Viability and glycosaminoglycan content in chondrocytes transfected with SRY (sex-determining region y)-BOX 9 and telomerase reverse transcriptase genes

Glycosaminoglycan (GAG) is a major component of cartilage matrix. This essential polysaccharide has the capacity to attract water molecules. Such capacity is important in cartilage primarily because it absorbs shock and provides lubrication in joint. Cartilage degeneration is often a normal ageing c...

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Main Authors:	Md Nazir, Noorhidayah, Zulkifly, Ahmad Hafiz, Khalid, Kamarul Ariffin, Zainol, Ismail, Zamli, Zaitunnatakhin, Sha'ban, Munirah
Format:	Conference or Workshop Item
Language:	English
Published:	2016
Subjects:	TA164 Bioengineering
Online Access:	http://irep.iium.edu.my/55035/ http://irep.iium.edu.my/55035/1/Noorhidayah%20PABSC2016%20Abstract.pdf

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recordtype	eprints
spelling	iium-550352018-05-18T07:00:55Z http://irep.iium.edu.my/55035/ Viability and glycosaminoglycan content in chondrocytes transfected with SRY (sex-determining region y)-BOX 9 and telomerase reverse transcriptase genes Md Nazir, Noorhidayah Zulkifly, Ahmad Hafiz Khalid, Kamarul Ariffin Zainol, Ismail Zamli, Zaitunnatakhin Sha'ban, Munirah TA164 Bioengineering Glycosaminoglycan (GAG) is a major component of cartilage matrix. This essential polysaccharide has the capacity to attract water molecules. Such capacity is important in cartilage primarily because it absorbs shock and provides lubrication in joint. Cartilage degeneration is often a normal ageing change. Along the process, the cartilage cell; chondrocyte may become less viable and thus, downregulate GAG production. This study evaluates cell viability and GAG content in chondrocytes using monolayer culture model. Chondrocyte usually loses its phenotype after several replications in culture. Hypothetically, replicative senescence contributes to ageing indirectly. Hence, this study compares SOX9 and/or TERT transfected and non-transfected chondrocytes to see whether or not the cells could remain viable and produce GAG after serial passages. While SOX9 is essential for chondrogenesis, TERT is responsible for cells longevity. Upon research approval (IIUM/IACUC/Approval/2015/[5]/[24]), rabbits’ articular cartilages (n=2) were harvested. Isolated chondrocytes were seeded in 6 well-plate with an initial seeding of 5,000 cells/cm2. At passage-1, the chondrocytes were transfected with SOX9 and/or TERT genes via lipofection. The four groups namely non-transfected (control), SOX9-, TERT- and SOX9/TERT-transfected chondrocytes were evaluated at passage-1, -2 and -3. It can be appreciated that the post-transfected chondrocytes have cells viability and GAG content comparable to that of the non-transfected group. They exhibited similar downregulation pattern for viability and GAG content as postulated in theory of ageing. The viability ranged from the highest 93.43% to the lowest 78.49% throughout the culture. The total GAG contents are 25.23μg/ml, 23.87μg/ml, 21.59μg/ml and 23.86μg/ml for SOX9-, TERT-, SOX9/TERT-post-transfected chondrocytes and control, respectively. While cells viability gives an indication that transfection may have non-toxic effect on cells, this study raises some significant questions whether or not SOX9 and TERT could maintain or enhance chondrocyte properties at structural and ultrastructural levels. Detailed investigations into molecular and protein levels are also needed. 2016-12-07 Conference or Workshop Item PeerReviewed application/pdf en http://irep.iium.edu.my/55035/1/Noorhidayah%20PABSC2016%20Abstract.pdf Md Nazir, Noorhidayah and Zulkifly, Ahmad Hafiz and Khalid, Kamarul Ariffin and Zainol, Ismail and Zamli, Zaitunnatakhin and Sha'ban, Munirah (2016) Viability and glycosaminoglycan content in chondrocytes transfected with SRY (sex-determining region y)-BOX 9 and telomerase reverse transcriptase genes. In: 3rd Pan-Asian Biomedical Science Conference 2016 Kuala Lumpur, 7 – 8 Dec 2016, Premiera Hotel Kuala Lumpur, Malaysia.
repository_type	Digital Repository
institution_category	Local University
institution	International Islamic University Malaysia
building	IIUM Repository
collection	Online Access
language	English
topic	TA164 Bioengineering
spellingShingle	TA164 Bioengineering Md Nazir, Noorhidayah Zulkifly, Ahmad Hafiz Khalid, Kamarul Ariffin Zainol, Ismail Zamli, Zaitunnatakhin Sha'ban, Munirah Viability and glycosaminoglycan content in chondrocytes transfected with SRY (sex-determining region y)-BOX 9 and telomerase reverse transcriptase genes
description	Glycosaminoglycan (GAG) is a major component of cartilage matrix. This essential polysaccharide has the capacity to attract water molecules. Such capacity is important in cartilage primarily because it absorbs shock and provides lubrication in joint. Cartilage degeneration is often a normal ageing change. Along the process, the cartilage cell; chondrocyte may become less viable and thus, downregulate GAG production. This study evaluates cell viability and GAG content in chondrocytes using monolayer culture model. Chondrocyte usually loses its phenotype after several replications in culture. Hypothetically, replicative senescence contributes to ageing indirectly. Hence, this study compares SOX9 and/or TERT transfected and non-transfected chondrocytes to see whether or not the cells could remain viable and produce GAG after serial passages. While SOX9 is essential for chondrogenesis, TERT is responsible for cells longevity. Upon research approval (IIUM/IACUC/Approval/2015/[5]/[24]), rabbits’ articular cartilages (n=2) were harvested. Isolated chondrocytes were seeded in 6 well-plate with an initial seeding of 5,000 cells/cm2. At passage-1, the chondrocytes were transfected with SOX9 and/or TERT genes via lipofection. The four groups namely non-transfected (control), SOX9-, TERT- and SOX9/TERT-transfected chondrocytes were evaluated at passage-1, -2 and -3. It can be appreciated that the post-transfected chondrocytes have cells viability and GAG content comparable to that of the non-transfected group. They exhibited similar downregulation pattern for viability and GAG content as postulated in theory of ageing. The viability ranged from the highest 93.43% to the lowest 78.49% throughout the culture. The total GAG contents are 25.23μg/ml, 23.87μg/ml, 21.59μg/ml and 23.86μg/ml for SOX9-, TERT-, SOX9/TERT-post-transfected chondrocytes and control, respectively. While cells viability gives an indication that transfection may have non-toxic effect on cells, this study raises some significant questions whether or not SOX9 and TERT could maintain or enhance chondrocyte properties at structural and ultrastructural levels. Detailed investigations into molecular and protein levels are also needed.
format	Conference or Workshop Item
author	Md Nazir, Noorhidayah Zulkifly, Ahmad Hafiz Khalid, Kamarul Ariffin Zainol, Ismail Zamli, Zaitunnatakhin Sha'ban, Munirah
author_facet	Md Nazir, Noorhidayah Zulkifly, Ahmad Hafiz Khalid, Kamarul Ariffin Zainol, Ismail Zamli, Zaitunnatakhin Sha'ban, Munirah
author_sort	Md Nazir, Noorhidayah
title	Viability and glycosaminoglycan content in chondrocytes transfected with SRY (sex-determining region y)-BOX 9 and telomerase reverse transcriptase genes
title_short	Viability and glycosaminoglycan content in chondrocytes transfected with SRY (sex-determining region y)-BOX 9 and telomerase reverse transcriptase genes
title_full	Viability and glycosaminoglycan content in chondrocytes transfected with SRY (sex-determining region y)-BOX 9 and telomerase reverse transcriptase genes
title_fullStr	Viability and glycosaminoglycan content in chondrocytes transfected with SRY (sex-determining region y)-BOX 9 and telomerase reverse transcriptase genes
title_full_unstemmed	Viability and glycosaminoglycan content in chondrocytes transfected with SRY (sex-determining region y)-BOX 9 and telomerase reverse transcriptase genes
title_sort	viability and glycosaminoglycan content in chondrocytes transfected with sry (sex-determining region y)-box 9 and telomerase reverse transcriptase genes
publishDate	2016
url	http://irep.iium.edu.my/55035/ http://irep.iium.edu.my/55035/1/Noorhidayah%20PABSC2016%20Abstract.pdf
first_indexed	2023-09-18T21:17:48Z
last_indexed	2023-09-18T21:17:48Z
_version_	1777411679587401728

Viability and glycosaminoglycan content in chondrocytes transfected with SRY (sex-determining region y)-BOX 9 and telomerase reverse transcriptase genes

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