多孔磷酸三钙-羟基磷灰石对人牙龈成纤维细胞黏附行为的影响

目的　研究多孔磷酸三钙 羟基磷灰石 (tricalcium phosphate hydroxyapatite ,TCP HA)复合材料对人牙龈成纤维细胞黏附行为的影响。方法　应用离子束辅助沉积法在纯钛试件表面制备多孔TCP HA复合涂层材料和羟基磷灰石 (hydroxyapatite,HA)涂层材料 ,定量对比人牙龈成纤维细胞在涂层和未涂层材料表面初期黏附、增殖、细胞铺展面积、细胞外基质和黏着斑形成的情况。结果　TCP HA和HA涂层材料表面黏附的细胞数、细胞铺展面积高于纯钛未涂层组 ,差异有显著性(P <0 0 5 ) ,黏着斑的形成早于未涂层组 ;TCP HA表面黏附的细胞数和Ⅰ型胶原的形成都高于纯钛未涂层组和HA涂层组。在培养 2 4h后TCP HA组表面黏附细胞数为 198 1± 2 7 7,Ⅰ型胶原形成的荧光强度为 15 4 10± 31 5 6 ,同纯钛组表面的细胞数 ( 12 5 1± 2 9 9)和Ⅰ型胶原荧光强度( 132 6 3± 35 2 6 )相比 ,差异有显著性 (P <0 0 5 )。结论　与纯钛未涂层和HA涂层材料相比 ,多孔TCP HA复合涂层材料更有利于人牙龈成纤维细胞的初期黏附 ,具有更良好的生物相容性

Effects of TCP/HA-coating titanium on the adhesion behavior of human gingival fibroblasts

OBJECTIVE: To investigate effects of hydroxyapatite (HA) and porous tricalcium phosphate/hydroxyapatite (TCP/HA)-coating titanium on the adhesion behavior of human gingival fibroblasts (HGFs). METHODS: Coatings of HA and duplex phases TCP/HA on titanium (Ti) were formed by ion beam assisted deposition (IBAD) method. Attachment, spreading, extracellular matrix (ECM) production, and focal adhesion plaque formation of HGFs were investigated on commercially pure (CP) titanium, HA-coated CP titanium and porous TCP/HA-coated CP titanium. After incubation of HGFs on these substrates, the number of attached cell, the area of cell spreading, immunostained ECM including fibronectin (FN) and type I collage, and vinculin (presenting the formation of focal adhesion plaque) were quantified by morphometric analysis using immunofluorescence microscope. RESULTS: TCP/HA and HA coatings exhibited that the attached cell number and cell spreading area were greater than those of CP titanium (P < 0.05), and the formation of focal adhesion plaque was earlier than that of uncoated substrate (P < 0.05). The number of attached cell and the formation of type I collagen on TCP/HA were more than those on Ti and HA. After 24-hour incubation on TCP/HA surface, the number of attached cell was 198.1 +/- 27.7 and the fluorescent intensity of type I collagen was 154.10 +/- 31.56. While under the same condition, the corresponding numbers for the CP titanium were 125.1 +/- 29.9 and 132.63 +/- 35.26. The differences between the two groups were significant (P < 0.05). CONCLUSIONS: In this study, the porous TCP/HA coating significantly facilitated the adherence of human gingival fibroblasts to Ti surface and could improve the biocompatibility of titanium.