Biocompatibility and biotoxicity of in-situ synthesized carboxylated nanodiamond-cobalt oxide nanocomposite

doi:10.1016/j.jmst.2017.03.016

Abstract

Abstract:

A nanocomposite that incorporates cobalt oxide (Co₃O₄) and nanodiamond (ND) can present both high magnetism (Co₃O₄) and high hardness (ND). ND particles have potential applications in a variety of fields such as protein immobilization, biosensors, therapeutic molecule delivery and bio-imaging. However, limited information is available in literature on the in-situ synthesis of biocompatible magnetic materials and also on their potential biotoxicity as a result of their entry into environmental compartments and subsequent interaction with biota. In this work, a new kind of bio-compatible magnetic material - carboxylated nanodiamond (cND) and Co₃O₄ was synthesized to obtain the cND-Co₃O₄ nanocomposite. The synthesis procedure involved in-situ and chemical reduction of cobalt chloride (CoCl₂ 6H₂O) and sodium borohydrate (NaBH₄). The synthesized cND-Co₃O₄ nanocomposite was characterized by X-ray diffraction, scanning electron microscopy, transmission electron microscopy and X-ray photoelectron spectroscopy. The cyto-genotoxicity of the synthesized nanocomposite material was studied by using onion (Allium cepa L.) as a test model with concentrations of 5, 10 and 20 μg/ml. The study was also extended to cND and Co₃O₄ materials for comparison purpose. Co₃O₄ and cND exhibited their contrasting effects on mitosis and other cyto-genotoxic indices studied herein. This work provided fundamental data on the synthesis and the bio-toxicity of the cND-Co₃O₄ nanocomposite, which, in turn, can help to expand their multidisciplinary applications.

Key words: Nanodiamond, Cobalt oxide, Nanodiamond, anocomposite, Bio-toxicity, Allium cepa

Syam Sundar L., A. Anjum Naser, Ferro M.C., Pereira Eduarda, K. Singh Manoj, Sousa A.C.M.. Biocompatibility and biotoxicity of in-situ synthesized carboxylated nanodiamond-cobalt oxide nanocomposite[J]. J. Mater. Sci. Technol., 2017, 33(8): 879-888.

Figures/Tables 10

Fig. 1. Optical images of bulk (a) Co3O4 (b) cND, and (c) cND-Co3O4 nanocomposite.

Fig. 2. XRD patterns of cND, Co3O4,and cND-Co3O4 nanocomposite.

Fig. 3. SEM analysis of cND-Co3O4 nanocomposite (a) general scans (b) all the elements of carbon, cobalt and oxygen present in the composite; individual elements (c) cobalt (d) oxygen (e) carbon; (f) EDX analysis of cND-Co3O4 nanocomposite.

Fig. 4. TEM analysis of cND-Co3O4 (a) general scan (b) selected area X-ray diffraction, and (c) high resolution-TEM.

Fig. 5. High resolution XPS spectra obtained for the composite. (a) Co 2p, (b) O 1 s and (c) C 1 s core levels.

Fig. 6. FT-IR spectrum of cND, and cND-Co3O4 nanocomposite.

Fig. 7. (a) Average particle size distribution (～16.95 μm) of cND-Co3O4 nanocomposite dispersed in water and the polydisperse Index is 0.451 from DLS analysis, and (b) synthesized cND-Co3O4 nanocomposite showing magnetic behaviour under magnetic field.

Table 1 Mitotic index and percent phase indices of prophase, metaphase, anaphase and telophase stages in Allium cepa root meristematic cells exposed to various concentration of cobalt oxide (Co3O4), carboxylated nanodiamond (cND) and cND-Co3O4. 1000 cells were scored per treatment group (n = 3).

Concentrations (&#x003bc;g/mL)	Nanoparticles	Mitotic Index (mean &#x000b1; SE)	Prophase (%)	Metaphase (%)	Anaphase (%)	Telophase (%)
Control	-	71.3 &#x000b1; 2.2	62.3	4.03	2.4	3.7
5	Co₃O₄	58.07 &#x000b1; 1.7^a	54.2	3.5	2.0	3.1
	cND	68.3 &#x000b1; 2.0^b	59.0	3.9	2.1	3.5
	cND-Co₃O₄	67.5 &#x000b1; 2.0^bc	58.7	3.6	2.4	3.7
10	Co₃O₄	37.8 &#x000b1; 1.2^a	43.8	3.0	1.7	2.7
	cND	65.7 &#x000b1; 1.9^ab	55.3	3.3	2.04	2.9
	cND-Co₃O₄	63.8 &#x000b1; 1.9^ab	56.1	2.8	2.3	3.04
20	Co₃O₄	28.6 &#x000b1; 0.8^a	32.6	1.7	1.77	2.5
	cND	59.0 &#x000b1; 1.7^ab	51.8	2.8	2.1	2.8
	cND-Co₃O₄	51.7 &#x000b1; 1.5^ab	44.07	3.05	1.2	3.2

Table 2 Quantitative estimates of occurrences of different chromosomal aberrations observed in Allium cepa root meristematic cells exposed to various concentration of cobalt oxide (Co3O4), carboxylated nanodiamond (cND) and cND-Co3O4. 1000 cells were scored per treatment group (n? = 3).

Concentrations (&#x003bc;g/mL)	Nanoparticles	Chromosomal breaks (%)	Chromosomal Bridge (%)	Sticky chromosomes (%)	Laggard chromosome (%)	Disturbed anaphase/ metaphase (%)
Control	-	0 &#x000b1; 0	0 &#x000b1; 0	0 &#x000b1; 0	0 &#x000b1; 0	0 &#x000b1; 0
5	Co₃O₄	35.5 &#x000b1; 2.5	41.2 &#x000b1; 2.8	26.7 &#x000b1; 1.8	8.8 &#x000b1; 0.6	7.4 &#x000b1; 0.5
	cND	3.2 &#x000b1; 0.2^a	4.6 &#x000b1; 0.3^a	0 &#x000b1; 0	0 &#x000b1; 0	1.3 &#x000b1; 0.1^a
	cND-Co₃O₄	8.5 &#x000b1; 0.6^abc	14.3 &#x000b1; 1.0^abc	4.4 &#x000b1; 0.3^abc	0 &#x000b1; 0	3.7 &#x000b1; 0.3^abc
10	Co₃O₄	44.7 &#x000b1; 3.1	57.7 &#x000b1; 4.0	33.7 &#x000b1; 2.3	14.5 &#x000b1; 1.0	13.5 &#x000b1; 0.9
	cND	5.5 &#x000b1; 0.4^a	6.6 &#x000b1; 0.4^a	1.4 &#x000b1; 0.1^a	1.3 &#x000b1; 0.1^a	3.5 &#x000b1; 0.3^a
	cND-Co₃O₄	17.3 &#x000b1; 1.2^abc	21.4 &#x000b1; 1.5^abc	5.7 &#x000b1; 0.4^abc	3.5 &#x000b1; 0.2^abc	6.0 &#x000b1; 0.4^abc
20	Co₃O₄	58.8 &#x000b1; 4.1	69.4 &#x000b1; 4.8	47.9 &#x000b1; 3.3	21.4 &#x000b1; 1.5	20.5 &#x000b1; 1.4
	cND	7.1 &#x000b1; 0.5^ab	8.5 &#x000b1; 0.6^a	2.6 &#x000b1; 0.2^a	4.0 &#x000b1; 0.3^a	5.3 &#x000b1; 0.4^a
	cND-Co₃O₄	26.4 &#x000b1; 1.8^abc	27.8 &#x000b1; 2.0^abc	11.5 &#x000b1; 0.8^abc	16.6 &#x000b1; 1.2^abc	9.7 &#x000b1; 0.7^abc

Fig. 8. Representative images showing different chromosomal aberrations observed in Allium cepa root meristematic cells under control (A-D) and exposed to various concentrations of cobalt (Co) (D-G) and Co and carboxylated nanodiamond (Co-cND) (H-K). A = Prophase; B = Metaphase; C = Anaphase; D = Telophase; E = Chromosomal break; F = Cytoplasmic bridge; G = Disturbed anaphase; H = Laggard; I = Sticky anaphase; J = Scattered anaphase; K = Prophase nuclei with micronucleus in interphase; L = Binucleate cells.

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