1dfc51a910c8ea23bc5ba1e4a693fbfc 327869.pdf 28b41647bc9a4a8a5b2c4ce99844f4d0c9e74b10 327869.pdf 7b671717834ef99cd25e8e629cd6f5204ea54669b2a81cc16d219f5369876d95 327869.pdf Title: Osmotic Fragility in Leukodepleted Stored Red Blood Cells: Implications for Neurocritical Care Transfusion Strategies Subject: Background: Anemia is frequent in critically ill patients with traumatic brain injury (TBI) and worsens neurological outcomes. Red blood cell (RBC) transfusion is a cornerstone of management, but storage-related biochemical and structural changes may impair oxygen delivery. This study examined the effect of storage duration on osmotic fragility (OF) and free hemoglobin (fHb) in leukodepleted packed RBCs (pRBCs) as indicators of membrane stability and hemolysis. Methods: Twenty-four leukodepleted pRBC units in SAGM (saline, adenine, glucose, and mannitol) solution were analyzed from Day 3 to Day 42. OF was assessed by Beutler’s method with H50 values derived from logistic models, and fHb was quantified spectrophotometrically. Flow cytometry with phosphate-buffered saline (PBS)-induced osmotic stress provided complementary OF data. Results: OF increased significantly beyond 28 days, with Week 6 H50 values exceeding those at Weeks 2 and 4 (p < 0.0001). fHb rose progressively with storage: 7.3 4.3 mol/L (Week 2), 14.6 7.9 (Week 4), and 25.7 12.1 (Week 6) (p < 0.0001). Hemolysis remained below the 0.8% threshold but increased from 0.09% to 0.29% (p < 0.0001). Conclusions: pRBC storage beyond 28 days leads to greater OF and fHb release, reflecting reduced membrane stability. These changes may compromise transfusion efficacy and oxygen delivery in neurocritical care. Keywords: traumatic brain injury; red blood cell storage; osmotic fragility; free hemoglobin; transfusion Author: Marta Peris, Maria A. Poca, Ana Ortuño, Verónica Pons, Nuria Rodríguez-Borrero, Desiree Jurado, Rafael Parra-López, Marina Rierola and Juan Sahuquillo Creator: LaTeX with hyperref Producer: pdfTeX-1.40.25; modified using OpenPDF 1.4.2 CreationDate: Fri Nov 7 11:18:13 2025 CET ModDate: Fri Nov 7 11:21:43 2025 CET Custom Metadata: yes Metadata Stream: no Tagged: no UserProperties: no Suspects: no Form: none JavaScript: no Pages: 22 Encrypted: no Page size: 595.276 x 841.89 pts (A4) Page rot: 0 File size: 3097396 bytes Optimized: no PDF version: 1.7 name type encoding emb sub uni object ID ------------------------------------ ----------------- ---------------- --- --- --- --------- ZRCKTB+CMSY10 Type 1 Builtin yes yes yes 136 0 QKZKVH+VnURWPalladioL Type 1 Custom yes yes yes 137 0 HPKNJR+URWPalladioL-Ital Type 1 Custom yes yes yes 138 0 XULDEY+URWPalladioL-Bold Type 1 Custom yes yes yes 139 0 WUTTHV+URWPalladioL-Roma Type 1 Custom yes yes yes 140 0 STVQSE+EURM10 Type 1 Builtin yes yes yes 141 0 BKMEMW+PalatinoLinotype-Bold TrueType WinAnsi yes yes yes 265 0 LHKMWC+PalatinoLinotype-Roman TrueType WinAnsi yes yes yes 266 0 VJIPGI+PalatinoLinotype-Italic TrueType WinAnsi yes yes yes 267 0 LHKMWC+PalatinoLinotype-Roman CID TrueType Identity-H yes yes yes 268 0 BKMEMW+PalatinoLinotype-Bold TrueType WinAnsi yes yes yes 309 0 LHKMWC+PalatinoLinotype-Roman TrueType WinAnsi yes yes yes 310 0 VJIPGI+PalatinoLinotype-Italic TrueType WinAnsi yes yes yes 311 0 LHKMWC+PalatinoLinotype-Roman CID TrueType Identity-H yes yes yes 312 0 BKMEMW+PalatinoLinotype-Bold TrueType WinAnsi yes yes yes 332 0 LHKMWC+PalatinoLinotype-Roman TrueType WinAnsi yes yes yes 333 0 VJIPGI+PalatinoLinotype-Italic TrueType WinAnsi yes yes yes 334 0 LHKMWC+PalatinoLinotype-Roman CID TrueType Identity-H yes yes yes 335 0 BKMEMW+PalatinoLinotype-Bold TrueType WinAnsi yes yes yes 396 0 LHKMWC+PalatinoLinotype-Roman TrueType WinAnsi yes yes yes 397 0 VJIPGI+PalatinoLinotype-Italic TrueType WinAnsi yes yes yes 398 0 LHKMWC+PalatinoLinotype-Roman CID TrueType Identity-H yes yes yes 399 0 BKMEMW+PalatinoLinotype-Bold TrueType WinAnsi yes yes yes 365 0 LHKMWC+PalatinoLinotype-Roman TrueType WinAnsi yes yes yes 366 0 VJIPGI+PalatinoLinotype-Italic TrueType WinAnsi yes yes yes 367 0 LHKMWC+PalatinoLinotype-Roman CID TrueType Identity-H yes yes yes 368 0 BKMEMW+PalatinoLinotype-Bold TrueType WinAnsi yes yes yes 406 0 LHKMWC+PalatinoLinotype-Roman TrueType WinAnsi yes yes yes 407 0 VJIPGI+PalatinoLinotype-Italic TrueType WinAnsi yes yes yes 408 0 LHKMWC+PalatinoLinotype-Roman CID TrueType Identity-H yes yes yes 409 0 BKMEMW+PalatinoLinotype-Bold TrueType WinAnsi yes yes yes 527 0 LHKMWC+PalatinoLinotype-Roman TrueType WinAnsi yes yes yes 528 0 VJIPGI+PalatinoLinotype-Italic TrueType WinAnsi yes yes yes 529 0 LHKMWC+PalatinoLinotype-Roman CID TrueType Identity-H yes yes yes 530 0 Jhove (Rel. 1.28.0, 2023-05-18) Date: 2026-04-23 04:05:32 CEST RepresentationInformation: 327869.pdf ReportingModule: PDF-hul, Rel. 1.12.4 (2023-03-16) LastModified: 2026-04-22 09:16:37 CEST Size: 3097396 Format: PDF Version: 1.7 Status: Well-Formed and valid SignatureMatches: PDF-hul MIMEtype: application/pdf PDFMetadata: Objects: 713 FreeObjects: 1 IncrementalUpdates: 0 DocumentCatalog: ViewerPreferences: HideToolbar: false HideMenubar: false HideWindowUI: false FitWindow: true CenterWindow: false DisplayDocTitle: false NonFullScreenPageMode: UseNone Direction: L2R ViewArea: CropBox ViewClip: CropBox PrintArea: CropBox PageClip: CropBox PageLayout: SinglePage PageMode: UseNone Outlines: Item: Title: Introduction Item: Title: Materials and Methods Children: Item: Title: Sample Size and Power Analysis Item: Title: Leukodepleted Red Blood Cell Concentrates (RBC) Item: Title: Beutler’s Method for Assessing OF Item: Title: Free Hemoglobin Accumulation and Storage-Induced Hemolysis Item: Title: Flow Cytometry Item: Title: Statistical Analysis Item: Title: Results Children: Item: Title: Osmotic Fragility: Beutler’s Method Item: Title: Osmotic Fragility: Flow Cytometry Item: Title: Free Hemoglobin in the Stored Bags Item: Title: Discussion Children: Item: Title: RBC Storage Lesion: A Missing Link in Transfusion Outcomes Item: Title: OF, Storage Lesions, and the Transfusion Dilemma Item: Title: Reevaluating Storage Duration and Quality Control in Transfusion Practice Item: Title: Study Limitations Item: Title: Clinical Implications and Future Directions Item: Title: Conclusions Item: Title: References Info: Title: Osmotic Fragility in Leukodepleted Stored Red Blood Cells: Implications for Neurocritical Care Transfusion Strategies Author: Marta Peris, Maria A. Poca, Ana Ortuño, Verónica Pons, Nuria Rodríguez-Borrero, Desiree Jurado, Rafael Parra-López, Marina Rierola and Juan Sahuquillo Subject: Background: Anemia is frequent in critically ill patients with traumatic brain injury (TBI) and worsens neurological outcomes. Red blood cell (RBC) transfusion is a cornerstone of management, but storage-related biochemical and structural changes may impair oxygen delivery. This study examined the effect of storage duration on osmotic fragility (OF) and free hemoglobin (fHb) in leukodepleted packed RBCs (pRBCs) as indicators of membrane stability and hemolysis. Methods: Twenty-four leukodepleted pRBC units in SAGM (saline, adenine, glucose, and mannitol) solution were analyzed from Day 3 to Day 42. OF was assessed by Beutler’s method with H50 values derived from logistic models, and fHb was quantified spectrophotometrically. Flow cytometry with phosphate-buffered saline (PBS)-induced osmotic stress provided complementary OF data. Results: OF increased significantly beyond 28 days, with Week 6 H50 values exceeding those at Weeks 2 and 4 (p < 0.0001). fHb rose progressively with storage: 7.3 4.3 mol/L (Week 2), 14.6 7.9 (Week 4), and 25.7 12.1 (Week 6) (p < 0.0001). Hemolysis remained below the 0.8% threshold but increased from 0.09% to 0.29% (p < 0.0001). Conclusions: pRBC storage beyond 28 days leads to greater OF and fHb release, reflecting reduced membrane stability. These changes may compromise transfusion efficacy and oxygen delivery in neurocritical care. Keywords: traumatic brain injury; red blood cell storage; osmotic fragility; free hemoglobin; transfusion Creator: LaTeX with hyperref Producer: pdfTeX-1.40.25; modified using OpenPDF 1.4.2 CreationDate: Fri Nov 07 11:18:13 CET 2025 ModDate: Fri Nov 07 11:21:43 CET 2025 ID: 0x9030429befffd37afac4615771e0eb80, 0xdced55c3ba95630d361955c21f598a25 Filters: FilterPipeline: FlateDecode Fonts: Type1: Font: BaseFont: ZRCKTB+CMSY10 FontSubset: true FirstChar: 0 LastChar: 48 FontDescriptor: FontName: ZRCKTB+CMSY10 Flags: Symbolic FontBBox: -29, -960, 1116, 775 FontFile: true ToUnicode: true Font: BaseFont: QKZKVH+VnURWPalladioL FontSubset: true FirstChar: 177 LastChar: 225 FontDescriptor: FontName: QKZKVH+VnURWPalladioL Flags: Symbolic FontBBox: -166, -283, 1021, 943 FontFile: true EncodingDictionary: Differences: true ToUnicode: true Font: BaseFont: HPKNJR+URWPalladioL-Ital FontSubset: true FirstChar: 39 LastChar: 122 FontDescriptor: FontName: HPKNJR+URWPalladioL-Ital Flags: Symbolic FontBBox: 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