The Effect of Stichopus chloronotus Aqueous Extract on Human Osteoarthritis Articular Chondrocytes in Three-Dimensional Collagen Type I Hydrogel in vitro

Research output: Contribution to journalArticle

Abstract

Autologous chondrocyte-seeded scaffolds have proved to be one of the most promising alternative therapies for articular cartilage defects. However, the chondrocytes have specific nutritional requirements and risk of dedifferentiation during in vitro expansion. Stichopus chloronotus aqueous extract (SCAE) was investigated as a medium supplement for three-dimensional (3D) collagen type I hydrogel scaffold seeded with chondrocytes to determine whether SCAE is capable of maintaining phenotype and sustaining extracellular matrix synthesis and deposition. Human osteoarthritis articular chondrocytes were isolated from the knee joint cartilage of patients underwent total knee replacement surgery. Human osteoarthritis articular chondrocytes were encapsulated in collagen type I hydrogel and cultured in basic medium with 0, 0.1 and 0.2% of SCAE. The chondrocytes in 3D culture were evaluated by means cell morphology and proliferation, quantitative phenotypic expression of collagen type I, II, aggrecan core protein and sox-9. H&E, toluidine blue staining and sulfated glycosaminoglycan (sGAG) production were analyzed after 7 days in culture. Chondrocytes cultured in 3D with various SCAE concentration appeared with polygonal morphology maintaining their chondrocytes characteristic. SCAE supplementation promoted chondrocytes proliferation and the ability of the cells to express gene encoding collagen type I decreased, whereas their ability to express collagen type II, aggrecan core protein and sox9 increased as compared to control. The cartilaginous matrices were positively stained toluidine blue concomitant with higher sGAG accumulation in SCAE-supplemented culture medium. This study shown that SCAE may be beneficial for in vitro development of 3D chondrocytes culture for use in cartilage tissue engineering therapies.

Original languageEnglish
Pages (from-to)1671-1683
Number of pages13
JournalSains Malaysiana
Volume48
Issue number8
DOIs
Publication statusPublished - 1 Jan 2019

Fingerprint

Stichopus
Hydrogel
Chondrocytes
Collagen Type I
Osteoarthritis
Joints
A73025
Aggrecans
Tolonium Chloride
Collagen Type II
Cartilage
Cell Proliferation
In Vitro Techniques
Nutritional Requirements
Knee Replacement Arthroplasties
Articular Cartilage
Tissue Engineering
Complementary Therapies
Cell- and Tissue-Based Therapy
Knee Joint

Keywords

  • Chondrocytes
  • Collagen Type I
  • Osteoarthritis
  • Stichopus chloronotus
  • Three dimensional culture

ASJC Scopus subject areas

  • General

Cite this

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title = "The Effect of Stichopus chloronotus Aqueous Extract on Human Osteoarthritis Articular Chondrocytes in Three-Dimensional Collagen Type I Hydrogel in vitro",
abstract = "Autologous chondrocyte-seeded scaffolds have proved to be one of the most promising alternative therapies for articular cartilage defects. However, the chondrocytes have specific nutritional requirements and risk of dedifferentiation during in vitro expansion. Stichopus chloronotus aqueous extract (SCAE) was investigated as a medium supplement for three-dimensional (3D) collagen type I hydrogel scaffold seeded with chondrocytes to determine whether SCAE is capable of maintaining phenotype and sustaining extracellular matrix synthesis and deposition. Human osteoarthritis articular chondrocytes were isolated from the knee joint cartilage of patients underwent total knee replacement surgery. Human osteoarthritis articular chondrocytes were encapsulated in collagen type I hydrogel and cultured in basic medium with 0, 0.1 and 0.2{\%} of SCAE. The chondrocytes in 3D culture were evaluated by means cell morphology and proliferation, quantitative phenotypic expression of collagen type I, II, aggrecan core protein and sox-9. H&E, toluidine blue staining and sulfated glycosaminoglycan (sGAG) production were analyzed after 7 days in culture. Chondrocytes cultured in 3D with various SCAE concentration appeared with polygonal morphology maintaining their chondrocytes characteristic. SCAE supplementation promoted chondrocytes proliferation and the ability of the cells to express gene encoding collagen type I decreased, whereas their ability to express collagen type II, aggrecan core protein and sox9 increased as compared to control. The cartilaginous matrices were positively stained toluidine blue concomitant with higher sGAG accumulation in SCAE-supplemented culture medium. This study shown that SCAE may be beneficial for in vitro development of 3D chondrocytes culture for use in cartilage tissue engineering therapies.",
keywords = "Chondrocytes, Collagen Type I, Osteoarthritis, Stichopus chloronotus, Three dimensional culture",
author = "Yunus, {Mohd Heikal Mohd} and Shuid, {Ahmad Nazrun} and Busra, {Mohd Fauzi} and Hui, {Chua Kien} and Ghafar, {Norzana Abdul} and Rani, {Rizal Abd}",
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T1 - The Effect of Stichopus chloronotus Aqueous Extract on Human Osteoarthritis Articular Chondrocytes in Three-Dimensional Collagen Type I Hydrogel in vitro

AU - Yunus, Mohd Heikal Mohd

AU - Shuid, Ahmad Nazrun

AU - Busra, Mohd Fauzi

AU - Hui, Chua Kien

AU - Ghafar, Norzana Abdul

AU - Rani, Rizal Abd

PY - 2019/1/1

Y1 - 2019/1/1

N2 - Autologous chondrocyte-seeded scaffolds have proved to be one of the most promising alternative therapies for articular cartilage defects. However, the chondrocytes have specific nutritional requirements and risk of dedifferentiation during in vitro expansion. Stichopus chloronotus aqueous extract (SCAE) was investigated as a medium supplement for three-dimensional (3D) collagen type I hydrogel scaffold seeded with chondrocytes to determine whether SCAE is capable of maintaining phenotype and sustaining extracellular matrix synthesis and deposition. Human osteoarthritis articular chondrocytes were isolated from the knee joint cartilage of patients underwent total knee replacement surgery. Human osteoarthritis articular chondrocytes were encapsulated in collagen type I hydrogel and cultured in basic medium with 0, 0.1 and 0.2% of SCAE. The chondrocytes in 3D culture were evaluated by means cell morphology and proliferation, quantitative phenotypic expression of collagen type I, II, aggrecan core protein and sox-9. H&E, toluidine blue staining and sulfated glycosaminoglycan (sGAG) production were analyzed after 7 days in culture. Chondrocytes cultured in 3D with various SCAE concentration appeared with polygonal morphology maintaining their chondrocytes characteristic. SCAE supplementation promoted chondrocytes proliferation and the ability of the cells to express gene encoding collagen type I decreased, whereas their ability to express collagen type II, aggrecan core protein and sox9 increased as compared to control. The cartilaginous matrices were positively stained toluidine blue concomitant with higher sGAG accumulation in SCAE-supplemented culture medium. This study shown that SCAE may be beneficial for in vitro development of 3D chondrocytes culture for use in cartilage tissue engineering therapies.

AB - Autologous chondrocyte-seeded scaffolds have proved to be one of the most promising alternative therapies for articular cartilage defects. However, the chondrocytes have specific nutritional requirements and risk of dedifferentiation during in vitro expansion. Stichopus chloronotus aqueous extract (SCAE) was investigated as a medium supplement for three-dimensional (3D) collagen type I hydrogel scaffold seeded with chondrocytes to determine whether SCAE is capable of maintaining phenotype and sustaining extracellular matrix synthesis and deposition. Human osteoarthritis articular chondrocytes were isolated from the knee joint cartilage of patients underwent total knee replacement surgery. Human osteoarthritis articular chondrocytes were encapsulated in collagen type I hydrogel and cultured in basic medium with 0, 0.1 and 0.2% of SCAE. The chondrocytes in 3D culture were evaluated by means cell morphology and proliferation, quantitative phenotypic expression of collagen type I, II, aggrecan core protein and sox-9. H&E, toluidine blue staining and sulfated glycosaminoglycan (sGAG) production were analyzed after 7 days in culture. Chondrocytes cultured in 3D with various SCAE concentration appeared with polygonal morphology maintaining their chondrocytes characteristic. SCAE supplementation promoted chondrocytes proliferation and the ability of the cells to express gene encoding collagen type I decreased, whereas their ability to express collagen type II, aggrecan core protein and sox9 increased as compared to control. The cartilaginous matrices were positively stained toluidine blue concomitant with higher sGAG accumulation in SCAE-supplemented culture medium. This study shown that SCAE may be beneficial for in vitro development of 3D chondrocytes culture for use in cartilage tissue engineering therapies.

KW - Chondrocytes

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KW - Osteoarthritis

KW - Stichopus chloronotus

KW - Three dimensional culture

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