Effect of Liposome Charge and Composition on the Delivery of Trehalose into Red Blood Cells
Bibliographic record
Abstract
Although the disaccharide trehalose has been shown to protect cells during freezing and desiccation when present intracellularly, a major obstacle to using intracellular sugars for biopreservation applications is the impermeability of plasma membranes. We are investigating the use of liposomes, which are synthetic, microscopic vesicles, for the intracellular delivery of stabilizing sugars into mammalian cells. Previous work has shown that the mechanism of red blood cell (RBC)–liposome interaction includes both liposome fusion with the RBC membranes, as well as tight adsorption of the vesicles onto the RBC surface. However, the fusion efficiency of liposomes was low, with only micromolar concentrations of trehalose delivered to the RBC cytosol. The purpose of this study is to enhance the efficacy of liposomes’ delivery of trehalose into RBCs with minimal detrimental effects on RBC membrane quality, by manipulating liposome physical properties and liposome–RBC incubation conditions. Charged and uncharged unilamellar liposomes were synthesized using an extrusion method to contain 300 mM trehalose in their aqueous core. Various concentrations of these liposomes (0.25–4.0 mM lipid) were incubated with RBCs at temperatures between 4°C and 37°C, for incubation times ranging from 15 min to 6 h. Red blood cell membrane quality was assessed using percent hemolysis, RBC morphology, phosphatidylserine (PS) and CD47 expression. The results of this study show that negatively charged liposomes containing PS in their membrane delivered about 100 times more trehalose (~15 mM) than neutral liposomes. Red blood cell–liposome incubation in an extracellular trehalose solution, as opposed to saline, results in a 34 ± 4 % increase of RBC intracellular trehalose concentration. In contrast to incubation time and temperature, liposome concentration and charge are not important predictors of RBC membrane injury. This study demonstrates that altering liposome physical properties and incubation conditions can cause an increase of RBC uptake of trehalose-containing liposomes, with minimal detrimental effects on RBC membrane quality.
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How this classification was reachedexpand
Full frame distilled prediction
Teacher imitationNot calibrated prevalence, not ground truth. Human validation pending. Learned from the 10,348 direct Codex labels and 10,348 direct Gemma labels. Candidate is the union of thresholded teacher heads; consensus is their intersection. These outputs are machine_predicted_unvalidated and are not human labels or direct frontier model labels.
Codex and Gemma teacher scores by category
| Category | Codex | Gemma |
|---|---|---|
| Metaresearch | 0.000 | 0.000 |
| Meta-epidemiology (narrow) | 0.000 | 0.000 |
| Meta-epidemiology (broad) | 0.000 | 0.000 |
| Bibliometrics | 0.000 | 0.000 |
| Science and technology studies | 0.000 | 0.000 |
| Scholarly communication | 0.000 | 0.000 |
| Open science | 0.000 | 0.000 |
| Research integrity | 0.000 | 0.000 |
| Insufficient payload (model declined to judge) | 0.000 | 0.000 |
Machine scores (provisional)
The two teacher heads of the student model, read on this work. A score orders the frame for review; it never asserts a category, and the validation status ships verbatim with every row.
Baseline scores from an immature model (maturity gate not passed, 7 training rounds). Scores rank; they never assert a category.
score_only:v0-immature-baseline · verbatim from the scoring run: score_only means the number may rank works, and no category label ships from itClassification
machine, unvalidatedMachine predicted; a candidate call from one teacher head, not a consensus.
How this classification was reached, model by model and score by score, is at the end of the page under "How this classification was reached".