Ex Vivo Microsphiltration Profile of Plasmodium Falciparum Infected Red Blood Cells in Patients with Malaria in Kéniéroba, Mali: Exploring the Spleen in Vivo Retention Function

Abstract Malaria pathophysiology is not still fully understood. The main mechanisms of malaria involve the synergistic interactions between host and parasite. Although, the role of the spleen has been mentioned in various clinical forms of malaria, a supportive clinical evidence is still needed. We conducted a pilot study to determine the impact of the spleen functional state in different clinical forms of malaria. Ex vivo microsphiltration was used to assess the splenic function in patients received during routine consultation with mild malaria at the Kéniéroba health center, a high malaria endemic area in Mali. A total of 25 patients were enrolled for ex vivo microsphiltration. Spleen was non-palpable (Hackett stage 0) in two patients, palpable with deep inspiration (Hackett stage 1) in 22 patients and without deep inspiration (Hackett stage 2) in one patient, parasitaemia ranged from 5360 trophozoites/µl to 342720 trophozoites/µl with a mean parasitemia of 50774 trophozoites/µl ± 65540 trophozoites/µl. The mean hemoglobin level was 11.2g/dl ± 1.2 [8.7-13.4]. The retention rate of the infected red blood cell ranged from 11.11% to 94.44% with 65.4% ± 23.7% on average. A higher ex vivo retention rate of infected red blood cells was observed in patients with Hackett stage other than 0 (p= 0.03). This pilot study proved that it was feasible to use the ex vivo microsphiltration to explore the spleen filtering function in malaria patients.

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Ex Vivo Microsphiltration Profile of Plasmodium Falciparum Infected Red Blood Cells in Patients with Malaria in Kéniéroba, Mali: Exploring the Spleen in Vivo Retention Function

10.21203/rs.3.rs-557105/v1 ◽

2021 ◽

Author(s):

Bourama KEITA ◽

Seidina A.S. Diakité ◽

Agnes M. Guindo ◽

Drissa S. Konaté ◽

Karim Traoré ◽

...

Keyword(s):

Pilot Study ◽

Red Blood Cells ◽

Blood Cells ◽

Ex Vivo ◽

Retention Rate ◽

Clinical Forms ◽

Filtering Function ◽

Stage 1 ◽

The Impact

Abstract Malaria pathophysiology is not still fully understood. The main mechanisms of malaria involve the synergistic interactions between host and parasite. Although, the role of the spleen has been mentioned in various clinical forms of malaria, a supportive clinical evidence is still needed. We conducted a pilot study to determine the impact of the spleen functional state in different clinical forms of malaria.Ex vivo microsphiltration was used to assess the splenic function in patients received during routine consultation with malaria at the Kéniéroba health center, a high malaria endemic area in Mali.A total of 25 patients were enrolled for microsphiltration. Two patients (8%) had a no palpable spleen (Hackett stage 0), 22 patients (88%) had a palpable spleen with at deep inspiration (Hackett stage 1) and only one patient (4%) presented a palpable spleen (Hackett stage 2). Parasitaemia ranged from 5360 trophozoites/µl to 342720 trophozoites/µl with a mean parasitemia of 50774 trophozoites/µl ± 65540 trophozoites/µl; the mean hemoglobin rate was 11.2 ± 1.2 g/dl with the extremes of 8.7 g/dl and 13.4 g/dl. The retention rate of the infected red blood cell ranged from 11.11% to 94.44% with an average of 65.4% ± 23.7%. A higher ex vivo retention rate of infected red blood cells was observed in patients with Hackett stages greater than or equal to 1 (p= 0.03). This pilot study proved the feasibility of the exploration of the spleen filtering function in malaria patients using the ex vivo microsphiltration.

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Human erythroid cells produced ex vivo at large scale differentiate into red blood cells in vivo

Nature Biotechnology ◽

10.1038/nbt0502-467 ◽

2002 ◽

Vol 20 (5) ◽

pp. 467-472 ◽

Cited By ~ 186

Author(s):

Thi My Anh Neildez-Nguyen ◽

Henri Wajcman ◽

Michael C. Marden ◽

Morad Bensidhoum ◽

Vincent Moncollin ◽

...

Keyword(s):

Red Blood Cells ◽

Blood Cells ◽

Large Scale ◽

Ex Vivo ◽

Erythroid Cells

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Ex vivo selective isolation of young red blood cells using the IBM-2991 cell washer

Blood ◽

10.1182/blood.v61.6.1068.1068 ◽

1983 ◽

Vol 61 (6) ◽

pp. 1068-1071 ◽

Cited By ~ 15

Author(s):

AW Bracey ◽

HG Klein ◽

S Chambers ◽

L Corash

Keyword(s):

Red Blood Cells ◽

Blood Cells ◽

Ex Vivo ◽

Major Complication ◽

Red Cell ◽

Transfusion Requirements ◽

Paired Measurement ◽

Time Required ◽

Cell Fractions

Abstract Transfusion-induced hemochromatosis is a major complication in the therapy of severe chronic anemia. Improvement of transfused cell survival with a reduction in transfusion frequency is one possible approach to this problem. Using continuous-flow centrifugation (CFC), young red blood cells (YRBC) with enhanced in vivo survival have been isolated, but the expense and donor time required with this technique prohibit its widespread use for patient support. We studied the use of the IBM 2991 cell washer (CW) to isolate YRBC ex vivo from previously collected donor blood. Age-dependent red cell separation could be achieved using this instrumentation. Autologous mean red cell half-life (RBC-T50) (n = 9) for the younger cell fractions was 43.9 +/- 7.8 days compared to 34.7 +/- 5.8 days for the older cell fractions (n = 6, p less than 0.05). Paired measurement of RBC-T50 for young and old fractions in three donors showed an average survival increase of 41% for the YRBC. Adequate quantities of YRBC with enhanced survival can be obtained with less cost and less donor stress using the CW system compared to CFC. This approach could improve the management of patients with chronic transfusion requirements and merits further examination.

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Vascular Drug Delivery Using Carrier Red Blood Cells: Focus on RBC Surface Loading and Pharmacokinetics

Pharmaceutics ◽

10.3390/pharmaceutics12050440 ◽

2020 ◽

Vol 12 (5) ◽

pp. 440 ◽

Cited By ~ 10

Author(s):

Patrick M. Glassman ◽

Carlos H. Villa ◽

Anvay Ukidve ◽

Zongmin Zhao ◽

Paige Smith ◽

...

Keyword(s):

Drug Delivery ◽

Drug Development ◽

Red Blood Cells ◽

Blood Cells ◽

Drug Delivery Systems ◽

Ex Vivo ◽

Delivery Systems ◽

Surface Loading ◽

Intended Application

Red blood cells (RBC) have great potential as drug delivery systems, capable of producing unprecedented changes in pharmacokinetics, pharmacodynamics, and immunogenicity. Despite this great potential and nearly 50 years of research, it is only recently that RBC-mediated drug delivery has begun to move out of the academic lab and into industrial drug development. RBC loading with drugs can be performed in several ways—either via encapsulation within the RBC or surface coupling, and either ex vivo or in vivo—depending on the intended application. In this review, we briefly summarize currently used technologies for RBC loading/coupling with an eye on how pharmacokinetics is impacted. Additionally, we provide a detailed description of key ADME (absorption, distribution, metabolism, elimination) changes that would be expected for RBC-associated drugs and address unique features of RBC pharmacokinetics. As thorough understanding of pharmacokinetics is critical in successful translation to the clinic, we expect that this review will provide a jumping off point for further investigations into this area.

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Ex vivo selective isolation of young red blood cells using the IBM-2991 cell washer

Blood ◽

10.1182/blood.v61.6.1068.bloodjournal6161068 ◽

1983 ◽

Vol 61 (6) ◽

pp. 1068-1071 ◽

Cited By ~ 1

Author(s):

AW Bracey ◽

HG Klein ◽

S Chambers ◽

L Corash

Keyword(s):

Red Blood Cells ◽

Blood Cells ◽

Ex Vivo ◽

Major Complication ◽

Red Cell ◽

Transfusion Requirements ◽

Paired Measurement ◽

Time Required ◽

Cell Fractions

Transfusion-induced hemochromatosis is a major complication in the therapy of severe chronic anemia. Improvement of transfused cell survival with a reduction in transfusion frequency is one possible approach to this problem. Using continuous-flow centrifugation (CFC), young red blood cells (YRBC) with enhanced in vivo survival have been isolated, but the expense and donor time required with this technique prohibit its widespread use for patient support. We studied the use of the IBM 2991 cell washer (CW) to isolate YRBC ex vivo from previously collected donor blood. Age-dependent red cell separation could be achieved using this instrumentation. Autologous mean red cell half-life (RBC-T50) (n = 9) for the younger cell fractions was 43.9 +/- 7.8 days compared to 34.7 +/- 5.8 days for the older cell fractions (n = 6, p less than 0.05). Paired measurement of RBC-T50 for young and old fractions in three donors showed an average survival increase of 41% for the YRBC. Adequate quantities of YRBC with enhanced survival can be obtained with less cost and less donor stress using the CW system compared to CFC. This approach could improve the management of patients with chronic transfusion requirements and merits further examination.

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Propensity of red blood cells to undergo P2X7 receptor-mediated phosphatidylserine exposure does not alter during in vivo or ex vivo aging

Transfusion ◽

10.1111/trf.13101 ◽

2015 ◽

Vol 55 (8) ◽

pp. 1946-1954 ◽

Cited By ~ 9

Author(s):

Reece A. Sophocleous ◽

Phillip R.F. Mullany ◽

Kelly M. Winter ◽

Denese C. Marks ◽

Ronald Sluyter

Keyword(s):

Red Blood Cells ◽

Blood Cells ◽

P2x7 Receptor ◽

Ex Vivo ◽

Phosphatidylserine Exposure

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Ultrastructural observations on the in vitro cytoadherence of Plasmodium falciparum infected red blood cells

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100162132 ◽

1990 ◽

Vol 48 (3) ◽

pp. 914-915

Author(s):

D.J.P. Ferguson ◽

A.R. Berendt ◽

J. Tansey ◽

K. Marsh ◽

C.I. Newbold

Keyword(s):

Plasmodium Falciparum ◽

Red Blood Cells ◽

Blood Cells ◽

Umbilical Vein ◽

Cell Types ◽

Amelanotic Melanoma ◽

Cytoplasmic Process ◽

Umbilical Vein Endothelial Cells

In human malaria, the most serious clinical manifestation is cerebral malaria (CM) due to infection with Plasmodium falciparum. The pathology of CM is thought to relate to the fact that red blood cells containing mature forms of the parasite (PRBC) cytoadhere or sequester to post capillary venules of various tissues including the brain. This in vivo phenomenon has been studied in vitro by examining the cytoadherence of PRBCs to various cell types and purified proteins. To date, three Ijiost receptor molecules have been identified; CD36, ICAM-1 and thrombospondin. The specific changes in the PRBC membrane which mediate cytoadherence are less well understood, but they include the sub-membranous deposition of electron-dense material resulting in surface deformations called knobs. Knobs were thought to be essential for cytoadherence, lput recent work has shown that certain knob-negative (K-) lines can cytoadhere. In the present study, we have used electron microscopy to re-examine the interactions between K+ PRBCs and both C32 amelanotic melanoma cells and human umbilical vein endothelial cells (HUVEC).We confirm previous data demonstrating that C32 cells possess numerous microvilli which adhere to the PRBC, mainly via the knobs (Fig. 1). In contrast, the HUVEC were relatively smooth and the PRBCs appeared partially flattened onto the cell surface (Fig. 2). Furthermore, many of the PRBCs exhibited an invagination of the limiting membrane in the attachment zone, often containing a cytoplasmic process from the endothelial cell (Fig. 2).

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The Ullrastructure of Platelet Participation in Hemostasis

Thrombosis and Haemostasis ◽

10.1055/s-0038-1656288 ◽

1965 ◽

Vol 13 (01) ◽

pp. 065-083 ◽

Cited By ~ 7

Author(s):

Shirley A. Johnson ◽

Ronaldo S. Balboa ◽

Harlan J. Pederson ◽

Monica Buckley

Keyword(s):

Platelet Aggregation ◽

Red Blood Cells ◽

Blood Cells ◽

Guinea Pigs ◽

Platelet Factor ◽

Mesenteric Vessels ◽

Platelet Aggregates ◽

Electron Lucent

SummaryThe ultrastructure of platelet aggregation in vivo in response to bleeding brought about by transection of small mesenteric vessels in rats and guinea pigs has been studied. Platelets aggregate, degranulate and separating membranes disappear in parallel with fibrin appearance which is first seen at several loci after 30 seconds of bleeding. About 40 per cent of the electron opaque granules, some of which contain platelet factor 3 have disappeared after one minute of bleeding while the electron lucent granules increase by 70 per cent suggesting that some of them may be empty vesicles. Most of the platelet aggregates of the random type disappear leaving clumped red blood cells entrapped by a network of fibrin fibers which emanate from the remains of platelet aggregates of the rosette type to maintain hemostasis.

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Cytotoxicity of chitosan ultrafine nanoshuttles on MCF-7 cell line as surrogate model for breast cancer

Current Drug Delivery ◽

10.2174/1567201817666200719005440 ◽

2020 ◽

Vol 17 ◽

Author(s):

Tarek Faris ◽

Gamaleldin I. Harisa ◽

Fars K. Alanazi ◽

Mohamed M. Badran ◽

Afraa Mohammad Alotaibi ◽

...

Keyword(s):

Red Blood Cells ◽

Factorial Design ◽

Cell Line ◽

Cell Lines ◽

Blood Cells ◽

Sodium Tripolyphosphate ◽

Physicochemical Characterization ◽

Spherical Shape ◽

Mcf 7

Aim: This study aimed to explore an affordable technique for the fabrication of Chitosan Nanoshuttles (CSNS) at the ultrafine nanoscale less than 100 nm with improved physicochemical properties, and cytotoxicity on the MCF-7 cell line. Background: Despite several studies reported that the antitumor effect of CS and CSNS could achieve intracellular compartment target ability, no enough available about this issue and further studies are required to address this assumption. Objectives: The objective of the current study was to investigate the potential processing variables for the production of ultrafine CSNS (> 100 nm) using Box-Benhken Design factorial design (BBD). This was achieved through a study of the effects of processing factors, such as CS concentration, CS/TPP ratio, and pH of the CS solution, on PS, PDI, and ZP. Moreover, the obtained CSNS was evaluated for physicochemical characteristics, morphology Also, hemocompatibility, and cytotoxicity using Red Blood Cells (RBCs) and MCF-7 cell lines were investigated. Methods: Box-Benhken Design factorial design (BBD) was used in the analysis of different selected variables. The effects of CS concentration, sodium tripolyphosphate (TPP) ratio, and pH on particle size, Polydispersity Index (PDI), and Zeta Potential (ZP) were measured. Subsequently, the prepared CS nanoshuttles were exposed to stability studies, physicochemical characterization, hemocompatibility, and cytotoxicity using red blood cells and MCF-7 cell lines as surrogate models for in vivo study. Result: The present results revealed that the optimized CSNS have ultrafine nanosize, (78.3±0.22 nm), homogenous with PDI (0.131±0.11), and ZP (31.9±0.25 mV). Moreover, CSNS have a spherical shape, amorphous in structure, and physically stable. Also, CSNS has biological safety as indicated by a gentle effect on red blood cell hemolysis, besides, the obtained nanoshuttles decrease MCF-7 viability. Conclusion: The present findings concluded that the developed ultrafine CSNS has unique properties with enhanced cytotoxicity. thus promising for use in intracellular organelles drug delivery.

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Histological, Biomechanical, and Biological Properties of Genipin-Crosslinked Decellularized Peripheral Nerves

International Journal of Molecular Sciences ◽

10.3390/ijms22020674 ◽

2021 ◽

Vol 22 (2) ◽

pp. 674

Author(s):

Óscar Darío García-García ◽

Marwa El Soury ◽

David González-Quevedo ◽

David Sánchez-Porras ◽

Jesús Chato-Astrain ◽

...

Keyword(s):

Ex Vivo ◽

Nerve Repair ◽

Wistar Rat ◽

Biomechanical Properties ◽

Biological Properties ◽

Suitable Alternative ◽

Promising Alternative ◽

Histological Pattern ◽

The Impact

Acellular nerve allografts (ANGs) represent a promising alternative in nerve repair. Our aim is to improve the structural and biomechanical properties of biocompatible Sondell (SD) and Roosens (RS) based ANGs using genipin (GP) as a crosslinker agent ex vivo. The impact of two concentrations of GP (0.10% and 0.25%) on Wistar rat sciatic nerve-derived ANGs was assessed at the histological, biomechanical, and biocompatibility levels. Histology confirmed the differences between SD and RS procedures, but not remarkable changes were induced by GP, which helped to preserve the nerve histological pattern. Tensile test revealed that GP enhanced the biomechanical properties of SD and RS ANGs, being the crosslinked RS ANGs more comparable to the native nerves used as control. The evaluation of the ANGs biocompatibility conducted with adipose-derived mesenchymal stem cells cultured within the ANGs confirmed a high degree of biocompatibility in all ANGs, especially in RS and RS-GP 0.10% ANGs. Finally, this study demonstrates that the use of GP could be an efficient alternative to improve the biomechanical properties of ANGs with a slight impact on the biocompatibility and histological pattern. For these reasons, we hypothesize that our novel crosslinked ANGs could be a suitable alternative for future in vivo preclinical studies.

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