Abstract
ObjectivesThe goal of this study was to prepare lyophilized platelet-rich fibrin (L-PRF) and analyze the combined use of L-PRF and osteogenic bone marrow mesenchymal stem cell (BMSC) sheet fragments for bone tissue engineering via in vivo injection. MethodsFirst, fresh PRF (F-PRF) was lyophilized to prepare L-PRF, the characteristics of which were examined through gross morphological, and histological and microstructural observations. In addition, the kinetics of growth factor release from L-PRF and F-PRF were also determined by enzyme-linked immunosorbent assay (ELISA). Subsequently, after assessing the proliferation and osteogenic differentiation of BMSCs exposed to L-PRF or F-PRF in vitro, we subcutaneously injected BMSC sheet fragments with L-PRF or F-PRF into nude mice and assessed bone formation through microcomputed tomography and histological analyses. ResultsWe observed that L-PRF released growth factors that favored BMSC proliferation and osteogenic differentiation in vitro. The combined use of L-PRF and osteogenic BMSC sheet fragments enabled bone tissue regeneration in vivo, and no significant difference between the F-PRF and L-PRF groups was observed (P = 0.24). ConclusionsThe results of this study demonstrate that the combined use of L-PRF and osteogenic BMSC sheets may have potential in the fabrication of engineered bone.
Highlights
The treatment of nonunion and bone defects remains a common clinical problem [1]
The Alkaline phosphatase (ALP) activity assays and reverse transcription polymerase chain reaction (RT-PCR) results confirmed that similar to fresh Platelet-rich fibrin (PRF) (F-PRF), lyophilized PRF (L-PRF) improved the osteogenic potential of bone marrow mesenchymal stem cell (BMSC) in vitro (P 1⁄4 0.13) (Fig. 5A–E). These results indicated that L-PRF is a novel biomaterial scaffold that is suitable for the sustained release of growth factors and the protection of fibrogenic factors that are essential for protecting engineered tissue against proteolytic degradation [37, 38]
The results of this study demonstrated that L-PRF derived from lyophilized F-PRF consists of numerous densely arranged fibrin fibers, releases growth factors and favors BMSC proliferation and osteogenic differentiation in vitro
Summary
Rapid developments in tissue engineering and regenerative medicine have provided promising strategies for the repair of bone defects [2]. Tissue-engineered bone therapies are commonly divided into two major categories, scaffold-based and scaffold-free cell therapies [7]. Scaffold-free cell therapies, such as cell sheet technologies without additional need of scaffold, were used as alternatives to scaffold-based cell therapies prior to the development of the ideal scaffold material [10, 11, 12]. Gruene et al used computer-assisted MSC deposition technology to print high-density MSC suspensions in vitro and form a bone-free structure without scaffolds [15]. The development of a scaffold-free cell transplantation technique based on cell sheets indicates that BMSC sheets can be used to generate injectable tissue-engineered bone [16, 17]. Compared to dispersed cells, cell aggregates have been shown to significantly improve cell utilization and ease of operation [18]
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