Avicenna journal of medical biotechnology最新文献

Background: Nanoparticles, as small extracellular vesicles, are considered promising tools in tissue engineering and regenerative medicine. This study aimed to investigate the effects of different processing and culture condition on the quality and quantity of extracts derived from human Adipose-Mesenchymal Stem Cells (AD-MSCs).

Methods: AD-MSCs were proliferated in both the experimental and control groups. Nanoparticles were extracted from AD-MSCs-extracts and analyzed using SEM, TEM, DLS, Zeta potential, FTIR and BCA analyses. The morphological characteristics (shape, size, distribution, surface topography, and agglomeration/aggregation), structural appearance (poly-disperse intensity, colloidal particle behavior, surface charge, and stability), chemical properties (functional groups and ionic interactions) and total protein concentration were detected in the extracted nanoparticles. Additionally, the morphological characteristics, apoptosis, mitochondrial oxidoreductase activity, and migration potential of AD-MSCs in both groups were evaluated using acridine orange staining, MTT, and scratch assays.

Results: In the experimental group, 100% of the nanoparticles had a diameter of 112.8± 25 nm, with the most frequency of 111.4 nm. However, in the control group, 72% of nanoparticles had a diameter of 350.2±43.6 nm with the highest frequency of 339.8 nm (p≤0.05). The Z-average, Poly-disperse intensity, and electrostatic stability of nanoparticles in the control and experimental groups were 171.9 nm, 0.727 and -0.000011 cm² /Vs vs. 103.7 nm, 0.205 and 0.000481 cm² /Vs, respectively (p≤0.05). In the experimental group, Zeta potential was -61.8 mV, which is in the range of ζ >-30mV. Although, Zeta potential in the control group was -1.5 mV, which is in the range of -30 mV <ζ <30 mV (p≤0.05). Total protein concentrations in the control and experimental groups were 11 and 41%, respectively (p≤0.05).

Conclusion: Nanoparticles derived from AD-MSCs have high therapeutic applications in tissue engineering and regenerative medicine.

Background: Immunohistochemistry (IHC) is a practical technique that utilizes the specific binding between an antigen and antibody to detect and localize specific antigens in tissue and cells. The optimal sensitivity in IHC is of utmost importance to achieve reliable results even when antigens are present at low abundance on the samples. Here, a dextran polymer labeled with Horseradish Peroxidase (HRP) and an antibody to improve the sensitivity of the IHC technique was synthesized.

Methods: To this end, free thiol groups were introduced on sodium periodate-activated 30 kDa dextran using cystamine, followed by attachment of sulfo-MBS-activated goat anti-mouse antibody and sulfo-MBS-activated HRP to the activated dextran.

Results: The production of Poly-HRP Antibody (PHA) was confirmed by the appearance of a new protein band exceeding 150 kDa on Sodium Dodecyl Sulfate-Polyacrylamide Gel Electrophoresis (SDS-PAGE). Additionally, Enzyme-Linked Immunosorbent Assay (ELISA) and IHC techniques were employed to characterize PHA's functionality. The data demonstrated that PHA effectively detected target antigens in these assays.

Conclusion: The newly synthesized PHA has the potential to provide a more sensitive platform for early detection of biomarkers in IHC. Further research is needed to evaluate its cost-effectiveness.

Artificial bone materials were synthesized using the "solvent casting method" using polylactide/hydroxyapatite and various organic-inorganic modifiers. The physicochemical properties of the materials were studied using modern methods. IR spectroscopy showed that interactions between polymer macromolecules and hydroxyapatite occurred. When the powder was studied by the X-ray diffraction method, it was found to have an average crystallinity of 50-60%. When the textural properties were examined using SEM analysis, it was found that the introduction of magnesium phosphate into the samples resulted in the formation of porous particles with dimensions of 100-250 μm. This in turn, leads to the improvement of metabolic processes when the samples are introduced into living tissues. When the microhardness was determined by the Vickers method, it was found to be close to the hardness of natural bone, i.e. 27-34 HV. In vitro resorption was also performed in Simulated Body Fluids (SBF). Non-toxicity was observed when cytotoxic properties were studied. When the resorption process was studied in vivo in the upper third of the femur of rabbits, it was found that the ossification process of the samples was satisfactory after 28 days.

Background: An innovative approach for tissue restoration using Umbilical Cord-derived Mesenchymal Stem Cells (UC-MSCs) is hindered by their poor survival rate due to the detrimental effects of the injured tissue microenvironment. Activation of NLRP3 inflammasome in an inflammatory environment which is followed by cellular impairment, has been reported. However, the expression of NLRP3 inflammasome in UC-MSCs in response to the inflammatory environment is not well understood. This study aims to investigate the impact of short-term exposure to an inflammatory environment induced by Lipopolysaccharide (LPS) on hUC-MSCs, focusing on cell viability, migration, differentiation, and the expression of NLRP3 inflammasome-related genes.

Methods: hUC-MSC were exposed to LPS at concentration of 10 and 50 μg/ml for 3 and 6 hr. Cell viability was assessed using CCK-8 assay, migration capacity was evaluated using a scratch test, and differentiation capacity and the expression of NLRP3 inflammasome-related genes were measured using qRT-PCR.

Results: Short-term LPS induction did not affect the viability of hUC-MSCs but reduced their migration and differentiation capacity, particularly at 50 μg/ml for both time points (p<0.05). The induction caused an increase in the mRNA levels of NLRP3, TLR-4, and RelA/p65, which correlated with elevated expression of caspase-1 and IL-1β.

Conclusion: Short-term exposure to LPS influences hUC-MSCs by upregulating NLRP3, TLR4/ReIA (p65), IL-1β, and caspase-1 mRNA levels, leading to impaired migration and differentiation ability. This study underscores the significant impact of short-term exposure to an inflammatory microenvironment on hUC-MSC, potentially compromising their migration and differentiation capacity through the NLRP3 pathway.

Background: Doxorubicin (DOX) is a widely used drug in cancer chemotherapy, but its cardiotoxicity limits its clinical applications. Combining exercise with chemotherapy offers a promising approach to mitigate the side effects of chemotherapy drugs. Limited information is available on the effects of swimming exercise on the molecular mechanisms related to DOX cardiotoxicity. This study aims to investigate the modulatory effect of swimming exercise on the apoptosis and miR-23a-dependent mitochondrial biogenesis and dynamics pathways in rat heart tissue treated with DOX.

Methods: In this experimental study, thirty-two adult male Wistar desert rats (200-220 g) were randomly divided into four groups, including control, Doxorubicin (DOX; intraperitoneal injection of 5 mg/kg of DOX, once a week, for five weeks), swimming exercise (SE; water exercise for 60 min/day, five days a week, for six weeks) and DOX group along with Swimming Exercise (DOX-SE). At the end of the study, the cardiac expression of proteins related to apoptosis and mitochondrial biogenesis and mir23-a were analyzed using western blot and real-time PCR methods, respectively. One-way analysis of variance (ANOVA)with Tukey's post hoc test was used to analyze the data.

Results: These findings revealed that DOX administration led to a significant decrease in the cardiac expression of PGC-1α and DRP-1 proteins and an increase in apoptotic proteins (caspase 3 and cytochrome C) compared to the control group (p<0.0001). Swimming exercise resulted in a significant increase expression in cardiac tissue of PGC-1α and DRP-1 proteins and a decrease in the expression of apoptotic proteins in the DOX-treated group (p<0.0001, p<0.01). Compared to the control group, the protein levels in the heart of the miR-23a were significantly increased in the DOX-treated group (p<0.001). However, exercise training attenuated the DOX-induced reduction in miR-23a expression gene in the cardiac muscle of DOX-treated mice (p<0.05).

Conclusion: These findings suggest that swimming exercise may protect against DOX-induced cardiotoxicity by regulating apoptosis and DRP1/PGC1α/miR-23a pathway. This highlights exercise as a potential non-pharmacological strategy to mitigate chemotherapy-related heart damage.

Background: Interleukin-6 plays an essential role in cytokine storm and cytokine release syndrome, which occur in response to pathogen infection or tissue injury and are associated with severe symptoms. Neutralizing IL-6 can help reduce symptom severity. Chicken eggs serve as an excellent alternative antibody source compared to mammalian serum. The immunoglobulin Y (IgY) in the chicken's blood is transferred to and deposited within the egg yolk in large amounts. Several IgY products have been developed for therapeutic applications in various diseases. This study focuses on producing anti-human IL-6 (IL-6) IgY antibodies to support therapeutic advancements.

Methods: Anti-IL-6 IgY was generated by immunizing three hens with recombinant IL-6 protein mixed with alum adjuvant, immunized four times at three-week intervals. Then, IgY was extracted from egg yolks. The specificity of IgY was determined by Western blot. The neutralizing activity against secreted IL-6 was demonstrated by Human Coronavirus OC43 (HCoV-OC43)-infected cells and Lipopolysaccharide (LPS)-stimulated human lung fibroblast MRC-5 cells.

Results: The specific anti-IL-6 was detected starting from day 10 to day 90 after immunization. The average yield of total IgY was 17.97±15.66 mg per egg. The extracted anti-IL6 IgY antibody exhibited efficient neutralizing effects against secreted IL-6 in the HCoV-OC43-infected or LPS-stimulated cells in a dose-dependent manner.

Conclusion: This study highlights the ease of production and the satisfactory yield of anti-IL-6 IgY derived from chicken eggs. The antibody demonstrates an in vitro inhibitory effect on IL-6, with potential applications in therapeutic development.

Background: Type 2 diabetes mellitus (T2DM) is accompanied by a significant risk of oxidative stress. While a link between T2DM and G6PD deficiency has been suggested, their interaction is not precisely understood. Furthermore, emerging evidence suggests an expression association between G6PD and miR-1, miR-122, and miR-206. Given the antioxidant and anti-inflammatory properties of Curcumin (Cur) and Quercetin (Q), This study aimed to assess the effects of curcumin and quercetin on G6PD expression and its correlation with the mentioned microRNA expression in liver, renal, heart, and muscle in rats with T2DM.

Methods: RT-qPCR was employed to determine miR-1, miR-122, miR-206, and G6PD expression.

Results: The findings revealed that curcumin and quercetin treatment elevated G6PD gene expression. Also, the treated groups exhibited down-regulation of miR-1, miR-122, and miR-206 (p<0.05). Furthermore, there was a significant inverse correlation between G6PD and miR-1 in heart, miR-122 in all tissues except renal and miR-206 expression in skeletal muscle and heart (p<0.05).

Conclusion: This study suggests that curcumin and quercetin may prevent the development of T2DM by effectively increasing G6PD expression and reducing miR-1, miR-122, and miR-206 expression.

Background: L-Asparaginase is a crucial enzyme to treat Acute Lymphoblastic Leukemia (ALL), as it depletes L-asparagine, an essential amino acid for cancer cell survival. However, its clinical use is often restricted due to hypersensitivity reactions. This study examined the anti-proliferative effects and hypersensitivity of fungal L-asparaginases (L-ASNases) from Trichosporon asahii isolate ChL11 (TaIChL11 L-ASNase) and Candida palmioleophila isolate JK12 (CpIJK12 L-ASNase).

Methods: The enzymes were produced and purified through ammonium sulfate precipitation, dialysis, and Sephadex G-100 chromatography, and tested on leukemia cells and BALB/c female mice to assess immune responses.

Results: TaIChL11 L-ASNase had a molecular weight of 40 kDa, Michaelis constant (K_M) of 1.66×10^-2 mM, and V_max of 37.23 mM/min, while CpIJK12 L-ASNase had a molecular weight of 135 kDa, K_M of 2.3×10^-2 mM, and V_max of 14.03 mM/min. Both enzymes exhibited significant anti-proliferative effects against CCRF-CEM cancer cells, with half-maximal inhibitory concentration (IC₅₀) values of 2.74 U/ml for TaIChL11 LASNase and 3.30 U/ml for CpIJK12 L-ASNase after 48 hr, improving further after 72 hr. They also showed low cytotoxicity toward normal Vero E6 cells. in vivo studies demonstrated that TaIChL11 ASNase-treated mice had significantly lower Immunoglobulin (Ig) G levels than those treated with commercial L-ASNase from Erwinia chrysanthemi (Owenism) (p<0.005), with no detectable IgE response.

Conclusion: These findings indicate that fungal L-ASNases, particularly TaIChL11 ASNase, with lower L-glutaminase activity and a favorable safety profile, could be promising alternatives to bacterial L-ASNases, potentially enhancing ALL treatment with fewer side effects.

Background: Biotechnology is a rapidly developing field, and Iran aims to enhance its position in biosimilars through improved educational frameworks. Successful nations, such as the U.S., UK, Switzerland, China, and India, exemplify the positive impact of targeted educational programs in producing skilled graduates who advance the biotech sector. Research indicates a significant connection between curriculum relevance and graduate employability, suggesting that comprehensive curriculum assessment and reform can enhance student success and faculty involvement. This study investigated the necessity of revising the Master's curriculum in Medical Biotechnology in Iran to align with industry demands and global advancements.

Methods: The current curriculum's effectiveness using the Delphi method was assessed to survey Master's students, professors, and industry stakeholders.

Results: Findings revealed that while the current curriculum moderately aligns with educational objectives, significant gaps exist in practical training and resource availability. Many students reported feeling inadequately prepared for employment, particularly in essential skills such as animal cell culture, vaccine and monoclonal antibody design and production, and human skills like effective communication and teamwork. To address these deficiencies, some new courses focusing on practical experience and interdisciplinary approaches were recommended. Recommendations included enhancing laboratory facilities, integrating internships, and adopting team-based learning methods to improve student engagement and skill acquisition.

Conclusion: Continuous investment in biotechnology education is crucial for maintaining competitive advantages in a globalized market. Overall, this research highlighted the importance of adapting educational programs to meet the dynamic needs of the biotechnology industry, ensuring graduates possess the necessary skills for successful careers in this field.