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Annals of the rheumatic diseases1992; 51(2); 214-219; doi: 10.1136/ard.51.2.214

Methylprednisolone acetate induced release of cartilage proteoglycans: determination by high performance liquid chromatography.

Abstract: A high performance liquid chromatography (HPLC) procedure suitable for the simultaneous determination of the molecular size and concentration of macromolecular hyaluronate and proteoglycans in synovial fluid has been developed. Irrigation of the equine tarsocrural joint with 20 ml physiological saline (PSS) caused a mild inflammation with an increase of proteoglycans in the synovial fluid over the baseline arthrocentesis control sample. Proteoglycan and hyaluronate in the synovial fluid did not interact to form hyaluronate-proteoglycan aggregates, but separated as distinct chromatographic peaks. This suggests that the cartilage derived proteoglycans in synovial fluid in the inflamed joint have been proteolytically cleaved from the non-covalent aggregates containing link protein and hyaluronate. Hyaluronidase digestion completely abolished the hyaluronate peak without affecting the proteoglycans. This seems to indicate that proteoglycan in synovial fluid is unable to interact with hyaluronate in synovial fluid to form cartilage type aggregates. Proteolytic degradation and the time dependent release into the synovial fluid of such digested proteoglycan also resulted from the intra-articular injection of methylprednisolone acetate into normal tarsocrural joints and joints irrigated with PSS. These proteoglycans were insensitive to hyaluronidase but may consist of a protein moiety with attached glycosaminoglycans, as suggested by their sensitivity to proteinase and keratanase/chondroitinase digestion. These observations with cartilage treated with methylprednisolone acetate and mildly stimulated articular cartilage are inconsistent with earlier work on osteoarthritic and rheumatoid articular cartilage and have interesting implications for the pathogenesis and for the therapeutic action of intraarticular corticosteroids. A rapid HPLC procedure applicable to unprocessed small volume samples of synovial fluid gives information simultaneously on hyaluronate and proteoglycan in synovial fluid which is not attainable with immunoradiometric or isotope tracer techniques. It therefore appears to be useful for the analysis of cartilage turnover and destruction in health and disease.
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Publication Date: 1992-02-01 PubMed ID: 1550406PubMed Central: PMC1005661DOI: 10.1136/ard.51.2.214Google Scholar: Lookup
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  • Comparative Study
  • Journal Article

Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

The study presents a developed procedure for high performance liquid chromatography (HPLC) suitable for assessing the size and concentration of hyaluronate and proteoglycans in synovial fluid, which is the key focus. The process is tested on joints of horses, and it’s found that both substances do not aggregate, but appear as separate peaks in the study. Additionally, the intra-articular injection of methylprednisolone acetate into joints yields results that are inconsistent with previous research on osteoarthritic and rheumatoid articular cartilage, offering new insight into therapeutic action of intraarticular corticosteroids.

Development of HPLC Procedure

  • The researchers have developed an HPLC procedure to simultaneously measure the size and concentration of macromolecular hyaluronate and proteoglycans in synovial fluid, a lubricating fluid found in joints.
  • This method was developed as an improvement over earlier techniques such as immunoradiometric or isotope tracing, which do not provide simultaneous information on the two compounds.

Application and Findings

  • The new procedure was applied to the study of equine tarsocrural joints (horse ankle joints).
  • It was observed that irrigating the joint with 20 ml physiological saline (PSS) caused mild inflammation and an increase in proteoglycans in the synovial fluid, compared to the control sample.
  • The study found that proteoglycan and hyaluronate in the inflamed joint do not interact to form aggregates but separate as distinct chromatographic peaks.

Proteolytic Degradation and Time-Dependent Release

  • Proteoglycans that had undergone proteolytic degradation and were released into the synovial fluid over time resulted from the injection of methylprednisolone acetate into the joints.
  • This suggested that these proteoglycans had been cleaved from aggregates containing link protein and hyaluronate and were not able to interact with hyaluronate to form cartilage type aggregates.
  • The researchers confirmed this by showing that digestion with hyaluronidase completely abolished the hyaluronate peak without affecting the proteoglycans.

Implications for Disease and Therapy

  • The results contradict previous research involving osteoarthritic and rheumatoid articular cartilage, indicating potential new insights into these conditions and the action of corticosteroids used in therapy.
  • The HPLC procedure could be useful for analyzing cartilage turnover and destruction in health and disease, as well as studying the therapeutic action of intraarticular corticosteroids.

Cite This Article

APA
Saari H, Tulamo RM, Konttinen YT, Sorsa T. (1992). Methylprednisolone acetate induced release of cartilage proteoglycans: determination by high performance liquid chromatography. Ann Rheum Dis, 51(2), 214-219. https://doi.org/10.1136/ard.51.2.214

Publication

Annals of the rheumatic diseases
ISSN: 0003-4967
NlmUniqueID: 0372355
Country: England
Language: English
Volume: 51
Issue: 2
Pages: 214-219

Researcher Affiliations

Saari, H
  • Fourth Department of Medicine, Helsinki University Central Hospital, Finland.
Tulamo, R M
    Konttinen, Y T
      Sorsa, T

        MeSH Terms

        • Animals
        • Anti-Inflammatory Agents / pharmacology
        • Cartilage, Articular / metabolism
        • Chromatography, High Pressure Liquid / methods
        • Disease Models, Animal
        • Horses
        • Hyaluronic Acid / biosynthesis
        • Methylprednisolone / analogs & derivatives
        • Methylprednisolone / pharmacology
        • Methylprednisolone Acetate
        • Proteoglycans / biosynthesis
        • Synovial Fluid / metabolism
        • Tarsal Joints

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        Citations

        This article has been cited 1 times.
        1. Ruckmani K, Shaikh SZ, Khalil P, Muneera MS, Thusleem OA. Determination of sodium hyaluronate in pharmaceutical formulations by HPLC-UV. J Pharm Anal 2013 Oct;3(5):324-329.
          doi: 10.1016/j.jpha.2013.02.001pubmed: 29403834google scholar: lookup
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