The Potential of Acellular Dermal Matrix Combined With Neural Stem Cells Induced From Human Adipose-Derived Stem Cells in Nerve Tissue Engineering

Abstract

Introduction Reconstruction of segmental peripheral nerve gap is still challenging when the autografts are unavailable owing to limited availability of donor site and functional recovery. The creation of artificial conduits composed of biological or synthetic materials is still developing. Acellular dermal matrix (ADM) has been widely studied and its extension and plasticity properties may become suitable nerve conduits under different forms of nerve gaps. Adipose-derived stem cells (ADSCs) have the potential to differentiate into various cell types of different germ layers including neural stem cells (NSCs). The purpose of this experiment is to use ADM as a scaffold combined with NSCs induced by ADSCs to establish neural tissue engineering. Methods The ADSCs were isolated from syringe-liposuction adipose tissue harvested from abdominal fat and then cultured in keratinocyte serum free media to trigger into neural stem cells. Stem cells were confirmed by the expression of surface markers nestin and SOX2 in NSCs with Western blot and immunofluorescent staining. Matrix enzyme treatment was used to obtain ADM to remove immunogenic cells while maintaining the integrity of the basement membrane complex and the extracellular matrix structure of the dermis. The NSCs were cocultured with ADM for 3 days, and survival markers Ki67 and neural stem cell markers nestin were detected. Results These NSCs can form neurospheres and express nestin and SOX2. The NSC can further coculture with ADM, and it will continue to express survivor markers and neural stem cell markers on ADM. Conclusions These findings provide evidence that the combination of ADM and NSC has the same potential as neural tissue engineering as other acellular sciatic nerve.

Adipose-Derived Neural Stem Cells Combined with Acellular Dermal Matrix as a Neural Conduit Enhances Peripheral Nerve Repair

“…The experimental procedures used in this study were reviewed and approved by the Institutional Animal Care and Use Committee (IACUC-16-114) of National Defense Medical Center, Taipei, Taiwan, ROC. Acellular tissue was prepared mainly as described previously [16][17][18]23 . For the preparation of ADM, patches (3 Â 5 cm 2 ) of full-thickness skin were removed from 8-to 12-week-old nude mice (BALB/c-nu; BioLASCO Taiwan Co, Ltd, Taipei, Taiwan, ROC) and immersed in trypsin for 24 h at 37 C to remove the epidermis and other cells.…”

“…Samples of ADM (1 cm 2 ) and ASN (1 cm) were placed in the wells of a 24-well plate 23 , after which 1 Â 10 5 NSCs were added to each well and cultured on the ADM or ASN in NSC induction medium. After incubation for 72 h, unattached cells and medium were washed away with HBSS.…”

“…The ductility and plasticity of ADM suggest that it is potentially useful for repair of nerve gaps of various sizes in nerves of different diameters, and that it could be combined with neuro-engineered materials for future benefits to patient. We previously showed that ADSCs can be induced into NSCs, which can grow on ADM 23 . In the present study, we present several lines of evidence that ADM combined with ADSC-derived NSCs can contribute to nerve gap repair in a rat model of sciatic nerve transection with a 10-mm gap.…”