Abstract

Implants for bone regeneration to remedy segmental bone defects, osteomyelitis, necrotic bone tissue and non-union fractures have worldwide appeal. Although biomaterials offer most of the advantages by improving tissue growth but developments are more commonly achieved via biologically derived molecules. To aid site specific bone tissue regeneration by synthetic scaffold, cell derived extracellular matrix (ECM) can be a crucial component. In this study, co-cultured bone marrow mesenchymal stem cell and osteoblastic cells derived ECM incorporated electrospun polycaprolactone (PCL) membranes were assessed for bone tissue engineering application. The preliminary experimental details indicated that, co-culture of cells supported enhanced in vitro ECM synthesis followed by successful deposition of osteoblastic ECM into electrospun membranes. The acellular samples revealed retention of ECM related biomacromolecules (collagen, glycosaminoglycan) and partial recovery of pores after decellularization. In vitro biocompatibility tests ensured improvement of proliferation and osteoblastic differentiation of MC3T3-E1 cells in decellularized ECM containing membrane (PCL-ECM) compared to bare membrane (PCL-B) which was further confirmed by osteogenic marker proteins expression analysis. The decellularized PCL-ECM membrane allowed great improvement of bone regeneration over the bare membrane (PCL-B) in 8 mm size critical sized rat skull defects at 2 months of post implantation. In short, the outcome of this study could be impactful in development and application of cell derived ECM based synthetic electrospun templates for bone tissue engineering application.

摘要

用于修复节段性骨缺损、骨髓炎、坏死骨组织和不愈合骨折的骨再生植入物在世界范围内具有吸引力。尽管生物材料通过改善组织生长提供了大部分优势，但更常见的是通过生物衍生分子实现发展。为了通过合成支架帮助特定部位的骨组织再生，细胞衍生的细胞外基质 (ECM) 可能是一个关键的组成部分。在这项研究中，对共培养的骨髓间充质干细胞和成骨细胞衍生的 ECM 并入电纺聚己内酯 (PCL) 膜进行了骨组织工程应用评估。初步实验细节表明，细胞共培养支持体外增强ECM 合成，然后成骨细胞 ECM 成功沉积到电纺膜中。脱细胞样品显示 ECM 相关生物大分子（胶原蛋白、糖胺聚糖）的保留和脱细胞后孔的部分恢复。体外与裸膜 (PCL-B) 相比，生物相容性测试确保 MC3T3-E1 细胞在含脱细胞 ECM 的膜 (PCL-ECM) 中的增殖和成骨细胞分化得到改善，这通过成骨标记蛋白表达分析得到进一步证实。在植入后 2 个月，脱细胞的 PCL-ECM 膜在 8 毫米大小的临界尺寸大鼠颅骨缺损中比裸膜 (PCL-B) 的骨再生有很大改善。简而言之，这项研究的结果可能会对用于骨组织工程应用的细胞衍生 ECM 合成电纺模板的开发和应用产生影响。