Abstract
Nerve reconstruction of extended nerve defect injuries still remains challenging with respect to therapeutic options. The gold standard in nerve surgery is the autologous nerve graft. Due to the limitation of adequate donor nerves, surgical alternatives are needed. Nerve grafts made out of either natural or artificial materials represent this alternative. Several biomaterials are being explored and preclinical and clinical applications are ongoing. Unfortunately, nerve conduits with successful enhancement of axonal regeneration for nerve defects measuring over 4.0 cm are sparse and no conduits are available for nerve defects extending to 10.0 cm. In this study, spider silk nerve conduits seeded with Schwann cells were investigated for in vitro regeneration on defects measuring 4.0 cm, 10.0 cm and 15.0 cm in length. Schwann cells (SCs) were isolated, cultured and purified. Cell purity was determined by immunofluorescence. Nerve grafts were constructed out of spider silk from Nephila edulis and decellularized ovine vessels. Finally, spider silk implants were seeded with purified Schwann cells. Cell attachment was observed within the first hour. After 7 and 21 days of culture, immunofluorescence for viability and determination of Schwann cell proliferation and migration throughout the conduits was performed. Analyses revealed that SCs maintained viable (>95%) throughout the conduits independent of construct length. SC proliferation on the spider silk was determined from day 7 to day 21 with a proliferation index of 49.42% arithmetically averaged over all conduits. This indicates that spider silk nerve conduits represent a favorable environment for SC attachment, proliferation and distribution over a distance of least 15.0 cm in vitro. Thus spider silk nerve implants are a highly adequate biomaterial for nerve reconstruction.
Highlights
More than 300,000 cases of peripheral nerve injuries are reported annually in the European Union [1]
The results demonstrated clearly superior regeneration in Schwann cell pre-seeded allografts compared to acellular allografts alone, which was inferior to the current gold standard of autologous nerve grafts [39]
We examined the Schwann cell survival in an artificial nerve graft filled with spider silk in three different construct sizes (4.0 cm, 10.0 cm and 15.0 cm) for nerve gap implantation
Summary
More than 300,000 cases of peripheral nerve injuries are reported annually in the European Union [1]. Current gold standard in peripheral nerve surgery is the tension-free end-to-end suture of injured nerve tissue. In cases with a remarkable loss of nerve tissue and a resulting nerve gap, where a tension free end-to-end-suture is not achievable, an autologous nerve transplantation has to be performed. Autologous nerves for transplantation have to be surgically removed and microsutured into the existing nerve defect [2]. Sensory nerves (e.g., sural nerves) are being used, resulting in a subsequent sensory loss at the donor site. Due to the limitation of donor nerves and the severe donor site morbidity with loss of sensitivity in the area of distribution, the development of surgical alternatives is desperately needed [3]
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