Novel Acellular Scaffold Made from Decellularized Schwann Cell Sheets for Peripheral Nerve Regeneration

Abstract

Extracellular matrix surrounding Schwann cells and neurons provides critical determinants of cellular phenotype during development as well as essential cues in stimulating and guiding regrowth. Using cell sheet technology, we developed a novel scaffold enriched with native extracellular matrix from Schwann cells. Schwann cells were grown into sheets and layered onto polycaprolactone fibers for support. Upon decellularization of these constructs, extracellular matrix remained with few traces of nucleic acids. This method of deposition of extracellular matrix provided more protein than traditional seeding method after decellularization. Additionally, the isolated matrix supported proliferation of Schwann cells better than covalently bound laminin. The proliferation and differentiation of Schwann cells grown on decellularized sheets were complemented by upregulation of Erbb2 and myelin protein zero. Laminin expression of β1 and γ1 chains was also elevated. PC12 cells grown on decellularized sheets produced longer neurite extensions than aligned polycaprolactone fibers alone, proving potential of these scaffolds to be used in future peripheral nerve regenerative studies. Lay Summary Peripheral nerve injuries present a serious clinical need with approximately 50 % of surgical cases achieving only some restoration of function. In order to better guide regenerating nerves, supporting cells of the nerve tissue were grown into sheets and subsequently decellularized, leaving a myriad of surrounding protein as a scaffold. Constructs have been shown to support cell growth and neurite extension in vitro. Future projects will combine various cell types present in the nerve tissue as well as stem cells to fully support and reconstruct architecture of the peripheral nerves.

Albumin-Coated Polycaprolactone (PCL)–Decellularized Extracellular Matrix (dECM) Scaffold for Bone Regeneration

“…Chondrocytes were seeded at a density of 10 decellularized according to a previously published protocol. 33 Briefly, scaffolds were washed three times with PBS and placed in a 10 mM phosphate-buffered 50 mM sodium solution containing 125 mM 3-(decyldimethylammonio)propanesulfonate (SB3-10) overnight at 4 °C under agitation. After another three PBS washes, scaffolds were placed in 10 mM phosphate-buffered 50 mM sodium solution containing 0.14% sodium deoxycholate and 0.6 mM 3-(N,Ndimethylpalmitylammonio)propanesulfonate (SB3-16) overnight at 4 °C under agitation.…”

“…Culture medium was changed every other day. The chondrocyte cultures were decellularized according to a previously published protocol . Briefly, scaffolds were washed three times with PBS and placed in a 10 mM phosphate-buffered 50 mM sodium solution containing 125 mM 3-(decyldimethylammonio)­propanesulfonate (SB3-10) overnight at 4 °C under agitation.…”

Recent Advances in Scaffolding from Natural-Based Polymers for Volumetric Muscle Injury

“…Biological scaffolds derived from de-cellularized tissues and organs are progressively applied in regenerative medicine and tissue engineering [ 119 ]. The technology uses a thermo-responsive substrate to generate artificial conduits containing sheets of cells embedded in a native ECM [ 120 ]. The biological conduits offered a non-immunogenic allogenic scaffold containing guidance cues present in the tissue regeneration.…”

Recent Advances in Scaffolding from Natural-Based Polymers for Volumetric Muscle Injury

“…Biological scaffolds derived from de-cellularized tissues and organs are progressively applied in regenerative medicine and tissue engineering [119]. The technology uses a thermo-responsive substrate to generate artificial conduits containing sheets of cells embedded in a native ECM [120]. The biological conduits offered a non-immunogenic allogenic scaffold containing guidance cues present in the tissue regeneration.…”