Open Access Original Research Article

Differential Viability in Alpha-MEM Culturing Media May Predict Alternative Media Responsiveness in Dental Pulp Stem Cell (DPSC)

Crystal Viss, Gavin Banning, Sarah Swanbeck, Karl Kingsley

Journal of Advances in Biology & Biotechnology, Page 1-13
DOI: 10.9734/jabb/2022/v25i330269

 

Objective: Dental pulp stem cells (DPSC) are the subjects of new and emerging fields of clinically applied biotechnology. However, much remains unknown regarding the most effective and appropriate methods for isolation, expansion and culture techniques for DPSC. To address these deficiencies, the primary objective of this study was to evaluate any effects of the major, commercially available cell culture media on DPSC phenotypes, such as growth, viability and biomarker expression.

Methods: This Institutional Review Board (IRB) approved study involved previously collected and cryopreserved DPSC isolates that were identified, thawed and cultured for this study (n=16). Each DPSC isolate was plated into 96-well assays under each of the experimental conditions (DMEM, DMEM:F12, RPMI, alpha-MEM) to determine any effects on cellular growth and viability. RNA was extracted from all DPSC isolates under the optimal growth conditions for screening using qPCR primers specific for DPSC biomarkers, such as Sox-2, Oct-4 and NANOG.

Results: Comparison of the standard DPSC cell culture media alpha-MEM to DMEM revealed differential results. Comparison of alpha-MEM to DMEM:F12 revealed no change among some DPSCs (n=3), decreased viability (n=8) or increased viability (n=5) - similar to the comparisons with RMPI demonstrating no change (n=5), decreased viability (n=6) or increased viability (n=5). Further analysis revealed that DPSC with low viability (<50%) in alpha-MEM responded positively to one or more of the culture media alternatives, while virtually none of DPSC with high viability (>50%) responded to any of the other experimental conditions. Screening of mRNA using qPCR revealed most DPSC isolates continued to express one or more of the pluripotent stem cell biomarkers (Oct4, Sox2, Nestin, NANOG), but no clear pattern of growth  with the optimal media type correlated with viability.

Conclusions: These results demonstrated that many DPSC isolates responded positively to one or more of these media, including DMEM, DMEM:F12, RPMI when viability was <50% using the standard DPSC culture media alpha-MEM, but not when viability was >50%. These findings may be broadly applicable and add significantly to the evidence regarding the potential culturing methods that may be employed in various ex vivo and in vitro DPSC studies.

   

Open Access Original Research Article

Screening of Carcinogenic and Non-carcinogenic Polycyclic Aromatic Hydrocarbon in a Typical Urban City Center, Owerri, Nigeria

Ukachukwu, Chidinma Ogochukwu, Udebuani, Angela Chika, Ugwu, Tochukwu Nicholas

Journal of Advances in Biology & Biotechnology, Page 14-21
DOI: 10.9734/jabb/2022/v25i330270

The study screened the carcinogenic and non-carcinogenic PAH in urban city center Owerri, Imo State. Soxhlet extraction-gas chromatography-flame ionization detector (SE-GC-FID) method was used to analyze PAH concentrations in the different environmental component to determine its impact. Result of the physicochemical properties of the spent engine oil polluted soil sample showed that exchangeable base (4.67g/kg), exchangeable acidity (1.25g/kg), organic carbon (6.32g/kg), organic matter (6.45g/kg), percentage nitrogen (0.46%), were higher in the polluted soil when compared to soil collected from a non-polluted agricultural farm. However, the effective cation exchange capacity (9.27g/kg), was lower in the polluted soil sample although, the pH value of the polluted soil was acidic. Result of the water sample collected from urban city center, showed that water sample properties of turbidity and magnesium, had mean values of 9.6±1.10mg/l and 26.7±0.08mg/l, respectively. Five carcinogenic PAH were detected in soil, water and sediment of the study area. The concentration of PAHs detected were within the range of 0.55±0.1 and 0.87±0.11 for sediment and water samples while, spent engine oil polluted sample and Abattoir soil sample were found in the range of 2.25±0.11 and 0.50±0.01 respectively. However, the detection of these carcinogenic PAH obtained from the different sampling point suggest the toxicity propensity of these pollutant.

Open Access Original Research Article

Effects of Helianthus annuus and Anthocleista djalonensis on Staphylococcus aureus and Escherichia coli

Olasupo John Ilori, Esther Chidima Okeke, `, Adesola Esther Adeneye, Temiloluwa Folasayo Afolayan, Abiola Hilder Emmanuel-Akerele, Omolola Abiola Bassey

Journal of Advances in Biology & Biotechnology, Page 22-27
DOI: 10.9734/jabb/2022/v25i330271

Plant based medicines have been a part of traditional healthcare in most parts of the world for thousands of years. The medicinal plants are of great interest to human health. It has been proven that antimicrobials of plant origin work more efficiently with fewer side effects. The study aimed to identify the phytochemicals in the extracts of Helianthus annus and Anthocleista djalonensis and examine the effects of extracts of these plants on the growth of clinical bacterial isolates. Leaves of Helianthus annus and Anthocleista djalonensis were extracted in methanol, distilled water and hot water. The extract solution (100%) was further diluted to various concentrations (75%, 50%, and 25%). The data obtained were analyzed by Analysis of Variance (ANOVA) to determine significant (P< 0.05) effects. Significant differences between means were determined using Duncan’s Multiple Range Test (DMRT). The phytochemical screening indicated the presence of alkaloids, phenols, glycosides, flavonoids, terpenoids, saponins and quinone. The extracts of the plants inhibited the growth of the bacteria tested with varied effectiveness. The maximum antibacterial activities were observed in the aqueous extracts. Thus the extracts of Helianthus annus and Anthocleista djalonensis can be used in the development of new pharmaceuticals that address unmet therapeutic use.

Open Access Original Research Article

Physicochemical Characterization of Cassava (Manihot esculenta) and Potato (Ipomea batatas) Varieties from Southeast of Nigeria in Comparison with Agarose for use in the Separation of Deoxyribonucleic Acid

Nkechi Eucharia Egbe, Ume Austine Okpaga, Vantsawa Philip Anthony, Nwankwo Henry Chizoba, Effiong Thomson Essien, Eze Godson Ofobuike, Ayuba-Buhari Sharifat

Journal of Advances in Biology & Biotechnology, Page 28-35
DOI: 10.9734/jabb/2022/v25i330272

Starch is an essential biomaterial and food product globally used for different purposes. This study determined the physicochemical composition of starch extracted from selected cassava (M.ExOboma I and M.ExOboma II) and sweet potato (I.ExOboma I and I.ExOboma II) varieties in comparison with agarose for use in the separation of deoxyribonucleic acid. Starch extraction was carried out using standard procedure. Granule morphology and amylose-amylopectin contents were determined using Polarized Light Microscopy and Dual Wavelength Iodine-Binding methods respectively. Relationships between starch properties were determined using correlation analysis. The result of the starch granule morphology revealed that there was no significant difference (p > 0.05) in the pore sizes of M.ExOboma I (5.527 ± 1.9346 µm) and M.ExOboma II (5.650 ± 1.0472 µm). Similarly, I.ExOboma I (8.275 ± 1.5064 µm) and I.ExOboma II (6.075 ± 2.3838 µm) showed no significant difference (p > 0.05) in their pore sizes. Meanwhile, the pore size of agarose was reported to be within the range of 6 to 35μm. No significant difference (p > 0.05) was also observed in the ratios of amylose and amylopectin (Am:Ap) contents between the varieties of cassava (M.ExOboma I 25:75 and M.ExOboma II 24:77) and sweet potato (I.ExOboma I 26:72 and I.ExOboma II 27:75). No significant difference (P > 0.05) between the pH range values (7.7 to 7.8) of cassava and sweet potato with that of agarose (7.5). The study also revealed that there were significant differences (p ≤ 0.05) in the clarity as well as the purity level of the starch samples compared to agarose. The results of this study are of great significance in predicting starch functionality and will form a basis for the improvement of the physicochemical attributes of starch for use in gel electrophoresis for the separation of biomolecules.

Open Access Original Research Article

Studies on Fungal Spoilage of Stored Zea mays L. (Maize) Grains in Two Markets in Lagos State, Nigeria

Adebayo O. Abdulrazak, Ewekeye S. Tolulope, Oyetunji S. Opeyemi, Oke A. Oyedamola, Alonge O. Faith

Journal of Advances in Biology & Biotechnology, Page 36-41
DOI: 10.9734/jabb/2022/v25i330273

Maize is one of the most essential staple foods consumed and its spoilage by fungi has been a serious concern to humans since the dawn of agriculture and food storage. Therefore, this research was aimed at isolating, identifying and determining the toxicity level of fungi causing spoilage of stored maize grains. Grain samples of maize were collected from Igando and Alaba markets in (Lagos State) from the top and bottom of maize bags from both market stores. Samples were cultured on Potato Dextrose Agar for fungal isolation. All isolates were identified using morphological and microscopic features. Also, the affordable qualitative method (ammonia vapour test) was carried out in order to identify some selected isolates that are aflatoxigenic. Petri- plates containing these isolates were flipped upside down and 2ml of concentrated ammonia solution (Extra Pure AR grade) was poured into the lid of inverted culture plates and observed for 10-15 minutes inside a desiccator for proper release of ammonia vapour. The genera of fungi isolated were; Aspergillus (57.15%), Fusarium (21.43%), Penicillium (7.14%) and Rhizopus (14.29%). The dominant genus was Aspergillus. The exposure of selected isolates to ammonia vapour led to varying degrees of colour changes which included pink, red and plum red. Similarly, isolates that were moderately poisonous were pink in colour, least poisonous showed red colour and very poisonous isolates indicated plum red colour. The findings from this study indicated that Aspergillus spp. are mostly responsible for spoilage of maize grains in storage and contamination with A. flavus can lead to poison production.