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Introduction <p>Platelet derived microparticles (PMPs) are small fragments (0.1–1 μm) shed from platelets during activation, shear stress, or apoptosis. They act as both biomarkers and mediators of chronic diseases. However, their detection is technically challenging, highlighting the need for simple and reliable methods.</p> Aim <p>To compare PMP levels isolated by sedimentation, low speed centrifugation [Platelet Rich Plasma (PRP)], and high speed centrifugation [Platelet Poor Plasma (PPP)] in patients with chronic diseases.</p> <b>Materials and Methods</b> <p>A comparative cross-sectional study was conducted enrolling 30 patients with chronic diseases. Complete haematological profiling and immunophenotyping by flow cytometry were performed for detection of PMPs. Three protocols—sedimentation, PRP and PPP were applied for PMP isolation. Antibodies against CD45, CD41, CD61, Annexin V, and CD42b were used to differentiate PMPs from platelets.</p> <b>Results</b> <p>Significant differences in PMP yield were observed. In sedimentation, PMP levels ranged from 0.11–11.48% (mean± SD: 3.34 ± 3.20; median [IQR]: 1.87 [4.11]). In PRP, PMP levels ranged from 0.32–24.03% (mean ± SD: 4.56 ± 5.00; median [IQR]: 3.00 [4.58]). In PPP, PMP levels were 0.62–35.79% (mean ± SD: 8.95 ± 8.67; median [IQR]: 6.44 [5.41]). The highest yield was obtained with PPP across mean, median, and range. A strong correlation was noted between Annexin V+CD42b+ PMP% in PRP and PPP ( Interclass Coefficient = 0.66, p&lt;0.001).</p> <b>Conclusion</b> <p>Centrifugation-based protocols yield higher PMP counts than sedimentation. PRP and PPP methods show comparable efficacy, with PPP providing the highest yield. Larger studies are required to confirm these findings.</p>

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A comparative study of isolation protocols for estimation of platelet derived microparticles in chronic diseases

  • 期刊论文

  • 发表日期：

    2026年3月11日

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  • Shagun Wadhwa,
  • Kaninika Sanyal,
  • Mrinalini Kotru,
  • Richa Gupta,
  • Priyanka Gogoi,
  • Shiva Narang

摘要

Introduction

Platelet derived microparticles (PMPs) are small fragments (0.1–1 μm) shed from platelets during activation, shear stress, or apoptosis. They act as both biomarkers and mediators of chronic diseases. However, their detection is technically challenging, highlighting the need for simple and reliable methods.

Aim

To compare PMP levels isolated by sedimentation, low speed centrifugation [Platelet Rich Plasma (PRP)], and high speed centrifugation [Platelet Poor Plasma (PPP)] in patients with chronic diseases.

Materials and Methods

A comparative cross-sectional study was conducted enrolling 30 patients with chronic diseases. Complete haematological profiling and immunophenotyping by flow cytometry were performed for detection of PMPs. Three protocols—sedimentation, PRP and PPP were applied for PMP isolation. Antibodies against CD45, CD41, CD61, Annexin V, and CD42b were used to differentiate PMPs from platelets.

Results

Significant differences in PMP yield were observed. In sedimentation, PMP levels ranged from 0.11–11.48% (mean± SD: 3.34 ± 3.20; median [IQR]: 1.87 [4.11]). In PRP, PMP levels ranged from 0.32–24.03% (mean ± SD: 4.56 ± 5.00; median [IQR]: 3.00 [4.58]). In PPP, PMP levels were 0.62–35.79% (mean ± SD: 8.95 ± 8.67; median [IQR]: 6.44 [5.41]). The highest yield was obtained with PPP across mean, median, and range. A strong correlation was noted between Annexin V+CD42b+ PMP% in PRP and PPP ( Interclass Coefficient = 0.66, p<0.001).

Conclusion

Centrifugation-based protocols yield higher PMP counts than sedimentation. PRP and PPP methods show comparable efficacy, with PPP providing the highest yield. Larger studies are required to confirm these findings.