骨细胞在力学信号感受过程中的作用

目的 综述骨细胞在力学信号感受过程中的作用.资料来源与选择国内外公开发表的相关学术论文及研究报告.资料引用引用文献资料57篇.资料综合主要对骨细胞的生物学特点,骨细胞与成骨细胞之间的差异,骨细胞的特殊结构在力传导中的作用,骨细胞是骨组织的力学感受器,骨细胞力学感受过程中的信息传递,骨细胞与失重性骨丢失等6个方面进行综述.结论 骨细胞是骨组织中的力学感受器,其感受机械刺激,通过细胞网络传递信号,在骨重塑过程中调节成骨细胞和破骨细胞的功能. Abstract： Objective To summarize the roles of osteocytes in bone mechanosensation. Literature resource and selection Related papers and reviews that published at home and abroad. Literature quotation Fifty-seven papers were cited. Literature synthesis Such subjects as biological characteristics of osteocytes, differences between osteocytes and osteoblasts, the roles of special structures of osteocytes in mechanotransduction, the mechanosensors of bone, signal transmission during mechanosensation, the relationship of osteocytes and weightless caused bone loss were reviewed and summarized. Conclusion Osteocytes are the mechanosensors in bone. When mechanical stimuli are sensed, osteocytes regulate the functions of osteoblasts and osteoclasts by the cellular network transmission in the process of bone remodeling.     

仙灵骨葆胶囊治疗绝经后骨质疏松症的生物信息学和网络药理学分析和实验验证

目的：基于生物信息学和网络药理学挖掘仙灵骨葆胶囊(XLGB)治疗绝经后骨质疏松症(PMOP)的活性成分和作用机制。方法：采用生物信息学挖掘PMOP相关靶点,采用网络药理学筛选XLGB的活性成分并预测作用靶点,阐明XLGB治疗PMOP的相关靶点,富集分析其信号通路以探究分子机制。动物实验采用去卵巢SD大鼠动物模型,XLGB灌胃3月,检测骨密度,血清活性氧水平,TUNEL染色检测骨组织凋亡情况,蛋白质免疫印迹法检测骨组织内凋亡相关蛋白B细胞淋巴瘤2(Bcl-2)、Bcl-2相关X蛋白(Bax)表达量。结果：筛选出差异表达基因883个,筛选出XLGB活性成分119种,共预测1 734个靶蛋白。通过对XLGB和PMOP共有的26个共同靶点分析显示,AKT1、NFKB1、BAX等靶点及细胞凋亡、破骨细胞分化、cAMP、AMPK等信号通路起重要作用。XLGB组骨密度明显改善,血清活性氧水平降低,TUNEL染色显示XLGB可减少骨组织凋亡情况,提高抗凋亡蛋白Bcl-2、降低促凋亡蛋白Bax的水平。结论：XLGB治疗PMOP具有多成分、多靶点的特点,与调控细胞凋亡密切相关。     

Cell-based resorption assays for bone graft substitutes

The clinical utilization of resorbable bone substitutes has been growing rapidly during the last decade, creating a rising demand for new resorbable biomaterials. An ideal resorbable bone substitute should not only function as a load-bearing material but also integrate into the local bone remodeling process. This means that these bone substitutes need to undergo controlled resorption and then be replaced by newly formed bone structures. Thus the assessment of resorbability is an important first step in predicting the in vivo clinical function of bone substitute biomaterials. Compared with in vivo assays, cell-based assays are relatively easy, reproducible, inexpensive and do not involve the suffering of animals. Moreover, the discovery of RANKL and M-CSF for osteoclastic differentiation has made the differentiation and cultivation of human osteoclasts possible and, as a result, human cell-based bone substitute resorption assays have been developed. In addition, the evolution of microscopy technology allows advanced analyses of the resorption pits on biomaterials. The aim of the current review is to give a concise update on in vitro cell-based resorption assays for analyzing bone substitute resorption. For this purpose models using different cells from different species are compared. Several popular two-dimensional and three-dimensional optical methods used for resorption assays are described. The limitations and advantages of the current ISO degradation assay in comparison with cell-based assays are discussed.     

Effects of neridronic acid on osteoclasts derived by physiological dual-cell cultures

Increased osteoclastic activity is observed in many osteopathic disorders - including postmenopausal osteoporosis, Paget's disease, primary bone tumours, lytic bone metastases, multiple myeloma and rheumatoid arthritis - that involve increased bone resorption and a loss of bone mass. Bisphosphonates are highly effective inhibitors of bone resorption that selectively affect the osteoclasts. The aim of this study was to obtain more information about the mechanism of action of bisphosphonates such as neridronic acid using a dual-cell culture model. As a model of osteoclastogenesis we used a murine monocyte/macrophage cell line RAW 264.7 type CRL 2278 co-cultured with murine osteoblasts. The monocyte-osteoblast system allows physiological experimentation of bone anti-resorption drugs, simulating bone turnover in pathologies such as osteoporosis. The direct actions of neridronic acid on cell proliferation and functionality in the co-culture model were examined using tartrate-resistant acid phosphatase (TRAP) assay, immunohistochemical localization of actin, and transmission and scanning electron microscopy (SEM). Results showed that the percentage of TRAP-positive cells, an early marker of osteoclastic differentiation, was significantly higher in control cultures than in co-cultures treated with variable concentrations of neridronic acid. Neridronic acid induced dramatic morphological changes, characterized by the loss of the ruffled border. The actin ring associated with the plasma membrane of the cells treated with neridronic acid was shown to break down. The tissue-specific targeting of neridronic acid to bone mineral suggests that it may inhibit bone resorption by direct effects on osteoclasts or other bone cells in the immediate microenvironment of the osteoclasts. From our study, we conclude that structural alterations induced by neridronic acid in our co-culture system lead to decreased osteoclast function. This may encourage the use of neridronic acid to reduce bone resorption in the therapy of demineralizing metabolic bone disorders.     

破骨细胞在类风湿关节炎骨破坏机制中作用的研究进展

破骨细胞是类风湿关节炎骨破坏机制研究的核心.NF-κB受体激活剂配体（RANKL）以及Th17细胞释放的白细胞介素（IL）-17与其分化及功能密切相关.破骨细胞（前体）内的受体、衔接子、激酶和转录因子在其融合、增殖与存活、细胞骨架重构和骨吸收等活动中起着至关重要作用.     

CaMKIIδ在破骨细胞分化不同阶段表达规律的研究

目的:研究钙离子/钙调蛋白依赖性蛋白激酶II(Ca MKII)δ在破骨细胞分化不同阶段的表达规律,以揭示其在破骨细胞分化中的作用。方法:应用50μg/L核因子κB受体激活因子配体(RANKL)诱导小鼠RAW264.7细胞向破骨细胞分化;通过抗酒石酸酸性磷酸酶(TRAP)染色及骨磨片吸收陷窝检测评价破骨细胞生成情况;同时于诱导第0、1、3、5天末通过免疫荧光细胞化学、RT-q PCR和Western blot法检测Ca MKIIδ的mRNA及蛋白表达水平。结果:TRAP染色及骨吸收陷窝检测显示于诱导第5天有多核破骨细胞生成。第0、1、3、5天Ca MKIIδ的mRNA表达水平分别为1.028±0.041、2.478±0.087、10.524±1.284和42.914±2.667,蛋白相对水平分别为0.762、0.963、1.802和3.136,免疫荧光细胞化学检测显示Ca MKIIδ的荧光6强度呈时间依赖性递增。结论:Ca MKIIδ的表达随破骨细胞分化逐步增高,提示Ca MKIIδ在破骨细胞分化中可能起着关键调控作用。     

Ultrastructural study of cell-cell interaction between osteoclasts and osteoblasts/stroma cells in vitro

Many biochemical reports support cell-cell interaction between osteoclasts and osteoblasts/stroma cells in vitro, however there have been few morphological studies supporting this. Details of cell-cell interaction between osteoclasts and osteoblasts/stroma cells remain unclear. The present study examined cell-cell interaction between osteoclasts and osteoblasts/stroma cells by scanning electron microscopy (SEM) and transmission electron microscopy (TEM). Osteoclasts, osteoblasts/stroma cells, and bone marrow cells obtained from 10-day-old ddY mice were cultured on dentin slices for 72 hr. Specimens were fixed, and some were examined by SEM. Specimens were decalcified, embedded in Epon after determination of the tartrate-resistant acid phosphatase activity (TRAP), and TRAP-positive cells for investigation were serially sectioned by alternating semithin and ultrathin sections, and then examined by TEM. By SEM, many cellular contacts were seen between the cells cultured on the dentin, but by TEM there were few special structures on the cell membranes between osteoclasts and osteoblasts/stroma cells, or between osteoclasts and bone marrow cells. A special structure on the cell membranes of osteoclasts was observed between an osteoclast and a cytoplasmic process of osteoblast/stroma cells, and this cell membrane was coated with electron dense or bristle-like structures. These bristle-like structures were very similar to those of coated pits. The present results show that the coated pit-like structure plays an important role in cell-cell interaction between osteoclasts and osteoblasts/stroma cells in vitro, and suggest that macromolecules binding to the osteoclast-surface receptor via ligands, accumulate in the coated pits, and enter the osteoclast as receptor-macromolecule complexes in endocytic vesicles.     

Moonlighting osteoclasts as undertakers of apoptotic cells

Rapid clearance of apoptotic cells, frequently referred to as efferocytosis, is crucial for the maintenance of tissue homeostasis and the prevention of autoimmunity. The common model of apoptotic cell clearance involves a system of released “Find me” and exposed “Eat me” signals on apoptotic cells, detected and recognized by matching receptors on macrophages or dendritic cells (DC), referred to as the phagocytic synapse. Osteoclasts share the monocyte lineage with these professional mononuclear phagocytes, thus raising the question if, in addition to bone resorption, osteoclasts can act as scavengers for apoptotic cells. Our qPCR data clearly show that osteoclasts express most of the genes required for dying cell clearance at mRNA levels similar to or even higher than those observed in M1-macrophages, M2-macrophages or DC. Our microscopical analyses reveal that osteoclasts in fact can bind and/or engulf apoptotic cells in an essentially serum-independent fashion. Together with our data on the abundance of the respective mRNAs, these results identify the vitronectin receptor (integrin α_νβ₃)/milk fat globule-EGF factor 8 protein (MFG-E8) axis, the scavenger receptors (CD36, CD68 and class A macrophage scavenger receptor (SR-A)), the complement/complement receptor axis, the Mer/Tyro3/Protein S axis, and the phosphatidylserine (PS) receptor brain-specific angiogenesis inhibitor 1 (BAI1) as the most promising candidates to be involved in osteoclast-mediated efferocytosis.     

An ultrastructural study of cartilage resorption at the site of initial endochondral bone formation in the fetal mouse mandibular condyle

An ultrastructural study was undertaken on cartilage resorption at the site of initial endochondral bone formation in the mouse mandibular condyle on d 16 of pregnancy. After resorbing the bone collar, the osteoclasts extended their cell processes into the cartilage matrix and made contact with hypertrophic chondrocytes. By means of cell processes or vacuolar structures, these osteoclasts entrapped the calcified cartilage matrices, cell debris, and the degraded uncalcified cartilage matrices. In particular, since the calcified cartilage matrices were sometimes seen to be disrupted within the osteoclastic vacuolar structures, they were probably disposed of by the osteoclasts. Invading endothelial cells giving rise to capillaries also directly surrounded the degraded uncalcified cartilage matrices and small deposits of cell debris. In addition, hypertrophic chondrocytes that had attached to or were in the process of attaching to the invading osteoclasts often enclosed the small calcified cartilage matrices. Other cell types that have often been reported in other regions of cartilage resorption were not seen at the site of initial endochondral bone formation in this study. Our findings in relation to cartilage resorption may therefore represent unique features of the site of initial endochondral bone formation site. We consider that the manner of cartilage resorption is likely to vary by site, age, and species.