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Determination of red blood cell deformability using centrifugal force in a three-dimensional-printed mini-disk (3D-PMD).

ABSTRACT: Measuring red blood cell (RBC) deformability has become important for clinical disease diagnostics. Various methods for measuring RBC deformability have been developed; however, they require costly and large instruments, long measuring time, and skilled personnel. In this study, we present a three-dimensional-printed mini-disk (3D-PMD) for measuring RBC deformability to overcome the previous limitations. For a miniaturized and low-cost setup, the 3D-PMD was fabricated by a 3D printing technique, which had not yet been used for fabricating a lab-on-a-compact disk (LOCD). Using a 3D printing technique, a multi-layered fluidic channel on the mini CD could be fabricated easily. During rotation by a spinning motor, the difference of the length of compressed RBCs in the fluidic channel was measured and analysed as compressibility indices (CIs) of normal and glutaraldehyde-treated hardened RBCs. The rotation speed and time were decided as 3000 rpm and 30 min, respectively, at which the difference of CI values between normal and hardened RBCs was largest (CInormal-CIhardened = 0.195).

SUBMITTER: Lim H

PROVIDER: S-EPMC5963765 | biostudies-literature | 2018

REPOSITORIES: biostudies-literature

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Determination of red blood cell deformability using centrifugal force in a three-dimensional-printed mini-disk (3D-PMD).

Lim Hyunjung H Back Seung Min SM Nam Jeonghun J Choi Hyuk H

PloS one 20180522 5

Measuring red blood cell (RBC) deformability has become important for clinical disease diagnostics. Various methods for measuring RBC deformability have been developed; however, they require costly and large instruments, long measuring time, and skilled personnel. In this study, we present a three-dimensional-printed mini-disk (3D-PMD) for measuring RBC deformability to overcome the previous limitations. For a miniaturized and low-cost setup, the 3D-PMD was fabricated by a 3D printing technique, ...[more]

PMID: 29787582

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Project description:Background/objectiveA novel porous scaffold poly (lactide-co-glycolide) and tricalcium phosphate (PLGA/TCP) was developed by three-dimensional printing technology for bone defect repair. As a Class 2 solvent with less severe toxicity, content of residual 1,4-dioxane in this newly developed scaffold should be rigorously controlled when it is translated to clinical use. In this study, a headspace gas chromatography-mass spectrometric (HS-GC-MS) method and related testing protocol were developed for quantitative determination of 1,4-dioxane in the PLGA/TCP composite scaffolds.MethodsMatrix effect analysis was used to optimise the pretreatment method of the scaffolds. Then, the procedure for testing 1,4-dioxane using HS-GC-MS was set up. The accuracy, precision, and robustness of this newly developed quantitative method were also validated before quantification of 1,4-dioxane in the scaffolds with different drying procedures.ResultsDimethyl formamide (DMF) was the optimal solvent for dissolving scaffolds for GC-MS with proper sensitivity and without matrix effect. Then, the optimised procedure was determined as: the scaffolds were dissolved in DMF and kept at 90°C for 40 minutes, separated on a HP-5MS column, and detected by mass spectroscopy. Recovery experiments gave 97.9-100.7% recovery for 1,4-dioxane. The linear range for 1,4-dioxane was determined as 1-40 ppm with linear correlation coefficient ≥ 0.9999. Intraday and interday precision was determined as being within relative standard deviation of below 0.68%. The passable drying procedure was related to lyophilising (-50°C, 50 Pa) the scaffolds for 2 days and drying in vacuum (50 Pa) for 7 days.ConclusionThis is the first quantitative method established to test 1,4-dixoane in a novel scaffold. This method was validated with good accuracy and reproducibility, and met the methodological requirements of the Guideline 9101 documented in the Chinese Pharmacopoeia 2015 Edition.The translational potential of this articleThis quantitative method for determination of residual 1,4-dioxane in the novel scaffolds is a key technical method during its translation into clinical use because this method is an important and indispensable file in the enterprise standard when the porous scaffold is registered as a Class III implanted medical device for bone defect repair, which is used to guarantee the safety of the scaffolds. It is also applied to optimise the drying process of scaffolds and to monitor the quality of scaffolds in the industrialisation process. Further, this method provides references for other solvents quantitative determination in porous scaffolds or materials.

Project description:BackgroundCentrifugation is routinely employed in handling the ejaculates of some species, but it is not part of the commonly used protocols in ram. However, the development and implementation of new assisted reproductive technologies, alternative preservation models based on washing sperm from a cellular ageing-accelerating substance such as the seminal plasma, and basic studies in spermatology is associated with the use of centrifugation. This requires a specific evaluation of the centrifugation protocols considering the species-specific relationship with the potential damage produced by this procedure. No previous studies have determined the effect of different centrifugation forces on ram sperm. Therefore, we aimed to assess the performance of three centrifugal forces (600×g, 3000×g, and 6000×g for 10 min at room temperature) and their effects on ram sperm motility and functionality.ResultsSperm motility and functionality parameters were assessed at 0 h and after 2 h of incubation at 37 °C. As expected, a higher cell packaging degree was obtained at high centrifugation forces (P ≤ 0.0001). Cell packaging was unstable at all centrifugal forces. Thus, there was a high cell resuspension rate after less than 2 min. Regarding sperm quality, there was a change in movement pattern of 3000×g and 6000×g centrifuged sperm after 2 h of incubation at 37 °C, characterized by an increase in rapid progressive motility, linearity, straightness, and beat frequency, and a decrease in medium progressive motility, curvilinear velocity, path velocity, and head lateral amplitude. Non-significant differences were obtained among the different treatments concerning the total viability. However, we observed a significant increase (P ≤ 0.05) in the percentage of viable apoptotic sperm in the samples centrifuged at 6000×g at 0 h.ConclusionsCentrifugal forces equal to or greater than 3000×g induced some deleterious effects in ram sperm quality, and lower forces did not provide a successful cell packaging degree.

Dataset Information

Determination of red blood cell deformability using centrifugal force in a three-dimensional-printed mini-disk (3D-PMD).

Publications

Determination of red blood cell deformability using centrifugal force in a three-dimensional-printed mini-disk (3D-PMD).

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