The ripening of new diamond based bio-mechanically active hybrid nano-structured scaffolds for cartilage cells tissue engineering are proposed in this study. Innovative tissue engineering biomimetic materials based on hydrogel have shown appealing physical, biological and technical properties in several biomedical applications A highly biocompatible romance hybrid akin based on nanodiamonds and hydrophilic poly-(hydroxyl-ethyl-methacrylate) (pHEMA) is proposed
Nano-Diamond Hybrid Materials for Structural Biomedical Application
Biomaterials are today playing a cash role in tissue engineering and regenerative medicine applications The fellowship 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 bob the diverse 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 command to profit in life expectancy and life level (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 further thermodynamically stable at ambient temperatures and pressures, while diamond, in these conditions, may exist only in its metastable area In fact, due to the high-energy hurdle that separated the graphitic sp2 and diamond sp3 configurations (Fig 1A and B), big temperatures and pressures in presence of catalysts are required to modify graphite in diamond
Nevertheless, a third parameter (surface area) becomes decisive at the nanoscale superiority and it become allied in the definition of the method equilibrium delectation levels: At this nano-dimensions, the Gibbs free liveliness becomes dependent on the contribution of the surface energy, highest to changes in the thermodynamic equilibrium phase diagram (Barnard et al., 2003; 2007; Viecelli et al, 2001) Tetrahedral hydrocarbons in the lair of nano-diamonds of 3 nm own been demonstrated by atomistic models to be more stable than poly-aromatics graphite (Fig 1C).
In addition, a fresh 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 eventuate even at room temperature and oblige paramount to the formation of diamond cores with graphitic shells (bucky-diamond) (Fig 1C) (Barnard and Sternberg, 2007)
These findings allowed us to surmise that the nanodiamond surfaces can be then soft modified through the chemistry of graphitic carbon in many different chemical methods, such are the DielsAlder cycloaddition reactions between conjugated diene and dienophile, to form functionalised cyclohexene systems (Jarre et al., 2011).
This new level of materials based on Carbon Sp2 and Sp3 nanocrystalline structures is very alluring for future nanotechnological incubation 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 posses been initially utilized in applications such as galvanic coatings, polishing systems, polymer nano-composities, lubricants New cranny applications, however, are recently developing; magnetic recording, adsorbents, diamond ceramics production, coatings in territory emission devices, catalyzes of heterogeneous catalysts and in fuel cells as proton-conducting nanocomposite membranes Preliminary appraisal demonstrated that detonation nanodiamonds are non-toxic and biocompatible, manufacture them extraordinary enticing for bio-medical applications considering its feasible controllable virile 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 tall surface chemical purity and uniformity surfaces are necessary for biomedical applications (Lai and Barnard, 2011a; 2011b) A unworldly cleansing fashion utilizes oxidation procedures Depending on the genus 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 donate as diamond can be rarefied up to 95% by responsibility by oxidation at big 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 naive air/ozone purification, then, produces carboxylated nano-diamond with highly reactive and hydrophilic surface OH terminations embezzle in biomedical applications (Krueger et al, 2008; Kruger et al, 2006)
Diamond and transparent carbon has been recognized in literature, however, the toxicity of nano-diamonds remains a TRUE concern (Schrand et al., 2009) In vitro and in vivo studies are still obligatory to evaluate characteristics such as in vivo practical 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 adaption activity
Previous investigations of our band own shown that big standard of biocompatibility and bioactivity has been practical for nano-composite materials made combining dim 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 posses shown a colossal inactive for usage in advanced creation models finalized to the knob of well-organized tissue engineered structures (Petrescu and Calautit, 2016 a-b)
Bone scaffolds retain been always a relevant debate for research since they should provide sufficiently strained but resilient trellis to be an ideal scaffold that momentarily improvised the damaged bone. Nevertheless, they should be able at the alike time to happily biodegrade after the formation of the new tissue in behest to quite 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 team hold investigated hydrogel hybrid composites, based on the partnership of pHEMA with Amorphous Pyrogenic Silica that were tested for the intake of water, the tally of knob in soak and in salted clue and for the cell passion with assays of adhesion, morphology, distribution, using fibroblasts and osteoblasts as cell-models The presence of the silica makes this biomaterials excellent, with duty to the pHEMA alone. Good properties of osteoinduction keep been also empirical for differentiation of dental blend originate 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; Tyrsa et al., 2001; Wolff, 1892)
Silica nano-composites synthesized in our laboratory, which contained highly-bioactive shadowy fumed, obtain been found to represent a new class of hybrid polymeric-ceramic scaffolding materials able to echo the scientific behavior of the bone Micro-foamed self assembled nanostructured composite posses been tested as scaffold that showed osteoblast develop knack and arise 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 thickness ranged between 3-5 nm and which specific surface state was of 400 m2g1, were utilized as bioactive filler. HEMA monomers (Fig 2) have been thermally polymerized in presence of an initiator for extreme polymerization, namely, the – azoisobutyrronitrile (AIBN), obtained from Fluka Milan, Italy In a preliminary check of nanocomposite preparation, the nanodiamond were diverse in the percentage of 5% by volume with the HEMA monomers and degassed The combination was then poured into 2.0 mm thick planar moulds before polymerization in the oven that was hug at the controlled temperature of 60C for 24 h. The nano-composite plates were subjected to a latter post-cure at 90C for 1 h
Results and Discussion
Nano-diamonds dispersion in the HEMA monomer resulted in a transparent and clear, illuminate grey colour, answer This behaviour testified the profit dispersion and need of nanofiller clusters The profit dispersion bent of the Oxidized Detonation nano-diamonds in the reacting concoction could be attributed to the strong interactions between the oxygen containing functional groups on the padding and the HEMA hydroxyl that led to the preferential self-assembly orientation of the monomers toward the nano-filler surface (Fig 3 detail upper left) The sequential polymerization of the HEMA resulted in a torpid decided and glassy transparent oppressive The wellbeing dispersion of the nano-diamond was quite 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; Tyrsa et al, 2001; Wolff, 1892)
A alike self gathering condition has been described by Aversa et al (2016; 2009) to befall between indistinct nanosilica particles, which are characterized by a disordered shelf containing many not average rings and not bridging Oxygen atoms (red in Fig. 4) and the alike HEMA monomer
The polymerization of HEMA/amorphous nanosilica mixtures leads to the formation of a hybrid nanostructured applicable with particularly outlandish and improved technical properties and biocompatibility (Aversa, 2016)
In the time of nono-diamond filled pHEMA, the identical improvement of the specialist properties and biocompatibility could be then expected. However, the expected technical properties enhancements could be much additional relevant due to diamond much higher rigidity and tightness (Azo tech spech)
The shear Modulus of synthetic diamond, which ranges from 440 to 470 GPa (Azo tech information), is nearly 15 times higher than that of Silica, which ranges from 27.9 to 32.3 (Azo tech spec) According to this news and considering the scientific 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 pertinent could be evaluated Fig 5.
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 cushioning words 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 colony at increasing cargo of nanosilica stuffing This episode confirmed the hybrid mood 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, reported as grey sector in Fig 5). Similar contact hold been described by Aversa et al (2016; 2009) to eventuate 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 quiescent improved bone scaffold mineralization and ossification properties hold been developed by sequential 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 wrapping concentration up to 5% by volume. Moreover, this transparent hybrid relevant swells to rubber in presence of aqueous physiological answer picking-up further than 40% of dampen At very low levels of nano-diamond loading, the specialist behaviour of the proposed hybrid materials could be comparable with that of bone when in the pellucid state, or to that of cartilage and ligaments when in the rubbery territory later moisten sorption
The use as scaffolds of these mechanically compatible hybrid hydrogels is expected to rectify the adaptation mechanisms of the bone by introducing an active interface that could renovate 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 profit of use of biomechanically compatible and bioactive scaffold biomaterials associated to new motif 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 friendly 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 besides appreciate the research technicians,Basim Al Turkiand 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://thescipubcom/abstract/103844/ajbbsp2017.34.41