关键词: 骨缺损, 腺病毒, 促红细胞生成素, 聚乳酸羟基乙酸共聚物, 纳米纤维, 冷冻干燥法

Abstract:

Objective To investigate the influence of erythropoietin(EPO) in the bone defect after localized on polylactic-co-glycolic acid(PLGA) surface. Methods The bone marrow stromal cells (BMSCs) were isolated and cultured,and the adenovirus vector AdCMV-EGFP was used for gene transfection. The lyophilization method was used to compound AdCMV-EGFP and PLGA nanofiber scaffold as experiment group(PLGA/Ad-EGFP group),and the equivalent Ad-EGFP as control group;the adenovirus viability was detected using the percentage of fluorescence cell area of BMSCs infected with EGFP.Quantitative Picogreen dsDNA kit was used to detect the release kinetics of adenovirus.The Ad-EPO and PLGA nanofiber scaffold was compounded as experiment group(PLGA/Ad-EPO group),and single bone defect as control group;the PLGA/Ad-EPO were implanted into the critical skull bone defect of the rats,and the percentages of new bone areas at the 4th week and 8th week were detected by muro-CT and HE staining. Results Compared with control group,the percentage of fluorescentce cell area of the rats in PLGA/Ad-EGFP group was increased(P<0.05).The release kinetics of adenovirus on PLGA was a burst release curve,the retention on PLGA of adenovirus in the first hour was 53.0%±5.6%,and in the 16th hour was 20.0%±3.3%. Compared with control group, the erythrocytopoiesis was promoted at the 4th week, and the percentages of new bone areas at the 4th and 8th week were increased to 25.0%±5.7% and 35.0%±6.3% in PLGA/Ad-EPO group(P<0.05). Conclusion The lyophilizaion method could reserve the adenovirus activity,and PLGA/Ad-EPO nanofiber scaffold could promote the bone defect repair in a certain extent.

Key words: bone defect, adenovirus, erythropoietin, polylactic-co-glycolic acid, nanofiber scaffold, lyophilization method