南方医科大学学报

南方医科大学学报 ›› 2021, Vol. 41 ›› Issue (8): 1277-1282.doi: 10.12122/j.issn.1673-4254.2021.08.22

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人骨形态发生蛋白-2/人胰岛素样生长因子-1对钛种植体表面细胞的成骨效率的影响

张 迪，邓 甜，罗 震，朱安棣，杨 勃，钟惠兰，李少萍，杨晓喻   

  1. 南方医科大学口腔医院种植科，广东 广州 510280；中山大学附属第七医院口腔科，广东 深圳 518107
  • 出版日期:2021-08-20 发布日期:2021-09-07

Surface modification of titanium implant with hBMP-2/hIGF-1 for promoting biocompatibility and osteogenesis

ZHANG Di, DENG Tian, LUO Zhen, ZHU Andi, YANG Bo, ZHONG Huilan, LI Shaoping, YANG Xiaoyu   

  1. Department of Implantology, Stomatological Hospital, Southern Medical University, Guangzhou 510280,China; Department of Stomatology, Seventh Affiliated Hospital of Sun Yat-sen University, Shenzhen 518107, China
  • Online:2021-08-20 Published:2021-09-07

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摘要： 目的 制备人骨形态发生蛋白-2（hBMP-2）/人胰岛素样生长因子-1（hIGF-1）涂层钛（Ti）片，探索其作为牙种植体的功效与机制。方法 将hBMP-2与hIGF-1两个生长因子涂覆到具有光滑表面的Ti上，比较两种生长因子包被的Ti与原始Ti在增强骨形成和骨整合的功效。通过扫描电子显微镜SEM和接触角测量，分别测定金属样品的表面特性。将MG63细胞种植于Ti上，MTT实验检测细胞增殖，茜素红实验检查细胞钙化结节，Elisa实验检测碱性磷酸酶，Western blot实验检测骨钙素与骨调素的表达。结果 SEM检测显示，包被了hBMP-2/hIGF-1的Ti，表面呈雪花辐射样。与原始Ti相比，hBMP-2/Ti、hIGF-1/Ti和hBMP-2/hIGF-1/Ti显示出较低的接触角（P=0.032，0.029，0.028），样本呈现良好的亲水性。MTT实验结果显示，MG63细胞能在hBMP-2/hIGF-1加持的钛片表面良好生长。茜素红实验显示，hBMP-2/hIGF-1的加持能提高MG63细胞的成骨效率（P<0.05）。Elisa实验显示，hBMP-2/hIGF-1能促进碱性磷酸酶的分泌（P=0.021，0.014）。Western blot 实验显示，hBMP-2/ hIGF-1的加持能诱导骨钙素与骨调素的表达（P<0.05）。结论 以hBMP-2和hIGF-1两种生长因子涂覆的Ti，能提高其表面细胞的成骨效率，可以作为一种种植材料进一步用于牙科实验。

关键词: hBMP-2；hIGF-1；肝素；骨形成；骨整合

Abstract: Objective To prepare the human bone morphogenetic protein-2 (hBMP-2)/ human insulin-like growth factor-1 (hIGF-1) coating titanium (Ti) and assess its performance as a dental implant material. Methods hBMP-2 and hIGF-1 were coated to the smooth surface of a Ti plate, and its efficacy for promoting bone formation and bone integration was compared with a pristine Ti plate. The surface characteristics of the metal samples were evaluated using scanning electron microscope (SEM) and by contact angle measurement. MG63 cells were seeded on the surface of the Ti plates, and MTT assay and alizarin red staining was used to examine the cell proliferation and formation of calcified nodules, respectively. Alkaline phosphatase (ALP) secretion of the cells was examined with ELISA, and cellular expressions of osteocalcin and osteopontin were detected with Western blotting for assessing osteogenesis. Results SEM examination showed that the surface of Ti with hBMP-2 and hIGF-1 coating presented with a radial pattern resembling snowflakes. The contact angles of non-coated Ti, hBMP-2-coated Ti, hIGF-1-coated, and hBMP-2/-hIGF-1-coated Ti samples were 83.2°, 54°, 56° and 54°, respectively. Compared with the non-coated Ti plate, the surface-modified Ti samples showed a significantly smaller contact angle (P=0.032, 0.029, and 0.028), indicating a good hydrophilicity of the samples. MTT assay showed that MG63 cells grew well on the surface of the coated Ti plates. The hBMP-2/IGF-1 coating significantly induced cellular secretion of ALP (P=0.021, 0.014) and obviously promoted osteogenesis of MG63 cells (P<0.05). Western blotting results showed that hBMP-2/IGF-1 coating significantly enhanced the expressions of osteocalcin and osteopontin in the seeded cells (P<0.05). Conclusion hBMP-2 and hIGF-1 coating of Ti material can promote osteogenesis of the cells seeded on its surface to improve the performance of such Ti material as dental implants.

Key words: human bone morphogenetic protein-2; insulin-like growth factor-1; heparin; osteogenesis; osseointegration