CAJ | 학술논문

目的：探讨使用小鼠基质细胞衍生因子1（murine stromal cell-derived factor 1， mSDF-1）结合辛伐他汀（simvastatin，SIM）以及骨胶原支架（Bio-Oss?）构建无外加种子细胞的组织工程化骨的可行性，并检验其体内异位成骨的效果。方法：将32只ICR小鼠随机分为4组，每组8只，于小鼠颅部做皮肤切口，各组小鼠分别植入：（1）1∶50（体积比）的二甲基亚砜（dimethyl sulfoxide，DMSO）／磷酸盐缓冲液（phosphate-buffered saline，PBS）混合液＋骨胶原支架（空白对照组）；（2）10－3mol／L SIM溶液＋骨胶原支架（SIM组）；（3）200 mg／L mSDF-1溶液＋骨胶原支架（mSDF-1组）；（4）10－3 mol／L SIM＋200 mg／L mSDF-1溶液＋骨胶原支架（SIM＋mSDF-1组）。植入1周后，连续2 d，每天分别在支架局部注射上述各组相应的溶液50μL。饲养6周后，取出支架及其周围组织，通过软X射线投射成像及灰度测定、HE染色、免疫组织化学染色的方法，定性及定量观察其成骨效果。结果：SIM＋mSDF-1组软X射线灰度值[（421836．5±65425．7）像素]明显高于空白对照组[（153345．6±45222．2）像素，P＜0．01]、SIM组[（158119．2±100284．2）像素，P＜0．01]以及mSDF-1组[（255529．5±152142．4）像素，P＜0．05]；在SIM＋mSDF-1组内可见明显的骨桥蛋白和骨钙素的表达；SIM＋mSDF-1组血管丛密度[（46±8）条／mm2]明显高于空白对照组[（23±7）条／mm2， P＜0．01]和SIM组[（24±6）条／mm2， P＜0．01]。结论：使用mSDF-1结合SIM以及骨胶原支架构建的无外加种子细胞的组织工程化骨可于小鼠颅部皮下异位成骨。
목적：탐토사용소서기질세포연생인자1（murine stromal cell-derived factor 1， mSDF-1）결합신벌타정（simvastatin，SIM）이급골효원지가（Bio-Oss?）구건무외가충자세포적조직공정화골적가행성，병검험기체내이위성골적효과。방법：장32지ICR소서수궤분위4조，매조8지，우소서로부주피부절구，각조소서분별식입：（1）1∶50（체적비）적이갑기아풍（dimethyl sulfoxide，DMSO）／린산염완충액（phosphate-buffered saline，PBS）혼합액＋골효원지가（공백대조조）；（2）10－3mol／L SIM용액＋골효원지가（SIM조）；（3）200 mg／L mSDF-1용액＋골효원지가（mSDF-1조）；（4）10－3 mol／L SIM＋200 mg／L mSDF-1용액＋골효원지가（SIM＋mSDF-1조）。식입1주후，련속2 d，매천분별재지가국부주사상술각조상응적용액50μL。사양6주후，취출지가급기주위조직，통과연X사선투사성상급회도측정、HE염색、면역조직화학염색적방법，정성급정량관찰기성골효과。결과：SIM＋mSDF-1조연X사선회도치[（421836．5±65425．7）상소]명현고우공백대조조[（153345．6±45222．2）상소，P＜0．01]、SIM조[（158119．2±100284．2）상소，P＜0．01]이급mSDF-1조[（255529．5±152142．4）상소，P＜0．05]；재SIM＋mSDF-1조내가견명현적골교단백화골개소적표체；SIM＋mSDF-1조혈관총밀도[（46±8）조／mm2]명현고우공백대조조[（23±7）조／mm2， P＜0．01]화SIM조[（24±6）조／mm2， P＜0．01]。결론：사용mSDF-1결합SIM이급골효원지가구건적무외가충자세포적조직공정화골가우소서로부피하이위성골。
Objective:To construct and evaluate a novel tissue-engineered bone composed of murine stromal cell-derived factor 1(mSDF-1), simvastatin (SIM) and collagen scaffold (Bio-Oss?), serving as a cell-homing approach for bone formation .Methods: In the study , 32 ICR mice were randomly divided into 4 groups,each group including 8 mice.The drug-loaded collagen scaffolds were implanted subcutaneously onto the cranium of each mouse according to the groups: ( 1 ) 1 ∶50 ( volume ratio ) dimethyl sulfoxide ( DMSO ) /phosphate-buffered saline ( PBS ) solution +collagen scaffold ( blank control group ); ( 2 ) 10 -3 mol/L SIM solution +collagen scaffold ( SIM group ); ( 3 ) 200 mg/L mSDF-1solution +collagen scaffold (mSDF-1 group); and (4) 10 -3mol/L SIM +200 mg/L mSDF-1 solution +collagen scaffold ( SIM +mSDF-1 group) .One week after implantation , the mice were trea-ted by injecting the same drug solution mentioned above around the scaffold once a day for two days .The specimens were harvested 6 weeks after implantation and the bone formation was evaluated by soft X-ray analysis , HE staining and immunohistochemical staining .Angiogenesis of each group was checked by calculation of vessels in each tissue section .Results:Six weeks after implantation , the collagen scaffolds were retrieved.The value of gray scale for the SIM +mSDF-1 group[(421 836.5 ±65 425.7) pixels] was significantly higher than that of the blank control group [(153 345.6 ±45 222.2)pixels, P<0.01], the SIM group [(158 119.2 ±100 284.2) pixels, P<0.01], and the mSDF-1 group[(255 529.5 ± 152 142.4) pixels, P <0.05 ]; HE staining analysis revealed that significant bone formation was achieved in the SIM +mSDF-1 group; The immunohistochemical staining showed the existence of os-teopontin and osteocalcin in the SIM +mSDF-1 group; There were more vessels in the SIM +mSDF-1 group[(46 ±8)vessels/mm2] than in the blank control group [(23 ±7) vessels/mm2, P<0.01], and the SIM group[(24 ±6) vessels/mm2 , P<0.01].Conclusion:The novel tissue-engineered bone com-posed of mSDF-1, SIM and collagen scaffolds has the potential to form bone subcutaneously in vivo.It re-presents a novel method of in vivo bone re-generation without seed cell delivery .