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Fulltext Current Insight of Printability Quality Improvement Strategies in Natural-Based Bioinks for Skin Regeneration and Wound Healing

Masri S, Fauzi MB

Polymers (Basel), 2021 Mar 25;13(7).
PMID: 33805995 DOI: 10.3390/polym13071011

Skin tissue engineering aimed to replace chronic tissue injury commonly occurred due to severe burn and chronic wound in diabetic ulcer patients. The normal skin is unable to be regenerated until the seriously injured tissue is disrupted and losing its function. 3D-bioprinting has been one of the effective methods for scaffold fabrication and is proven to replace the conventional method, which reported several drawbacks. In light of this, researchers have developed a new fabrication approach via 3D-bioprinting by combining biomaterials (bioinks) with cells and biomolecules followed by a suitable crosslinking approach. This advanced technology has been subcategorised into three different printing techniques including inject-based, laser-based, and extrusion-based printing. However, the printable quality of the currently available bioinks demonstrated shortcomings in the physicochemical and mechanical properties. This review aims to identify the limitations raised by using natural-based bioinks and the optimum temperature for various applied printing techniques. It is essential to ensure maintaining the acceptable printed scaffold property such as the optimum pore sizes and porosity that allow cell migration activity. In addition, the properties required for an ideal bioinks design for better scaffold printability were also summarised.
Fulltext Field bioefficacy of deltamethrin residual spraying against dengue vectors

Rozilawati H, Lee HL, Mohd Masri S, Mohd Noor I, Rosman S

Trop Biomed, 2005 Dec;22(2):143-8.
PMID: 16883280 MyJurnal

Field bioefficacy of residual-sprayed deltamethrin against Aedes vectors was evaluated in an urban residential area in Kuala Lumpur. The trial area consisted of single storey wood-brick houses and a block of flat. The houses were treated with outdoor residual spraying while the flat was used as an untreated control. Initial pre-survey using ovitrap surveillance indicated high Aedes population in the area. Deltamethrin WG was sprayed at a dosage of 25mg/m2 using a compression sprayer. The effectiveness of deltamethrin was determined by wall bioassay and ovitrap surveillance. The residual activity of 25mg/m2 deltamethrin was still effective for 6 weeks after treatment, based on biweekly bioassay results. Bioassay also indicated that both Aedes aegypti and Aedes albopictus were more susceptible on the wooden surfaces than on brick. Aedes aegypti was more susceptible than Ae. albopictus against deltamethrin. Residual spraying of deltamethrin was not very effective against Aedes in this study since the Aedes population in the study area did not reduce as indicated by the total number of larvae collected using the ovitrap (Wilcoxon Sign Test, p> 0.05). Further studies are required to improve the effectiveness of residual spraying against Aedes vectors.
Fulltext Hyaluronatelyase production by Streptococcus pneumoniae isolated from patients and carriers

Yusof HA, Desa M NM, Masri SN, Malina O, Jamal F

Trop Biomed, 2015 Sep;32(3):413-8.
PMID: 26695201 MyJurnal

Hyaluronatelyase produced by various microorganisms are capable of degrading hyaluronic acid in connective tissues and initiating the spread of infection by opening an access for the pathogen into host tissues. The present study attempts to determine the distribution of hyaluronatelyase-producing Streptococcus pneumoniae among invasive, non invasive and carriage isolates, and correlate it with the clinical sources, year of isolation, colonial morphology and their serotypes. A total of 100 isolates from various clinical samples were selected and screened for hyaluronatelyase production and presence of the encoding SpnHyl gene. All isolates possessed SpnHyl gene. Ninety-six isolates including 34 carriage isolates were positive for production of hyaluronatelyase. Four hyaluronatelyase-negative isolates were from blood (2 isolates) and sputum (2 isolates). No significant association was detected among hyaluronatelyase production and bacterial characteristics except for colonial morphology (p = 0.040). High percentages of hyaluronatelyase production in these isolates suggest their possible role as human pathogens.
Fulltext Impact of larviciding with a Bacillus thuringiensis israelensis formulation, VectoBac WG, on dengue mosquito vectors in a dengue endemic site in Selangor State, Malaysia

Lee HL, Chen CD, Masri SM, Chiang YF, Chooi KH, Benjamin S

Southeast Asian J. Trop. Med. Public Health, 2008 Jul;39(4):601-9.
PMID: 19058596

The field bioefficacy of a wettable granule (WG) formulation of Bacillus thuringiensis israelensis (Bti), VectoBac WG (Bti strain AM65-52) against dengue vectors, Aedes aegypti and Ae albopictus; was evaluated in a suburban residential area (TST) and in a temporary settlement site (KB) in the state of Selangor, Malaysia. Pre-control ovitrap surveillance of the trial sites indicated a high population of both types of Aedes mosquitoes. The populations were monitored continuously by weekly ovitrapping. Bti was sprayed biweekly at a dosage of 500 g/ha by using a mist-blower. The spray application was targeted into outdoor larval habitats. If required, Bti formulation was also applied directly into indoor water-holding containers at 8 g/1,000 l. Based on ovitrap surveillance, a significant reduction in Aedes populations was evident 4 weeks after initiating the first Bti treatment. The ovitrap index (OI) and the larvae density decreased drastically in both trial sites. In TST, the indoor OI was significantly reduced from 57.50 +/- 7.50% to 19.13 +/- 5.49% (p<0.05), while the outdoor OI decreased from 38.89 +/- 11.11% to 15.36 +/- 5.93%. In KB, similarly, the OI was significantly reduced by more than half, from 66.66 +/- 6.67% to 30.26 +/- 2.99% (p< 0.05). In all cases, the reduction in OI was paralleled by reduction in larval density.
Distribution of virulence genes and the molecular epidemiology of Streptococcus pyogenes clinical isolates by emm and multilocus sequence typing methods

Hamzah SNA, Mohd Desa MN, Jasni AS, Mohd Taib N, Masri SN, Hamat RA

Med J Malaysia, 2021 03;76(2):164-170.
PMID: 33742623

BACKGROUND: Streptococcus pyogenes has a variety of virulence factors and the predominant invasive strains differ according to specific emm types and geographical orientation. Although emm typing is commonly used as the gold standard method for the molecular characterisation, multilocus sequence typing (MLST) has become an important tool for comparing the genetic profiles globally. This study aimed to screen selected virulence genes from invasive and non-invasive clinical samples and to characterise the molecular epidemiology by emm typing and MLST methods.
MATERIALS AND METHODS: A total of 42 S. pyogenes isolates from invasive and non-invasive samples collected from two different tertiary hospitals were investigated for the distribution of virulence factors and their molecular epidemiology by emm and multilocus sequence typing methods. Detection of five virulence genes (speA, speB, speJ, ssa and sdaB) was performed using multiplex polymerase chain reaction (PCR) using the standard primers and established protocol. Phylogenetic tree branches were constructed from sequence analysis utilised by neighbour joining method generated from seven housekeeping genes using MEGA X software.
RESULTS: Multiplex PCR analysis revealed that sdaB/speF (78.6%) and speB (61.9%) were the predominant virulence genes. Regardless of the type of invasiveness, diverse distribution of emm types/subtypes was noted which comprised of 27 different emm types/subtypes. The predominant emm types/subtypes were emm63 and emm18 with each gene accounted for 11.8% whereas 12% for each gene was noted for emm28, emm97.4 and emm91. The MLST revealed that the main sequence type (ST) in invasive samples was ST402 (17.7%) while ST473 and ST318 (12% for each ST) were the major types in non-invasive samples. Out of 18 virulotypes, Virulotype A (five genes, 55.6%) and Virulotype B (two genes, 27.8%) were the major virulotypes found in this study. Phylogenetic analysis indicated the presence of seven different clusters of S. pyogenes. Interestingly, Cluster VI showed that selected emm/ST types such as emm71/ST318 (n=2), emm70.1/ST318 (n=1), emm44/ST31 (n=1) and emm18/ST442 (n=1) have clustered within a common group (Virulotype A) for both hospitals studied.
CONCLUSION: The present study showed that group A streptococcci (GAS) are genetically diverse and possess virulence genes regardless of their invasiveness. Majority of the GAS exhibited no restricted pattern of virulotypes except for a few distinct clusters. Therefore, it can be concluded that virulotyping is partially useful for characterising a heterogeneous population of GAS in hospitals.
Fulltext Surveillance of Aedes albopictus Skuse breeding preference in selected dengue outbreak localities, peninsular Malaysia

Rozilawati H, Tanaselvi K, Nazni WA, Mohd Masri S, Zairi J, Adanan CR, et al.

Trop Biomed, 2015 Mar;32(1):49-64.
PMID: 25801254 MyJurnal

Entomological surveillance was conducted in order to determine the abundance and to evaluate any changes of biological vectors or ecology, especially in the dengue outbreak areas. The abundance and breeding preference of Aedes albopictus and Aedes aegypti were conducted in selected dengue outbreak localities in three states of peninsular Malaysia namely Selangor, Federal Territory of Kuala Lumpur, and Penang Island using ovitraps and larval survey method. It was determined that Ae. albopictus was predominant in most of the localities and found to breed more outdoor than indoor. A wide range of breeding foci were recorded in this study. It was also determined that ovitrap method was more effective to detect the presence of Aedes mosquitoes when the larval survey was at low rate of infestation. The abundance of Ae. albopictus in dengue outbreak localities emphasis that the vector control programme should also target this species together with the primary dengue vector, Ae. aegypti.
Fulltext Sterilisation of Lucilia cuprina Wiedemann maggots used in therapy of intractable wounds

Mohd Masri S, Nazni WA, Lee HL, T Rogayah TA, Subramaniam S

Trop Biomed, 2005 Dec;22(2):185-9.
PMID: 16883286 MyJurnal

Three new techniques of sterilising maggots of Lucilia cuprina for the purpose of debriding intractable wounds were studied. These techniques were utilisation of ultra-violet C (UVC) and maggot sterilisation with disinfectants. The status of sterility was checked on nutrient agar and blood agar and confirmed with staining. The indicators for the effectiveness of the methods were sterility and survival rate of the eggs or larvae. Egg sterilisation with UVC had the lowest hatching rate (16+/-0.00%) while egg sterilisation with disinfectants showed high hatching rate (36.67+/-4.41%) but low maggot survival rate (31.67+/-1.67%). Sterilisation of the maggots was the most suitable, since the survival rate was the highest (88.67+/-0.88%). Complete sterility was achieved in all cases, except that Proteus mirabilis was consistently found. However, the presence of this microorganism was considered beneficial.
Fulltext Mixed breeding of Aedes aegypti (L.) and Aedes albopictus Skuse in four dengue endemic areas in Kuala Lumpur and Selangor, Malaysia

Chen CD, Nazni WA, Lee HL, Seleena B, Mohd Masri S, Chiang YF, et al.

Trop Biomed, 2006 Dec;23(2):224-7.
PMID: 17322826 MyJurnal

Ovitrap surveillance was conducted in four dengue endemic areas in Kuala Lumpur and Selangor, Malaysia to determine the distribution and percentage of mixed breeding of both Aedes aegypti and Aedes albopictus. The percentage of mixed breeding in all study sites both indoors and outdoors accounted for 10 to 32 % from the total ovitraps collected. Ae. aegypti was found at a higher frequency than Ae. albopictus in these ovitraps. This study again indicates that ovitrap is a sensitive tool to attract gravid females of more than one mosquito species to oviposit in the container.
The importance of using a right test method in diagnosing leptospirosis

Ilham NE, Joseph NSM, Bahtiar Affendy N, Mohd Taib N, Vasantha KN, Masri SN

Trop Biomed, 2020 Jun 01;37(2):357-362.
PMID: 33612804

Leptospirosis is a common febrile illness in Malaysia. The disease is caused by pathogenic bacteria called leptospires that are transmitted directly or indirectly from animals to humans via contaminated water or soil. It is a potentially serious but treatable disease. Its symptoms may mimic those of other unrelated febrile illnesses such as dengue, influenza, meningitis, hepatitis or viral haemorrhagic fevers. The spectrum of the disease is extremely wide, ranging from subclinical infection to a severe syndrome of multiorgan infection with high mortality. The diagnosis requires high suspicion with history of exposure to water or environment possibly contaminated with infected animal urine. This is a case of a 13 year-oldgirl with no known medical illness, and a history of exposure to outdoor activities. However, paired sera for leptospirosis serology was not diagnostic. She then developed septic shock on day 14 of illness. But due to high suspicion of leptospirosis, antibiotic therapy was upgraded to ceftriaxone and samples were sent for further testing which revealed that leptospires were detected in the urine, using molecular technique. She improved after treated as leptospirosis.
Evaluation of Tween 80 incorporated media to increase pathogen isolation from peritoneal fluid of CAPD patients at Hospital Kuala Lumpur

Ganapathy Pillay S, Mohd Amin SDW, Masri SN, Joseph NM, Amran F, Van Belkum A, et al.

Malays J Pathol, 2021 Aug;43(2):261-268.
PMID: 34448790

INTRODUCTION: Continuous ambulatory peritoneal dialysis (CAPD)-associated peritonitis remains a major complication in patients on CAPD leading to increased morbidity and mortality. Successful therapy of peritonitis is highly dependent on a positive microbiological culture because narrow spectrum antibiotics are essential to efficiently combat infection. Therefore, this study evaluated the performance of Tween 80 containing media at three different concentrations (0.1%, 1.0% and 2.0%) to increase the pathogen yield from peritoneal fluid in comparison with the standard culture media.
MATERIALS AND METHODS: Peritoneal fluid samples (n=121) obtained from CAPD patients suspected of peritonitis at Hospital Kuala Lumpur were analysed macroscopically and microscopically prior to culture. All samples were cultured on seven different culture media, including sheep blood agar, MacConkey agar, Sabouraud dextrose agar, brain heart infusion agar and Tween 80 incorporated blood agar. All plates were incubated at an optimum temperature up to 48 hours.
RESULTS AND CONCLUSION: Among all the culture media investigated, 0.1% to 2.0% Tween 80 incorporated blood agar yielded the highest positive culture (23/121) in comparison with all other standard media, thus lowering the negative culture rate among CAPD patients. Statistical analysis by Chi Square revealed significant differences (p <0.001) between the three concentrations of Tween 80 tested in this study. Among the three different concentrations of Tween 80 optimised in this study, blood agar containing 0.1% Tween 80 generated the best results, achieved by optimum growth of all Gram-positive organisms, Gram-negative organisms and yeast cells simultaneously. Using a small amount of detergent at low cost significantly increased the pathogen yield during CAPD-associated peritonitis.
Fulltext Injectable Crosslinked Genipin Hybrid Gelatin-PVA Hydrogels for Future Use as Bioinks in Expediting Cutaneous Healing Capacity: Physicochemical Characterisation and Cytotoxicity Evaluation

Masri S, Maarof M, Mohd NF, Hiraoka Y, Tabata Y, Fauzi MB

Biomedicines, 2022 Oct 20;10(10).
PMID: 36289912 DOI: 10.3390/biomedicines10102651

The irregular shape and depth of wounds could be the major hurdles in wound healing for the common three-dimensional foam, sheet, or film treatment design. The injectable hydrogel is a splendid alternate technique to enhance healing efficiency post-implantation via injectable or 3D-bioprinting technologies. The authentic combination of natural and synthetic polymers could potentially enhance the injectability and biocompatibility properties. Thus, the purpose of this study was to characterise a hybrid gelatin−PVA hydrogel crosslinked with genipin (GNP; natural crosslinker). In brief, gelatin (GE) and PVA were prepared in various concentrations (w/v): GE, GPVA3 (3% PVA), and GPVA5 (5% PVA), followed by a 0.1% (w/v) genipin (GNP) crosslink, to achieve polymerisation in three minutes. The physicochemical and biocompatibility properties were further evaluated. GPVA3_GNP and GPVA5_GNP with GNP demonstrated excellent physicochemical properties compared to GE_GNP and non-crosslinked hydrogels. GPVA5_GNP significantly displayed the optimum swelling ratio (621.1 ± 93.18%) and excellent hydrophilicity (38.51 ± 2.58°). In addition, GPVA5_GNP showed an optimum biodegradation rate (0.02 ± 0.005 mg/h) and the highest mechanical strength with the highest compression modulus (2.14 ± 0.06 MPa). In addition, the surface and cross-sectional view for scanning electron microscopy (SEM) displayed that all of the GPVA hydrogels have optimum average pore sizes (100−199 μm) with interconnected pores. There were no substantial changes in chemical analysis, including FTIR, XRD, and EDX, after PVA and GNP intervention. Furthermore, GPVA hydrogels influenced the cell biocompatibility, which successfully indicated >85% of cell viability. In conclusion, gelatin−PVA hydrogels crosslinked with GNP were proven to have excellent physicochemical, mechanical, and biocompatibility properties, as required for potential bioinks for chronic wound healing.
Fulltext Human epidermal keratinocytes and human dermal fibroblasts interactions seeded on gelatin hydrogel for future application in skin in vitro 3-dimensional model

Tahri S, Maarof M, Masri S, Che Man R, Masmoudi H, Fauzi MB

Front Bioeng Biotechnol, 2023;11:1200618.
PMID: 37425369 DOI: 10.3389/fbioe.2023.1200618

Introduction: Plenty of biomaterials have been studied for their application in skin tissue engineering. Currently, gelatin-hydrogel is used to support three-dimensional (3D) skin in vitro models. However, mimicking the human body conditions and properties remains a challenge and gelatin-hydrogels have low mechanical properties and undergo rapid degradation rendering them not suitable for 3D in vitro cell culture. Nevertheless, changing the concentration of hydrogels could overcome this issue. Thus, we aim to investigate the potential of gelatin hydrogel with different concentrations crosslinked with genipin to promote human epidermal keratinocytes and human dermal fibroblasts culture to develop a 3D-in vitro skin model replacing animal models. Methods: Briefly, the composite gelatin hydrogels were fabricated using different concentrations as follows 3%, 5%, 8%, and 10% crosslinked with 0.1% genipin or non-crosslinked. Both physical and chemical properties were evaluated. Results and discussion: The crosslinked scaffolds showed better properties, including porosity and hydrophilicity, and genipin was found to enhance the physical properties. Furthermore, no alteration was prominent in both formulations of CL_GEL 5% and CL_GEL8% after genipin modification. The biocompatibility assays showed that all groups promoted cell attachment, cell viability, and cell migration except for the CL_GEL10% group. The CL_GEL5% and CL_GEL8% groups were selected to develop a bi-layer 3D-in vitro skin model. The immunohistochemistry (IHC) and hematoxylin and eosin staining (H&E) were performed on day 7, 14, and 21 to evaluate the reepithelization of the skin constructs. However, despite satisfactory biocompatibility properties, neither of the selected formulations, CL_GEL 5% and CL_GEL 8%, proved adequate for creating a bi-layer 3D in-vitro skin model. While this study provides valuable insights into the potential of gelatin hydrogels, further research is needed to address the challenges associated with their use in developing 3D skin models for testing and biomedical applications.
Multifunctionalised skin substitute of hybrid gelatin-polyvinyl alcohol bioinks for chronic wound: injectable vs. 3D bioprinting

Masri S, Fadilah NIM, Hao LQ, Maarof M, Tabata Y, Hiraoka Y, et al.

Drug Deliv Transl Res, 2023 Nov 08.
PMID: 37938542 DOI: 10.1007/s13346-023-01447-z

Chronic wounds are challenging to heal and increase global mortality. The effectiveness of skin graft is limited by rejection, fibrosis, and inadequate donor site. Multifunctionalised-hydrogel skin substitutes promoted higher wound healing by maintaining the moisture microenvironment and permit gas exchange/nourishment in prolong cell viability/activity. The purpose of this study was to evaluate a skin substitute using two strategies; via injectable and 3D bioprinting technique. New hydrogel formulations that composed of gelatin (GE) and polyvinyl-alcohol (PVA) were constructed using a pre-mix crosslinking approach with genipin (GNP) to generate the biodegradable and biocompatible skin substitute with reduced secondary traumatic wound. GPVA5_GNP (6% GE: 5% PVA crosslinked with GNP) was the most stable hydrogel for wound healing application with the longest enzymatic degradation and stable hydrogel for absorption of excess wound exudates. Primary human dermal fibroblasts (HDFs) migrated extensively through 3D bioprinted hydrogels with larger average pore sizes and interconnected pores than injectable hydrogels. Moreover, 3D bioprinted GPVA hydrogels were biocompatible with HDFs and demonstrated > 90% cell viability. HDFs maintained their phenotype and positively expressed collagen type-I, vinculin, short and dense F-actin, alpha-smooth muscle actin, and Ki67. Additionally, the presence of GNP demonstrated antioxidant capacity and high-ability of angiogenesis. The utilisation of the 3D bioprinting (layer-by-layer) approach did not compromise the HDFs' growth capacity and biocompatibility with selected bioinks. In conclusion, it allows the cell encapsulation sustainability in a hydrogel matrix for a longer period, in promoting tissue regeneration and accelerating healing capacity, especially for difficult or chronic wound.
Fulltext Evaluation of PCR-based approach for serotype determination of Streptococcus pneumoniae

Shakrin NN, Balasubramaniam SD, Yusof HA, Mastuki MF, Masri SN, Taib NM, et al.

Trop Biomed, 2013 Jun;30(2):338-44.
PMID: 23959499 MyJurnal

Determination of Streptococcus pneumoniae serotypes is essential for epidemiological surveillance. Therefore accurate, reliable and cost effective serotyping method is crucial. In this study, we determined the serotypes of 41 pneumococcal isolates recovered from human anterior nares by multiplex Polymerase Chain Reaction (PCR) utilizing published primers. The data was then compared with conventional serology using latex agglutination (LA) and the Quellung reaction. Based on the PCR-approach, 8 different serogroups/serotypes were detected with one isolate classified as non-typeable (cpsA-negative). In reference to the serology-based data, the results were in agreement except for one isolate. For the latter isolate, the LA and Quellung tests failed to show a reaction but the PCR-approach and sequencing identified the isolate as serogroup 15B/C. Based on this experimental setting, we found that the PCR-approach for pneumococcal serotypes determination is reliable to serve as the alternative for determining the pneumococcal serotyping.
Seroprevalence of leptospiral antibodies among market workers and food handlers in the central state of Malaysia

Samsudin S, Sakinah SNS, Malina O, Norliza BA, Noh MA, Fairuz A, et al.

Trop Med Int Health, 2018 03;23(3):327-333.
PMID: 29356240 DOI: 10.1111/tmi.13033

OBJECTIVE: The high prevalence of leptospirosis in humans is of great public health concern, particularly in tropical and subtropical regions. This study aimed to determine the seroprevalence of leptospiral antibodies and distribution of serovars, and to assess the usefulness of enzyme-linked immunosorbent assay (ELISA) as a screening method for leptospiral antibodies in a high-risk healthy community.
METHODS: Cross-sectional study of 231 market workers and food handlers in wet markets and food premises from two localities in central Malaysia. Respondents' background information was obtained using a questionnaire. Serum samples were tested for leptospiral antibodies using ELISA and microscopic agglutination test (MAT).
RESULTS: Seroprevalence of leptospirosis among healthy workers was 46.3%. Detection of seropositivity was higher by MAT (46%) than ELISA (15%). We observed high seropositivity among local workers (49%), food handlers (49.5%), females (60.8%) and those aged 34 years and older (46.3%). Local strain LEP175 was the predominant serovar, followed by WHO strain Patoc.
CONCLUSION: Overall seroprevalence among healthy food handlers and market workers was high in this study. The workplace places susceptible individuals at risk of leptospirosis.

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