J Orthop Surg Res. 2008 Apr 25;3:17. doi: 10.1186/1749-799X-3-17.
Fibrin and poly(lactic-co-glycolic acid) hybrid scaffold promotes early chondrogenesis of articular chondrocytes: an in vitro study.
Journal of orthopaedic surgery and research
Munirah Sha'ban, Soon Hee Kim, Ruszymah Bh Idrus, Gilson Khang
Affiliations
Affiliations
- BK-21 Polymer BIN Fusion Research Team, Department of Polymer Science and Technology, Chonbuk National University, 664-14 Dukjin, Jeonju 561-756, Seoul, Korea.
PMID: 18435862
PMCID: PMC2405772 DOI: 10.1186/1749-799X-3-17
Abstract
BACKGROUND: Synthetic- and naturally derived- biodegradable polymers have been widely used to construct scaffolds for cartilage tissue engineering. Poly(lactic-co-glycolic acid) (PLGA) are bioresorbable and biocompatible, rendering them as a promising tool for clinical application. To minimize cells lost during the seeding procedure, we used the natural polymer fibrin to immobilize cells and to provide homogenous cells distribution in PLGA scaffolds. We evaluated in vitro chondrogenesis of rabbit articular chondrocytes in PLGA scaffolds using fibrin as cell transplantation matrix.
METHODS: PLGA scaffolds were soaked in chondrocytes-fibrin suspension (1 x 10(6) cells/scaffold) and polymerized by dropping thrombin-calcium chloride (CaCl2) solution. PLGA-seeded chondrocytes was used as control. All constructs were cultured for a maximum of 21 days. Cell proliferation activity was measured at 1, 3, 7, 14 and 21 days in vitro using 3-(4,5-dimethylthiazole-2-yl)-2-, 5-diphenyltetrazolium-bromide (MTT) assay. Morphological observation, histology, immunohistochemistry (IHC), gene expression and sulphated-glycosaminoglycan (sGAG) analyses were performed at each time point of 1, 2 and 3 weeks to elucidate in vitro cartilage development and deposition of cartilage-specific extracellular matrix (ECM).
RESULTS: Cell proliferation activity was gradually increased from day-1 until day-14 and declined by day-21. A significant cartilaginous tissue formation was detected as early as 2-week in fibrin/PLGA hybrid construct as confirmed by the presence of cartilage-isolated cells and lacunae embedded within basophilic ECM. Cartilage formation was remarkably evidenced after 3 weeks. Presence of cartilage-specific proteoglycan and glycosaminoglycan (GAG) in fibrin/PLGA hybrid constructs were confirmed by positive Safranin O and Alcian Blue staining. Collagen type II exhibited intense immunopositivity at the pericellular matrix. Chondrogenic properties were further demonstrated by the expression of genes encoded for cartilage-specific markers, collagen type II and aggrecan core protein. Interestingly, suppression of cartilage dedifferentiation marker; collagen type I was observed after 2 and 3 weeks of in vitro culture. The sulphated-glycosaminoglycan (sGAG) production in fibrin/PLGA was significantly higher than in PLGA.
CONCLUSION: Fibrin/PLGA promotes early in vitro chondrogenesis of rabbit articular chondrocytes. This study suggests that fibrin/PLGA may serve as a potential cell delivery vehicle and a structural basis for in vitro tissue-engineered articular cartilage.
References
- Med J Malaysia. 2004 May;59 Suppl B:13-4 - PubMed
- Med J Malaysia. 2004 May;59 Suppl B:30-1 - PubMed
- Tissue Eng. 2005 Jul-Aug;11(7-8):1065-76 - PubMed
- Biomaterials. 2000 Jun;21(11):1145-53 - PubMed
- Biomaterials. 1999 Jul;20(13):1167-75 - PubMed
- J Bone Joint Surg Br. 2007 Aug;89(8):1099-109 - PubMed
- Biomaterials. 2005 Jun;26(16):3075-82 - PubMed
- Biomaterials. 2007 Jan;28(1):55-65 - PubMed
- Med J Malaysia. 2004 May;59 Suppl B:11-2 - PubMed
- J Biomater Sci Polym Ed. 2004;15(8):1003-17 - PubMed
- Biomaterials. 2000 Dec;21(24):2561-74 - PubMed
- Adv Exp Med Biol. 2006;585:167-81 - PubMed
- Biomaterials. 2002 Jun;23(11):2303-10 - PubMed
- Med J Malaysia. 2004 May;59 Suppl B:9-10 - PubMed
- N Engl J Med. 1994 Oct 6;331(14):889-95 - PubMed
- Arthroscopy. 2005 Oct;21(10):1155-63 - PubMed
- J Orthop Surg (Hong Kong). 2008 Aug;16(2):220-9 - PubMed
- J Biomater Sci Polym Ed. 2007;18(5):609-22 - PubMed
- J Zhejiang Univ Sci B. 2006 Oct;7(10):817-24 - PubMed
- Tissue Eng. 2004 Mar-Apr;10(3-4):323-30 - PubMed
- Clin Chem. 1989 Mar;35(3):374-9 - PubMed
- Biomaterials. 2006 Mar;27(7):1071-80 - PubMed
- Tissue Eng. 2005 Sep-Oct;11(9-10):1562-73 - PubMed
- Tissue Cell. 2007 Oct;39(5):293-301 - PubMed
- J Orthop Res. 2001 May;19(3):482-92 - PubMed
- Adv Exp Med Biol. 2003;534:235-45 - PubMed
- Spine J. 2002 Nov-Dec;2(6):415-20 - PubMed
- Med J Malaysia. 2004 May;59 Suppl B:15-6 - PubMed
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