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,
1.5×106
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.) Urban (CA) is a well- known plant used to improve brain and memory functions in traditional
medicine. Scientifically it was proven to show neurogenic effect on neural cell lines and in rat’s hippocampus. Its effect
on spinal cord (SC) neurons, however, have not been studied. Aim of this study was to investigate the effects of raw
extract of CA (RECA) on neurite outgrowths in an organotypic model of SC injury (OMSCI). OMSCI was prepared using SC
slices obtained from postnatal-day 8 rat pups. Spinal cord tissues were embedded in gelatine gel and sliced to produce
300 µm thick slices. These slices were 100% viable for 8 days in culture. RECA, in concentrations of 0-800 µg/mL was
added to the OMSCI media for 7 days, followed by immunostaining for TUJ-1 and GFAP. The investigated parameters
were mean neurite count, mean neurite length, mean longest neurite and growth ratio. The tested RECA concentrations
showed no cytotoxicity. ANOVA and Kruskal-Wallis tests showed no significant difference between groups in all the tested
parameters. This may be due to low content of neurotrophic bioactive compounds content in the extract, which probably
due to differences in geographical location, extraction method and absence of neurotrophic factors in the media. In
conclusion, the tested RECA concentration were found to be safe; but without notable neurotrophic effects on the spinal
cord organotypic model as demonstrated in this study.
Although electrospun poly(methyl methacrylate) (PMMA) may mimic structural features of extracellular matrix, its highly
hydrophobic nature causes reduced cell attachment. This study analysed the physicochemical and structural changes
of the surface modified PMMA nanofiber. The electrospun PMMA nanofibers (PM) were surface-treated as follows: PM
alone, collagen coated-PM (PM-C), UV-irradiated PM (PM-UV), collagen coated UV-irradiated PM (PM-C-UV) and collagen
coated-PM crosslinked with genipin (PM-C-GEN). They were subjected to scanning electron microscopy, Fourier transform
infrared (FTIR), cell attachment analysis, X-ray photoelectron spectroscopy (XPS), atomic force microscopy and X-ray
powder diffraction (XRD). The surface roughness was lower in PM-C-UV group compared to others. Based on FTIR
results, all expected functional group were present in all groups. XPS result showed that there are changes in the mass
concentration of UV-treated surfaces and in the collagen coated surfaces. All PM groups showed amorphous nature through
XRD. UV irradiation and collagen coating were shown to increase PM’s functional groups and modify its surface, which
contributed to the increased attachment of cells onto the inert PM scaffold. As conclusion, collagen coated UV irradiated
PMMA provided a better surface for cell to attach hence are suitable to be used further as scaffold for in vitro model.