Use of citrate to minimize neutrophil matrix metalloproteinase-9 in human plasma

Makowski, Gregory S.; Ramsby, Melinda L.

doi:10.1016/j.ab.2003.07.030

Cited by 55 publications

(58 citation statements)

References 18 publications

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“…The use of blood samples collected with sodium citrate was suggested recently as material of choice for measurement of MMP-9 and MMP-2 to optimize the diagnostic validity in peripheral blood. 11,12,17 So far, no such systematic studies for TIMP were carried out.…”

Section: Dear Sirmentioning

confidence: 99%

Blood sampling as critical preanalytical determinant to use circulating MMP and TIMP as surrogate markers for pathological processes

Jung

Meisser

Bischof

2005

Intl Journal of Cancer

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Dear Sir,It has been suggested that circulating matrix metalloproteinases (MMP) and tissue inhibitors of matrix metalloproteinases (TIMP) may be useful diagnostic and prognostic indicators in malignant and nonmalignant diseases.1 Numerous reports seem to confirm that assumption although it is unknown whether increased MMP concentration are based on the initial step of tissue damage or the following repairing process.2 There are also contrasting results when similar patients were studied, i.e., increased plasma MMP-9 concentrations but unchanged serum concentrations were found in patients with colon carcinoma. 3,4 We believe that in addition to the analytical and clinical reasons to explain these differences between various studies, more attention should be given to the preanalytical conditions of sampling. In this respect, we read with great interest a recent article published in this journal.5 Gianelli et al. 5 found no difference in serum concentrations of MMP-9 and TIMP-1 between healthy women and patients with breast cancer, but decreased MMP-9 and increased TIMP-1 after surgery in comparison with initial concentrations. The authors concluded that these changes might serve as prognostic indices in the follow-up of breast cancer patients. It should be remembered that, however, that using serum material as samples for the determination of those analytes, the authors had rather inadequately considered the impact of blood collection as an important preanalytical determinant of MMP and TIMP results. There is rising evidence that blood sampling and handling markedly determine the concentrations of circulating MMP and TIMP.6-12 Thus, to interpret that data correctly and to avoid wrong expectations in future studies using these markers, we want to draw the attention of interested clinicians to these facts that were particularly discussed in analytical journals. [6][7][8]12 Although the pitfalls in the measurement of circulating growth factors and cytokines for diagnostic purposes have become generally accepted, 13 similar evidences for MMP and TIMP have been widely ignored.Results of own experiments (summarized in Fig. 1) demonstrate the differences of MMP-9 and TIMP-1 in serum and plasma as the analytes come into question. Briefly, for MMP-9 measurements, blood samples from 12 healthy volunteers were collected in Vacutainer tubes (Becton Dickinson, Franklin Lakes, NJ) containing a clot activator for serum preparations or lithium heparinate, dipotassium EDTA or sodium citrate for plasma preparations. Corresponding prepared plastic tubes (Monovettes; Sarstedt, N€ urtingen, Germany) were used for blood sampling for TIMP-1 measurements. For the MMP-9 experiments, the tubes were either centrifuged (1,600g, 15 min) immediately (considered as t 0 ) or after 0.5 or 2 hr storage at room temperature and after 24 hr storage at 4°C, respectively. Within 5 days of sampling while the samples were stored at 4°C MMP-9 measurements were carried out using our own enzyme immunoassay described previously.14 Briefly, samples (and sta...

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Section: Dear Sirmentioning

confidence: 99%

Blood sampling as critical preanalytical determinant to use circulating MMP and TIMP as surrogate markers for pathological processes

Jung

Meisser

Bischof

2005

Intl Journal of Cancer

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“…10 No question is of greater interest to the clinicians dealing with breast diseases than the differentiation of BC subtypes through the determination of biomolecular markers useful for BC diagnosis and prognosis. Although the identification of the best source between plasma and serum for surrogate biomarkers is a matter of strenuous efforts and debate, [11][12][13][14][15][16][17] the scientific interest in the measurement of MMP in plasma/serum continues to mount, enhancing their promising development as reliable biomarkers.…”

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confidence: 99%

Gelatinase concentrations and zymographic profiles in human breast cancer: Matrix metalloproteinases circulating in plasma are better markers for the subclassification and early prediction of cancer: The coagulation/fibrinolysis pathways alter the release, activation and recovery of different gelatinases in serum

Mannello

Tonti

2007

Intl Journal of Cancer

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“…4,5 When evaluating the immunoreactive protein by ELISA assay, it should not equate with the assay of measuring the activities of MMP-9. [6][7][8] The source of circulating MMP-9 whether it is measured from serum or plasma is not easy to define. In our study, however, we were also able to show that there is a statistically significant correlation between the serum level of MMP-9 immunoreactive protein and MMP-9 immunohistochemical staining in primary tumors of HNSCC.…”

Section: Dear Sirmentioning

confidence: 99%

Reply to: Serum samples should not be used to access circulating matrix metalloproteinase‐9 levels as a prognostic marker of disease

Ruokolainen

Turpeenniemi‐Hujanen

2005

Intl Journal of Cancer

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Dear Sir,We thank Tanus-Santos and Gerlach for the special interest to our work regarding the serum matrix metalloproteinase-9 as a prognostic marker in head and neck squamous cell carcinoma 1 .In our study, we demonstrated that high preoperative serum MMP-9 levels were linked with poor cause-specific as well as poor relapse-free survival in head and neck squamous cell carcinoma patients.1 The median serum level for MMP-9 immunoreactive protein of HNSCC patients was 89.9 ng/ml and the mean value was 115.2 ng/ml. Additionally, we demonstrated that the MMP-9 levels in 44 healthy controls were lower, median 55.6 ng/ml, mean 69.7 ng/ml (p < 0.001). The levels of MMP-9 immunoreactive protein in our study are thus clearly in line with studies published previously concerning MMP-9 serum levels in HNSCC patients 2,3 and the MMP-9 levels in controls equal the control plasma levels in study by Jung et al. 4 Moreover, Jung et al.4 have compared the serum and plasma levels of MMP-9 immunoreactive protein among controls using serum samples collected with or without tubes based on additives with clot activator. In our study, the serum samples were all so-called native serum samples i.e., collected with tubes with no additives. We also want to point out the fact that in our study all samples have been handled in consistent method. This fact would have been very important to add to the Material and Methods section and we are very grateful to have this opportunity to clarify the issue. In general, the pre-analytical conditions and methodological issues are essential when assessing MMP-9 in clinical samples. 4,5 When evaluating the immunoreactive protein by ELISA assay, it should not equate with the assay of measuring the activities of MMP-9. [6][7][8] The source of circulating MMP-9 whether it is measured from serum or plasma is not easy to define. In our study, however, we were also able to show that there is a statistically significant correlation between the serum level of MMP-9 immunoreactive protein and MMP-9 immunohistochemical staining in primary tumors of HNSCC.

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Use of citrate to minimize neutrophil matrix metalloproteinase-9 in human plasma

Cited by 55 publications

References 18 publications

Blood sampling as critical preanalytical determinant to use circulating MMP and TIMP as surrogate markers for pathological processes

Blood sampling as critical preanalytical determinant to use circulating MMP and TIMP as surrogate markers for pathological processes

Reply to: Serum samples should not be used to access circulating matrix metalloproteinase‐9 levels as a prognostic marker of disease

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