BDNF blended chitosan scaffolds for human umbilical cord MSC transplants in traumatic brain injury therapy

摘要

This study tested the cytotoxicity of a BDNF blended chitosan scaffold with human umbilical cord mesenchymal stem cells (hUC-MSCs), and the invitro effect of BDNF blended chitosan scaffolds on neural stem cell differentiation with the aim of contributing alternative methods in tissue engineering for the treatment of traumatic brain injury (TBI). The chitosan scaffold based on immobilization of BDNF by genipin (GP) as a crosslinking agent referred to hereafter as a CGB scaffold was prepared by freezing-drying technique. hUC-MSCs were co-cultured with the CGB scaffold. Fluorescent nuclear staining (Hoechst 33342) was employed to determine the attachment of the hUC-MSCs to CGB scaffolds on the 1st, 3rd, 7th and 10th day of co-culture. The viability of hUC-MSCs adhered to the CGB scaffold was determined by digesting with 0.25% trypsin and evaluating with the cell counting kit-8 (CCK-8). Prior to this, the diameter and porosity of CGB scaffolds were measured. The amount of BDNF released from CGB over a 30 day period was determined by ELISA. Finally, we investigated whether the released BDNF can induce NSC to differentiate into neurons. There were no significant differences in diameter and porosity of individual CGB scaffolds (P>0.05). There were on average more cells on the CGB scaffold on the first day than on any other day sampled (P<0.05). The CGB scaffolds released BDNF in a uniform profile, whereas the CB scaffolds only released BDNF during the first 3 days. BDNF released from CGB scaffold promoted neuronal differentiation of NSCs and led to significant differences in differentiation rate and average neuron perimeter compared with the control group. The results of this study demonstrate that CGB scaffolds are biocompatible with hUC-MSCs and that granular CGB scaffolds covered with hUC-MSCs are expected to generate new advances for future treatment of traumatic brain injury.