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Fulltext Nanofiber Scaffold Based on Polylactic Acid-Polycaprolactone for Anterior Cruciate Ligament Injury

Aminatun, Huriah R, Hikmawati D, Hadi S, Amrillah T, Abdullah CAC

Polymers (Basel), 2022 Jul 23;14(15).
PMID: 35893947 DOI: 10.3390/polym14152983

Anterior Cruciate Ligament (ACL) injuries are becoming more prevalent in athletes. Anterior Cruciatum Ligament Reconstruction (ACLR) surgery was used to treat ACL injuries and resulted in a recurrence rate of 94% due to the biomechanically repaired tissue being weaker than the original tissue. As a result, biodegradable artificial ligaments must be developed that can withstand mechanical stress during neoligament formation and stabilize the ACL. The purpose of this study is to determine the effect of composition variations in polylactic acid (PLA) and polycaprolactone (PCL) used as ACL nanofiber scaffolds on ultimate tensile strength (UTS) and modulus of elasticity, fiber diameter, cytotoxicity level, and degradation level, as well as the PLA-PCL concentration that provides the best value as an ACL scaffold. Electrospinning was used to fabricate the nanofiber scaffold with the following PLA-PCL compositions: A (100:0), B (85:15), C (80:20), D (70:30), and E (0:100) (wt%). The functional group test revealed no new peaks in any of the samples, and the ester group could be identified in the C-O bond at wave numbers 1300-1100 cm-1 and in the C=O bond at wave numbers 1750-1730 cm-1. The average fiber diameter, as determined by SEM morphology, is between 1000 and 2000 nm. The unbraided sample had a UTS range of 1.578-4.387 MPa and an elastic modulus range of 8.351-141.901 MPa, respectively, whereas the braided sample had a range of 0.879-1.863 MPa and 2.739-4.746 MPa. The higher the PCL composition, the lower the percentage of viable cells and the faster the sample degrades. All samples had a cell viability percentage greater than 60%, and samples C, D, and E had a complete degradation period greater than six months. The ideal scaffold, Sample C, was composed of PLA-PCL 80:20 (wt%), had an average fiber diameter of 827 ± 271 nm, a living cell percentage of 97.416 ± 5.079, and a degradation time of approximately 219 days.
Fulltext Multiple gene targeting siRNAs for down regulation of Immediate Early-2 (Ie2) and DNA polymerase genes mediated inhibition of novel rat Cytomegalovirus (strain All-03)

Balakrishnan KN, Abdullah AA, Bala JA, Jesse FFA, Abdullah CAC, Noordin MM, et al.

Virol J, 2020 Oct 27;17(1):164.
PMID: 33109247 DOI: 10.1186/s12985-020-01436-5

BACKGROUND: Cytomegalovirus (CMV) is an opportunistic pathogen that causes severe complications in congenitally infected newborns and non-immunocompetent individuals. Developing an effective vaccine is a major public health priority and current drugs are fronting resistance and side effects on recipients. In the present study, with the aim of exploring new strategies to counteract CMV replication, several anti-CMV siRNAs targeting IE2 and DNA polymerase gene regions were characterized and used as in combinations for antiviral therapy.
METHODS: The rat embryo fibroblast (REF) cells were transfected with multi siRNA before infecting with CMV strain ALL-03. Viral growth inhibition was measured by tissue culture infectious dose (TCID50), cytopathic effect (CPE) and droplet digital PCR (ddPCR) while IE2 and DNA polymerase gene knockdown was determined by real-time PCR. Ganciclovir was deployed as a control to benchmark the efficacy of antiviral activities of respective individual siRNAs.
RESULTS: There was no significant cytotoxicity encountered for all the combinations of siRNAs on REF cells analyzed by MTT colorimetric assay (P > 0.05). Cytopathic effects (CPE) in cells infected by RCMV ALL-03 had developed significantly less and at much slower rate compared to control group. The expression of targeted genes was downregulated successfully resulted in significant reduction (P
Fulltext Synthesis and Characterization of Gefitinib and Paclitaxel Mono and Dual Drug-Loaded Blood Cockle Shells (Anadara granosa)-Derived Aragonite CaCO3 Nanoparticles

Chemmalar S, Intan-Shameha AR, Abdullah CAC, Ab Razak NA, Yusof LM, Ajat M, et al.

Nanomaterials (Basel), 2021 Aug 02;11(8).
PMID: 34443820 DOI: 10.3390/nano11081988

Calcium carbonate has slowly paved its way into the field of nanomaterial research due to its inherent properties: biocompatibility, pH-sensitivity, and slow biodegradability. In our efforts to synthesize calcium carbonate nanoparticles (CSCaCO3NP) from blood cockle shells (Anadara granosa), we developed a simple method to synthesize CSCaCO3NP, and loaded them with gefitinib (GEF) and paclitaxel (PTXL) to produce mono drug-loaded GEF-CSCaCO3NP, PTXL-CSCaCO3NP, and dual drug-loaded GEF-PTXL-CSCaCO3NP without usage of toxic chemicals. Fourier-transform infrared spectroscopy (FTIR) results reveal that the drugs are bound to CSCaCO3NP. Scanning electron microscopy studies reveal that the CSCaCO3NP, GEF-CSCaCO3NP, PTXL-CSCaCO3NP, and GEF-PTXL-CSCaCO3NP are almost spherical nanoparticles, with a diameter of 63.9 ± 22.3, 83.9 ± 28.2, 78.2 ± 26.4, and 87.2 ± 26.7 (nm), respectively. Dynamic light scattering (DLS) and N2 adsorption-desorption experiments revealed that the synthesized nanoparticles are negatively charged and mesoporous, with surface areas ranging from ~8 to 10 (m2/g). Powder X-ray diffraction (PXRD) confirms that the synthesized nanoparticles are aragonite. The CSCaCO3NP show excellent alkalinization property in plasma simulating conditions and greater solubility in a moderately acidic pH medium. The release of drugs from the nanoparticles showed zero order kinetics with a slow and sustained release. Therefore, the physico-chemical characteristics and in vitro findings suggest that the drug loaded CSCaCO3NP represent a promising drug delivery system to deliver GEF and PTXL against breast cancer.
Immediately early 2 (IE-2) and DNA polymerase SiRNA as virus-specific antiviral against novel transplacental cytomegalovirus strain ALL-03 in vitro

Balakrishnan KN, Abdullah AA, Bala JA, Jesse FFA, Abdullah CAC, Noordin MM, et al.

Infect Genet Evol, 2021 06;90:104783.
PMID: 33640483 DOI: 10.1016/j.meegid.2021.104783

OBJECTIVE: This study investigated the suitability of siRNA targeting specific genes that cause inhibition of virus replication in vitro especially for the virus that capable of crossing placenta and we employed a novel transplacental rat cytomegalovirus that mimics infection in human.
METHODS: Six unique siRNAs with three each targeting different regions of IE2 (ie2a, ie2b and ie2c) and DNA polymerase (dpa, dpb and dpc) were prepared and tested for antiviral activities. The efficacy as an antiviral was determined in in-vitro by measuring TCID50 virus titer, severity of virus-induced cytopathic effect (CPE), intracellular viral genome loads by droplet digital PCR, the degree of apoptosis in siRNA-treated cells and relative expression of viral mRNA in infected Rat Embryo Fibroblast (REF) cells.
FINDINGS: Remarkably, the siRNAs: dpa, dpb and IE2b, significantly reduced virus yield (approximately >90%) compared to control group at day 18 post infection (p.i). Changes in CPE indicated that DNA polymerase siRNAs were capable of protecting cells against CMV infection at day 14 p.i with higher efficiency than GCV (at the concentration of 300 pmol). Gene expression analysis revealed a marked down regulation of the targeted DNA polymerase gene (73.9%, 96.0% and 90.7% for dpa, dpb and dpc siRNA, respectively) and IE2 gene (50.8%, 49.9% and 15.8% for ie2a, ie2b and ie2c siRNA, respectively) when measured by RT-qPCR. Intracellular viral DNA loads showed a significant reduction for all the DNA polymerase siRNAs (dpa: 96%, dpb: 98% and dpc:92) compared to control group (P
Time dependent effect of chronic embryonic exposure to ethanol on zebrafish: Morphology, biochemical and anxiety alterations

Ramlan NF, Sata NSAM, Hassan SN, Bakar NA, Ahmad S, Zulkifli SZ, et al.

Behav Brain Res, 2017 08 14;332:40-49.
PMID: 28559182 DOI: 10.1016/j.bbr.2017.05.048

Exposure to ethanol during critical period of development can cause severe impairments in the central nervous system (CNS). This study was conducted to assess the neurotoxic effects of chronic embryonic exposure to ethanol in the zebrafish, taking into consideration the time dependent effect. Two types of exposure regimen were applied in this study. Withdrawal exposure group received daily exposure starting from gastrulation until hatching, while continuous exposure group received daily exposure from gastrulation until behavioural assessment at 6dpf (days post fertilization). Chronic embryonic exposure to ethanol decreased spontaneous tail coiling at 24hpf (hour post fertilization), heart rate at 48hpf and increased mortality rate at 72hpf. The number of apoptotic cells in the embryos treated with ethanol was significantly increased as compared to the control. We also measured the morphological abnormalities and the most prominent effects can be observed in the treated embryos exposed to 1.50% and 2.00%. The treated embryos showed shorter body length, larger egg yolk, smaller eye diameter and heart edema as compared to the control. Larvae received 0.75% continuous ethanol exposure exhibited decreased swimming activity and increased anxiety related behavior, while withdrawal ethanol exposure showed increased swimming activity and decreased anxiety related behavior as compared to the respective control. Biochemical analysis exhibited that ethanol exposure for both exposure regimens altered proteins, lipids, carbohydrates and nucleic acids of the zebrafish larvae. Our results indicated that time dependent effect of ethanol exposure during development could target the biochemical processes thus leading to induction of apoptosis and neurobehavioral deficits in the zebrafish larvae. Thus it raised our concern about the safe limit of alcohol consumption for pregnant mother especially during critical periods of vulnerability for developing nervous system.
Crafting two-dimensional materials for contrast agents, drug, and heat delivery applications through green technologies

Setyawan D, Amrillah T, Abdullah CAC, Ilhami FB, Dewi DMM, Mumtazah Z, et al.

J Drug Target, 2023 Apr;31(4):369-389.
PMID: 36721905 DOI: 10.1080/1061186X.2023.2175833

The development of two-dimensional (2D) materials for biomedical applications has accelerated exponentially. Contrary to their bulk counterparts, the exceptional properties of 2D materials make them highly prospective for contrast agents for bioimage, drug, and heat delivery in biomedical treatment. Nevertheless, empty space in the integration and utilisation of 2D materials in living biological systems, potential toxicity, as well as required complicated synthesis and high-cost production limit the real application of 2D materials in those advance medical treatments. On the other hand, green technology appears to be one of strategy to shed a light on the blurred employment of 2D in medical applications, thus, with the increasing reports of green technology that promote advanced technologies, here, we compile, summarise, and synthesise information on the biomedical technology of 2D materials through green technology point of view. Beginning with a fundamental understanding, of crystal structures, the working mechanism, and novel properties, this article examines the recent development of 2D materials. As well as 2D materials made from natural and biogenic resources, a recent development in green-related synthesis was also discussed. The biotechnology and biomedical-related application constraints are also discussed. The challenges, solutions, and prospects of the so-called green 2D materials are outlined.
Fulltext Down-top nanofabrication of binary (CdO)x (ZnO)1-x nanoparticles and their antibacterial activity

Al-Hada NM, Mohamed Kamari H, Abdullah CAC, Saion E, Shaari AH, Talib ZA, et al.

Int J Nanomedicine, 2017;12:8309-8323.
PMID: 29200844 DOI: 10.2147/IJN.S150405

In the present study, binary oxide (cadmium oxide [CdO])x (zinc oxide [ZnO])1-x nanoparticles (NPs) at different concentrations of precursor in calcination temperature were prepared using thermal treatment technique. Cadmium and zinc nitrates (source of cadmium and zinc) with polyvinylpyrrolidone (capping agent) have been used to prepare (CdO)x (ZnO)1-x NPs samples. The sample was characterized by X-ray diffraction (XRD), scanning electron microscopy, energy-dispersive X-ray (EDX), transmission electron microscopy (TEM), and Fourier transform infrared (FTIR) spectroscopy. XRD patterns analysis revealed that NPs were formed after calcination, which showed a cubic and hexagonal crystalline structure of (CdO)x (ZnO)1-x NPs. The phase analysis using EDX spectroscopy and FTIR spectroscopy confirmed the presence of Cd and Zn as the original compounds of prepared (CdO)x (ZnO)1-x NP samples. The average particle size of the samples increased from 14 to 33 nm as the concentration of precursor increased from x=0.20 to x=0.80, as observed by TEM results. The surface composition and valance state of the prepared product NPs were determined by X-ray photoelectron spectroscopy (XPS) analyses. Diffuse UV-visible reflectance spectra were used to determine the optical band gap through the Kubelka-Munk equation; the energy band gap was found to decrease for CdO from 2.92 to 2.82 eV and for ZnO from 3.22 to 3.11 eV with increasing x value. Additionally, photoluminescence (PL) spectra revealed that the intensity in PL increased with an increase in particle size. In addition, the antibacterial activity of binary oxide NP was carried out in vitro against Escherichia coli ATCC 25922 Gram (-ve), Salmonella choleraesuis ATCC 10708, and Bacillus subtilis UPMC 1175 Gram (+ve). This study indicated that the zone of inhibition of 21 mm has good antibacterial activity toward the Gram-positive B. subtilis UPMC 1175.