sentative quantitative RT-PCR prod- ucts are shown. Expression levels of ACP5 (98 bp) determined were nor- malized against the internal reference control -actin (108 bp). The ACP5 expression in the HCC tumors (H36, H40, H42 and H46) was found to be lower than that of the adjacent nonma- lignant normal liver tissues (TN36, TN40, TN42 and TN46).

Article Snippet: Plasmid transfection and effect on in vitro cell growth A plasmid clone encoding a full-length human ACP5 cDNA (TC119151) was acquired from OriGene (OriGene Technologies, Rockville, MD, USA).

Techniques: Expressing, Quantitative RT-PCR, Control

Journal: International journal of cancer

Article Title: Transcriptional profiling on chromosome 19p indicated frequent downregulation of ACP5 expression in hepatocellular carcinoma.

doi: 10.1002/ijc.20684

Figure Lengend Snippet: FIGURE 4 – Effect of ectopic expression of ACP5 on HCC cell growth. (a) Growth curve of HKCI-4 cells expressing empty vector (HKCI-4/Vec). A similar result was also ob- served in the experiment with mock lipo- fectamine alone (p 0.05). (b) HKCI-4 cells expressing ACP5 (HKCI-4/ACP5) displayed a significant growth reduction at day 6 compared to both control and mock experiments (p 0.001). The values are expressed as a percent- age of maximum and each point represents the average of 5 replicates derived from 2 inde- pendent experiments (mean SD).

Article Snippet: Plasmid transfection and effect on in vitro cell growth A plasmid clone encoding a full-length human ACP5 cDNA (TC119151) was acquired from OriGene (OriGene Technologies, Rockville, MD, USA).

Techniques: Expressing, Plasmid Preparation, Control, Derivative Assay

Journal: Molecular Medicine Reports

Article Title: Carnosol inhibits osteoclastogenesis in vivo and in vitro by blocking the RANKL-induced NF-κB signaling pathway

doi: 10.3892/mmr.2022.12741

Figure Lengend Snippet: Carnosol attenuates OVX-induced bone loss in vivo . (A) H&E staining of femoral sections from the sham, OVX and OVX + carnosol groups 6 weeks following OVX. Scale bar, 500 µm. (B) Micro-CT of femoral sections from the sham, OVX and OVX + carnosol groups 6 weeks following OVX. Scale bar, 1 mm. (C) Quantitative analysis of femoral sections. (D) Serum CTX-1, TRAcp5b and IL-6 levels in the sham, OVX and OVX + carnosol groups 6 weeks following OVX. (E) Serum OCN levels in the sham, OVX and OVX + carnosol groups. **P<0.01, ***P<0.001. OVX, ovariectomy; CTX-1, C-terminal telopeptide; TRAcp5b, tartrate-resistant acid phosphatase type 5b; OCN, osteocalcin; BMD, bone mineral density; BV/TV, bone volume/total volume; Tb.N, trabecular number; BS/TV, bone surface area/total volume.

Article Snippet: IL-6 ELISA Kit (cat. no. BMS603-2, Invitrogen, USA), C-terminal telopeptide (CTX-1) ELISA Kit (cat. no. MBS458686, Mybiosource, USA), tartrate-resistant acid phosphatase type 5b (TRAcp5b) ELISA kit (cat. no. MBS163341, Mybiosource, USA) and osteocalcin (OCN) ELISA Kit (cat. no. MBS725134, Mybiosource, USA) were used for ELISA according to the manufacturer's protocol.

Techniques: In Vivo, Staining, Micro-CT

Journal: Evidence-based Complementary and Alternative Medicine : eCAM

Article Title: Gujiansan Ameliorates Avascular Necrosis of the Femoral Head by Regulating Autophagy via the HIF-1 α /BNIP3 Pathway

doi: 10.1155/2021/6683007

Figure Lengend Snippet: Concentrations of OC, BAP, TRACP-5b, NTX-1, and CTX-1 in each group on the 2nd, 4th, and 8th weeks ( n = 6).

Article Snippet: A rat OC ELISA kit (Cloud-Clone Corp, #SEA471Ra, China), rat BAP ELISA kit (Jiancheng, #A059-1, China), rat TRACP-5b ELISA kit (Cloud-Clone Corp, #SEA902Ra, China), rat NTX-1 ELISA kit (Cloud-Clone Corp. #CEA639Ra, China), rat CTX-1 ELISA kit (Cloud-Clone Corp, #CEA665Ra, China), rat interleukin-2 (IL-2) ELISA kit (CUSABIO, #CSB-E04628r, China), rat tumor necrosis factor- α (TNF- α ) ELISA kit (CUSABIO, #CSB-E11987r, China), rat interleukin-6 (IL-6) ELISA kit (CUSABIO, #CSB-E04640r, China), and rat C-reactive protein (CRP) ELISA kit (CUSABIO, #CSB-E07922r, China) were used to measure the concentrations of bone formation- and bone resorption-specific markers and cytokines in this study.

Techniques: