title: Proliferation and differentiation of osteoblasts from the mandible of osteoporotic rats.
abstract: The objective of this study was to identify the differences between osteoblasts derived from normal adult rat mandibles and osteoporotic adult rats. An osteoporotic animal model was established by performing a bilateral ovariectomy (ovx group). The proliferation and differentiation abilities of osteoblasts were determined by MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl-2H-tetrazolium bromide), alkaline phosphatase (ALP) and osteocalcin release (OC) assays. Transmission electron microscopy (TEM) was performed to assess differences in the ultrastructure. Proliferating cell nuclear antigen (PCNA) and uncoupling protein 2 (UCP2) protein concentrations were analyzed by Western blot. In addition, UCP2 protein in osteoblasts was assessed by immunohistochemistry staining. ATP and reactive oxygen species (ROS) concentrations were analyzed separately with ATP and ROS quantification kits. At four and 12 weeks after the operation, osteoblasts of the ovx group showed earlier attachment, fewer dead cells and faster growth compared with cells in the sham group. TEM showed that osteoblasts of the ovx group had fewer folds, lysosomes, peroxisomes and less rough endoplasmic reticulum. The results of the MTT, ALP activity and OC assays were all higher in osteoblasts from the ovx group at four or 12 weeks postsurgery than osteoblasts from the sham group. PCNA protein concentrations in the ovx group increased significantly compared with those of the sham group at four or 12 weeks after the operation, but UCP2 concentrations decreased over the same time period. UCP2 immunohistochemical staining of osteoblasts showed that the protein was concentrated in the cytoplasm and that the osteoblasts from the sham group had higher expression than those from the ovx group. The ATP and ROS concentrations of the ovx groups were significantly higher than the sham groups at four or 12 weeks postsurgery. Therefore, we concluded that there are differences in cell ultrastructure, proliferation, differentiation, ATP and ROS concentrations, and PCNA and UCP2 protein expression levels in osteoblasts from the mandibles of rats of the ovx group compared with those from the sham group.
authors.affiliation: Department of Stomatology, The General Hospital of Jinan Military Region of PLA, Jinan 250031, China.
authors.forename: Shu-Juan
authors.initials: SJ
authors.lastname: Yu
authors.forename: Hong-Chen
authors.initials: HC
authors.lastname: Liu
authors.forename: E
authors.initials: E
authors.lastname: Ling-Ling
authors.forename: Dong-Sheng
authors.initials: DS
authors.lastname: Wang
authors.forename: Guo-Xiong
authors.initials: GX
authors.lastname: Zhu
mesh_terms: D000818:Animals; D002454:Cell Differentiation; D049109:Cell Proliferation; D005260:Female; D007150:Immunohistochemistry; D008334:Mandible / Q000166:cytology / Q000378:metabolism*; D046529:Microscopy, Electron, Transmission; D010006:Osteoblasts / Q000166:cytology* / Q000378:metabolism; D015675:Osteocalcin / Q000378:metabolism; D010024:Osteoporosis / Q000378:metabolism / Q000473:pathology; D051381:Rats; D017207:Rats, Sprague-Dawley
chemical_list: D015675:Osteocalcin