Abstract
BackgroundAutologous transplantation of modified mesenchymal stem cells (MSCs) is a promising candidate for the treatment of the refractory clinical disease, avascular necrosis of the femoral head (ANFH). Our previous attempts by compounding MSCs with medical fibrin glue to treat ANFH in animal model have achieved excellent effects. However, the underlying molecular mechanism is unclear, especially on the transgenic gene expression.MethodsRabbit MSCs were isolated and compounded with fibrin glue. Following degrading of fibrin glue, proliferation, viability, expression of transgenic hepatocyte growth factor gene as well as osteogenic differentiation of MSCs were evaluated together with that of uncompounded MSCs. Fibrin glue-compounded MSCs were transplanted into the lesion of ANFH model, and the therapeutic efficacy was compared with uncompounded MSCs. One-Way ANOVA was used to determine the statistical significance among treatment groups.ResultsFibrin glue compounding will not affect molecular activities of MSCs, including hepatocyte growth factor (HGF) secretion, cell proliferation and viability, and osteogenic differentiation in vitro. When applying fibrin glue-compounded MSCs for the therapy of ANFH in vivo, fibrin glue functioned as a drug delivery system and provided a sustaining microenvironment for MSCs which helped the relatively long-term secretion of HGF in the femoral head lesion and resulted in improved therapeutic efficacy when compared with uncompounded MSCs as indicated by hematoxylin-eosin staining and immunohistochemistry of osteocalcin, CD105 and HGF.ConclusionTransplantation of fibrin glue-compounding MSCs is a promising novel method for ANFH therapy.
Highlights
Autologous transplantation of modified mesenchymal stem cells (MSCs) is a promising candidate for the treatment of the refractory clinical disease, avascular necrosis of the femoral head (ANFH)
Fibrin glue-compounded transgenic MSCs showed similar biological activities as uncompounded transgenic MSCs in short-term in vitro culture To assess the effects of MSCs compounded with fibrin glue, we evaluated the proliferation and osteogenic differentiation in vitro
The results of EdU incorporation and WST-8 assays showed that MSCs from the MSC-fibrin glue complex have a high proliferation rate similar as uncompounded MSCs resulting from the expression of hepatocyte growth factor (HGF) transgene (Figure 2A-C)
Summary
Autologous transplantation of modified mesenchymal stem cells (MSCs) is a promising candidate for the treatment of the refractory clinical disease, avascular necrosis of the femoral head (ANFH). Fibrin glue is commonly used for surgical hemostasis and tissue sealing as a sealant [10,11,12]. In recent years it has been applied as supporting material for cell growth, migration, and differentiation [13,14]. The advantages of clinical application of fibrin glue were widely studied, yet the detailed underlying molecular mechanism still need further exploration, such as long-term sustainable effects in MSCsmedicated treatments
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