Collagen I and collagen II immunohistochemistry analyses on in vitro 3D poly(lactic-co-glycolic acid) seeded with intervertebral disc cells with and without fibrin scaffold

Objectives/Research Problem: Successful formation of 3D tissue constructs requires appropriate combination of the three established tissue engineering principles. The principles include quality cell source, biocompatible material scaffold and suitable biological signalling factors. This preliminary...

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Main Authors:	Mohamed Amin, Muhammad Azri Ifwat, Shuib, Noor Faizatul Husna, Md Nazir, Noorhidayah, Mohamad, Mohd Yusof, Ahmad Radzi, Muhammad Aa'zamuddin, Sha'ban, Munirah
Format:	Conference or Workshop Item
Language:	English
Published:	2016
Subjects:	TA164 Bioengineering
Online Access:	http://irep.iium.edu.my/54739/ http://irep.iium.edu.my/54739/1/Azri%20KRW2016%20Abstract.pdf

id	iium-54739
recordtype	eprints
spelling	iium-547392017-02-11T07:00:21Z http://irep.iium.edu.my/54739/ Collagen I and collagen II immunohistochemistry analyses on in vitro 3D poly(lactic-co-glycolic acid) seeded with intervertebral disc cells with and without fibrin scaffold Mohamed Amin, Muhammad Azri Ifwat Shuib, Noor Faizatul Husna Md Nazir, Noorhidayah Mohamad, Mohd Yusof Ahmad Radzi, Muhammad Aa'zamuddin Sha'ban, Munirah TA164 Bioengineering Objectives/Research Problem: Successful formation of 3D tissue constructs requires appropriate combination of the three established tissue engineering principles. The principles include quality cell source, biocompatible material scaffold and suitable biological signalling factors. This preliminary study aims to evaluate in vitro tissue constructs engineered from poly(lactic-co-glycolic acid) (PLGA) seeded with intervertebral disc (IVD) cells namely annulus fibrosus (AF), nucleus pulposus (NP), and a combination of AF:NP (1:1) with and without fibrin using specific cartilaginous markers i.e. collagen I and II immunohistochemistry staining. Materials and Method: Porous PLGA discs (7.0mm dia. X 3.0mm height) were fabricated using solvent casting and salt leaching method. The cells were harvested from rabbits’ IVD, cultured and seeded onto the pre-fabricated PLGA-based scaffolds. The resulting six “cells-scaffolds” construct groups were cultured for 3-weeks. The immunohistochemistry procedure and microscopic observation were performed at week 1, 2 and 3. Results and Discussion: Minimal cartilaginous tissue formation is noted in all constructs at week 1 until week 3. This can be appreciated by the presence of cartilage-isolated cells in lacunae embedded within extracellular matrix (ECM) ground substance. Cellular and ECM distribution are better in PLGA+Fibrin+AF:NP group than the other groups. Presence of brownish precipitation in most of the constructs after the immunolocalization of collagen I and II indicates positive results. It demonstrates that certain constructs have cartilaginous properties. Conclusion: The combination of PLGA and fibrin has the potential in facilitating early chondrogenesis of in vitro constructs engineered from AF, NP, and the combination of AF:NP (1:1) cells. The minimal cartilaginous tissue formation may be due to inefficient cells seeding. While it is apparent that human factor is unavoidable, there is much evidence in biomedical research and other fields that certain methodology can be refined to prevent errors before one’s experiment is compromised. In this case, the cells seeding method should be observed and improved for future IVD tissue engineering research. 2016-11-21 Conference or Workshop Item PeerReviewed application/pdf en http://irep.iium.edu.my/54739/1/Azri%20KRW2016%20Abstract.pdf Mohamed Amin, Muhammad Azri Ifwat and Shuib, Noor Faizatul Husna and Md Nazir, Noorhidayah and Mohamad, Mohd Yusof and Ahmad Radzi, Muhammad Aa'zamuddin and Sha'ban, Munirah (2016) Collagen I and collagen II immunohistochemistry analyses on in vitro 3D poly(lactic-co-glycolic acid) seeded with intervertebral disc cells with and without fibrin scaffold. In: The 2nd KAHS Research Week 2016 and the 1st Allied Health Scientific Colloquium 2016, 21-25 Nov 2016, Kuantan.
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 Mohamed Amin, Muhammad Azri Ifwat Shuib, Noor Faizatul Husna Md Nazir, Noorhidayah Mohamad, Mohd Yusof Ahmad Radzi, Muhammad Aa'zamuddin Sha'ban, Munirah Collagen I and collagen II immunohistochemistry analyses on in vitro 3D poly(lactic-co-glycolic acid) seeded with intervertebral disc cells with and without fibrin scaffold
description	Objectives/Research Problem: Successful formation of 3D tissue constructs requires appropriate combination of the three established tissue engineering principles. The principles include quality cell source, biocompatible material scaffold and suitable biological signalling factors. This preliminary study aims to evaluate in vitro tissue constructs engineered from poly(lactic-co-glycolic acid) (PLGA) seeded with intervertebral disc (IVD) cells namely annulus fibrosus (AF), nucleus pulposus (NP), and a combination of AF:NP (1:1) with and without fibrin using specific cartilaginous markers i.e. collagen I and II immunohistochemistry staining. Materials and Method: Porous PLGA discs (7.0mm dia. X 3.0mm height) were fabricated using solvent casting and salt leaching method. The cells were harvested from rabbits’ IVD, cultured and seeded onto the pre-fabricated PLGA-based scaffolds. The resulting six “cells-scaffolds” construct groups were cultured for 3-weeks. The immunohistochemistry procedure and microscopic observation were performed at week 1, 2 and 3. Results and Discussion: Minimal cartilaginous tissue formation is noted in all constructs at week 1 until week 3. This can be appreciated by the presence of cartilage-isolated cells in lacunae embedded within extracellular matrix (ECM) ground substance. Cellular and ECM distribution are better in PLGA+Fibrin+AF:NP group than the other groups. Presence of brownish precipitation in most of the constructs after the immunolocalization of collagen I and II indicates positive results. It demonstrates that certain constructs have cartilaginous properties. Conclusion: The combination of PLGA and fibrin has the potential in facilitating early chondrogenesis of in vitro constructs engineered from AF, NP, and the combination of AF:NP (1:1) cells. The minimal cartilaginous tissue formation may be due to inefficient cells seeding. While it is apparent that human factor is unavoidable, there is much evidence in biomedical research and other fields that certain methodology can be refined to prevent errors before one’s experiment is compromised. In this case, the cells seeding method should be observed and improved for future IVD tissue engineering research.
format	Conference or Workshop Item
author	Mohamed Amin, Muhammad Azri Ifwat Shuib, Noor Faizatul Husna Md Nazir, Noorhidayah Mohamad, Mohd Yusof Ahmad Radzi, Muhammad Aa'zamuddin Sha'ban, Munirah
author_facet	Mohamed Amin, Muhammad Azri Ifwat Shuib, Noor Faizatul Husna Md Nazir, Noorhidayah Mohamad, Mohd Yusof Ahmad Radzi, Muhammad Aa'zamuddin Sha'ban, Munirah
author_sort	Mohamed Amin, Muhammad Azri Ifwat
title	Collagen I and collagen II immunohistochemistry analyses on in vitro 3D poly(lactic-co-glycolic acid) seeded with intervertebral disc cells with and without fibrin scaffold
title_short	Collagen I and collagen II immunohistochemistry analyses on in vitro 3D poly(lactic-co-glycolic acid) seeded with intervertebral disc cells with and without fibrin scaffold
title_full	Collagen I and collagen II immunohistochemistry analyses on in vitro 3D poly(lactic-co-glycolic acid) seeded with intervertebral disc cells with and without fibrin scaffold
title_fullStr	Collagen I and collagen II immunohistochemistry analyses on in vitro 3D poly(lactic-co-glycolic acid) seeded with intervertebral disc cells with and without fibrin scaffold
title_full_unstemmed	Collagen I and collagen II immunohistochemistry analyses on in vitro 3D poly(lactic-co-glycolic acid) seeded with intervertebral disc cells with and without fibrin scaffold
title_sort	collagen i and collagen ii immunohistochemistry analyses on in vitro 3d poly(lactic-co-glycolic acid) seeded with intervertebral disc cells with and without fibrin scaffold
publishDate	2016
url	http://irep.iium.edu.my/54739/ http://irep.iium.edu.my/54739/1/Azri%20KRW2016%20Abstract.pdf
first_indexed	2023-09-18T21:17:25Z
last_indexed	2023-09-18T21:17:25Z
_version_	1777411655228981248

Collagen I and collagen II immunohistochemistry analyses on in vitro 3D poly(lactic-co-glycolic acid) seeded with intervertebral disc cells with and without fibrin scaffold

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