Poudel Sher Bahadur, Bhattarai Govinda, Kook Sung-Ho, Shin Yun-Ji, Kwon Tae-Ho, Lee Seung-Youp, Lee Jeong-Chae
Cluster for Craniofacial Development & Regeneration Research, Institute of Oral Biosciences, Chonbuk National University, Jeonju 54896, South Korea.
Cluster for Craniofacial Development & Regeneration Research, Institute of Oral Biosciences, Chonbuk National University, Jeonju 54896, South Korea; Department of Bioactive Material Sciences, Research Center of Bioactive Materials, Chonbuk National University, Jeonju 54896, South Korea.
Growth Horm IGF Res. 2017 Oct;36:1-10. doi: 10.1016/j.ghir.2017.07.003. Epub 2017 Jul 30.
Transgenic plant cell suspension culture systems have been utilized extensively as convenient and efficient expression systems for the production of recombinant human growth factors. We produced insulin-like growth factor-1 using a plant suspension culture system (p-IGF-1) and explored its effect on new bone formation in calvarial defects. We also compared the bone regenerating potential of p-IGF-1 with commercial IGF-1 derived from Escherichia coli (e-IGF-1). Male C57BL/6 mice underwent calvarial defect surgery, and the defects were loaded with absorbable collagen sponge (ACS) only (ACS group) or ACS impregnated with 13μg of p-IGF-1 (p-IGF-1 group) or e-IGF-1 (e-IGF-1 group). The sham group did not receive any treatment with ACS or IGFs after surgery. Live μCT and histological analyses showed critical-sized bone defects in the sham group, whereas greater bone formation was observed in the p-IGF-1 and e-IGF-1 groups than the ACS group both 5 and 10weeks after surgery. Bone mineral density, bone volume, and bone surface values were also higher in the IGF groups than in the ACS group. Local delivery of p-IGF-1 or e-IGF-1 more greatly enhanced the expression of osteoblast-specific markers, but inhibited osteoclast formation, in newly formed bone compared with ACS control group. Specifically, p-IGF-1 treatment induced higher expression of alkaline phosphatase, osteocalcin, and osteopontin in the defect site than did e-IGF-1. Furthermore, treatment with p-IGF-1, but not e-IGF-1, increased mineralization of MC3T3-E1 cells, with the attendant upregulation of osteogenic marker genes. Collectively, our findings suggest the potential of p-IGF-1 in promoting the processes required for bone regeneration.
转基因植物细胞悬浮培养系统已被广泛用作生产重组人生长因子的便捷高效表达系统。我们利用植物悬浮培养系统生产了胰岛素样生长因子-1（p-IGF-1），并探讨了其对颅骨缺损中新骨形成的影响。我们还比较了p-IGF-1与源自大肠杆菌的商业IGF-1（e-IGF-1）的骨再生潜力。雄性C57BL/6小鼠接受颅骨缺损手术，缺损处仅填充可吸收胶原海绵（ACS组），或填充含有13μg p-IGF-1的ACS（p-IGF-1组）或e-IGF-1（e-IGF-1组）。假手术组术后未接受任何ACS或IGF治疗。活体μCT和组织学分析显示，假手术组存在临界尺寸的骨缺损，而在术后5周和10周时，p-IGF-1组和e-IGF-1组的骨形成均比ACS组更多。IGF组的骨矿物质密度、骨体积和骨表面积值也高于ACS组。与ACS对照组相比，局部递送p-IGF-1或e-IGF-1在新形成的骨中更显著地增强了成骨细胞特异性标志物的表达，但抑制了破骨细胞的形成。具体而言，p-IGF-1治疗在缺损部位诱导的碱性磷酸酶、骨钙素和骨桥蛋白表达高于e-IGF-1。此外，p-IGF-1治疗而非e-IGF-1治疗增加了MC3T3-E1细胞的矿化，并伴随成骨标志物基因的上调。总的来说，我们的研究结果表明p-IGF-1在促进骨再生所需过程方面具有潜力。
