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Detergent-free Nerve Decellularization For Long-gap Peripheral Nerve Reconstruction
Srikanth Vasudevan, MS; Edward W. Keefer, PhD; Barry R. Botterman, PhD; Jonathan Cheng, MD
University of Texas Southwestern Medical Center, Dallas, TX, USA
Hypothesis: For long gap peripheral nerve defects, autografts and allografts are the only available treatment modalities. Autografts have donor site morbidity and limited supply, while allografts require transient immunosuppression. As an alternative to overcome these limitations, a detergent-free nerve decellularization method was investigated by our laboratory for reconstructing long gap defects in a rat model. We reseeded these decellularized constructs in order to investigate the contribution of exogenous cells to nerve regeneration.
Methods: Donor sciatic nerves were harvested from 36 rats (300-350 g, male, Lewis). Recipient rats (250-300 g, male, Lewis) were divided into 5 groups (6 animals/group/time point): (1) Nerve graft (NG, positive control), (2) Silicone tube (ST, negative control), (3) Decellularized nerve graft (DE), (4)
Results: Schwann cell-seeded decellularized nerve graft (DE-SC) and (5) Skin derived progenitor-seeded decellularized nerve graft (DE-SKPs). A 3.5 cm long nerve defect was reconstructed with the construct in a reversed tension-free manner. Six animals from each group were assessed at 6 wk, 8 wk and 12 wk after implantation. Gastrocnemius muscle twitch and tetanic force, gastrocnemius muscle mass, and semi-automated quantitative histomorphometry were used to compare nerve regeneration between groups.
Conclusion: No nerve regeneration was observed in ST groups which served as the negative control. Muscle force data indicated maximum functional recovery in NG groups, followed by DE and DE-SC groups. Muscle masses were consistent with force data. Histomorphometry indicated maximum axonal growth in NG as compared with other groups. DE and DE-SC groups showed comparable regeneration at 12 wk. In conclusion, detergent-free nerve decellularization was sufficient to promote nerve regeneration across long gap injuries, which provides a potential alternative to current strategies. Addition of Schwann cells did not enhance nerve regeneration, while undifferentiated SKPs were found to inhibit nerve regeneration. Our detergent-free decellularization will also be compared against existing decellularization methods.
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