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1.
J Orthop Res ; 32(4): 557-65, 2014 Apr.
Article in English | MEDLINE | ID: mdl-24302463

ABSTRACT

The objective of the study is to understand the therapeutic effects of lipophilic (simvastatin) and hydrophilic statins (pravastatin) combined with/without hyaluronic acid for osteoarthritis by an in vitro LPS-induced inflammatory model of articular chondrocytes. HA in combination with different doses of simvastatin or pravastatin were used. Beside cytotoxicity, the influence of statins on NO production, pro-inflammatory cytokine, inflammatory mediators, and NF-κB p50 protein were analyzed. Finally, TUNEL assay was performed to detect DNA strand breakage. Two statins were less able to lower NF-κB activity when they were administrated along without HA. The gene expression demonstrates that simvastatin and pravastatin had the ability to decrease pro-inflammatory and inflammatory mediator levels. High dose simvastatin with or without HA down regulated inflammatory cytokines, but resulted in higher cytotoxicity. TUNEL assay confirms the regulatory effect of statins with or without HA over the apoptosis of chondrocytes, especially in hydrophilic statins. The significant down-regulation of inflammatory mediators suggests that intra-articular injection of HA in combination with statins might feasibly slow the progress of osteoarthritis. Administration of simvastatin or pravastatin with hyaluronic acid may produce beneficial effects for OA treatment, but with better results when hydrophilic statin was used.


Subject(s)
Chondrocytes/drug effects , Hydroxymethylglutaryl-CoA Reductase Inhibitors/therapeutic use , Osteoarthritis/drug therapy , Pravastatin/therapeutic use , Simvastatin/therapeutic use , Animals , Chondrocytes/metabolism , Collagen Type II/metabolism , Hyaluronic Acid/therapeutic use , Hydroxymethylglutaryl-CoA Reductase Inhibitors/pharmacology , In Situ Nick-End Labeling , Injections, Intra-Articular , Lipopolysaccharides/toxicity , NF-kappa B p50 Subunit/metabolism , Nitric Oxide Synthase Type II/metabolism , Osteoarthritis/pathology , Pravastatin/pharmacology , Real-Time Polymerase Chain Reaction , Simvastatin/pharmacology , Swine , Viscosupplements/therapeutic use
2.
J Biomed Mater Res A ; 92(3): 843-51, 2010 Mar 01.
Article in English | MEDLINE | ID: mdl-19280637

ABSTRACT

To understand whether a low-frequency pulsed electromagnetic field (EMF) could help cartilage tissue repair in the scope of tissue engineering, we tested how EMF affected collagen gel properties and the behaviors of chondrocyte cells embedded in collagen constructs. Collagen gel and primary chondrocytes embedded in collagen were exposed to EMF for 24 h. Gel and cells that were not exposed to EMF served as controls. Collagen gel exposed to EMF was more hydrophobic and less susceptible to enzymatic degradation (both p < 0.05) than control. Three weeks after EMF exposure, chondrocytes showed higher proliferation and lower glycosaminoglycan (GAG) production (both p < 0.05) than control. By the end of the third week, aggrecan, type I, II, and X collagen mRNA expressions in the EMF group were 1.8 times higher (p < 0.05), except for type II collagen) than control. The increase in gene expression did not show up in aggrecan histological staining and type II and type X collagen immunohistochemical staining. Cells from both groups kept a normal polygonal shape through out the test period. Our results suggested that one-time EMF exposure could promote collagen-embedded chondrocytes proliferation and their gene expressions. It also promoted short-term (week 1) GAG production and lacuna formation. No apparent GAG and type II collagen production was seen in histological staining three weeks after the EMF exposure.


Subject(s)
Cartilage/growth & development , Electromagnetic Fields , Tissue Engineering , Animals , Cartilage/cytology , Cartilage/metabolism , Cell Proliferation , Chondrocytes/cytology , Chondrocytes/metabolism , Collagen/genetics , Collagen/metabolism , Glycosaminoglycans/metabolism , Immunohistochemistry , Reverse Transcriptase Polymerase Chain Reaction , Swine
3.
Artif Organs ; 32(7): 561-6, 2008 Jul.
Article in English | MEDLINE | ID: mdl-18638311

ABSTRACT

Human mesenchymal stem cells (hMSCs) can differentiate into cells of connective tissue lineages, including cartilage. To overcome the limiting autogenous chondrocyte populations available in cartilage repair, various methods have been developed to maximize chondrogenesis of hMSCs in vitro, most of which use cells derived from primary culture. In this study, we compared chondrogenesis of immortalized hMSCs embedded in collagen gel to those grown in pellet culture. The hMSCs in collagen scaffolds expressed more glycosaminoglycan than those in pellet culture. Real-time reverse transcriptase-polymerase chain reaction (RT-PCR) analysis demonstrated that the expression of genes encoding sox-9, aggrecan, and types I and II collagen increased in pellet culture over time. However, in the collagen cultures, only type II collagen and aggrecan expression increased over time, whereas sox-9 expression remained unchanged and type I collagen expression decreased. These results indicate that the immortalized hMSC line is a promising tool for further in vitro chondrogenic studies.


Subject(s)
Chondrogenesis/physiology , Collagen Type II/genetics , Mesenchymal Stem Cells/physiology , Tissue Engineering/methods , Aggrecans/genetics , Cell Culture Techniques , Cell Line , Collagen Type I/genetics , DNA/analysis , Gene Expression , Glycosaminoglycans/genetics , High Mobility Group Proteins/genetics , Humans , Immunohistochemistry , Reverse Transcriptase Polymerase Chain Reaction , SOX9 Transcription Factor , Tissue Scaffolds , Transcription Factors/genetics
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