中华骨科杂志
中華骨科雜誌
중화골과잡지
CHINESE JOURNAL OF ORTHOPAEDICS
2013年
1期
82-88
,共7页
俞索静%童培建%吴承亮%金红婷%单乐天
俞索靜%童培建%吳承亮%金紅婷%單樂天
유색정%동배건%오승량%금홍정%단악천
破骨细胞%细胞融合%细胞分裂
破骨細胞%細胞融閤%細胞分裂
파골세포%세포융합%세포분렬
Osteoclasts%Cell fusion%Cell division
目的 研究破骨细胞的形成及其特殊细胞生物学行为.方法 采用活细胞成像技术连续动态观察大鼠周围血单核细胞在核因子κB受体激活物配体(receptor activator of NF-κB ligand,RANKL)和巨噬细胞集落刺激因子(macrophage colony stimulating factor,M-CSF)诱导下向破骨细胞分化及形成具有骨吸收功能的多核细胞的全过程.采用倒置相差显微镜观察、抗酒石酸酸性磷酸酶染色、骨磨片扫描电镜观察法对破骨细胞进行鉴定.结果 细胞诱导培养2周后,倒置相差显微镜观察可见大量多核细胞形成；抗酒石酸酸性磷酸酶染色显示绝大部分多核细胞与单核细胞均呈阳性反应；骨磨片扫描电镜观察可见较多的骨吸收陷窝、坑洼、沟道及破骨细胞.活细胞成像观察表明多核破骨细胞是由单核细胞、单核细胞与多核细胞及多核细胞之间相互融合而成,其细胞间的融合均发生在贴壁状态；显微缩时电影观察显示破骨细胞形态复杂多变,多核破骨细胞可以发生分裂.结论 大鼠周围血单核细胞在RANKL和M-CSF诱导下可向破骨细胞分化,形成具有骨吸收功能的破骨细胞.破骨细胞能够通过多种方式融合形成巨大的多核细胞,使其核数增加、体积增大、形态伸缩多变、质膜贴附面积广泛扩展,同时破骨细胞还可通过分裂来缩小体积、减少核数,以适应局部形态学、生物力学及骨吸收动力学的需求.这提示破骨细胞的融合及非有丝分裂方式可能是其发挥功能效应与骨吸收效率的一种特殊细胞生物学行为.
目的 研究破骨細胞的形成及其特殊細胞生物學行為.方法 採用活細胞成像技術連續動態觀察大鼠週圍血單覈細胞在覈因子κB受體激活物配體(receptor activator of NF-κB ligand,RANKL)和巨噬細胞集落刺激因子(macrophage colony stimulating factor,M-CSF)誘導下嚮破骨細胞分化及形成具有骨吸收功能的多覈細胞的全過程.採用倒置相差顯微鏡觀察、抗酒石痠痠性燐痠酶染色、骨磨片掃描電鏡觀察法對破骨細胞進行鑒定.結果 細胞誘導培養2週後,倒置相差顯微鏡觀察可見大量多覈細胞形成；抗酒石痠痠性燐痠酶染色顯示絕大部分多覈細胞與單覈細胞均呈暘性反應；骨磨片掃描電鏡觀察可見較多的骨吸收陷窩、坑窪、溝道及破骨細胞.活細胞成像觀察錶明多覈破骨細胞是由單覈細胞、單覈細胞與多覈細胞及多覈細胞之間相互融閤而成,其細胞間的融閤均髮生在貼壁狀態；顯微縮時電影觀察顯示破骨細胞形態複雜多變,多覈破骨細胞可以髮生分裂.結論 大鼠週圍血單覈細胞在RANKL和M-CSF誘導下可嚮破骨細胞分化,形成具有骨吸收功能的破骨細胞.破骨細胞能夠通過多種方式融閤形成巨大的多覈細胞,使其覈數增加、體積增大、形態伸縮多變、質膜貼附麵積廣汎擴展,同時破骨細胞還可通過分裂來縮小體積、減少覈數,以適應跼部形態學、生物力學及骨吸收動力學的需求.這提示破骨細胞的融閤及非有絲分裂方式可能是其髮揮功能效應與骨吸收效率的一種特殊細胞生物學行為.
목적 연구파골세포적형성급기특수세포생물학행위.방법 채용활세포성상기술련속동태관찰대서주위혈단핵세포재핵인자κB수체격활물배체(receptor activator of NF-κB ligand,RANKL)화거서세포집락자격인자(macrophage colony stimulating factor,M-CSF)유도하향파골세포분화급형성구유골흡수공능적다핵세포적전과정.채용도치상차현미경관찰、항주석산산성린산매염색、골마편소묘전경관찰법대파골세포진행감정.결과 세포유도배양2주후,도치상차현미경관찰가견대량다핵세포형성；항주석산산성린산매염색현시절대부분다핵세포여단핵세포균정양성반응；골마편소묘전경관찰가견교다적골흡수함와、갱와、구도급파골세포.활세포성상관찰표명다핵파골세포시유단핵세포、단핵세포여다핵세포급다핵세포지간상호융합이성,기세포간적융합균발생재첩벽상태；현미축시전영관찰현시파골세포형태복잡다변,다핵파골세포가이발생분렬.결론 대서주위혈단핵세포재RANKL화M-CSF유도하가향파골세포분화,형성구유골흡수공능적파골세포.파골세포능구통과다충방식융합형성거대적다핵세포,사기핵수증가、체적증대、형태신축다변、질막첩부면적엄범확전,동시파골세포환가통과분렬래축소체적、감소핵수,이괄응국부형태학、생물역학급골흡수동역학적수구.저제시파골세포적융합급비유사분렬방식가능시기발휘공능효응여골흡수효솔적일충특수세포생물학행위.
Objective To investigate the formation and special cell biological behavior of osteoclasts.Methods The live-cell imaging technology was adopted to consecutively and dynamically observe the whole process of multinuclear osteoclast formation induced by RANKL and M-CSF from rat peripheral blood monocyte.Meanwhile,the inverted phase contrast microscopy,TRAP staining,and scanning electron microscopy were also applied to identify the osteoclast.Results After 2-week cultivation,a great number of apocytes were found by the inverted phase contrast microscopy,and most apocytes and monocytes had positive reaction after TRAP staining.Moreover,many bone resorption lacunae in which osteoclasts were perhrming bone resorption function could be found in the bone slice under the scanning electron microscope.Live-cell imaging observation showed that the multinuclear osteoclasts were generated through self-fusion of monocytes,fusion of monocytes and apocytes,as well as fusion between apocytes.All fusion processes occurred under the condition of cell adherence.Time-lapse Microcinematography observation showed diverse shapes of osteoclasts and the cell division of multinuclear osteoclasts.Conclusion Rat peripheral blood monocyte can develop into osteoclast under induction of RANKL and M-CSF.Osteoclast can form gigantic apocyte via various types of cell fusion to increase its nucleus number and cell volume,vary its shape,and increase the area of plasma membrane.On the other hand,osteoclast can decrease its cell volume and nucleus number via cell division to adapt the needs of local morphology,biomechanics and bone resorption dynamics.It suggests that this non-mitosis cell division is a special cell biological behavior of osteoclast,which may be the basis of exerting its function and improving bone resorption efficiency.
