Annals of Medicine
ISSN: 0785-3890 (Print) 1365-2060 (Online) Journal homepage: http://www.tandfonline.com/loi/iann20
Effect of Joint Mobilization on Serum Hyaluronate Herkko Saari, Yrjö T. Konttinen & Dan Nordstrom To cite this article: Herkko Saari, Yrjö T. Konttinen & Dan Nordstrom (1991) Effect of Joint Mobilization on Serum Hyaluronate, Annals of Medicine, 23:1, 29-32, DOI: 10.3109/07853899109147927 To link to this article: http://dx.doi.org/10.3109/07853899109147927
Published online: 08 Jul 2009.
Submit your article to this journal
Article views: 6
View related articles
Citing articles: 2 View citing articles
Full Terms & Conditions of access and use can be found at http://www.tandfonline.com/action/journalInformation?journalCode=iann20 Download by: [RMIT University Library]
Date: 23 March 2016, At: 19:37
Brief ReDort
Effect of Joint Mobilization on Serum Hyaluronate
Downloaded by [RMIT University Library] at 19:37 23 March 2016
Herkko Saari’, Yrjo T. Konttinen2 and Dan Nordstroml
Serum hyaluronate was monitored in volunteer patients with rheumatoid arthritis who were mobilized on day 1 at 7 am, but stayed at rest in bed on day 2 until mobilized at 11:30 am. 1251-hyaluronatebinding protein assay disclosed a normal morning peak on day 1 but not on day 2. Instead, on day 2 transiently high hyaluronate levels were seen after 11:30 am. This suggests that joint mobilization affects serum hyaluronate. The morning peak may depend on the accumulation of hyaluronate in immobilized joints at night and its mobilization in the morning. High performance liquid chromatography with a size exclusion column used for fraction collection with 1251-hyaluronatebinding protein assay showed that the serum hyaluronate in the morning peak i s of relatively low molecular weight when compared to synovial fluid hyaluronate. This difference may be due to degradation of synovial fluid hyaluronate before its clearance into the blood stream. Key words: serum hyaluronate; rheumatoid arthritis; mobilization. (Annals of Medicine 23: 29-32,
1991)
Introduction Baseline concentrations of serum hyaluronate (S-HA) correlate positively with that of acute phase plasma proteins (1). S-HA levels are higher in peripheral polyarthritis than in ankylosing spondylitis with few or no peripheral affected joints (1). Furthermore, there is a significant positive correlation between S-HA levels and the number of swollen joints in rheumatoid arthritis (RA)
morning activities (3). It is also well known from clinical observation that morning stiffness does not disappear until the joints are actively used. So we reasoned that if HA accumulates in immobilized joints at night it should be possible to change the profile of S-HA by hifting the mobilization from morning to later during the day. If such a change could be proved, the result would accord with the hypothesis put forward by Anna EngstromLaurent (3).
(2).
Anna Engstrom-Laurent and her coworkers have also described a diurnal variation in S-HA (3), reporting a slight increase in healthy subjects after morning activities but a marked increase in RA patients. A significant correlation was observed between the morning increase in S-HA and morning stiffness. They concluded, therefore, that accumulation of hydrophilic HA in the immobilized joints at night might explain morning stiffness, which disappears upon mobilization of synovial HA into the blood stream during normal
From the ’Fourth Department of Medicine, Helsinki University Central Hospital, Helsinki, and 21nstitute of Molecular Immunology, Hospital for Joint Diseases, NYU Medical School, New York, USA. This study was financially supported by the Sigrid Juselius Foundation, the Paulo Foundation, the Finnish Medical Society Duodecim, Farmoksen tiede- ja tutkimussaatio and Arthritis Foundation in Finland. Address and reprint requests: Herkko Saari, M.D., Fourth Department of Medicine, Helsinki University Central Hospital, Unioninkatu 38, SF-001 70, Helsinki, Finland. Received: April 10, 1990; revision accepted December 11, 1990.
Patients and Methods Patients Four patients with RA as defined by the 1987 revised American Rheumatism Association criteria (4) and one healthy person (HS) were studied. The research followed the principles of the Declaration of Helsinki, was approved by Helsinki University’s ethics committee and conducted with informed consent of all subjects. Some clinical data on the patients are given in Table 1. All patients belonged to Steinbrocker’s functional class II (5) and in order to characterize the extent of the rheumatoid involement, a joint score index was recorded using Kaarela’s method (6) Table 1). To test for any liver involvement, which affects the half-life of S-HA, transaminases were studied and found to be normal in all subjects. Mobilization was standardized to include walking along the 25 meter long hospital corridor ten times, squatting ten times and having every joint group mobilized through its maximum range without
Saari Konttinen Nordstrom
30
Table 1. Clinical data on the R A patients studied. Pat
Sex
Age Yr
Year of diagnosis
1
F
52
1979
2 3
F F
44 41
1987 1971
4
F
64
1985
Medication' NSAID Hydroxychloroquine Prednisolon NSAID Prednisolon NSAID Aurotiomalate Prednison
ESR mmlh
CUP mall
Joint scoret (Kaarela)
14
18
11
58 110
13 47
18 13
86
77
10
* NSAID=non-steroidal anti-inflammatory drug; disease modifying antirheumatid drugs were used in standard dosages;
Downloaded by [RMIT University Library] at 19:37 23 March 2016
Corticosteroid dose was less than 10 mg per day for all patients; tnumber of swollen joints on a scale from 0 to 24 counted according to Kaarela (Ref. 6).
resistance ten times. Blood was drawn, 2 ml at a time, as shown on the xaxis in Figure 1; serum was recovered after clotting and centrifugation and preserved at -20°C before l Z 5 l HABP assay.
Radiometrid Detection of S-HA 1251-hyal~r~nate binding protein assay (Pharmacia Diagnostics AB, Uppsala, Sweden) is a sensitive radiometric assay for detecting hyaluronate from serum (7) and was used as described in detail elsewhere (8).
High Performance Liquid Chromatography HPLC chromatography was performed using an LKB2150 HPLC pump equipped with Toyo Soda TSK
Patient
The S-HA concentration increased after mobilization during the first experimental day in the morning and after 11.30 am on the second experimental day (Fig. 1). Although different individuals showed a different diurnal S-HA profile, all showed a shift in the position of the HA peak, its location depending on the time of
Patient
2
t A I
%1500
Results
1
1
500
g
6000PW (60 cm x 7.5 mm) size exclusion columns (9, 10). An LKB2150 variable monitor with an LKB2220 recording integrator was used to detect S-HA, at wave length 206 nm. The elution buffer was 50 mM sodium phospahte, pH 6.5, and the continuous flow rate 1.O ml/ min. Molecular weight calibration of the size exclusion column was performed as described (8).
I
I
Patient 3
10oot
10.
I
7.00
10.00
12.00
16.00
. ".."...
Pntirnt A,
1 7.00
10.00 12.00
16.00
Time
Figure 1. The effect of standardized mobilization on diurnal variation in the serum hyaluronate (S-HA) content in four patients with rheumatoid arthritis (RA). Shifting the mobilization from 7 am to 11.30 am also changed the localization of the S-HA peak. Circle=day 1, i.e. mobilization after 7 am; cross=day 2, i.e. mobilization after 113 0 am.
Ann Mnd 73
Downloaded by [RMIT University Library] at 19:37 23 March 2016
Serum Hyaluronate and Mobilization
31
Figure 2. Molecular weight of serum hyaluronate (S-HA) peak hyaluronate in two patients with rheumatoid arthritis RA (patients 2 and 3), two representative experirnetns. High performance liquid chromatography (HPLC) with a size exclusion column was used for fractionation, and hyaluronate (HA) in various fractions was measured using HABP assay.
where various parameters of disease activity and different medication were used as variables, showed that the total number of swollen joints is the most important predictive clinical parameter for S-HA content in RA (2). Normally, S-HA is removed from the Molecular Weight of Hyaluronate of the S-HA circulation by a specific, receptor mediated uptake to Peak liver endothelial cells (1 1). In the present study all Two serum samples obtained at the top of the S-HA patients included were screened for liver involvement peak were further analysed for molecular weight to exclude such secondary effects on the S-HA content. HA has many unusual physicochemical properties (patients 2 and 3). HPLC with a size exclusion column was used for fractionation, and the sensitive lZ51-HABP (1 1, 14) - for example, its exclusion properties give it assay was used to measure the HA concentration in an important part in immune complex precipitation in different fractions. S-HA from the HA peak in RA was of synovial membrane. One of its most remarkable relatively low molecular weight (Fig. 2). The retention properties is its hydrophilic nature, so accumulation of time of 16.4 minutes corresponded to the molecular HA in synovial tissue proper, containing many activated weight of 490 x lo3 and retention time 17.6 minutes to fibroblasts, may cause interstitial edema and stiffness. the molecular weight of 200 x lo3 daltons, respectively. Normally, synovial HA is mobilized via the lymphatic The elution profile of S-HA in HPLC was somewhat vessels into the bloodstream, ( 1 1 ) a process that polydispersed (Fig. 2 ) when compared to hyaluronate depends on the pumping effect of joint movement. Immobilization of the joint, such as occurs during a in synovial fluid (10). night’s rest, could lead to local accumulation of HA (3). This effect has been described previously, and is now confirmed. Even so, to demonstrate more convincingly Discussion that variation in the S-HA content is indeed regulated HA is a non-sulfated glycosaminoglycan of large by mobilization of the joints, it is necessary to show the molecular weight consisting of D-glucuronic acid and effect of different times of mobilization on the S-HA N-acetyl-0-glucosamine disacharide repeat units. HA is peak. The present study shows that S-HA in RA is found in various connective tissues, skin, vitreous body, regulated by mobilization, probably via its effect on the cartilage and synovial fluid (1 1). The physiological lymphatic drainage of synovial HA, so it also seems regulation of HA synthesis in synovial tissue is reasonable to suggest that patients should be mobilized imperfectly known. Agents which activate adenylate at least five hours before a sample is taken. Many researchers have reported that synovial fluid cyclase usually seem to activate the production of HA (11). Proper anchoring of the glucuronic acid- HA in RA has a relatively low molecular weight (9, 10, glucosamine polymer to the cell membrane HA- 15, 16). This might be caused by 1) synthesis of low synthetase (1 2, 13)is also necessary for HA synthesis. molecular weight HA by fibroblasts in arthritis; 2 ) Many clinical parameters, including acute phase degradation of high molecular weight HA by oxygen proteins, show a correlation with the serum baseline derived free radicals produced by activated neutrophils HA concentration in RA ( 2 ) , but multifactorial analysis, or an ischaemia reperfusion effects; or 3) preferential mobilization. The normal healthy volunteer did not have any morning stiffness and had no changes in S-HA concentration induced by mobilization (data not shown).
3
Saari Konttinen Nordstrom
32
mobilization of the high molecular weight HA from the synovial tissue via the lymphatic vessels into the blood stream ( 8 ) . We therefore studied the molecular weight of HA in the hyaluronate peak following mobilization. The S-HA peak did not contain high molecular weight hyaluronate. It is possible that this had been removed from the serum by a first pass metabolism (17). Further work is necessary to study the value of determinations of the concentration and molecular weight of S-HA as an index of processes such as oxidative injury (18).
Downloaded by [RMIT University Library] at 19:37 23 March 2016
We thank Professor Heikki Isomaki, Dr. Pentti Jarvinen and Dr. Kari Lehtinen for their generous help in finding volunteers for this study. Yrjo T. Konttinen is a Senior Investigator of the Arthritis Foundation in Finland. We are also grateful to the staff of the Research Department of Pharmacia in Uppsala for providing us with the HA standards for calibration of the HPLC columns.
References 1. Engstrom-Laurent A, Hallgren R. Circulating hyaluronate in rheumatoid arthritis: relationship to inflammatory activity and the effect of cortiosteroid therapy. Ann Rheum Dis 1985; 44: 83-8. 2. Konttinen YT, Saari H, Honkanen V, Szocsik K, Mussaio-Rauhamaa H, Friman C. Serum baseline hyaluronate and disease activity in rheumatoid arthritis. Clin Chim Acta (in press). 3. Engstrbm-Laurent A, Hlllgren R. Circulating hyaluronic acid levels vary with physical activity in healthy subjects and in rheumatoid arthritis patients. Arthritis Rheum 1987; 30: 1333-8. 4. Arnett FC, Edworthy SM, Bloch DA, et at. The American Rheumatism Association 1987 revised criteria for classification of rheumatoid arthritis. Arthritis Rheum 1988; 31 : 315-27. 5. Steinbrocker 0 , Traeger CH, Batterman EC.
Ann lAad 31
Therapeutic criteria in rheumatoid arthritis. JAMA 1949; 140: 659-62. 6. Kaarela K. Prognostic factors and diagnostic criteria in early rheumatoid arthritis. Scand J Rheumatol 1985; SUPPI57: 1-54, 7. Brandt R, Hedlbf E, Asman I, Bucht A, Tengblad A. A convinient radiometric assay for hyaluronate. Acta Otolaryngol 1987; Suppl 442: 31-5. 8. Saarl H. Oxygen derived free radicals and synovial fluid hyaluronate. Ann Rheum Dis (in press). 9. Saari HI Konttinen YT, Santavirta S. Synovial fluid hyaluronate: a study using high performance liquid chromatography with size ecxlusion column. Med Sci Res 1989; 17: 99-1 01. 10. Saari H, Konttinen YT. Determination of the concentration and polymerization of synovial fluid hyaluronate using high performance liquid chromatography. Ann Rheum Dis 1989; 48: 565-70. 11. Laurent TC, Fraser JRE. The properties and turnover of hvaluronate. Functions of the proteoqlvcans. Ciba Found -_ Simp 1986; 124: 9-24. 12. Prehm P. Hyaluronate is synthesized at plasma membranes. Biochem J 1984; 220: 597-600. 13. Philipson LH, Schwartz NB. Subcellular localization of hyaluronate synthetase in oligodendroglioma cells. J Biol Chem 1984; 259: 5017-23. 14. Konttinen YT, Saari H, Santavirta S, Anttl-Polka I, Sorsa T, Nykanen P, Kempplnen P. Synovial fibroblasts. Scand J Rheumatol 1988; Suppl 74: 95-103. 15. Bjelle A, Andersson T, Granath K. Molecular weight distribution of hyaluronic acid of human synovial fluid in rheumatic diseases. Scand J Rheumatol 1982; 12: 1338. 16. Dahl LB, Dahl IMS, Engstrbm-Laurent A, Granath K. Concentration and molecular weight of sodium hvaluronate in svnovial fluid from patients with rheumatoid arthritis-and other arthropathies. Ann Rheum Dis 1985; 44: 817-22. 17. Tengblad A, Laurent UBG, Lilja K, et al. Concentration and relative molecular mass of hyaluronate in lymph and blood. Biochem J 1986; 236: 521-5. 18. Saari H, Sorsa T, Konttinen YT. Reactive oxygen species and hyaluronate in serum and synovial fluid in arthritis. Int J Tissue React (in press).
ISSN: 0785-3890 (Print) 1365-2060 (Online) Journal homepage: http://www.tandfonline.com/loi/iann20
Effect of Joint Mobilization on Serum Hyaluronate Herkko Saari, Yrjö T. Konttinen & Dan Nordstrom To cite this article: Herkko Saari, Yrjö T. Konttinen & Dan Nordstrom (1991) Effect of Joint Mobilization on Serum Hyaluronate, Annals of Medicine, 23:1, 29-32, DOI: 10.3109/07853899109147927 To link to this article: http://dx.doi.org/10.3109/07853899109147927
Published online: 08 Jul 2009.
Submit your article to this journal
Article views: 6
View related articles
Citing articles: 2 View citing articles
Full Terms & Conditions of access and use can be found at http://www.tandfonline.com/action/journalInformation?journalCode=iann20 Download by: [RMIT University Library]
Date: 23 March 2016, At: 19:37
Brief ReDort
Effect of Joint Mobilization on Serum Hyaluronate
Downloaded by [RMIT University Library] at 19:37 23 March 2016
Herkko Saari’, Yrjo T. Konttinen2 and Dan Nordstroml
Serum hyaluronate was monitored in volunteer patients with rheumatoid arthritis who were mobilized on day 1 at 7 am, but stayed at rest in bed on day 2 until mobilized at 11:30 am. 1251-hyaluronatebinding protein assay disclosed a normal morning peak on day 1 but not on day 2. Instead, on day 2 transiently high hyaluronate levels were seen after 11:30 am. This suggests that joint mobilization affects serum hyaluronate. The morning peak may depend on the accumulation of hyaluronate in immobilized joints at night and its mobilization in the morning. High performance liquid chromatography with a size exclusion column used for fraction collection with 1251-hyaluronatebinding protein assay showed that the serum hyaluronate in the morning peak i s of relatively low molecular weight when compared to synovial fluid hyaluronate. This difference may be due to degradation of synovial fluid hyaluronate before its clearance into the blood stream. Key words: serum hyaluronate; rheumatoid arthritis; mobilization. (Annals of Medicine 23: 29-32,
1991)
Introduction Baseline concentrations of serum hyaluronate (S-HA) correlate positively with that of acute phase plasma proteins (1). S-HA levels are higher in peripheral polyarthritis than in ankylosing spondylitis with few or no peripheral affected joints (1). Furthermore, there is a significant positive correlation between S-HA levels and the number of swollen joints in rheumatoid arthritis (RA)
morning activities (3). It is also well known from clinical observation that morning stiffness does not disappear until the joints are actively used. So we reasoned that if HA accumulates in immobilized joints at night it should be possible to change the profile of S-HA by hifting the mobilization from morning to later during the day. If such a change could be proved, the result would accord with the hypothesis put forward by Anna EngstromLaurent (3).
(2).
Anna Engstrom-Laurent and her coworkers have also described a diurnal variation in S-HA (3), reporting a slight increase in healthy subjects after morning activities but a marked increase in RA patients. A significant correlation was observed between the morning increase in S-HA and morning stiffness. They concluded, therefore, that accumulation of hydrophilic HA in the immobilized joints at night might explain morning stiffness, which disappears upon mobilization of synovial HA into the blood stream during normal
From the ’Fourth Department of Medicine, Helsinki University Central Hospital, Helsinki, and 21nstitute of Molecular Immunology, Hospital for Joint Diseases, NYU Medical School, New York, USA. This study was financially supported by the Sigrid Juselius Foundation, the Paulo Foundation, the Finnish Medical Society Duodecim, Farmoksen tiede- ja tutkimussaatio and Arthritis Foundation in Finland. Address and reprint requests: Herkko Saari, M.D., Fourth Department of Medicine, Helsinki University Central Hospital, Unioninkatu 38, SF-001 70, Helsinki, Finland. Received: April 10, 1990; revision accepted December 11, 1990.
Patients and Methods Patients Four patients with RA as defined by the 1987 revised American Rheumatism Association criteria (4) and one healthy person (HS) were studied. The research followed the principles of the Declaration of Helsinki, was approved by Helsinki University’s ethics committee and conducted with informed consent of all subjects. Some clinical data on the patients are given in Table 1. All patients belonged to Steinbrocker’s functional class II (5) and in order to characterize the extent of the rheumatoid involement, a joint score index was recorded using Kaarela’s method (6) Table 1). To test for any liver involvement, which affects the half-life of S-HA, transaminases were studied and found to be normal in all subjects. Mobilization was standardized to include walking along the 25 meter long hospital corridor ten times, squatting ten times and having every joint group mobilized through its maximum range without
Saari Konttinen Nordstrom
30
Table 1. Clinical data on the R A patients studied. Pat
Sex
Age Yr
Year of diagnosis
1
F
52
1979
2 3
F F
44 41
1987 1971
4
F
64
1985
Medication' NSAID Hydroxychloroquine Prednisolon NSAID Prednisolon NSAID Aurotiomalate Prednison
ESR mmlh
CUP mall
Joint scoret (Kaarela)
14
18
11
58 110
13 47
18 13
86
77
10
* NSAID=non-steroidal anti-inflammatory drug; disease modifying antirheumatid drugs were used in standard dosages;
Downloaded by [RMIT University Library] at 19:37 23 March 2016
Corticosteroid dose was less than 10 mg per day for all patients; tnumber of swollen joints on a scale from 0 to 24 counted according to Kaarela (Ref. 6).
resistance ten times. Blood was drawn, 2 ml at a time, as shown on the xaxis in Figure 1; serum was recovered after clotting and centrifugation and preserved at -20°C before l Z 5 l HABP assay.
Radiometrid Detection of S-HA 1251-hyal~r~nate binding protein assay (Pharmacia Diagnostics AB, Uppsala, Sweden) is a sensitive radiometric assay for detecting hyaluronate from serum (7) and was used as described in detail elsewhere (8).
High Performance Liquid Chromatography HPLC chromatography was performed using an LKB2150 HPLC pump equipped with Toyo Soda TSK
Patient
The S-HA concentration increased after mobilization during the first experimental day in the morning and after 11.30 am on the second experimental day (Fig. 1). Although different individuals showed a different diurnal S-HA profile, all showed a shift in the position of the HA peak, its location depending on the time of
Patient
2
t A I
%1500
Results
1
1
500
g
6000PW (60 cm x 7.5 mm) size exclusion columns (9, 10). An LKB2150 variable monitor with an LKB2220 recording integrator was used to detect S-HA, at wave length 206 nm. The elution buffer was 50 mM sodium phospahte, pH 6.5, and the continuous flow rate 1.O ml/ min. Molecular weight calibration of the size exclusion column was performed as described (8).
I
I
Patient 3
10oot
10.
I
7.00
10.00
12.00
16.00
. ".."...
Pntirnt A,
1 7.00
10.00 12.00
16.00
Time
Figure 1. The effect of standardized mobilization on diurnal variation in the serum hyaluronate (S-HA) content in four patients with rheumatoid arthritis (RA). Shifting the mobilization from 7 am to 11.30 am also changed the localization of the S-HA peak. Circle=day 1, i.e. mobilization after 7 am; cross=day 2, i.e. mobilization after 113 0 am.
Ann Mnd 73
Downloaded by [RMIT University Library] at 19:37 23 March 2016
Serum Hyaluronate and Mobilization
31
Figure 2. Molecular weight of serum hyaluronate (S-HA) peak hyaluronate in two patients with rheumatoid arthritis RA (patients 2 and 3), two representative experirnetns. High performance liquid chromatography (HPLC) with a size exclusion column was used for fractionation, and hyaluronate (HA) in various fractions was measured using HABP assay.
where various parameters of disease activity and different medication were used as variables, showed that the total number of swollen joints is the most important predictive clinical parameter for S-HA content in RA (2). Normally, S-HA is removed from the Molecular Weight of Hyaluronate of the S-HA circulation by a specific, receptor mediated uptake to Peak liver endothelial cells (1 1). In the present study all Two serum samples obtained at the top of the S-HA patients included were screened for liver involvement peak were further analysed for molecular weight to exclude such secondary effects on the S-HA content. HA has many unusual physicochemical properties (patients 2 and 3). HPLC with a size exclusion column was used for fractionation, and the sensitive lZ51-HABP (1 1, 14) - for example, its exclusion properties give it assay was used to measure the HA concentration in an important part in immune complex precipitation in different fractions. S-HA from the HA peak in RA was of synovial membrane. One of its most remarkable relatively low molecular weight (Fig. 2). The retention properties is its hydrophilic nature, so accumulation of time of 16.4 minutes corresponded to the molecular HA in synovial tissue proper, containing many activated weight of 490 x lo3 and retention time 17.6 minutes to fibroblasts, may cause interstitial edema and stiffness. the molecular weight of 200 x lo3 daltons, respectively. Normally, synovial HA is mobilized via the lymphatic The elution profile of S-HA in HPLC was somewhat vessels into the bloodstream, ( 1 1 ) a process that polydispersed (Fig. 2 ) when compared to hyaluronate depends on the pumping effect of joint movement. Immobilization of the joint, such as occurs during a in synovial fluid (10). night’s rest, could lead to local accumulation of HA (3). This effect has been described previously, and is now confirmed. Even so, to demonstrate more convincingly Discussion that variation in the S-HA content is indeed regulated HA is a non-sulfated glycosaminoglycan of large by mobilization of the joints, it is necessary to show the molecular weight consisting of D-glucuronic acid and effect of different times of mobilization on the S-HA N-acetyl-0-glucosamine disacharide repeat units. HA is peak. The present study shows that S-HA in RA is found in various connective tissues, skin, vitreous body, regulated by mobilization, probably via its effect on the cartilage and synovial fluid (1 1). The physiological lymphatic drainage of synovial HA, so it also seems regulation of HA synthesis in synovial tissue is reasonable to suggest that patients should be mobilized imperfectly known. Agents which activate adenylate at least five hours before a sample is taken. Many researchers have reported that synovial fluid cyclase usually seem to activate the production of HA (11). Proper anchoring of the glucuronic acid- HA in RA has a relatively low molecular weight (9, 10, glucosamine polymer to the cell membrane HA- 15, 16). This might be caused by 1) synthesis of low synthetase (1 2, 13)is also necessary for HA synthesis. molecular weight HA by fibroblasts in arthritis; 2 ) Many clinical parameters, including acute phase degradation of high molecular weight HA by oxygen proteins, show a correlation with the serum baseline derived free radicals produced by activated neutrophils HA concentration in RA ( 2 ) , but multifactorial analysis, or an ischaemia reperfusion effects; or 3) preferential mobilization. The normal healthy volunteer did not have any morning stiffness and had no changes in S-HA concentration induced by mobilization (data not shown).
3
Saari Konttinen Nordstrom
32
mobilization of the high molecular weight HA from the synovial tissue via the lymphatic vessels into the blood stream ( 8 ) . We therefore studied the molecular weight of HA in the hyaluronate peak following mobilization. The S-HA peak did not contain high molecular weight hyaluronate. It is possible that this had been removed from the serum by a first pass metabolism (17). Further work is necessary to study the value of determinations of the concentration and molecular weight of S-HA as an index of processes such as oxidative injury (18).
Downloaded by [RMIT University Library] at 19:37 23 March 2016
We thank Professor Heikki Isomaki, Dr. Pentti Jarvinen and Dr. Kari Lehtinen for their generous help in finding volunteers for this study. Yrjo T. Konttinen is a Senior Investigator of the Arthritis Foundation in Finland. We are also grateful to the staff of the Research Department of Pharmacia in Uppsala for providing us with the HA standards for calibration of the HPLC columns.
References 1. Engstrom-Laurent A, Hallgren R. Circulating hyaluronate in rheumatoid arthritis: relationship to inflammatory activity and the effect of cortiosteroid therapy. Ann Rheum Dis 1985; 44: 83-8. 2. Konttinen YT, Saari H, Honkanen V, Szocsik K, Mussaio-Rauhamaa H, Friman C. Serum baseline hyaluronate and disease activity in rheumatoid arthritis. Clin Chim Acta (in press). 3. Engstrbm-Laurent A, Hlllgren R. Circulating hyaluronic acid levels vary with physical activity in healthy subjects and in rheumatoid arthritis patients. Arthritis Rheum 1987; 30: 1333-8. 4. Arnett FC, Edworthy SM, Bloch DA, et at. The American Rheumatism Association 1987 revised criteria for classification of rheumatoid arthritis. Arthritis Rheum 1988; 31 : 315-27. 5. Steinbrocker 0 , Traeger CH, Batterman EC.
Ann lAad 31
Therapeutic criteria in rheumatoid arthritis. JAMA 1949; 140: 659-62. 6. Kaarela K. Prognostic factors and diagnostic criteria in early rheumatoid arthritis. Scand J Rheumatol 1985; SUPPI57: 1-54, 7. Brandt R, Hedlbf E, Asman I, Bucht A, Tengblad A. A convinient radiometric assay for hyaluronate. Acta Otolaryngol 1987; Suppl 442: 31-5. 8. Saarl H. Oxygen derived free radicals and synovial fluid hyaluronate. Ann Rheum Dis (in press). 9. Saari HI Konttinen YT, Santavirta S. Synovial fluid hyaluronate: a study using high performance liquid chromatography with size ecxlusion column. Med Sci Res 1989; 17: 99-1 01. 10. Saari H, Konttinen YT. Determination of the concentration and polymerization of synovial fluid hyaluronate using high performance liquid chromatography. Ann Rheum Dis 1989; 48: 565-70. 11. Laurent TC, Fraser JRE. The properties and turnover of hvaluronate. Functions of the proteoqlvcans. Ciba Found -_ Simp 1986; 124: 9-24. 12. Prehm P. Hyaluronate is synthesized at plasma membranes. Biochem J 1984; 220: 597-600. 13. Philipson LH, Schwartz NB. Subcellular localization of hyaluronate synthetase in oligodendroglioma cells. J Biol Chem 1984; 259: 5017-23. 14. Konttinen YT, Saari H, Santavirta S, Anttl-Polka I, Sorsa T, Nykanen P, Kempplnen P. Synovial fibroblasts. Scand J Rheumatol 1988; Suppl 74: 95-103. 15. Bjelle A, Andersson T, Granath K. Molecular weight distribution of hyaluronic acid of human synovial fluid in rheumatic diseases. Scand J Rheumatol 1982; 12: 1338. 16. Dahl LB, Dahl IMS, Engstrbm-Laurent A, Granath K. Concentration and molecular weight of sodium hvaluronate in svnovial fluid from patients with rheumatoid arthritis-and other arthropathies. Ann Rheum Dis 1985; 44: 817-22. 17. Tengblad A, Laurent UBG, Lilja K, et al. Concentration and relative molecular mass of hyaluronate in lymph and blood. Biochem J 1986; 236: 521-5. 18. Saari H, Sorsa T, Konttinen YT. Reactive oxygen species and hyaluronate in serum and synovial fluid in arthritis. Int J Tissue React (in press).
