Effect Of Melatonin On Osteoblast Biological Behaviour In Pure Titanium Four Surfaces

Objective:(1)The purpose of this study was to find and compare surface topography?roughness and wettability of four different surfaces treatment which affected the attachment and spread of MC3T3-E1 osteoblast in vitro.(2)Observation of effect melatonin on osteoblast proliferation,migration and calcification in pure titanium surface.Methodes:(1)Discs were divided into 4 groups :discs-grooved(G),acid-etching(G-S,for S),micro-arc oxidation(G-S-MAO,for MAO),alkali(G-S-MAO-OH,for OH).The surface morphology,roughness and wettability were determined by scanning electron microscope,roughness measuring instrument and surface contact angle meter.The adhension amount of 30 min,60min,120 min were determined by CCK-8.Cell morphology and F-actin cy-toskeleton were measured at 3h,6h,12 h by laser confocal microscope.(2)The effect of different concentration MT on osteoblast proliferation were detected by CCK-8;On the basis of the above four groups to add the MT and were divided into eight groups :G,S,MAO,OH,G + MT,S + MT,MAO + MT,OH + MT.Osteoblast proliferation of each group were tested by using CCK-8 at 24 h,48 h,72 h;The laser confocal observed osteoblast migration of each group;Alizarin red staining detected calcification of each group.Results:(1)Ra degree:MAO(0.526±0,017um)? OH(0.460±0.019um)? S(0.120±0.018um)?G(0.056±0.005um);(2)Wettability angle: G(66.333°±2.073°)?S(61.518°±0.791°)?MAO(21.707°±1.555°)?OH(5.363°±0.669°);(3)Attachment value of osteoblast: 30 min,S?G?MAO?OH,60 min and 120 min,MAO?S?OH?G;(4)Cell morphology change:3h cells of G group were arranged regularly along the surface of the horizontal grain regularity and microfilament along horizontal grain distribution;S group were the circular partly and had not yet spread,MAO group were spread out short pseudopodia;6h OH group were enlongated along the horizontal grain elongation,S group were fusiform or irregular quadrilateral,MAO group set to extend filiform pseudopodia,OH group were spread out multiple pseudopodia all around;12 h G group were narrow strip along horizontal grain,S group had not changed partly and spread out filiform pseudopodia partly,MAO group were spread out tabular pseudopodia partly,OH group had been completely spread out tabular pseudopodia all around.(5)The effect different concentration melatonin on osteoblast proliferation: 50 umol/L>100 umol/L>10 umol/L>0 umol/L;(6)Osteoblast proliferation of every group: in addition to the OH group and OH+MT were not Significant difference(P > 0.05)at 72 h,other time-point three groups were statistically different comparing with corresponding adding MT groups(P < 0.05).(7)The degree of osteoblast migration: Osteoblast migration degree of four groups were increased after adding MT.(8)Calcification of Osteoblast each group: Four groups improve calcification degrees after adding MT and nano rough surface calcification degrees was increased obviously.Conclusion:(1)Nanometer rough surface was more beneficial for the initial attachment of the osteoblas;(2)Alkali treatment surface promoted the extension of cell,but reduced the cell attachment and proliferation;Porous MAO and OH group were in favor of early cellular pseudopod extend;(3)MT can promote osteoblast proliferation,migration and calcification on the different specimen surface;It was relatively weak for the effect of MT on osteoblast proliferation on the alkali treatment surface;MT promoted osteoblast calcification obviously on the nano rough surface.