Effects Of Alkaline Heat Treatment On Integrins Of Osteoblasts On Titanium Surface

[Objective] To evaluate the effects of alkali-and heat-treated surface of titanium discs on integrin αx2β1, integrin α5β1of osteoblasts in vitro.[Materials and Methods] In order to harvest abundant vital osteoblast, this research use the traditional tissue piece culture, and applied the continuous piece method to culture primary osteoblast of rat calvaria. The growth condition of primary osteoblast was observed under inverse microscope, identified by hematoxylin-eosinstaining, alkaline phosphatase staining and mineralized nodule alizarin red staining. Medical pure titanium which15mm in diameter, lmm in thick will grouping prepared before test. G surface were prepared by hand grinding to240grits,360grits,600grits with SiC paper. SB surface were treated by TiO2particles with356-411μm diameter. SLA surface were attacked with acidic solution(H2SO4/HCL) for10min at60°C. Alkali-and heat-treated(AHl, AH2, AH3) surface were prepared by G, SB, SLA surface respectively. G, SB, SLA surface were heated to600°C for1h after surfaces were soaked in5moL/L NaOH aqueous solution at60°C for24h. Scanning electron microscopy, contour measuring instrument were used to test surface properties. Osteoblasts were planted on the6titanium disks. At1d,3d and5d after planting, osteoblasts were harvested. mRNA expression of integrin subunit α2, α5, β1were examined by immunofluorescent quantitative real-time polymerase chain reaction. Datum were analyzed by one-way ANOVA and paired t-test(a=0.05).[Results] The result indicated that pieces primary osteoblasts are abundant and pure, which met the experimental need. The cultured cells by this method were identified to accord to the osteoblastic bionomics and had favorable vitality. The result have showed that integrin α2, α5and β1mRNA expressed in the osteoblasts continuously. The mRNA expression of integrin β1is most abundant and the mRNA expressions of a2, a5are relatively lower than integrin β1. In the study the mRNA expression of integrina2, integrina5, integrinβ1in group of SB surface is most abundant. The mRNA expression of integrina2, integrina5, integrinβ1in group of AH1surface is more abundant than that in group of G surface, same as group of AH3surface than group of SLA surface. The mRNA expression of integrina2, integrina5, integrinβ1in group of AH1surface, AH2surface, AH3surface are close. In the observation period, expression levels of integrina2, integrina5, integrinβ1mRNA increased to some extent when time went by.[Conclusions]1. Method of pieces is relative simple and consumed smaller amount of bone pieces, and the primary osteoblasts are abundant and pure, which met the experimental need. The cultured cells by this method were identified to accord to the osteoblastic bionomics and had favorable vitality.2. Specificity and efficacy of primers are qualified the primer expectation, expected hands are got,3target integrin existing are detected by PCR.3. The mRNA expression of integrina2, integrina5, integrinβ1in group of SB surface is most abundant.The group of SB surface could conducive osteoblasts adhesion.4. Alkali-heat treatment on integrina2, integrina5, integrinβof osteoblasts expression have a role in promoting, Alkali-heat treatment on the surface play a role in osteoblasts adhesion.