Development of Nerve Conduit using Decellularized Human Umbilical Cord Artery Seeded with Centella asiatica Induced-Neurodiferrentiated Human Mesenchymal Stem Cell

Abstract

Various natural biological conduits have been investigated to bridge peripheral nerve injury especially in critical gap (greater than 3 cm in human). Autograft, the current gold standard, has several drawbacks including limited availability of donor graft, donor-site morbidity and mismatch in size in clinical practices. The aim of this study was to analyze the development of nerve conduit using decellularized human umbilical cord (HUC) artery seeded with neurodifferentiated human MSCs (ndMSCs) in bridging peripheral nerve gap. Artery conduits obtained from HUC were decellularized to remove native cells (n=3), then characterized by Hematoxylin and Eosin (H&E) staining and nuclei counterstaining with DAPI. The decellularized artery conduit was measured for every 2 weeks until 12 weeks. Next, mesenchymal stem cells (MSCs) were differentiated into neural lineage using 400 µg/mL of Centella asiatica. Then, 1.5×10 6 of MSCs or ndMSCs were seeded into decellularized artery conduit to study cell attachment. H&E staining and nuclei counterstaining with DAPI showed that all cellular components were removed from the HUC arteries. The decellularized artery conduit did not collapse and the lumen remained rigid for 12 weeks. Immunocytochemistry analysis with neural markers namely S100β, P75 NGFR, MBP and GFAP showed that MSCs had differentiated into neural lineage cells. H&E staining showed that the seeded MSCs and ndMSCs attached to the lumen of the conduits as early as 2 days. In conclusion, this study showed that nerve conduit using decellularized HUC artery seeded with neurodifferentiated human MSCs was successfully developed and have the potential to bridge critical nerve gap.

Centella asiatica (L.)-Neurodifferentiated Mesenchymal Stem Cells Promote the Regeneration of Peripheral Nerve

“…H&E staining after the 2-day incubation of the seeded conduits showed that MSCs or ndMSCs were successfully seeded into the decellularized HUC artery conduit. The seeded cells, MSCs or ndMSCs, adhered, spread well in the inner wall of the decellularized artery conduit (Data not shown) and did not clump or aggregate, same as in our previous study [16]. These results showed the biocompatibility between the HUC artery conduit and the seeded cells.…”

“…Following our success in constructing a nerve conduit by using HUC artery seeded with C. asiatica ndMSCs [16], this conduit was further tested by bridging PNI in rats and utilising this nerve graft. Given the 12 weeks of recovery period, the study on a 15 mm transectional sciatic nerve injury in SD rat model revealed that (1) the HUC artery Fig.…”

“…The nerve conduit was developed as described previously [16]. For the isolation of human MSCs, the written informed consents were obtained from healthy women who delivered full-term infants (38-40 weeks) by elective caesarean delivery prior to the collection of umbilical cord samples.…”

“…During 12 weeks of incubation and observation, the HUC arteries did not degrade or swell. The ndMSCs were then seeded into the HUC artery [16]. Thus, the present study aimed to investigate the effect of ndMSCs seeded into decellularized HUC on the regeneration of peripheral nerve in critical-size defect animal model.…”

Nerve regeneration using decellularized tissues: challenges and opportunities

“…The treatment of PNS, particularly in the crucial gap of more than 3 cm, remains a significant clinical issue despite the tremendous advancements in medicine ( Hussin et al, 2018 ). According to reports, many variables, such as the extent of the injury, the presence of neurotrophic factors, the existence of Schwann cells, etc., affect the restricted regeneration of peripheral nerves.…”