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99Tc-MDP-induced human osteoblast proliferation, differentiation and expression of osteoprotegerin
- Source :
- Molecular Medicine Reports
- Publication Year :
- 2017
- Publisher :
- Spandidos Publications, 2017.
-
Abstract
- The aim of the present study was to examine the influence of technetium methylenediphosphonate (99Tc-MDP) on the proliferation and differentiation of human osteoblasts. Human iliac cancellous bone was isolated and cultured with either 99Tc‑MDP, β fibroblast growth factor (as a positive control) or medium only (as a negative control). Proliferation was assessed by direct cell counting, CCK‑8 assay and bromodeoxyuridine staining. The cell cycle and rate of apoptosis was assessed by propidium iodide staining and flow cytometry. Alkaline phosphatase (ALP) activity was assessed by the p‑nitrophenyl phosphate method and mineralized nodules were stained with Alizarin Red. Expression of osteocalcin (OCN) and bone morphogenetic protein‑2 (BMP‑2) was assessed by reverse transcription‑quantitative polymerase chain reaction (RT‑qPCR), and expression levels of osteoprotegerin (OPG) and receptor activator of NF‑κB ligand (RANKL) were assessed by RT‑qPCR and ELISA. Isolated human osteoblasts stained positively for ALP and developed mineralized nodules. Treatment with 10‑5‑10‑10 M 99Tc‑MDP enhanced proliferation and 48 h incubation with 10‑8 M 99Tc‑MDP increased the proportion of cells in S‑phase, decreased the proportion in G0/G1 phase, and increased the cell proliferation index. The rate of apoptosis also increased, but the increase was not significant. Cells incubated with 10‑6‑10‑9 M 99Tc‑MDP for 3‑9 days exhibited increased ALP activity and mineralized nodule development. 10‑8 M 99Tc‑MDP increased BMP‑2 and OPG expression levels and OPG secretion, but OCN mRNA expression levels and RANKL secretion were not significantly altered at 72 h. 99Tc‑MDP treatment induced osteoblast proliferation and differentiation without affecting apoptosis. These findings provide proof of concept for the future use of 99Tc‑MDP in the treatment of bone‑destructive diseases.
- Subjects :
- 0301 basic medicine
Cancer Research
Cellular differentiation
Bone Morphogenetic Protein 2
Technetium Tc 99m Medronate
Biochemistry
chemistry.chemical_compound
0302 clinical medicine
Cells, Cultured
biology
Cell Cycle
Cell Differentiation
Osteoblast
Articles
Middle Aged
Flow Cytometry
medicine.anatomical_structure
Oncology
RANKL
030220 oncology & carcinogenesis
Osteocalcin
Molecular Medicine
Alkaline phosphatase
Adult
musculoskeletal diseases
medicine.medical_specialty
Bone morphogenetic protein 2
03 medical and health sciences
Calcification, Physiologic
Osteoprotegerin
Internal medicine
Genetics
medicine
Humans
RNA, Messenger
Propidium iodide
Cell Shape
Molecular Biology
Cell Proliferation
receptor activator of NF-κB ligand
Osteoblasts
RANK Ligand
Alkaline Phosphatase
Molecular biology
030104 developmental biology
Endocrinology
technetium methylenediphosphonate
Gene Expression Regulation
chemistry
biology.protein
Subjects
Details
- ISSN :
- 17913004 and 17912997
- Volume :
- 16
- Database :
- OpenAIRE
- Journal :
- Molecular Medicine Reports
- Accession number :
- edsair.doi.dedup.....3cf38b655afe89a0edf34fbb2b76f0a7