Biotransfomation products from Clarius batrachus oil

Biotransformation can be defined as an application that utilized natural and recombinant organisms’ enzymes secreted by yeast, fungi and bacteria or whole cells as catalyst in synthesis of organic compound. Therefore, Pseudomonas aeruginosa bacterium has been chosen to be the biocatalyst for biotransformation of ω fatty acids extracted from Malaysian catfish, Clarias batrachus emphasizing on bioconversion of arachidonic acid. In addition, arachidonic acid was one of prostaglandin precursor which exerts a variety of pharmacological effects on human and animals. In this study, the fatty acids were extracted from the catfish using modified Folch method where the fish flesh was freeze dried prior to homogenization with the chloroform and methanol system. Then, the crude lipid extract was added to the bacterial culture and incubated for 4 days. After incubation, the biotransformation product was extracted and analyzed by using gas chromatography and mass spectrometer (GC-MS) to identify the fatty acids and other compounds. It was found that several fatty acids, especially ω-fatty acids were converted to cholesterol. This indicates that ω-fatty acids can be used as starting materials for other bioactive metabolites for pharmaceutical purposes

Establishment of growth kinetics profile and measurement of Sulphated Glycosaminoglycans (sGAG) production in monolayer cultured chondrocytes following Qur’anic recitation exposure

Objectives/Research Problem: Cartilage is an avascular tissue made of one cell type only which is the chondrocyte. Due to the low mitotic property, the chondrocytes have very limited self-repair capacity. As a result, once the cartilage is injured and left untreated, degeneration changes will precede ageing and can be progressive. Limitation in the currently available treatments is noted. Seeking alternative ways to facilitate the cartilage repair and regeneration have become crucial. This study aims to identify the potential effects of the Qur’anic recitation, particularly Surah Al-Fatihah on the sGAG production in the monolayer cultured chondrocytes derived from rabbit articular cartilage. Materials and Method: A cellular model based on a serially cultured and expanded chondrocytes is established in vitro and divided into four groups. The first group is exposed to the recitation of Surah Al-Fatihah. The second and third groups are exposed to the recitation of Arabic poem and Western poem respectively. The fourth group is not exposed to any sound and serves as control. Any significant changes are recorded and presented as photomicrographs. Growth kinetics assessment is performed to study the cell proliferation activities within each group. After reaching 80-90% confluency, the cells are harvested and pelleted through centrifugation step. The cell pellet is subjected to sGAG assay at different passages (P0, P1, P2, and P3). Results and Discussion: The cells exposed to Surah Al-Fatihah is expected to increase the proliferation and sGAG production of the chondrocytes better than the control group as well as the cells exposed to Arabic and Western poem recitation. Conclusion: Initial findings suggest that the Qur’anic recitation promotes cells proliferation and sGAG production. The Qur’anic recitation may serve as one of the potential signalling factors in tissue engineering studies and facilitate for cartilage repair and regeneration

A review on radiation effects towards cell culture

Effects of radiation on tissues and organs are well documented. However recently, radiation was used as an evaluation tool to assess newly formed engineered tissue reconstruction and regeneration. This raises questions on the positive application of radiation, amidst the usual discussions pertaining to the risks of radiation. Furthermore, the effects of radiation have yet to be established in primary cell culture. The objective of this review is to identify and evaluate previous works that examined the possible effects of radiation on cells. Emphasis is placed upon the effects of radiation on primary cell cultures in tissue engineering (TE). The SCOPUS databases were searched for articles published from 2001 to 2016. “Radiation” and “cell culture” were used as search terms either in the title, abstract or keywords of the articles. Thematic analysis was performed on the 5031 retrieved articles. Filtering for “radiation effects” in the title, 4139 articles were identified. Within these results the key search term was further narrowed down to “tissue engineering area”. 1164 articles were identified which were then subjected to key search term within “primary cell culture”. A total of 334 articles were abridged that were subjected to further thematic classifications. Within these classifications, 291 articles were summarized to fulfil the objectives of the review. The results show a certain trend towards the incompatible effect of radiation on primary cell culture. Work is in progress in filtering the keywords further to narrow the scope to radiation effects on chondrocytes in the title, abstract or keywords of the articles. This review will present the possible effects of radiation on primary cell culture. It is hoped that this review will be able to outline the current knowledge on the effects of radiation on primary cell cultures and to improve the understanding of those effects on cells in general

Microbial transformation of Clarias gariepinus oil by psuedomonas aeruginosa

Biotransformation of fatty acid from Malaysian catfish, Clarias gariepinus oil was carried out using Pseudomonas aeruginosa. The lipid from freeze-dried catfish flesh was extracted using a modified Folch method with chloroform-methanol mixture as an extracting solvent. The crude lipid substrate was added to P. aeruginosa culture and incubated for 4 days. After conversion, the products were analyzed by using GC-MS instrument. The result showed that 7,10-dihydroxy-8(E)-octadecenoic acid (DHOD) were abundantly found in the product. The hydroxyl derivative increased while fatty acid contents decreased after biotransformation process. It can be concluded that the bacterial cells had transformed the fatty acids to yield hydroxyl metabolite which can be utilized as starting materials for the bioconversion to pharmaceutical materials

Biotransformation of ω-fatty acid from Clarias gariepinus using Psuedomonas aeruginosa

Study on biotransformation of �-fatty acid from fish oil was carried out. Pseudomonas aeruginosa has been chosen to be the biocatalyst for biotransformation of w fatty acids extracted from Malaysian catfish, Clarias gariepinus. The lipid from freeze-dried catfish flesh was extracted using a modified Folch method and the mixture of chloroform and methanol was used as a solvent. The crude lipid substrate was added to the bacterial culture and incubated for 4 days. After conversion, the products were analyzed by using gas chromatography and mass spectrometer (GC-MS). The analytical result showed that several fatty acids and cholesterol were found in the product. However, the fatty acids and cholesterol contents before and after biotransformation were different. The cholesterol content increased while �-fatty acid contents decreased after biotransformation process. It can be concluded that the bacterial cells had oxidized the �-fatty acids to yield precursors which can be utilized as starting materials for the bioconversion to cholesterol

Early response in antibacterial activity of orthopaedic metal implant coated with silver composite as future implant related infection treatment – Preliminary result

INTRODUCTION: The prevalence of implant associated with infection remains high. The management of this incidence poses a major challenge in orthopaedic field since these scenarios are chronic and biofilm based. Colonization of bacterial and biofilm adhesion on implant may lead to infection at underlying bone and surrounding tissues. Surgically device removal and addition with prolonged used of antimicrobial theraphy are leads to patient suffering, substantial morbidity and even mortality. Hence, to overcome this, alternative treatment strategy using orthopaedic metallic implant coated with 3% silver was introduced. The 3% silver composite exhibits good bactericidal properties. It inhibits biofilm formation particularly, at the adhesion stage of the relevant bacteria. Hence, the aim of this study is to investigate the potential effect of silver coated orthopaedic metal implant against biofilm-producing Staphylococcus aureus through in vitro experimental setting. METHODS: The antibacterial property of orthopaedic metal implants was investigated using Staphylococcus aureus ATCC 25923. The sterile nutrient agar media was prepared. The bacterial colony then was smeared on the surface of the agar media. The coated six holes mini plate and non coated six holes plate which served as control were placed on the surface of agar media. The samples then were incubated for culture under aerobic condition with temperature 37ºC for 24 hours and continued incubated up to 48 hours for assessment. In this study, the antibacterial effect was evaluated based on the diameter of inhibition zone using in situ inhibitory test. RESULTS: The in situ test showed that the colony was noted after 24 hours of incubation. After 48 hours of incubation, the inhibition zone was noted and the number colony did not increase compared to control sample. DISCUSSIONS: Considering the prolonged used of antibiotic as theraphy which leads to resistance, silver coating on orthopaedic implant become very attractive alternative as antibacterial agent. According to Chen et al., (2006) silver is the well known agent compared to other dopants as antibacterial since its character not only can inhibit the bacterial adhesion with broad antibacterial spectrum, but also consists long lasting antibacterial effect. This was proved from the outcome of this study which revealed positive response of the silver as antibacterial agent. The experiment works is still ongoing. CONCLUSION: This result suggested that test material of orthopaedic metal implant coated with silver composite has the intended potential as antibacterial and provide protection against medical device-related infection

The in vivo study of open fracture in New Zealand white rabbit using orthopaedic metal implant coated with silver composite – A pilot study

INTRODUCTION: Bacterial infection during implant placement is a serious complication. The reason for this infection is mainly due to contamination during surgical implantation with the formation of a resistant biofilm at the implant surface. The infection risks may be overcome by application of orthopaedic metallic implant coated with 3% silver composite as alternative treatment of implant-infection related. The purpose of this study was to investigate the potential effect of test material against infection through in vivo experimental setting. METHODS: The effect of silver coated as antibacterial agent on orthopaedic metal implant was evaluated in New Zealand White Rabbit. The study was performed by surgically implanted at right tibia (silver coated plate) and non-coated implant at left tibia (non-coated plate) after six hours of exposure at surgical site. The subject was observed daily for infection effect up to three weeks. Microbial analyze was performed after euthanasia for assessment day. No antibiotics were administered. Animal was sacrificed at 3 weeks and evaluated for infection. Culture for the rabbit was taken at sacrifice after 3 week assessment duration. The organism on the swab on the implanted plate was inoculated on the sterile nutrient agar. After that, it was incubated for 24 hours before reading the colony. RESULT: The culture result on the nutrient agar showed that there is no colony bacteria growth on the silver coated implant meanwhile the non-coated implant has a few colony bacteria at the nutrient agar plate in 3 weeks observation period after 24 hours incubation. DISCUSSIONS: The outcome of this study revealed positive response of the silver as antibacterial agent. The study is still ongoing. CONCLUSION: This result suggested that orthopaedic metal implant coated with silver composite has the intended antibacterial properties in 3 weeks period and may provide potential as bacteriostatic or bactericidal properties against infection

Early response in antibacterial activity of orthopaedic metal implant coated with silver composite as future implant related infection treatment – Preliminary result

INTRODUCTION: The prevalence of implant associated with infection remains high. The management of this incidence poses a major challenge in orthopaedic field since these scenarios are chronic and biofilm based. Colonization of bacterial and biofilm adhesion on implant may lead to infection at underlying bone and surrounding tissues. Surgically device removal and addition with prolonged used of antimicrobial theraphy are leads to patient suffering, substantial morbidity and even mortality. Hence, to overcome this, alternative treatment strategy using orthopaedic metallic implant coated with 3% silver was introduced. The 3% silver composite exhibits good bactericidal properties. It inhibits biofilm formation particularly, at the adhesion stage of the relevant bacteria. Hence, the aim of this study is to investigate the potential effect of silver coated orthopaedic metal implant against biofilm-producing Staphylococcus aureus through in vitro experimental setting. METHODS: The antibacterial property of orthopaedic metal implants was investigated using Staphylococcus aureus ATCC 25923. The sterile nutrient agar media was prepared. The bacterial colony then was smeared on the surface of the agar media. The coated six holes mini plate and non coated six holes plate which served as control were placed on the surface of agar media. The samples then were incubated for culture under aerobic condition with temperature 37ºC for 24 hours and continued incubated up to 48 hours for assessment. In this study, the antibacterial effect was evaluated based on the diameter of inhibition zone using in situ inhibitory test. RESULTS: The in situ test showed that the colony was noted after 24 hours of incubation. After 48 hours of incubation, the inhibition zone was noted and the number colony did not increase compared to control sample. DISCUSSIONS: Considering the prolonged used of antibiotic as theraphy which leads to resistance, silver coating on orthopaedic implant become very attractive alternative as antibacterial agent. According to Chen et al., (2006) silver is the well known agent compared to other dopants as antibacterial since its character not only can inhibit the bacterial adhesion with broad antibacterial spectrum, but also consists long lasting antibacterial effect. This was proved from the outcome of this study which revealed positive response of the silver as antibacterial agent. The experiment works is still ongoing. CONCLUSION: This result suggested that test material of orthopaedic metal implant coated with silver composite has the intended potential as antibacterial and provide protection against medical device-related infection

The in vitro release study of orthopaedic metal implant coated with silver composite – Preliminary result

INTRODUCTION: The management of implant-infection poses a major challenge in orthopaedic field. Colonization of bacterial and biofilm adhesion on implant may lead to infection at underlying bone and surrounding tissues. Prolonged used of antimicrobial treatment with high dosage can lead to systemic toxicity. Hence, to overcome this, alternative treatment strategy using orthopaedic metallic implant coated with silver composite as antibacterial agent have been introduced. It exhibits good bactericidal properties for biomedical application. Hence, the aim of this study is to investigate the potential of silver coated release of orthopaedic metal implant through direct current magnetron sputtering technique against biofilm-producing Staphylococcus aureus through in vitro experimental setting. METHODS: This method was employed to determine the release of silver as antibacterial agent. The coated plate was soaked with 200ml of Phosphate Buffer Saline (PBS) in shaking waterbath at 370C for silver extract preparation. A total of 5.0ml of solution in each assessment at 1 hour, 3 hours, 24 hours, 48 hours, 168 hours, 336 hours and 504 hours of time point were withdrawn for analyze. The bacterial inoculation was smeared on sterile nutrient agar media. A total of 20µl of 5.0ml of silver release solution in each assessment was loaded on the disc. A total of 20µl of Phosphate Buffer Saline (PBS) was loaded on the disk which served as control. The samples then were incubated with 370C for 24 hours for bacterial culture under aerobic condition. The sample then was continued incubated up to 168 hours with 370C under aerobic condition for assessment. The antibacterial effect was evaluated based on the diameter of inhibition zone using disk diffusion test. RESULTS: The colony was noted on the surface on nutrient agar after 24 hours of incubation. However, the inhibition zone was noted with diameter more than 20mm at 168 hours time point of silver extraction after 168 hours of incubation. The colonies number increase was noted at other silver extraction of time points. DISCUSSIONS: The use of orthopaedic implants has increase dramatically in all subspecialties in orthopaedic field. Recently, the coating of implants has engendered much interest in order to prevent adverse tissue reactions such as infection, inflammation, the foreign body response. Ewald A et al stated that silver coatings can inhibit the bacterial growth effectively. The early outcome of this study revealed positive indicator of the silver as antibacterial agent. The experiment work is still ongoing. CONCLUSION: This result suggested that orthopaedic metal implant coated with silver composite may provide antibacterial protection against medical device-related infection