63 Developing a 3D in Vitro Schwann Cell Model of Myelination for Peripheral Nerve Injury

Abstract

Abstract Aim This study investigated the effectiveness of fabricating engineered neural tissue (EngNT) to develop a 3D in vitro model of myelination for peripheral nerve injury. Additionally, the effect of metformin and sulindac sulfide in nerve regeneration was evaluated by assessing neurite growth in both monolayer and 3D co-culture systems. Method Neuronal cells (NG108-15 and SH-SY5Y cell lines) were treated with metformin for 72 hrs in an Incucyte® live-cell analysis system. EngNT cellular collagen gels were fabricated using Schwann cells (SCL4.1/F7), SCL4.1/F7:NG108-15 (1:1) cells or harvested rat dorsal root ganglion cells. NG108–15 cells were co-cultured with SCL4.1/F7 cellular gels. 3D EngNT co-cultures were treated with metformin and sulindac sulfide for 72 hrs. EngNT cellular gels were analysed with transmission electron microscopy (TEM). Results TEM evaluation of EngNT co-cultures demonstrated the enfoldment of neurones by Schwann cells resulting in cell-cell contacts and myelination was identified. NG108-15 and SH-SY5Y cells exhibited significant neurite growth with 100 μM metformin being the optimal dose (151.4 ± 3 μM and 108.0 ± 12 μM respectively). Increased neurite growth of NG108-15 neurones was demonstrated with 100 μM (179.3 ± 2 μM) metformin treatment in 3D EngNT. Additionally, significant neurite growth was demonstrated following treatment with 100 μM (203.9 ± 32 μM) sulindac sulfide. Conclusions A 3D EngNT in vitro co-culture model of Schwann cell myelination was developed illustrating the cell-cell interaction between Schwann cells and neurones and the formation of myelin in the peripheral nervous system. Metformin elicits significant positive results on neurite growth in vitro highlighting its therapeutic potential in PNI.