Effect Of 3D Printed Titanium Alloy With Different Surface Roughness On The Early Osteogenic Activity Of HBMSCs

Objective:To explore the effect of 3D printing titanium alloy materials with different surface roughness on the early osteogenic activity of hBMSC.Method:A solid titanium alloy sample(diameter 30mm*height 3mm)was prepared by using Huashu Hi-Tech FS-121M selective laser melting(SLM)equipment.By setting the printing direction of the sample,the rough titanium block was obtained and the smooth titanium block was obtained by polishing;Culture human bone marrow mesenchymal stem cells(hBMSC)to the third generation.The above two materials and hBMSC were placed in the same six-well plate hole to construct a 3D printed titanium alloy-hBMSC co-culture system with different roughness surfaces.Cultured in vitro for 48h,using cignal-45-pathway-reporter-array detection method for transcription factor excitability screening.Then Western blot technology was used to detect the proteins(MEF2C,BMP2,TGFβ1,COL X,VEGF)related to early osteogenic indicators,as well as qRT-PCR technology and IF technology to detect the RNA transcription and protein expression of co-cultured cells.Results:The results of transcription factor activity screening in co-culture system suggested that MEF2C expression in smooth group cells was highly excited.Western blot results showed that compared with the blank group,the early expression of various osteogenic indicators(MEF2C,BMP2,TGFβ-1,VEGF,COL X)in hBMSCs in the smooth group increased(p<0.05),and in the rough group(MEF2C The early expression of BMP2,TGFβ-1)increased(p<0.05),and the early expression of various osteogenic indicators(MEF2C,BMP2,VEGF,COL X)in hBMSC in the relatively smooth group and rough group decreased(p<0.05);Immunofluorescence results:compared with the blank group,the various hBMSCs in the smooth group The early fluorescent expression intensity of osteogenic indicators(MEF2C,BMP2,VEGF,COL X)all increased(p<0.05).In the rough group,the early fluorescent expression intensity of various osteogenic indicators(BMP2,TGFβ-1,COL X)in hBMSC were enhanced(p<0.05).In the relatively smooth group,in the rough group,the early fluorescent expression intensity of various osteogenic indicators(MEF2C,VEGF)in hBMSC were all weakened(p<0.05),and the early fluorescent expression intensity of BMP2 was up-regulated(p<0.05);the QPCR results showed:relatively blank The early expression of various osteogenic indicators(MEF2C,BMP2,TGFβ-1,VEGF,COL X)in hBMSC in the smooth group and rough group were all up-regulated(p<0.05).In the relatively smooth group,the early expression of various osteogenic indicators(MEF2C,TGFβ-1,VEGF,COL X)in hBMSCs in the rough group were all down-regulated(p<0.05).Conclusions:3D printed titanium alloys with smooth surfaces and rough surfaces have better early osteogenic activity...