The preservation of synovial fluid using dimethyl sulfoxide.
Author(s): Pavic K, McGill V, D'Souza M, McGill N
Publication: Ann Clin Biochem, 2022, Vol. , Page 45632221076349
PubMed ID: 35044280 PubMed Review Paper? No
Purpose of Paper
This paper compared cellular morphology and the presence of clumping and artifacts among matched synovial fluid specimens stored at room temperature or -80°C for up to 8 weeks with and without dimethyl sulfoxide (DMSO). Analysis of synovial specimens was performed by two people, although only one was blinded to storage temperature and duration.
Conclusion of Paper
Overall, the authors observed a significant benefit to preserving synovial specimens with DMSO prior to frozen storage. Cellular morphology was best preserved when DMSO was added and specimens were stored at -80°C, but no differences in the presence of clumping or artifacts were noted between preservation methods (with and without DMSO) or storage temperature in the combined dataset. The duration of storage did not significantly affect cellular morphology, but significant differences in the presence of clumping and artifacts were noted between storage timepoints when data was combined. The kappa agreement in findings between the blinded and non-blinded assessor was only 0.1045 for morphology, 0.1474 for the presence of artifacts and 0.0523 for the presence of clumping.
Studies
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Study Purpose
This study compared cellular morphology and the presence of clumping and artifacts among matched synovial fluid specimens stored at room temperature or -80°C for up to 8 weeks with and without DMSO. Synovial fluid was collected from 15 patients who needed synovial fluid aspiration for either diagnostic or condition management, but no additional details were provided. Synovial fluid was immediately divided into 24 aliquots and DMSO was added to half the aliquots. An aliquot of each specimen was analyzed immediately, and the remaining aliquots of each specimen were stored at room temperature and -80°C for 1, 2, 3, 6, 7 and 8 weeks. Cellular morphology, clumping and artifact presence were graded by a blinded and a non-blinded assessor. Cellular morphology was graded and scored as the following: (1) intact membranes and appearance like fresh specimens, (2) membranes were partially ruptured, (3) membranes severely ruptured, but identifiable cells present, or (4) only cellular debris was observed. Clumping and the presence of artifacts were graded as yes/no.
Summary of Findings:
Both assessors observed a significant benefit to preserving synovial specimens with DMSO prior to frozen storage (P=0.007, both individually, P=0.001 combined). Findings from the assessor that was blinded to preservation method and storage conditions indicate that cellular morphology was significantly better preserved when synovial specimens were stored at -80°Cthan when stored at room temperature regardless of whether DMSO was added (P<0.05 all). Results recorded by the non-blinded assessor (who was aware of the preservation method and storage condition of each specimen), indicate that cellular morphology was significantly better preserved when specimens were stored at -80°C with DMSO than without DMSO or at room temperature regardless of the addition of DMSO (P<0.05 all). When the data from the two assessors were combined, synovial specimens stored at -80°C with DMSO resulted in significantly better preserved morphology than any other condition (P<0.001, all); and a non-significant trend toward better morphology was observed in specimens stored at -80°C without DMSO than those stored at room temperature. Importantly, neither assessor found an effect of storage duration on cellular morphology. While the blinded assessor found no difference in the presence of clumping among the preservation/storage groups, the non-blinded assessor found less clumping in specimens stored at room temperature when DMSO was added than when it was not (P=0.011) and less clumping was observed in specimens stored for 1 week than 6 or 8 weeks (P=0.037 and P=0.024, respectively), and stored for 3 weeks than 6, 7 or 8 weeks (P=0.040, P=0.023, and P<0.001, respectively). When data from the two assessors were combined, the presence of clumping was not significantly affected by preservation with DMSO or by storage temperature, but specimens stored for 3 weeks had less clumping than those stored for 7 weeks (P=0.024). While neither assessor individually observed a difference in the presence of artifacts among experimental groups, combined analysis revealed a significant effect of storage duration on the presence of artifacts with more artifacts observed in specimens stored for 7 weeks than stored for 1, 2, 3, or 8 weeks (P<0.05, all). The kappa agreement between the two assessors was 0.1045 for morphology, 0.1474 for the presence of artifacts and 0.0523 for the presence of clumping.
Biospecimens
Preservative Types
- Frozen
- None (Fresh)
- Other Preservative
Diagnoses:
- Not specified
Platform:
Analyte Technology Platform Morphology Light microscopy Pre-analytical Factors:
Classification Pre-analytical Factor Value(s) Biospecimen Preservation Type of fixation/preservation Frozen
None (fresh)
Dimethyl sulfoxide
Storage Storage temperature Room temperature
-80°C
Storage Storage duration 0 weeks
1 week
2 weeks
3 weeks
6 weeks
7 weeks
8 weeks
Light microscopy Specific Experimental bias Blinded assessment
Unblinded assessment