Biomaterials are today playing a central role in tissue engineering and regenerative medicine applications. The collaboration between engineers, chemists, physicists, biologists and physicians speeded up research in this field, allowing a faster development of new biomaterials and technologies to overcome the challenges and to front the various needs of each specific tissue-engineering field. Although many biomaterials applications are far from clinical translation, regenerative medicine has greatly advanced during the last years and it bodes well for future translation of research discoveries from bench-to-the-bedside, in order to gain in life expectancy and life quality (Montheard et al., 1992; Filmon et al., 2002; Davis et al., 1991; Kabra et al., 1991; Apicella et al., 1993; Peluso et al., 1997; Petrescu et al., 2016 a-e).
Among the different allotropic forms of Carbon, graphite is the more thermodynamically stable at ambient temperatures and pressures, while diamond, in these conditions, may exist only in its metastable state. In fact, due to the high-energy barrier that separated the graphitic sp2 and diamond sp3 configurations, high temperatures and pressures in presence of catalysts are needed to transform graphite in diamond.
Nevertheless, a third parameter (surface area) becomes crucial at the nanoscale level and it become relevant in the definition of the system equilibrium energy levels: At this nano-dimensions, the Gibbs free energy becomes dependent on the contribution of the surface energy, leading to changes in the thermodynamic equilibrium phase diagram (Barnard et al., 2003; 2007; Viecelli et al., 2001). Tetrahedral hydrocarbons in the form of nano-diamonds of 3 nm have been demonstrated by atomistic models to be more stable than poly-aromatics graphite.
In addition, a more complex morphological structure is generated at the nanodiamond interface; Barnard and Sternberg (2007) reported that cuboctahedral clusters presented a transition from Sp3 to Sp2 carbons at the surfaces of aggregations of 1.0-3.0 nm (Aversa et al., 2016 a-o, 2017 a-e; Mirsayar et al., 2017).
On this morphological transition at the interface, it has been recently demonstrated by Xiao et al. (2014) that reversible nanodiamond-graphitic carbon onion like phase transformation can occur even at room temperature and pressure leading to the formation of diamond cores with graphitic shells (bucky-diamond) (Barnard and Sternberg, 2007).
These findings allowed us to understand that the nanodiamond surfaces can be then easily modified through the chemistry of graphitic carbon in many different chemical methods, such are the Diels–Alder cycloaddition reactions between conjugated diene and dienophile, to form functionalised cyclohexene systems (Jarre et al., 2011).
This new class of materials based on Carbon Sp2 and Sp3 nanocrystalline structures is very attractive for future nanotechnological development in biomedical structural applications. Nanocrystalline particles, which are often named detonation nanodiamond and characterized by sizes of 3-6 nm, are produced by detonation of carbon explosive materials (Danilenko, 2004; Greiner et al., 1988; Ozawa et al., 2007; Chang et al., 2008).
Detonation nanodiamond have been initially utilized in applications such as galvanic coatings, polishing systems, polymer nano-composities, lubricants. New niche applications, however, are recently developing; magnetic recording, adsorbents, diamond ceramics production, coatings in field emission devices, catalyzes of heterogeneous catalysts and in fuel cells as proton-conducting nanocomposite membranes. Preliminary investigation demonstrated that detonation nanodiamonds are non-toxic and biocompatible, making them very attractive for bio-medical applications considering its easy controllable rich surface chemistry.
However, it has been reported that detonation nanodiamonds may be characterized by different levels of purity and by the presence of several undesired functional groups/elements at the diamond particles surface, while high surface chemical purity and uniformity surfaces are needed for biomedical applications (Lai and Barnard, 2011a; 2011b). A simple purification method utilizes oxidation procedures. Depending on the type of procedure, the detonation powder of different levels of purities and specific surface characteristics can be obtained. The fraction of the Carbon that is not present as diamond can be purified up to 95% by weight by oxidation at high temperatures in air/Ozone atmosphere (Osswald et al., 2006; Shenderova et al., 2011).
Oxidation, while removing undesired processing functional compunds at nanodiamond surfaces, forms oxygen-containing groups, such are anhydrides and carboxylic acids (Shenderova et al., 2011).
The simple air/ozone purification, then, produces carboxylated nano-diamond with highly reactive and hydrophilic surface OH terminations appropriate in biomedical applications (Krueger et al., 2008; Kruger et al., 2006).
Diamond and glassy carbon has been recognized in literature, however, the toxicity of nano-diamonds remains a real concern (Schrand et al., 2009). In vitro and in vivo studies are still needed to evaluate characteristics such as in vivo mechanical and physiological behaviours (Zhang et al., 2011; Schrand et al., 2009a; 2009b; Yuan et al., 2010; Mohan et al., 2010) as well as cell viability or undesired gene modification activity.
Previous investigations of our group have shown that high level of biocompatibility and bioactivity has been observed for nano-composite materials made combining amorphous silica nanoparticles of about 7 nm.
Bioengineering and nanotechnology applied to micro and nano-materials are being progressively adopted as emerging solutions in 2D (coatings) and 3D applicatons (scaffolds) (Sorrentino et al., 2007; Aversa et al., 2016a). Conclusively, such micro and nano-technologies have shown a high potential for usage in advanced manufacture models finalized to the growth of well-organized tissue engineered structures (Petrescu and Calautit, 2016 a-b).
Bone scaffolds have been always a relevant matter for research since they should provide sufficiently rigid but resilient network to be an ideal scaffold that momentarily substitute the damaged bone. Nevertheless, they should be able at the same time to readily biodegrade after the formation of the new tissue in order to fully integrate with it (Kabra et al., 1991; Montheard et al., 1992; Peluso et al., 1997; Schiraldi et al., 2004; Buzea et al., 2015; Aversa et al., 2016 a-o).
Our research group have investigated hydrogel hybrid composites, based on the association of pHEMA with Amorphous Pyrogenic Silica that were tested for the intake of water, the balance of swelling in water and in saline solution and for the cell response with assays of adhesion, morphology, distribution, using fibroblasts and osteoblasts as cell-models. The presence of the silica makes this biomaterials excellent, with respect to the pHEMA alone. Good properties of osteoinduction have been also observed for differentiation of dental pulp stem cells (Abdul-Razzak et al., 2012; Ajith et al., 2009; Ahmed et al., 2011; Apicella and Hopfenberg, 1982; Atasayar et al., 2009; Babaev et al., 2010; Chow et al., 2010; Comerun, 1986; Covic et al., 2007; Frost, 1964, 1990, 1994, 2003; Gramanzini et al., 2016; Holley et al., 1970; Krueger and Boedeker, 2008; Nicolais et al., 1984; Petrescu et al., 2015; Prashantha et al., 2001; Raffaella and Antonio, 2016; Raffaella et al., 2016; Sorrentino et al., 2009; Töyräsa et al., 2001; Wolff, 1892).
Silica nano-composites synthesized in our laboratory, which contained highly-bioactive amorphous fumed, have been found to represent a new class of hybrid polymeric-ceramic scaffolding materials able to mimic the mechanical behavior of the bone. Micro-foamed self assembled nanostructured composite have been tested as scaffold that showed osteoblast grow ability and stem cells differentiation (Marrelli et al., 2015).
Materials and Methods
The monomer 2-hydroxyethylmethacrylate (HEMA), obtained from Sigma-Aldrich Chemicals Co., St. Louis, MO, USA, has been used for the polymerization of a hydrophilic composite matrix. Raw detonation nanodiamonds (Aldrich, ≥ 97%), which mean diameter ranged between 3-5 nm and which specific surface area was of 400 m2⋅g−1, were utilized as bioactive filler. HEMA monomers have been thermally polymerized in presence of an initiator for radical polymerization, namely, the α-α’ azoisobutyrronitrile (AIBN), obtained from Fluka Milan, Italy. In a preliminary test of nanocomposite preparation, the nanodiamond were mixed in the ratio of 5% by volume with the HEMA monomers and degassed. The mixture was then poured into 2.0 mm thick planar moulds before polymerization in the oven that was hold at the controlled temperature of 60°C for 24 h. The nano-composite plates were subjected to a final post-cure at 90°C for 1 h.
Results and Discussion
Nano-diamonds dispersion in the HEMA monomer resulted in a transparent and clear, light grey colour, solution. This behaviour testified the good dispersion and absence of nanofiller clusters. The good dispersion ability of the Oxidized Detonation nano-diamonds in the reacting mixture could be attributed to the strong interactions between the oxygen containing functional groups on the filler and the HEMA hydroxyl that led to the preferential self-assembly orientation of the monomers toward the nano-filler surface. The successive polymerization of the HEMA resulted in a still clear and transparent glassy solid. The good dispersion of the nano-diamond was reasonably preserved after the polymerization (Abdul-Razzak et al., 2012; Ajith et al., 2009; Ahmed et al., 2011; Apicella and Hopfenberg, 1982; Atasayar et al., 2009; Babaev et al., 2010; Chow et al., 2010; Comerun, 1986; Covic et al., 2007; Frost, 1964, 1990, 1994, 2003; Gramanzini et al., 2016; Holley et al., 1970; Krueger and Boedeker, 2008; Nicolais et al., 1984; Petrescu et al., 2015; Prashantha et al., 2001; Raffaella and Antonio, 2016; Raffaella et al., 2016; Sorrentino et al., 2009; Töyräsa et al., 2001; Wolff, 1892).
A similar self assembly condition has been described by Aversa et al. (2016; 2009) to occur between amorphous nanosilica particles, which are characterized by a disordered structure containing many not regular rings and not bridging Oxygen atoms and the same HEMA monomer.
The polymerization of HEMA/amorphous nanosilica mixtures leads to the formation of a hybrid nanostructured material with particularly peculiar and improved mechanical properties and biocompatibility (Aversa, 2016).
In the case of nono-diamond filled pHEMA, the same improvement of the mechanical properties and biocompatibility could be then expected. However, the expected mechanical properties enhancements could be much more relevant due to diamond much higher rigidity and strength (Azo tech spech).
The shear Modulus of synthetic diamond, which ranges from 440 to 470 GPa (Azo tech information), is almost 15 times higher than that of Silica, which ranges from 27.9 to 32.3 (Azo tech spec.). According to this information and considering the mechanical shear behaviour of the analogous hybrid materials based on silica nanoparticles (Aversa et al., 2016), the behaviour of the variation of the shear modulus as a function of the diamond nanoparticles volume fraction in the hybrid material could be evaluated.
According to Aversa et al. (2016), strong plasticization is induced by the physiological solutions sorption in the hybrid pHEMA-nanosilica composite.
It has been described by Aversa et al. (2016; 2009) that the measured shear modulus of the Nanosilica hybrid composites at different filler content was not described by the classical Halpin and Kardos (1976) equation that is commonly utilized for the particulate composites. The hybrid nano-composites showed a linear dependency at increasing contents of nanosilica filler. This occurrence confirmed the hybrid nature of the nanosilica filled pHEMA.
At nano-diamond volumetric fractions ranging from 2 to and 5%, the shear moduli were comparable to those of the cortical bone (10-20 GPa). Similar results have been described by Aversa et al. (2016; 2009) to occur for nanosilica hybrids at higher loading ranging from 15 to 30% by volume.
New bioactive nanodiamond-polymeric hybrid materials to be used as biomechanical active scaffold materials showing potential improved bone scaffold mineralization and ossification properties have been developed by following a biomimetic approach.
The new nanocomposites based on poly-Hydroxyl-Ethyl-Methacrylate (pHEMA) filled with detonation nanodiamonds could be identified as a biomimetic biomaterial at filler concentration up to 5% by volume. Moreover, this glassy hybrid material swells to rubber in presence of aqueous physiological solution picking-up more than 40% of water. At very low levels of nano-diamond loading, the mechanical behaviour of the proposed hybrid materials could be comparable with that of bone when in the glassy state, or to that of cartilage and ligaments when in the rubbery state following water sorption.
The use as scaffolds of these mechanically compatible hybrid hydrogels is expected to improve the adaptation mechanisms of the bone by introducing an active interface that could ameliorate biomimetics by correctly reproducing cartilage and ligaments biomechanical functions (Schwartz-Dabney and Dechow, 2003; Perillo et al., 2010; Apicella et al., 2010; 2011; 2015; Aversa et al., 2016; 2009).
Adaptive properties of bone could benefit of use of biomechanically compatible and bioactive scaffold biomaterials associated to new design odontostomatological prostheses.
We acknowledge and thank Mr Taher M. Abu-Lebdeh, Associate Prof at North Carolina A and T State Univesity, United States and Mr Muftah H. El-Naas PhD MCIC FICCE QAFCO Chair Professor in Chemical Process Engineering Gas Processing Center College of Engineering Qatar University and Ms Shweta Agarwala, Senior Research Scientist at Singapore Center for 3D Printing Nanyang Technological University Singapore for their suggestions and comments. The Authors acknowledge Liquid Metals Technologies Inc, Ca USAthat kindly supply the samples for the characterization and Dr Francesco Tatti (FEI Company Application Specialist SEM-SDB) for its contribut in the preparation of this paper experiments and analyses. The authors would like to appreciate the facilities and assistance provided by the Advanced Technology Dental Research Laboratory, Faculty of dentistry, King Abdul Aziz University. The authors would also appreciate the research technicians, Basim Al Turki and Fahad Al Othaibi for their cooperation.
This research was partially funded by Italian Ministry of University and Research with the project FIRB “Future in Research 2008”, # RBFR08T83J.
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See the article with Figures at: http://thescipub.com/abstract/10.3844/ajbbsp.2017.34.41
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ABOUT THE AUTHOR
Ph.D. Eng. Relly Victoria V. PETRESCU
Senior Lecturer at UPB (Bucharest Polytechnic University), Transport, Traffic and Logistics department,
Date of birth: March.13.1958;
Higher education: Polytechnic University of Bucharest, Faculty of Transport, Road Vehicles Department, graduated in 1982, with overall average 9.50;
Doctoral Thesis: “Contributions to analysis and synthesis of mechanisms with bars and sprocket”.
Expert in Industrial Design, Engineering Mechanical Design, Engines Design, Mechanical Transmissions, Projective and descriptive geometry, Technical drawing, CAD, Automotive engineering, Vehicles, Transportations.
Member ARoTMM, IFToMM, SIAR, FISITA, SRR, SORGING, AGIR.प्रकाशित मिति २२ भाद्र २०७७, सोमबार ०७:४९