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Fulltext Effect of perivitelline fluid from horseshoe crab on the expression of COL1A1 in dental pulp stem cells

Amanina Fatinah Kamarudin, Najian Ibrahim, Thirumulu Ponnuraj Kannan, Ahmad Aizat Abdul Aziz

Archives of Orofacial Sciences, 2016;11(2):26-30.
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Perivitelline fluid, extracted from the fertilized eggs of horseshoe crabs, has been reported to play a
vital role in supporting embryogenesis as well as cell proliferation. The present study aims to evaluate the effect
of PVF on the expression of COL1A1 in human dental pulp stem cells (DPSCs). The cells were grouped into two;
untreated (control) and treated with a single dose of PVF (0.019 mg/ml). Gene expression was quantified for
COL1A1 on day 1, 3 and 7 using reverse transcriptase PCR. The expression of COL1A1 on day 3 of treated
group with PVF was the highest though there was a decline of COL1A1 expression on day 7. Mann Whitney test
was utilized to determine the significance of COL1A1 expression between treated and untreated groups.
Significant difference in the expression of COL1A1 was observed between the treated and untreated groups on
day 3 though there was no significance in the expression on day 7. The present study indicates that PVF may
have the potential to increase cell proliferation in human DPSCs.

Matched MeSH terms: Horseshoe Crabs
Fulltext Structural Characterization and Antioxidant Potential of Chitosan by γ-Irradiation from the Carapace of Horseshoe Crab

Pati S, Chatterji A, Dash BP, Raveen Nelson B, Sarkar T, Shahimi S, et al.

Polymers (Basel), 2020 Oct 15;12(10).
PMID: 33076234 DOI: 10.3390/polym12102361

Natural product extraction is ingenuity that permits the mass manufacturing of specific products in a cost-effective manner. With the aim of obtaining an alternative chitosan supply, the carapace of dead horseshoe crabs seemed feasible. This sparked an investigation of the structural changes and antioxidant capacity of horseshoe crab chitosan (HCH) by γ-irradiation using 60Co source. Chitosan was extracted from the horseshoe crab (Tachypleus gigas; Müller) carapace using heterogeneous chemical N-deacetylation of chitin, followed by the irradiation of HCH using 60Co at a dose-dependent rate of 10 kGy/hour. The average molecular weight was determined by the viscosimetric method. Regarding the chemical properties, the crystal-like structures obtained from γ-irradiated chitosan powders were determined using Fourier transfer infrared (FTIR) spectroscopy and X-ray diffraction (XRD) analyses. The change in chitosan structure was evident with dose-dependent rates between 10 and 20 kGy/hour. The antioxidant properties of horseshoe crab-derived chitosan were evaluated in vitro. The 20 kGy γ-irradiation applied to chitosan changed the structure and reduced the molecular weight, providing sufficient degradation for an increase in antioxidant activity. Our findings indicate that horseshoe crab chitosan can be employed for both scald-wound healing and long-term food preservation due to its buffer-like and radical ion scavenging ability.

Matched MeSH terms: Horseshoe Crabs
Fulltext Effect of perivitelline fluid from horseshoe crab on the expression of cell cycle regulatory genes in human dental pulp stem cells

Abdul Qawee Rani, Thirumulu Ponnuraj Kannan, Nur Izyan Azmi, Najian Binti Ibrahim, Nor Shamsuria Omar, Ahmad Azlina, et al.

Archives of Orofacial Sciences, 2016;11(1):7-14.
MyJurnal

Perivitelline fluid (PVF) of the horseshoe crab embryo has been reported to possess an important role
during embryogenesis by promoting cell proliferation. This study aims to evaluate the effect of PVF on the
expression of cell cycle regulatory genes from human dental pulp stem cells (DPSCs) between different cell
passages viz. 4, 5, 6. The cells were treated with a single dose of PVF (26.89 mg/ml) PVF. Gene expression was
quantified for CDKNA2A, PTEN, MDM2 and TP53 genes using reverse transcriptase PCR. CDKN2A and MDM2
expression for treated and untreated DPSCs, expressed a similar pattern of expression. The higher expression of
CDKN2A showed that the treatment increased cell proliferation and prevented cell senescence. DPSCs with PVF
treatment showed increased expression of MDM2 at passage 4 and drastically declined expression at passage 5
and slightly increased at passage 6. TP53 expression of DPSCs treated group showed a higher expression
compared to untreated group. On the other hand, the expression of PTEN in DPSCs treated group started to
increase from passage 5 to 6. However, on the whole, the PTEN expression was higher than the untreated group
in all the passages studied here. The results showed that PVF could enhance cell cycle regulatory gene
expression in DPSCs as indicated by the higher expression of all the genes considered in this study at different
cell passages in the treated group compared to the untreated group. Mann Whitney test was utilized to determine
the significance of cell cycle regulatory genes expression between treated and untreated group. Significant
difference in expression of genes between the treated and untreated groups were found at all passages except
for CDKN2A gene whereby, its expression was not significantly different at passage 5 though it did express
slightly higher in PVF treated DPSCs.

Matched MeSH terms: Horseshoe Crabs
Effects of Perivitelline Fluid Obtained from Horseshoe Crab on The Proliferation and Genotoxicity of Dental Pulp Stem Cells

Musa M, Mohd Ali K, Kannan TP, Azlina A, Omar NS, Chatterji A, et al.

Cell J, 2015;17(2):253-63.
PMID: 26199904

OBJECTIVE: Perivitelline fluid (PVF) of the horseshoe crab embryo has been reported to possess an important role during embryogenesis by promoting cell proliferation. This study aims to evaluate the effect of PVF on the proliferation, chromosome aberration (CA) and mutagenicity of the dental pulp stem cells (DPSCs).
MATERIALS AND METHODS: This is an in vitro experimental study. PVF samples were collected from horseshoe crabs from beaches in Malaysia and the crude extract was prepared. DPSCs were treated with different concentrations of PVF crude extract in an 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay (cytotoxicity test). We choose two inhibitory concentrations (IC50 and IC25) and two PVF concentrations which produced more cell viability compared to a negative control (100%) for further tests. Quantitative analysis of the proliferation activity of PVF was studied using the AlamarBlue®assay for 10 days. Population doubling times (PDTs) of the treatment groups were calculated from this assay. Genotoxicity was evaluated based on the CA and Ames tests. Statistical analysis was carried out using independent t test to calculate significant differences in the PDT and mitotic indices in the CA test between the treatment and negative control groups. Significant differences in the data were P<0.05.
RESULTS: A total of four PVF concentrations retrieved from the MTT assay were 26.887 mg/ml (IC50), 14.093 mg/ml (IC25), 0.278 mg/ml (102% cell viability) and 0.019 mg/ml (102.5% cell viability). According to the AlamarBlue®assay, these PVF groups produced comparable proliferation activities compared to the negative (untreated) control. PDTs between PVF groups and the negative control were insignificantly different (P>0.05). No significant aberrations in chromosomes were observed in the PVF groups and the Ames test on the PVF showed the absence of significant positive results.
CONCLUSION: PVF from horseshoe crabs produced insignificant proliferative activity on treated DPSCs. The PVF was non-genotoxic based on the CA and Ames tests.

Matched MeSH terms: Horseshoe Crabs