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Incorporation of zinc oxide nanoparticles into chitosan-collagen 3D porous scaffolds: Effect on morphology, mechanical properties and cytocompatibility of 3D porous scaffolds

Ullah S, Zainol I, Idrus RH

Int J Biol Macromol, 2017 Nov;104(Pt A):1020-1029.
PMID: 28668615 DOI: 10.1016/j.ijbiomac.2017.06.080

The zinc oxide nanoparticles (particles size <50nm) incorporated into chitosan-collagen 3D porous scaffolds and investigated the effect of zinc oxide nanoparticles incorporation on microstructure, mechanical properties, biodegradation and cytocompatibility of 3D porous scaffolds. The 0.5%, 1.0%, 2.0% and 4.0% zinc oxide nanoparticles chitosan-collagen 3D porous scaffolds were fabricated via freeze-drying technique. The zinc oxide nanoparticles incorporation effects consisting in chitosan-collagen 3D porous scaffolds were investigated by mechanical and swelling tests, and effect on the morphology of scaffolds examined microscopically. The biodegradation and cytocompatibility tests were used to investigate the effects of zinc oxide nanoparticles incorporation on the ability of scaffolds to use for tissue engineering application. The mean pore size and swelling ratio of scaffolds were decreased upon incorporation of zinc oxide nanoparticles however, the porosity, tensile modulus and biodegradation rate were increased upon incorporation of zinc oxide nanoparticles. In vitro culture of human fibroblasts and keratinocytes showed that the zinc oxide nanoparticles facilitated cell adhesion, proliferation and infiltration of chitosan-collagen 3D porous scaffolds. It was found that the zinc oxide nanoparticles incorporation enhanced porosity, tensile modulus and cytocompatibility of chitosan-collagen 3D porous scaffolds.
Development of various composition multicomponent chitosan/fish collagen/glycerin 3D porous scaffolds: Effect on morphology, mechanical strength, biostability and cytocompatibility

Ullah S, Zainol I, Chowdhury SR, Fauzi MB

Int J Biol Macromol, 2018 May;111:158-168.
PMID: 29305219 DOI: 10.1016/j.ijbiomac.2017.12.136

The various composition multicomponent chitosan/fish collagen/glycerin 3D porous scaffolds were developed and investigated the effect of various composition chitosan/fish collagen/glycerin on scaffolds morphology, mechanical strength, biostability and cytocompatibility. The scaffolds were fabricated via freeze-drying technique. The effects of various compositions consisting in 3D scaffolds were investigated via FT-IR analysis, porosity, swelling and mechanical tests, and effect on the morphology of scaffolds investigated microscopically. The biostability and cytocompatibility tests were used to explore the ability of scaffolds to use for tissue engineering application. The average pore sizes of scaffolds were in range of 100.73±27.62-116.01±52.06, porosity 71.72±3.46-91.17±2.42%, tensile modulus in dry environment 1.47±0.08-0.17±0.03MPa, tensile modulus in wet environment 0.32±0.03-0.14±0.04MPa and biodegradation rate (at day 30) 60.38±0.70-83.48±0.28%. In vitro culture of human fibroblasts and keratinocytes showed that the various composition multicomponent 3D scaffolds were good cytocompatibility however, the scaffolds contained high amount of fish collagen excellently facilitated cell proliferation and adhesion. It was found that the high amount fish collagen and glycerin scaffolds have high porosity, enough mechanical strength and biostability, and excellent cytocompatibility.
Fulltext The Effects of Silver Nanoparticles Compositions on the Mechanical, Physiochemical, Antibacterial, and Morphology Properties of Sugar Palm Starch Biocomposites for Antibacterial Coating

Rozilah A, Jaafar CNA, Sapuan SM, Zainol I, Ilyas RA

Polymers (Basel), 2020 Nov 06;12(11).
PMID: 33171913 DOI: 10.3390/polym12112605

Antibacterial sugar palm starch biopolymer composite films were developed and derived from renewable sources and inorganic silver nanoparticles (AgNPs) as main ingredients for antibacterial coatings. The composite films were produced by solution casting method and the mechanical and physicochemical properties were determined by tensile test, Fourier Transform Infrared (FTIR) analysis, thermal gravimetric analysis (TGA), antibacterial screening test and field emission scanning electron microscopy (FESEM) images. It was found that mechanical and antibacterial properties of biocomposite films were improved after the addition of AgNPs compared with the film without active metals. The weakness of neat biocomposite films was improved by incorporating inorganic AgNPs as a nanofiller in the films' matrix to avoid bacterial growth. The results showed that the tensile strength ranged between 8 kPa and 408 kPa and the elasticity modulus was between 5.72 kPa and 9.86 kPa. The addition of AgNPs in FTIR analysis decreased the transmittance value, caused small changes in the chemical structure, caused small differences in the intensity peaks, and produced longer wavelengths. These active films increased the degradation weight and decomposition temperature due to the more heat-stable AgNPs. Meanwhile, the average inhibited areas measured were between 7.66 and 7.83 mm (Escherichia coli), 7.5 and 8.0 mm (Salmonella cholerasuis), and 0.1 and 0.5 mm for Staphylococcus aureus. From the microscopic analysis, it was observed that the average size of all microbes for 1 wt% and 4 wt% AgNPs ranged from 0.57 to 2.90 mm. Overall, 3 wt% AgNP nanofiller was found to be the best composition that fulfilled all the mechanical properties and had better antimicrobial properties. Thus, the development of an organic-inorganic hybrid of antibacterial biopolymer composite films is suitable for antibacterial coatings.
Fulltext In vitro characterization of a chitosan skin regenerating template as a scaffold for cells cultivation

Hilmi AB, Halim AS, Hassan A, Lim CK, Noorsal K, Zainol I

Springerplus, 2013 Dec;2(1):79.
PMID: 23503998 DOI: 10.1186/2193-1801-2-79

Chitosan is a marine-derived product that has been widely used in clinical applications, especially in skin reconstruction. The mammalian scaffolds derived from bovine and porcine material have many limitations, for example, prion transmission and religious concerns. Therefore, we created a chitosan skin regenerating template (SRT) and investigated the behavior of fibroblast cell-scaffold constructs. Primary human dermal fibroblasts (HDF) were isolated and then characterized using vimentin and versican. HDF were seeded into chitosan SRT at a density of 3×10(6) cells/cm(2) for fourteen days. Histological analysis and live cells imaging revealed that the cell-chitosan constructs within interconnected porous chitosan showed significant interaction between the cells as well as between the cells and the chitosan. Scanning electron microscopy (SEM) analysis revealed cells spreading and covering the pores. As the pore sizes of the chitosan SRT range between 40-140 μm, an average porosity is about 93 ± 12.57% and water uptake ratio of chitosan SRT is 536.02 ± 14.29%, it is a supportive template for fibroblast attachment and has potential in applications as a dermal substitute.
Keloid pathogenesis via Drosophila similar to mothers against decapentaplegic (SMAD) signaling in a primary epithelial-mesenchymal in vitro model treated with biomedical-grade chitosan porous skin regenerating template

Lim CK, Halim AS, Yaacob NS, Zainol I, Noorsal K

J Biosci Bioeng, 2013 Apr;115(4):453-8.
PMID: 23177217 DOI: 10.1016/j.jbiosc.2012.10.010

The effects of locally produced chitosan (CPSRT-NC-bicarbonate) in the intervention of keloid pathogenesis were investigated in vitro. A human keratinocyte-fibroblast co-culture model was established to investigate the protein levels of human collagen type-I, III and V in a western blotting analysis, the secreted transforming growth factor-β1 (TGF-β1) in an enzyme-linked immunosorbent assay (ELISA) and the mRNA levels of TGF-β1's intracellular signaling molecules (SMAD2, 3, 4 and 7) in a real-time PCR analysis. Keratinocyte-fibroblast co-cultures were maintained in DKSFM:DMEM:F12 (2:2:1) medium. Collagen type-I was found to be the dominant form in primary normal human dermal fibroblast (pNHDF) co-cultures, whereas collagen type-III was more abundant in primary keloid-derived human dermal fibroblast (pKHDF) co-cultures. Collagen type-V was present as a minor component in the skin. TGF-β1, SMAD2 and SMAD4 were expressed more in the pKHDF than the pNHDF co-cultures. Co-cultures with normal keratinocytes suppressed collagen type-III, SMAD2, SMAD4 and TGF-β1 expressions and CPSRT-NC-bicarbonate enhanced this effect. In conclusion, the CPSRT-NC-bicarbonate in association with normal-derived keratinocytes demonstrated an ability to reduce TGF-β1, SMAD2 and SMAD4 expressions in keloid-derived fibroblast cultures, which may be useful in keloid intervention.
Matrix Production in Chondrocytes Transfected with Sex Determining Region Y-Box 9 and Telomerase Reverse Transcriptase Genes: An In Vitro Evaluation from Monolayer Culture to Three-Dimensional Culture

Md Nazir N, Zulkifly AH, Khalid KA, Zainol I, Zamli Z, Sha'ban M

Tissue Eng Regen Med, 2019 06;16(3):285-299.
PMID: 31205857 DOI: 10.1007/s13770-019-00191-1

Background: This study aimed to observe the cartilaginous matrix production in SRY (sex determining region Y)-box 9 (SOX9)- and/or telomerase reverse transcriptase (TERT)-transfected chondrocytes from monolayer to three-dimensional (3D) culture.
Methods: The genes were transferred into chondrocytes at passage-1 (P1) via lipofection. The post-transfected chondrocytes (SOX9-, TERT- and SOX9/TERT) were analysed at P1, P2 and P3. The non-transfected group was used as control. The 3D culture was established using the chondrocytes seeded in a disc-shaped PLGA/fibrin and PLGA scaffolds. The resulting 3D "cells-scaffolds" constructs were analysed at week-1, -2 and -3. The histoarchitecture was evaluated using haematoxylin and eosin, alcian blue and safranin o stains. The quantitative sulphated glycosaminoglycan (sGAG) content was measured using biochemical assay. The cartilage-specific markers expression were analysed via real-time polymerase chain reaction.
Results: All monolayer cultured chondrocytes showed flattened, fibroblast-like appearance throughout passages. Proteoglycan and sGAG were not detected at the pericellular matrix region of the chondrocytes. The sGAG content assay indicated the matrix production depletion in the culture. The cartilage-specific markers, COL2A1 and ACAN, were downregulated. However, the dedifferentiation marker, COL1A1 was upregulated. In 3D "cells-scaffolds" constructs, regardless of transfection groups, chondrocytes seeded in PLGA/fibrin showed a more uniform distribution and produced denser matrix than the PLGA group especially at week-3. Both sGAG and proteoglycan were clearly visualised in the constructs, supported by the increment of sGAG content, quantitatively. Both COL2A1 and ACAN were upregulated in SOX9/TERT-PLGA and SOX9/TERT-PLGA/fibrin respectively. While, COL1A1 was downregulated in SOX9/TERT-PLGA.
Conclusion: These findings indicated that the SOX9/TERT-transfected chondrocytes incorporation into 3D scaffolds facilitates the cartilage regeneration which is viable structurally and functionally.
Fulltext Risk Assessment of Fishing Trawl Activities to Subsea Pipelines of Sabah and Labuan Waters

Ahmad Fuad AF, Said MH, Samo K, Rahman MAA, Mohd MH, Zainol I

ScientificWorldJournal, 2020;2020:6957171.
PMID: 33414690 DOI: 10.1155/2020/6957171

Introduction. Trawling is a method of catching fish in a large volume where fish nets are pulled through water using one or two boats. Bottom trawling is where the nets are pulled over on the seabed. The gear of the bottom trawling would impact the exposed subsea pipeline, on the seabed. Subsea pipelines transport crude oil and gas from the offshore platform to shore facility. This study assesses the risk of fish trawling activities to the subsea pipelines at Sabah and Labuan offshore. The specification of trawl equipment used by local trawlers in Sabah was determined by the on-site survey. The frequency of a fish trawler crossing over the pipelines was calculated based interview on operation and site survey. The calculation of the pull-over load of the otter board was calculated using the DNVGL algorithm. The severity and frequency index of the risk matrix was developed based on literature review. Results showed that the pull-over load of the otter board would not damage the pipelines. The risk posed by the fish trawler activity to the pipelines is low and moderate.
Fulltext Preparation and characterization of FsHA/FsCol beads: Cell attachment and cytotoxicity studies

Mudhafar M, Zainol I, Alsailawi HA, Zorah M, Karhib MM, Mahmood Mahdi N

Heliyon, 2023 May;9(5):e15838.
PMID: 37206015 DOI: 10.1016/j.heliyon.2023.e15838

The present study was conducted to prepare the fish scales' hydroxyapatite/collagen beads (FsHA/FsCol) and characterize their biological, physical, and chemical properties. A new method was used to prepare FsHA/FsCol composite beads by infiltrating the beads of FsHA in the solution of FsCol as a green method. X-ray diffraction analysis (XRD), scanning electron microscopy (SEM) analysis, Fourier-transform infrared (FTIR) spectroscopy analysis and energy dispersive X-ray analysis (EDX), used to evaluate the physical-chemical properties of the synthesized samples. Meanwhile, the cytotoxic and attachment studies of the FsHA/FsCol beads were used to investigate the biological features against the MG-63 human cell line. The results specified the efficiency of the new method, functional groups of FsCol were indicated to be present inside the beads of FsHA according to the XRD analysis which shows the functional peaks of FsCol. The SEM image were conformed successfully use starch as a porous agent to increasing the porous of the FsHA beads after adding 20 wt% of it. Alamar Blue assay has been used to evaluate the cytotoxicity of FsHA/FsCol beads the results were shown 87% average cell viability of the MG-63 human cell line on the beads and attached very well to the surface of the composites, indicating no toxicity being exerted by all the composites at high concentrations.
Fulltext Corrigendum to "Risk Assessment of Fishing Trawl Activities to Subsea Pipelines of Sabah and Labuan Waters"

Fuad AFA, Said MH, Samo K, Rahman MAA, Mohd MH, Zainol I

ScientificWorldJournal, 2020;2020:7014928.
PMID: 33522519 DOI: 10.1155/2020/7014928

[This corrects the article DOI: 10.1155/2020/6957171.].
The effect of fish collagen on the silver nanoparticles sizes and shapes using modified microwave-assisted green synthesis method and their antibacterial activities

Mudhafar M, Zainol I, A J A, Abd MY, Alsailawi HA, Ghazaly NM, et al.

Heliyon, 2024 Jun 30;10(12):e32837.
PMID: 39022059 DOI: 10.1016/j.heliyon.2024.e32837

This work aimed to produce silver nanoparticles (AgNPs) by efficient green synthesis techniques, namely rapid green synthesis and modified microwave-assisted green synthesis methods. The study used fish scale collagen (FsCol) as a stabilizer to assess its impact on the dimensions and configurations of AgNPs. Four samples were prepared with varying concentrations of FsCol. The synthesized AgNPs were characterized using Ultraviolet-visible (UV-vis) spectroscopy, scanning electron microscope (SEM), energy dispersive X-ray analysis (EDX), Fourier Transform Infrared Spectroscopy (FTIR), X-Ray diffraction analysis (XRD), Dynamic Light Scattering (DLS), and Transmission electron microscopy (TEM) techniques. The obtained sizes are as follows: 85 ± 15 nm, 70 ± 10 nm, 50 ± 10 nm, and 28-40 nm. The UV-vis spectroscopy revealed a shift in the absorbance peaks from 400 to 446 nm. The SEM method showed a spherical form in all of the samples. The element silver was detected in the EDX examination, along with the presence of oxygen (O) and carbon (C). The FTIR analysis revealed that the peaks seen at 3307 cm-1 were attributed to the stretching of O-H bonds, while the mountain at 1638 cm-1 belonged to the extension of N-H bonds (amide A). Additionally, the band observed at 1638 cm-1 indicated the presence of CO bonds (amide I).The 2140 cm-1 and 1302 cm-1 peaks may be attributed to the C2H2 group present in the plant components and the N-H bending (Amide III), respectively. The XRD pattern indicates that the synthesis process resulted in the formation of crystalline AgNPs. The particle sizes measured using DLS were 121 nm, 96.36 nm, 82.3 nm, and 48.50 nm. The TEM approach revealed that all samples had a spherical morphology with varying sizes: 80-100 nm, 50-80 nm, 40-60 nm, and 28-42 nm. The synthesized AgNPs were tested for their antibacterial properties against the pathogenic pathogens Escherichia coli (E.coli) and Staphylococcus aureus (S. aureus). The influence of AgNPs on bacteria was amplified as the particle size decreased, resulting in a larger inhibitory zone for the smaller particles.

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