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These are physical, chemical and combination methods
for de-aggregation of NDs. Physical approaches are selected when NDs
aggregation is arbitrate through graphitic layering, while chemical methods are
base upon the task of the surface. Chemical approaches use the conjugation of
different organic or inorganic molecules on the surface of NDs to control
aggregation and to transmit them specific characteristics. De-aggregation of ND
in suspensions is produced by milling the ceramic microbeads (ZrO2, SiO2) or ultrasonic disintegration
with microbeads, dry milling with sodium
chloride or coarse sucrose, high-temperature hydrogen treatment, ultrasonic
treatment in borane presence . There was a data that allow the purification and
oxidation in air to isolate a stable hydrosol of particles 4-5 nm in diameter
by centrifugation. The first method yield the colloidal solutions of individual
NDs of size 4-5 nm in diameter, but during the bead milling graphitic layer is
formed around the primary particles15,70–74.

oxidants are used to remove it, following of the formation of new aggregates.
The second method is cheap that allows the particles and small aggregates of  size 5- 20 nm to be achieved without  any additional contamination. 2-4 nm size NDs are
produced by high-temperature hydrogen treatment. Ultrasonic treatment in borane
presence, decrease the aggregates size to ~20-nm. The possible re-aggregation
of ND particles is obstructed by ultrasound-assisted treatment in the presence
of sodium chloride. It is assumed that Na+ repels each other when they are
attached to the surface of the individual particles. Functionalization of the
surface can also oblige the reduction of the size of the aggregates. The above mentioned
method with borane, lice combined method showed the greatest reduction of
aggregates after functionalization. Functionalization can be used for
biomedical and pharmaceutical utilization, thus allows loading of drug

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surface improvement can be attained through physical adsorption or chemical
interaction. Physical adsorption of NDs has been widely accomplished using
proteins, drugs, and nucleic acid. The unification of the surface groups of the
NDs is an important step before the chemical functionalization of NDs, in order
to conform the similar behavior
conjugating from the entire surface. Oxidation or reduction of NDs is selected which
is based on the terminal functional groups required on the surface34,71,72. After oxidation, different oxidative agents give variable
functional group distribution on the surface. For example, nitric acid and
sulphuric acid result in carboxylate (COO-) rich surface, potassium
permanganate and sulphuric acid result in SO3- or O- derivatives of
phenol. Reduction transform most of the functional groups into hydrogen or
hydroxyl groups. Therefore, reduction of NDs through hydrogenation can create
either positive surface or negative surface through hydroxylation of ND
surfaces. There are three different types of surface chemistries: wet
chemistry, gas phase methods or atmospheric plasma treatments. In Wet chemistry
treatment, there is use of appropiate solvent systems to introduce the
functional groups. Depending on attached functional on the surface, oxidized
carboxylated NDs or reduced hydroxylated NDs can be used. Carboxylate
functionalized NDs can form acyl chloride functionalities on reacting with
thionyl chloride which can be further attached to amine-containing chemical moieties7,34,75. The treatment of NDs in gas or a vapor reactive medium is other different approach. The gas phases
can include the use of hydrogen, ammonia, carbon tetrachloride or argon. On
treating the NDs with ammonia yield carbonyl, amine or cyano groups on the surface
of ND, while on treatment with chlorine results in the production of chloro-NDs
or acylchloride functionalized NDs.
Functionalized with amino acids and alkyl chains via covalent bonding or
alkyl-, amino-, and amino acid-functionalized diamonds have been created by
chemical modification of fluorinated NDs with alkyl lithium, ethylenediamine,
or glycine ethyl ester hydrochloride, respectively. Another approach for
improvig the surface is atom transfers radical polymerization,
when radical initiators (benzoyl peroxides, hydroxyethyl-2-bromoisobutyrate or 2, 2, 2-trichloroethanol) are attached covalently to
oxidized NDs through esterification. Chemical groups are then introduced in the
system which polymerize and arrange as brush arrays on the surface15,70. This process can produce hydrophilic or hydrophobic surface
depending of the nature of polymer. Radical generation mechanism is used for
successful grafting of carboxylic groups onto NDs. Different functional groups
on the surface of NDs give possibilities for their conjugation with different
moieties without compromising the useful properties of the diamond core. Since the 1960s all traditional deaggregation techniques known in colloidal
science/materials have consistently failed to yield single-digit NDs
from DND aggregates. The problem was summarized by E Osawa: ‘The aqueous slurry of micron nanodiamond aggregates can be disintegrated into 60 nm
aggregates but never beyond by means of powerful 400 W
ultrasound’5 (Table2). Intense
aggregation is mainly explained by richsurface chemistry and small size of DNDs. The existence of various functional groups on the ND
surface, such as carboxyl, hydroxyl, lactone, etc, may result in the production
of multiple hydrogen bonds and even covalent bonds between the adjacent DND
particles, making it difficult to separate them. DND
primary particles are?5 nm in diameter, thus
many biological studies lead with DND aggregates of 100–200
nm can hardly reflect the performance of single-digit ND particles.

shape of nanomaterials also has a great impact on their application as a
therapeutic platform . For example, the use of spherical shape graphene oxide
is more advantageous for photothermal ablation of tumors as compared to needle-like
shape graphene oxide. And in this respect too, nearly spherical shaped primary DND
particles are beneficial as compared to the elongated, thin and sharp shapes of
other nanocarbons. The problem of ND deaggregation into single-digit particles was solved only in 2005 via
bead-assisted ball milling (table2) and its successor
bead-assisted sonic disintegration (BASD), (
table 2). A few years later, salt-assisted dry attrition milling (,
table 2) became available. Most recently, the salt-assisted
ultrasonic deaggregation has been added
to the arsenal, opening avenues to easily produced, inexpensive, ultra-pure
single-digit ND colloids for a multitude of applications (table

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