Li Xin-Yu, Ji Cai-Ni, Xu Ling-Juan, Hu Wei-Kun, Zhou Bin, Li Gui-Gang
Department of Ophthalmology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, Hubei Province, China.
Department of Ophthalmology, the Third Hospital of Wuhan, Wuhan 430060, Hubei Province, China.
Int J Ophthalmol. 2014 Feb 18;7(1):22-6. doi: 10.3980/j.issn.2222-3959.2014.01.04. eCollection 2014.
To investigate the influence of minTBP-1-PRGDN on the attachment, proliferation and collagen I synthesis of human keratocyte on titanium (Ti) surface.
The chimeric peptide RKLPDAPRGDN (minTBP-1-PRGDN) was synthesized by connecting RKLPDA (minTBP-1) to the N-terminal of PRGDN, the influence of minTBP-1-PRGDN on the attachment, proliferation and collagen I synthesis of human keratocyte on Ti surface were tested using PRGDN and minTBP-1as controls. The keratocytes attached to the surface of Ti were either stained with FITC-labeled phalloidin and viewed with fluorescence microscope or quantified with alamar Blue method. The proliferation of keratocytes on Ti were quantified with 3-(4,5-dim- ethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide up-taking methods. The secretion of type I collagen were determined using an ELISA kit.
The results showed that minTBP-1-PRGDN at a concentration of 100ng/mL was the most potent peptide to enhance the attachment of human keratocytes to the surface of Ti (1.40±0.03 folds, P=0.003), to promote the proliferation (1.26±0.05 folds, P=0.014) and the synthesis of type I collagen (1.530±0.128, P=0.008). MinTBP-1 at the same concentration could only promote the attachment (1.13±0.04 folds, P=0.020) and proliferation(1.15±0.06 folds, P=0.021), while PRGDN had no significant influence (P>0.05).
Our data shows that the novel chimeric peptide minTBP-1-PRGDN could promote the attachment, proliferation and type I collagen synthesis of human keratocytes on the surface of Ti.
研究微型TBP-1-PRGDN对人角膜细胞在钛（Ti）表面附着、增殖及I型胶原合成的影响。
通过将RKLPDA（微型TBP-1）连接到PRGDN的N端合成嵌合肽RKLPDAPRGDN（微型TBP-1-PRGDN），以PRGDN和微型TBP-1作为对照，检测微型TBP-1-PRGDN对人角膜细胞在Ti表面附着、增殖及I型胶原合成的影响。附着在Ti表面的角膜细胞用异硫氰酸荧光素标记的鬼笔环肽染色后用荧光显微镜观察，或用alamar Blue法进行定量。用3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐摄取法对Ti表面角膜细胞的增殖进行定量。用ELISA试剂盒测定I型胶原的分泌。
结果显示，浓度为100ng/mL的微型TBP-1-PRGDN是增强人角膜细胞在Ti表面附着（1.40±0.03倍，P=0.003）、促进增殖（1.26±0.05倍，P=0.014）及I型胶原合成（1.530±0.128，P=0.008）作用最强的肽。相同浓度的微型TBP-1仅能促进附着（1.13±0.04倍，P=0.020）和增殖（1.15±0.06倍，P=0.021），而PRGDN无显著影响（P>0.05）。
我们的数据表明，新型嵌合肽微型TBP-1-PRGDN可促进人角膜细胞在Ti表面的附着、增殖及I型胶原合成。
