同种异基因组织工程骨修复骨缺损IFN-γ和IL-10的表达

IFN-γ and IL-10 expression in serum and spleen after treatment of bone defect with allogeneic tissue engineering bone

[目的]通过检测同种异基因组织工程骨(tissue engineering bone,TEB)修复骨缺损血清IFN-γ、IL-10水平和脾内IFN-γmRNA、IL-10mRNA的表达,探讨同种异基因TEB的免疫原性。[方法]Ficoll-Hypaque梯度密度离心法分离出成年兔骨髓间充质干细胞(MSCs),培养扩增。取第3代细胞,化学法诱导成成骨细胞(ODC)。将ODC按1×109/L密度接种至羟基磷灰石/磷酸三钙(HA/TCP),共同培养,构建TEB。动物随机分三组(TEB,HA/TCP组,自体骨组),每组24只,人为造成兔左桡骨处1.0 cm左右长创伤性骨缺损,植入TEB、HA/TCP、自体骨。观察伤口愈合情况;于术后第4、10 d、6周,麻醉下取下腔静脉血ELISA法测血清IFN-γ、IL-10水平;取脾行HE染色观察;RT-PCR测IFN-γ、IL-10 mRNA表达。[结果]各组动物IL-10、IL-10 mRNA比IFN-γ和IFN-γ mRNA高,其中TEB组IFN-γ和IFN-γmRNA较其他两组稍高,IL-10和IL-10 mRNA较其他两组稍低,IFN-γ、IL-10水平和IFN-γmRNA、IL-10 mRNA表达同组前后比较有统计学差异(P<0.05),组间比较没有差异(P>0.05)。[结论]由同种异基因MSCs诱导而成的ODC与HA/TCP构建的TEB免疫原性极低,可以用于修复骨缺损。

[ Objective] To investigate immunogenicity of allogeneic tissue engineering bone （TEB） by checking IFN -γ, IL - 10 levels in serum and IFN - γmRNA, IL - 10mRNA expression in spleen after treatment bone defects with TEB. [ Meth- ods] Marrow mesenchymal stem cells （MSCs） were obtained from adult rabbit bone marrow by Ficoll -Hypaque density gradient centrifugation. Osteoblasts （ODC） were obtained by inducing of the MSCs of Passage 3 with the chemical method. TEB was constructed by culturing ODC which had been inoculated to hydroxyapatite/tri - calcium phosphate （HA/TCP） at density of 1× 109/ L together with HA/TCP. Animals were randomly divided into three groups （TEB, HA/TCP, selfbone ） , each groupincluded 24 animals with man - made traumatic bone defect at left radius were implanted TEB, HA/TCP and self - bone. After operation, animals were observed wound healing and IFN -γ, IL- 10 level in serum, IFN -γ mRNA IL- 10 mRNA in spleen were measured at 4, 10 days and 6 weeks. [Results] The level of IL- 10 and expression of IL- 10 mRNA were higher than that of IFN - γ and IFN -γ, mRNA in all groups and IFN - γ, IFN - γ mRNA of the group of TEB was higher than those in the other two groups. IL - 10, IL - 10 mRNA of the group of TEB were lower than those in the other two groups. With Statistical analysis there was difference between the level of IFN -γ, IL - 10 and the expression of IFN - γmRNA, IL - 10 mRNA （ P 〈 0. 05） and There was no difference among each groups （P 〉 0. 05 ） . [ Conclusion ] The allogeneic TEB constructed by the ODC can be ued to renair hone defects.