Mahmud, Khadijah (2018) Polymeric Nano-encapsulation Of Наемоglobin Isolated From Expired Human Redblood Cells For Use As Blood Substitute. Masters thesis, Universiti Sains Malaysia.
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Abstract
To date, blood scarcity in transfusion service demand vigorous scientific works looking for possible red blood cells alternatives. In this study, we aim to isolate and purify human haemoglobin (hb) from expired red cells by multiple washing, sonication, organic extraction and dialysis method. Analysis of the UV-Vis spectrum comparing hb and red blood cells (RBC) showed that the protein has been isolated successfully. Subsequently, the hb was encapsulated by polymeric micelle upon hydrophobic interaction (micellisation) of the methoxy poly (ethylene glycol)-b-poly (E-caprolactone) (PEG-PCL), a diblock copolymer in an aqueous buffer. The hb-micelle complex was then further analysed for half-life, oxygen reversibility, and cell-viability assay. Average encapsulation efficiency of the hb within the polymeric micelle achieved 45%. This is due to the high solubility of free hb in aqueous solution limiting the entry of hb into the polymer. However, the result indicated that the average of two units hb were successfully encapsulated by each polymer micelle. The hbmicelle recorded a higher half-life of 112,242 mins as compared to 30 mins of the free hb alone during oxygen stability study. The hb-micelle system also showed the ability to bind oxygen reversibly. Oxygenation and deoxygenation processes recorded average of six cycles before the sample was evaporated due to oxygenation, in contrast with only one cycle at average for free hb as a control. In cell viability study, the hb-micelle sample with concentrations of 1.89 x10³mg/mL in two preparations of micelles at 1.0 and 2.0 mg/mL while as a control, free hb sample from human and free bovine at 0.1 mg/mL and 0.2 mg/mL concentrations and also bovine hb-micelle (hb concentration at 2.21 ×105 mg/mL) have been tested on murine macrophage cells (RAW 264.7), incubated for 24 hours before treatments. Treated cells were then incubated further for 48 hours where the cell viability analysed at time points of 4, 8, 24 and 48 h. Generally, the free hb recorded reduction in cell growth approximately by 10% and 18% as early as 4h compared to hb-micelle which showed no toxic effects on the cells until 24 hours onwards. This result probably due to the free hb from human used in the test may have contained lipid debris from the membrane lysis during extraction process. However, it showed that hb-micelle system can safely interact with normal cells for potential future use compared to free hb without encapsulation by polymer. These findings indicated the potential of the hb 'harvested' from expired human red cells encapsulated by the diblock copolymer for development as red blood cells substitute.
| Item Type: | Thesis (Masters) |
|---|---|
| Subjects: | R Medicine |
| Divisions: | Institut Perubatan & Pergigian Termaju (Advanced Medical & Dental Institute (AMDI)) > Thesis |
| Depositing User: | NOR HASHIMY BIN ABDULLAH |
| Date Deposited: | 17 Jul 2026 01:42 |
| Last Modified: | 17 Jul 2026 01:42 |
| URI: | http://eprints.usm.my/id/eprint/64671 |
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