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B. Boro et al.
2.2 Preparation of RBCs for the in Vitro Experiment
Blood samples were freshly collected from the medical laboratory technician course
laboratory of Pub Kamrup College, Assam, India from five healthy human subjects.
Erythrocytes were separated using standard procedure and kept in a special buffer as
described in previous literature (Devi et al. 2016).
2.3 In Vitro RBCs Cytotoxic Evaluation of MgO NPs
RBC cells are separated by dissolving PBS buffer (pH = 7.4) with equal ratio and
centrifuged at 3000 rpm for 5 min. Three different concentrations of MgO NPs,
viz., 5, 15, and 25 mg/ml were treated with 500 μl of RBCs in equal volume and
incubated for 24 h at 37 °C. Water in PBS buffer is considered as negative control and
dH 2 O remains blank. The reaction mixture was centrifuged at 900 rpm for 5 min.
and the supernatant was used for spectrophotometric measurement using UV-Vis
spectrophotometer (Eppendorf Biospectrometer).
2.4 Microscopic Observation
After 24 h of incubation, precipitated erythrocytes were stained with Leishman stain
using standard protocol for further microscopic observation. Briefly, treated blood
smear was dried and working Leishman stain was used to stain the blood smear.
Excess stain was washed out with distilled water and slides were dried and observed
and microphotographs were taken by using a CCD camera attached microscope
(Labomed LX300) (Devi et al. 2016).
3 Results and Discussions
3.1 Spectrophotometric Characterization of Prepared MgO
Nanoparticles
Absorption of the synthesized MgO nanoparticles was measured in the range of 250–
600 nm in the UV-Vis spectrophotometer and observed as shown in Fig. 1. A sharp
absorption peak was observed at wavelength 275 nm and the linear curve indicates
the uniformity of the synthesized nanoparticles. The band gap is found to be 3.89 eV
and is shown in Fig. 1.
B. Boro et al.
2.2 Preparation of RBCs for the in Vitro Experiment
Blood samples were freshly collected from the medical laboratory technician course
laboratory of Pub Kamrup College, Assam, India from five healthy human subjects.
Erythrocytes were separated using standard procedure and kept in a special buffer as
described in previous literature (Devi et al. 2016).
2.3 In Vitro RBCs Cytotoxic Evaluation of MgO NPs
RBC cells are separated by dissolving PBS buffer (pH = 7.4) with equal ratio and
centrifuged at 3000 rpm for 5 min. Three different concentrations of MgO NPs,
viz., 5, 15, and 25 mg/ml were treated with 500 μl of RBCs in equal volume and
incubated for 24 h at 37 °C. Water in PBS buffer is considered as negative control and
dH 2 O remains blank. The reaction mixture was centrifuged at 900 rpm for 5 min.
and the supernatant was used for spectrophotometric measurement using UV-Vis
spectrophotometer (Eppendorf Biospectrometer).
2.4 Microscopic Observation
After 24 h of incubation, precipitated erythrocytes were stained with Leishman stain
using standard protocol for further microscopic observation. Briefly, treated blood
smear was dried and working Leishman stain was used to stain the blood smear.
Excess stain was washed out with distilled water and slides were dried and observed
and microphotographs were taken by using a CCD camera attached microscope
(Labomed LX300) (Devi et al. 2016).
3 Results and Discussions
3.1 Spectrophotometric Characterization of Prepared MgO
Nanoparticles
Absorption of the synthesized MgO nanoparticles was measured in the range of 250–
600 nm in the UV-Vis spectrophotometer and observed as shown in Fig. 1. A sharp
absorption peak was observed at wavelength 275 nm and the linear curve indicates
the uniformity of the synthesized nanoparticles. The band gap is found to be 3.89 eV
and is shown in Fig. 1.