Dynamic Bioreactor Culture for Infiltration of Bone Mesenchymal Stem Cells within Electrospun Nanofibrous Scaffolds for Annulus Fibrosus Repair

Abstract

Objective To compare the ability of three culture strategies of static culture, intermittent centrifugal culture and dynamic bioreactor culture in promoting the infiltration of bone marrow mesenchymal stem cells (BMSCs) throughout electrospun nanoporous aligned nanoyarn scaffold (AYS). Methods AYS was constructed by the method of conjugated electrospinning, using the blended solution of poly (L‐lactide‐co‐caprolactone) (P (LLA‐CL)) and gelatin. Then the bone marrow mesenchymal stem cells (BMSCs) were transplanted on the scaffolds. Culture the scaffold‐cells using three methods of static culture, intermittent centrifugal culture and dynamic bioreactor culture. After 7 and 14 days in culture, the infiltration depth of the cells were observed and measured by hematoxylin and eosin (HE) or 4′, 6‐diamidino‐2‐phenylindole (DAPI) staining. Result In the current study, on the 7th day, the BMSCs in the scaffolds of static culture group, intermittent centrifugal culture group, and dynamic bioreactor culture group infiltrated to an average depth of 11.88 ± 1.82 μm, 21.17 ± 13.17 μm, and 26.27 ± 7.42 μm, respectively. There were differences between the bioreactor culture group with the static culture group and the intermittent centrifugal culture group. On the time point of 14 days, the depth of infiltration of BMSCs in dynamic bioreactor culture was the most (115.13 ± 25.44 μm, P < 0.05), and the infiltration of the cells in the intermittent centrifugal culture group was 42.53 ± 13.07 μm, deeper than that of the static culture group (24.53 ± 6.06, P < 0.05). Conclusion Dynamic bioreactor culture may be a preferred method for tissue engineering approaches involving scaffolds with a low porosity, such as those needed for repair of the annulus fibrosus (AF).

Keywords

• Annulus fibrosus
• Cell culture techniques
• Mesenchymal stem cells
• Tissue engineering
• Tissue scaffolds