EDTA对人牙周膜细胞在病变根面附着和增殖影响

The effects of 240 g/L EDTA gel on the attachment and proliferation of human periodontal ligment cells onto the root surface of teeth with periodontitis

目的:评价240g/LEDTA凝胶处理病变根面不同时间对人牙周膜细胞附着、增殖的影响。方法:收集因晚期牙周炎拔除的单根牙44颗,经刮治、根面平整后,制备5mm×4mm大小的根片,随机分成EDTA2min组、EDTA3min组、EDTA4min组和对照组,在根片上接种细胞。分别于接种1、3、5d后,每组各取3个根片固定、染色,计数,分析细胞附着和增殖程度。另外每组2个根片做扫描电镜观察其形态学变化。结果:接种第1天发现240g/LEDTA凝胶处理不同时间的根片的细胞附着无差异(P>0.05);而和对照组之间都有差异(P<0.05);随着接种时间的延长,各组根片上的细胞数明显增加。240g/LEDTA凝胶处理3min、4min的根片上的细胞数无差异(P>0.05),与其他2组有差异(P<0.05);EDTA2min组与对照组之间亦有差异(P<0.05)。EDTA处理根片3min、4min,均能去除玷污层,暴露完整的胶原;处理2min的根片上有残留的玷污层。结论:240g/LEDTA处理不同时间后的根面能促进人牙周膜细胞的附着和增殖,并推荐临床处理时间为3min。

Objective:To study attachment and proliferation of human periodontal ligment cells(HPDLCs) on demineralized root surface of teeth with periodontitis following EDTA gel application. Methods:44 single-rooted teeth extracted because of severe periodontitis were treated by scaling and root planing(SRP). Then the fragments(5 mm×4 mm) of experimental surfaces were prepared. The fragments were randomly divided into four groups.The root surfaces in group 1,2,3 and 4 were treated by 240 g/L of EDTA gel for 2,3 and 4 min respectively or without treatment(control). Then 1×104 cells were seeded on the surface of each fragment and routinely cultured for 1,3 and 5 days respectively after seeding, the specimens were prepared for microscopic observasion, the cells on the root surfaces were counted. Moreover the other eight fragments were prepared for SEM observasion. Results:HPDLCs attached to and proliferated on all treated surfaces.Group 1, 2 and 3 presented significantly more cells on the root surfaces than the control group did at day 1,3 and 5. The cells on the root surfaces in group 2 and 3 were more than those in group 1 at day 1, 3 and 5. There was no evident difference between group 2 and 3 at any time. The root surfaces in group 2 and 3 had a fibrillar texture associated with an increased diameter of the exposed dentinal tubule openings.Smear layer were observed on root surfaces in group 1. Conclusion:EDTA gel(at 240 g/L) application may enhance attachment and proliferation on root surface of HPDLCs.