应用新型生物墨水和聚己内酯3D生物打印骨-软骨复合组织块修复关节软骨缺损  

Three-dimensional bioprinting of osteochondral composite tissue with innovative bio-ink and PCL to repair articular cartilage defects

作　　者：左强[1] 张啸[1] 刘杨[1] 罗春阳 范卫民[1] 刘锋[1] Zuo Qiang;Zhang Xiao;Liu Yang;Luo Chunyang;Fan Weimin;Liu Feng(Department of Orthopaedics,First Affiliated Hospital of Nanjing Medical University,Nanjing 210029,China)

机构地区：[1]南京医科大学第一附属医院骨科,210029

出　　处：《中华骨科杂志》2021年第16期1073-1080,共8页Chinese Journal of Orthopaedics

基　　金：国家自然基金项目(81974332,81902206)。

摘　　要：目的应用新型生物墨水和聚己内酯(polycaprolactone,PCL)通过3D生物打印构建骨-软骨复合组织块,并评估其修复关节软骨缺损的效果。方法实验分为三组:①丝素蛋白(silk fibroin,SF)-磷酸三钙冻干支架组(冻干组),应用冻干法制作的SF-TCP复合支架修复软骨缺损,作为对照;②3D生物打印组(3D打印组),以PCL为原料通过3D生物打印机打印一个中空的多层圆柱体框架,后以软骨细胞外基质、SF和骨髓间充质干细胞为原料配置新型生物墨水,并在PCL框架上方继续3D生物打印含有活细胞的软骨层,最后于底层的PCL框架中充填自体松质骨,以此构建骨-软骨复合组织块修复关节软骨缺损;③自体骨软骨移植组(马赛克组),通过自体骨软骨移植术修复软骨缺损,作为对照。术后3个月通过国际软骨修复协会(International Cartilage Repair Society,ICRS)软骨修复组织学评分系统对修复组织进行评分。通过测定硫化糖胺聚糖(sulfated glycosaminoglycan,sGAG)和胶原蛋白含量可分析软骨细胞外基质分泌的程度。通过测定缺损修复区域组织的压缩模量评价修复组织的性状。结果3D打印组新生软骨的压缩模量(2.56±0.30)MPa和马赛克组(2.51±0.13)MPa相接近(P>0.05),明显高于冻干组(1.37±0.14)MPa且差异具有统计学意义(F=11.058,P<0.05)。3D打印组中sGAG的含量(14.49±0.7)μg/mg和马赛克组(14.98±0.81)μg/mg接近(P>0.05),明显高于冻干组(8.72±0.73)μg/mg且差异有统计学意义(F=20.973,P<0.05)。三组的胶原含量并无明显统计学差异(P>0.05)。ICRS软骨修复组织学评分结果表明,在基质、细胞分布、细胞群活力和软骨下骨4项评分中,3D打印组与马赛克组的得分接近(P>0.05),明显高于冻干组且差异有统计学意义(F=19.544,P<0.05);在表面和软骨矿化2项评分中,组间差异无统计学意义。结论应用新型生物墨水和聚己内酯通过3D生物打印构建骨-软骨复合组织块能够简化组织�Objective A new type of bio-ink and polycaprolactone(PCL)were used to construct an integrated osteochon-dral composite tissue block by multi-nozzle 3D bioprinter.And the repair results to osteochondral defects were evaluated.Methods In freeze-drying group:Freeze-dried composite scaffold made by silk fibroin(SF)and β-tricalcium phosphate was used to repair osteochondral defects,as control.In the 3D printing group:PCL was used to printed a hollow multi-layer cylinder frame by 3D biological printer.Extracellular matrix,SF and bone marrow mesenchymal stem cells were used as chondral bio-ink.Then,chon-dral bio-ink was used to print tissue-engineered cartilage on top of PCL frame.Before implantation of cartilage defect,autogenous cancellous bone was filled in PCL frame,then the tissue-engineered osteochondral composite was used to repair osteochondral defects.In mosaic group:Autologous osteochondral transplantation was performed.The repair results of the above three groups were compared by histological score,biochemical analysis and biomechanical test to evaluate the effect of repairing rabbit cartilage defects.Results The compression modulus of neo-cartilage in the 3D print group 2.56±0.30 MPa was close to that of the mosaic group 2.51±0.13 MPa(P>0.05),and significantly higher than that of freeze-dried group 1.37±0.14 MPa(F=11.058,P<0.05).The sGAG contents in the 3D print group 14.49±0.7μg/mg was close to that of the mosaic group 14.98±0.81μg/mg(P>0.05),and significantly higher than that of freeze-dried group 8.72±0.73μg/mg(F=20.973,P<0.05).However,there was no significant difference in collagen content between the three groups(P>0.05).The results of ICRS cartilage repair histology score showed that the scores of the 3D print group were close to those of the mosaic group in the matrix,cell distribution,cell viability and subchondral bone(P>0.05),and were significantly higher than those of freeze-dried group in the surface and cartilage mineralization scores(F=19.544,P<0.05).Conclusion Using the new bio-ink t

关 键 词：生物打印 打印 三维 细胞外基质 羧甲基纤维素钠 丝素蛋白质类 

分 类 号：R318.08[医药卫生—生物医学工程] R681.3[医药卫生—基础医学]