Characterization of the F-box Proteins FBXW2 and FBXL14 in the Initiation of Bone Regeneration in Transplants given to Nude Mice

Abstract

Background: Cultured bovine-periosteum-derived cells can form three-dimensional structures on tissue culture dishes without artificial scaffolding material, can induce bone regeneration in vivo. The utility of cultured bovine-periosteum-derived cells for bone tissue regeneration after their transplantation into nude mice has been reported, the precise F-box molecular mechanism was unclear. Objective: The aim of this study was to investigate the specific F-box proteins required for bone regeneration by cultured bovine-periosteum-derived cells in vitro. Methods: In the present study, periosteum tissue and cultured periosteum-derived cells were cultured for 5 weeks in vitro and then embedded in collagen gel with a green tissue-marking dye. Electrophoresis and immunohistochemistry were used to identify the specific F-box proteins required for tissue bone regeneration. Results: The bovine-periosteum-derived cells were observed to form bone shortly after the expression of F-box proteins. After the initial phase of bone formation, the expression of the F-box proteins ceased. FBXW2 was shown to be expressed in the periosteum, but not in cultured periosteum-derived cells. Furthermore, FBXL14 disappeared during bone formation. Conclusions: Bone regeneration requires progenitor cells, such as bovine-periosteum-derived cells and the activation of the F-box Proteins FBXW2 and FBXL14, over time the expression of these proteins ceases. Further scientific and clinical trials are needed to investigate how the F-box Proteins can be used therapeutically to treat osteoporosis and osteonecrosis. 背景： 培养的牛骨膜来源的细胞可以在组织培养皿上形成三维结构，而无需人工支架材料，可以在体内诱导骨骼再生。已经报道了将培养的牛骨膜来源的细胞移植到裸鼠中后用于骨组织再生的效用，尚不清楚精确的F-box分子机制。 客观的： 这项研究的目的是研究体外培养的牛骨膜来源的细胞进行骨再生所需的特定F-box蛋白。 方法： 在本研究中，将骨膜组织和培养的骨膜来源的细胞体外培养5周，然后用绿色组织标记染料将其包埋在胶原蛋白凝胶中。电泳和免疫组织化学用于鉴定组织骨再生所需的特定F-box蛋白。 结果： 在表达F-box蛋白后不久，观察到牛骨膜来源的细胞形成骨。在骨骼形成的初始阶段之后，F-box蛋白的表达停止了。FBXW2显示在骨膜中表达，但在培养的骨膜来源的细胞中不表达。此外，FBXL14在骨形成过程中消失。 结论： 骨再生需要祖细胞（例如牛骨膜来源的细胞）和F-box蛋白FBXW2和FBXL14的激活，随着时间的流逝，这些蛋白的表达将停止。需要进一步的科学和临床试验来研究如何将F-box蛋白用于治疗性骨质疏松和骨坏死。 关键词：牛骨膜培养细胞FBXW2 FBXL14骨再生骨钙素TRAP染色