Delivery of demineralized bone matrix powder using a salt-leached silk fibroin carrier for bone regeneration

Abstract

Demineralized bone matrix (DBM) has been widely used for bone regeneration due to its osteoinductivity and osteoconductivity. However, the use of DBM powder is limited due to the difficulties in handling, the tendency to migrate from graft sites and the lack of stability after surgery. In this study, a mechanically stable, salt-leached porous silk fibroin carrier was used to improve the handling properties of DBM powder and to support the attachment, proliferation and osteogenic differentiation of rat bone marrow derived mesenchymal stem cells (rBMSCs). The DBM-silk fibroin (DBM/SF) scaffolds were fabricated with different contents of DBM powder (0%, 10%, 20%, 40% and 80% DBM/SF scaffolds). It was found that the DBM/SF scaffolds could form a stable composite preventing the migration of DBM powder. Moreover, the microarchitecture and mechanical properties of the scaffolds were influenced by the DBM powder. rBMSCs were seeded on the DBM/SF scaffolds and cultured for 14 days. Cell proliferation assays and cell morphology observations indicated that 20% DBM/SF scaffolds exhibited good cell attachment and proliferation. In addition, compared with the other groups, the cellular function was more actively exhibited on 20% DBM/SF scaffolds, as evident by the real-time reverse transcriptase-polymerase chain reaction (RT-PCR) analysis for osteoblast-related gene markers (e.g. COL1A1, ALP and cbfa1), the immunocytochemical evaluations of osteoblast-related extracellular matrix components (e.g. COL1A1, OCN and ONN) and the ALP activities. All the data suggested that DBM powder could be delivered using a silk fibroin carrier with improved handling characteristics and that 20% DBM/SF scaffolds had great potential as osteogenesis promoting scaffolds for successful applications in bone regeneration.