Plast Reconstr Surg. 2018 May 22. doi: 10.1097/PRS.0000000000004614. [Epub ahead of print]

A simple dynamic strategy to deliver stem cells to decellularized nerve allografts.

Rbia N, Bulstra LF, Bishop AT, van Wijnen AJ, Shin AY.

Abstract

BACKGROUND:

The addition of adipose-derived Mesenchymal Stromal Cells (MSCs) to decellularized nerve allografts may improve outcomes of nerve reconstruction. Prior techniques used for cell seeding are traumatic to both the MSCs and nerve graft. An adequate, reliable and validated cell seeding technique is an essential step for evaluating the translational utility of MSC-enhanced decellularized nerve grafts. The purpose of this study was to develop a simple seeding strategy with an optimal seeding duration.

METHODS:

A dynamic bioreactor was used to seed rat and human MSCs separately onto rat and human decellularized nerve allografts. Cell viability was evaluated by MTS assays and cellular topology after seeding was determined by SEM microscopy. Cell density and distribution were determined by LIVE/DEAD assays and Hoechst staining at 4 different time points (6, 12, 24 and 72 hours). The validity and reliability of the seeding method were calculated.

RESULTS:

Cells remained viable at all time points, and MSCs exhibited exponential growth in the first 12 hours of seeding. Seeding efficiency increased significantly from 79.5% at 6 hours to 89.2% after 12 hours of seeding (p = 0.004). Both intra-rater (r = 0.97) and inter-rater reliability (r = 0.92) of the technique were high.

CONCLUSIONS:

This study describes and validates a new method to effectively seed decellularized nerve allografts with MSCs. This method is reproducible, distributes cells homogenously over the graft and does not traumatize the intra-neural architecture of the allograft. Utilization of this validated seeding technique will permit critical comparison of graft outcomes.