In vitro and in vivo gene expression studies of cartilage-like tissue engineered construct using a combination of transiently transfected human osteoarthritic chondrocytes and tissue engineering technique

In vitro and in vivo gene expression studies of cartilage-like tissue engineered construct using a combination of transiently transfected human osteoarthritic chondrocytes and tissue engineering technique

Purpose: To evaluate the formation of cartilage-like tissue engineered constructs (TECs) using human osteoarthritic chondrocytes overexpressed with SOX9 gene seeded on poly(lactic-co-glycolic acid) (PLGA) with and without fibrin scaffolds through cartilaginous genes expression analysis. Methods:...

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Main Authors: Sha'ban, Munirah, Mohamad Sukri, Norhamiza, Zulkifly, Ahmad Hafiz
Format: Conference or Workshop Item
Language:English
English
Published: Frontiers Media 2016
Subjects:
TA164 Bioengineering
Online Access:http://irep.iium.edu.my/54740/
http://irep.iium.edu.my/54740/
http://irep.iium.edu.my/54740/1/Munirah%206thMTERMS2016%20Abstract.pdf
http://irep.iium.edu.my/54740/7/54740_In%20vitro%20and%20in%20vivo%20gene%20expression.pdf
id iium-54740
recordtype eprints
spelling iium-547402017-03-21T02:28:08Z http://irep.iium.edu.my/54740/ In vitro and in vivo gene expression studies of cartilage-like tissue engineered construct using a combination of transiently transfected human osteoarthritic chondrocytes and tissue engineering technique Sha'ban, Munirah Mohamad Sukri, Norhamiza Zulkifly, Ahmad Hafiz TA164 Bioengineering Purpose: To evaluate the formation of cartilage-like tissue engineered constructs (TECs) using human osteoarthritic chondrocytes overexpressed with SOX9 gene seeded on poly(lactic-co-glycolic acid) (PLGA) with and without fibrin scaffolds through cartilaginous genes expression analysis. Methods: Upon approval by the IREC18; NMRR-12-1383-14531 and IIUM/IACUCApproval/2015/[5][22], six cartilage samples were obtained from consented patients after joint replacement surgery. The cells were isolated, cultured and transfected with pcDNA3-SOX9 using Lipofectamine 2000™. Prefabricated disc-shaped porous PLGA with and without fibrin were used as scaffolds. The ‘cells-scaffolds’ TECs were formed and cultured for 3-week and implanted subcutaneously at the dorsum of athymic mice for 4-week. Collagens I, II, IX, X, XI, SOX9 and aggrecan expression were evaluated using a qualitative two-step reverse-transcriptase PCR. GAPDH and β-actin genes were used as internal controls. Results: Presence of cartilaginous markers can be detected in all TECs with various expression intensity. Collagen II, the cartilage-specific marker was down-regulated in vitro but re-expressed in vivo. Collagen I, X, SOX9 and aggrecan were steadily expressed in all TECs. Although collagen IX and XI are closely associated with collagen II, their expressions were almost untraceable except for few cases. Presence of GAPDH and β-actin genes indicated the reliability of the analysis. Conclusion: Cartilage-like TECs have been successfully formed based on genes expression analysis. Frontiers Media 2016-11-21 Conference or Workshop Item PeerReviewed application/pdf en http://irep.iium.edu.my/54740/1/Munirah%206thMTERMS2016%20Abstract.pdf application/pdf en http://irep.iium.edu.my/54740/7/54740_In%20vitro%20and%20in%20vivo%20gene%20expression.pdf Sha'ban, Munirah and Mohamad Sukri, Norhamiza and Zulkifly, Ahmad Hafiz (2016) In vitro and in vivo gene expression studies of cartilage-like tissue engineered construct using a combination of transiently transfected human osteoarthritic chondrocytes and tissue engineering technique. In: 6th MTERMS 2016 in conjunction with 2nd Malaysian Stem Cells Meeting, 21-25 Nov 2016, Pulau Pinang. 10.3389/conf.FBIOE.2016.02.00010
repository_type Digital Repository
institution_category Local University
institution International Islamic University Malaysia
building IIUM Repository
collection Online Access
language English
English
topic TA164 Bioengineering
spellingShingle TA164 Bioengineering
Sha'ban, Munirah
Mohamad Sukri, Norhamiza
Zulkifly, Ahmad Hafiz
In vitro and in vivo gene expression studies of cartilage-like tissue engineered construct using a combination of transiently transfected human osteoarthritic chondrocytes and tissue engineering technique
description Purpose: To evaluate the formation of cartilage-like tissue engineered constructs (TECs) using human osteoarthritic chondrocytes overexpressed with SOX9 gene seeded on poly(lactic-co-glycolic acid) (PLGA) with and without fibrin scaffolds through cartilaginous genes expression analysis. Methods: Upon approval by the IREC18; NMRR-12-1383-14531 and IIUM/IACUCApproval/2015/[5][22], six cartilage samples were obtained from consented patients after joint replacement surgery. The cells were isolated, cultured and transfected with pcDNA3-SOX9 using Lipofectamine 2000™. Prefabricated disc-shaped porous PLGA with and without fibrin were used as scaffolds. The ‘cells-scaffolds’ TECs were formed and cultured for 3-week and implanted subcutaneously at the dorsum of athymic mice for 4-week. Collagens I, II, IX, X, XI, SOX9 and aggrecan expression were evaluated using a qualitative two-step reverse-transcriptase PCR. GAPDH and β-actin genes were used as internal controls. Results: Presence of cartilaginous markers can be detected in all TECs with various expression intensity. Collagen II, the cartilage-specific marker was down-regulated in vitro but re-expressed in vivo. Collagen I, X, SOX9 and aggrecan were steadily expressed in all TECs. Although collagen IX and XI are closely associated with collagen II, their expressions were almost untraceable except for few cases. Presence of GAPDH and β-actin genes indicated the reliability of the analysis. Conclusion: Cartilage-like TECs have been successfully formed based on genes expression analysis.
format Conference or Workshop Item
author Sha'ban, Munirah
Mohamad Sukri, Norhamiza
Zulkifly, Ahmad Hafiz
author_facet Sha'ban, Munirah
Mohamad Sukri, Norhamiza
Zulkifly, Ahmad Hafiz
author_sort Sha'ban, Munirah
title In vitro and in vivo gene expression studies of cartilage-like tissue engineered construct using a combination of transiently transfected human osteoarthritic chondrocytes and tissue engineering technique
title_short In vitro and in vivo gene expression studies of cartilage-like tissue engineered construct using a combination of transiently transfected human osteoarthritic chondrocytes and tissue engineering technique
title_full In vitro and in vivo gene expression studies of cartilage-like tissue engineered construct using a combination of transiently transfected human osteoarthritic chondrocytes and tissue engineering technique
title_fullStr In vitro and in vivo gene expression studies of cartilage-like tissue engineered construct using a combination of transiently transfected human osteoarthritic chondrocytes and tissue engineering technique
title_full_unstemmed In vitro and in vivo gene expression studies of cartilage-like tissue engineered construct using a combination of transiently transfected human osteoarthritic chondrocytes and tissue engineering technique
title_sort in vitro and in vivo gene expression studies of cartilage-like tissue engineered construct using a combination of transiently transfected human osteoarthritic chondrocytes and tissue engineering technique
publisher Frontiers Media
publishDate 2016
url http://irep.iium.edu.my/54740/
http://irep.iium.edu.my/54740/
http://irep.iium.edu.my/54740/1/Munirah%206thMTERMS2016%20Abstract.pdf
http://irep.iium.edu.my/54740/7/54740_In%20vitro%20and%20in%20vivo%20gene%20expression.pdf
first_indexed 2023-09-18T21:17:25Z
last_indexed 2023-09-18T21:17:25Z
_version_ 1777411655377879040

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