Constitutes of cement.
\r\n\t
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He has carried out a great deal of research and technical survey work as well as several studies in the aforementioned areas with over 170 published international academic works in different languages.",coeditorOneBiosketch:null,coeditorTwoBiosketch:null,coeditorThreeBiosketch:null,coeditorFourBiosketch:null,coeditorFiveBiosketch:null,editors:[{id:"110471",title:"Dr.",name:"Amjad",middleName:"Zaki",surname:"Almusaed",slug:"amjad-almusaed",fullName:"Amjad Almusaed",profilePictureURL:"https://mts.intechopen.com/storage/users/110471/images/system/110471.png",biography:"Amjad Almusaed was born in 1967. He holds a PhD degree in Architecture (Environmental Design) from Ion Mincu University, Bucharest, Romania. He completed postdoctoral research in 2004 on sustainable and bioclimatic houses, from the School of Architecture in Aarhus, Denmark. His research expertise is sustainability in architecture and urban planning and design. He has carried out a great deal of research and technical survey work, and has performed several studies in the above-mentioned areas. He has edited many international books and is an active member of many worldwide architectural associations. He has published more than 170 international academic works (papers, research, books, and book chapters) in different languages.",institutionString:"Jönköping University",position:null,outsideEditionCount:0,totalCites:0,totalAuthoredChapters:"8",totalChapterViews:"0",totalEditedBooks:"9",institution:{name:"Jönköping University",institutionURL:null,country:{name:"Sweden"}}}],coeditorOne:null,coeditorTwo:null,coeditorThree:null,coeditorFour:null,coeditorFive:null,topics:[{id:"11",title:"Engineering",slug:"engineering"}],chapters:null,productType:{id:"1",title:"Edited Volume",chapterContentType:"chapter",authoredCaption:"Edited by"},personalPublishingAssistant:{id:"259492",firstName:"Sara",lastName:"Gojević-Zrnić",middleName:null,title:"Mrs.",imageUrl:"https://mts.intechopen.com/storage/users/259492/images/7469_n.png",email:"sara.p@intechopen.com",biography:"As an Author Service Manager my responsibilities include monitoring and facilitating all publishing activities for authors and editors. 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Theophanides",coverURL:"https://cdn.intechopen.com/books/images_new/1591.jpg",editedByType:"Edited by",editors:[{id:"37194",title:"Dr.",name:"Theophanides",surname:"Theophile",slug:"theophanides-theophile",fullName:"Theophanides Theophile"}],productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}}]},chapter:{item:{type:"chapter",id:"39539",title:"Lipoprotein Structure and Dynamics: Low Density Lipoprotein Viewed as a Highly Dynamic and Flexible Nanoparticle",doi:"10.5772/48145",slug:"lipoprotein-structure-and-dynamics-low-density-lipoprotein-viewed-as-a-highly-dynamic-and-flexible-n",body:'Low density lipoproteins (LDLs) are the principal transporter of cholesterol and fat in human blood. Circulating LDLs guarantee a constant supply of cholesterol for tissues and cells, whereas cholesterol is required for membrane synthesis, modulation of membrane fluidity and the regulation of cell signaling pathways. The function of LDL in metabolism is mediating by cellular uptake via receptor-mediated endocytosis followed by lysosomal degradation [1,2], and is strongly dependent on the lipid distribution, the structure of LDL particles and on the proper conformational orientation of apolipoprotein B100 (apo-B100). Apo-B100 is the sole protein component of LDL being mainly located on the surface of LDL. Apart from their well established role as lipid transporter, LDL particles are intimately involved in the progression of cardiovascular diseases such as atherosclerosis or stroke, which are among the most prevalent causes of death in developed countries [3]. In particular, raised plasma levels of LDL are linked to an increased risk for disease. Moreover, dysregulations of LDL due to abnormalities in LDL structure have been identified as independent predictors of risk for coronary heart diseases [4,5]. LDL particles by themselves are highly heterogeneous in nature, varying in their buoyant densities, size, surface charge and chemical composition [6,7], and these biochemical characteristics determine the fate of LDL in the subendothelial space [8,9]. For example, small, dense LDL subclasses are more atherogenic than their light counterparts, which are more susceptible to modifications [5,10]. Modifications of LDL, primarily through oxidation, enzymatic degradation or lipolysis are the initiating factors in early atherosclerosis. In that case, LDL particles accumulate in the intima of the arterial wall where apo-B100 binds to proteoglycans of the extracellular matrix through ionic interactions. As a consequence, LDL becomes trapped in the subendothelium, where it is prone to oxidation processes, aggregation and fusion. Bioactive lipids, such as oxidized phospholipids, lysolipids or oxidized cholesterylester, are released from LDL particles, which are simultaneously non-specifically altered. A broad spectrum of diverse LDL particles with non-defined physicochemical properties is generated that, in turn, promotes a rapid uptake of these particles by macrophages to form foam cells [11]. This is one of the key steps in the progression of atherosclerosis. Today, atherosclerosis is known to be a chronic inflammatory disorder of the blood vessels and recognized as a prevailing cause of cardiovascular disorders, the leading causes of morbidity and mortality worldwide [12]. Since the early initiation of atherosclerosis strongly depends on the metabolism of LDL, which is predominantly triggered by molecular characteristics of LDL, it is of paramount biomedical importance to explore structural features of LDL particles in great detail. However, mostly due to the complex nature of LDL particles many questions concerning molecular details are still unanswered.
This article will review our current knowledge on the structure and dynamics of LDL particles. In fact, several recent studies revealed that the molecular organisation and dynamics of LDL core lipids, in close relationship to the intrinsic dynamics of LDL surface components, control not only the metabolism of lipids in humans, but determine the role of LDL in the pathogenesis of cardiovascular diseases. In this article, we will give a short historical review on LDL structure and then present prevailing concepts on the self-organisation of LDL. Special emphasis will be paid to dynamic features of LDL particles. In particular, we will discuss the interplay between structure and dynamics in more detail. Finally, we will give an outlook to promising future strategies to clarify the molecular structural details of LDL and how to exploit LDL nanoparticles for medical needs.
LDLs are composed of lipids and protein, which assemble to form a supramolecular complex with a molecular mass exceeding 2.5 - 3.0 million Da and involving 2000 to 3000 lipid molecules. Thus, LDL particles are commonly described as micellar complexes, macromolecular assemblies, self-organized nanoparticles or microemulsions. Regardless of diverse definitions, it is generally accepted that assembled LDL particles are organized into two major compartments, namely an apolar core, comprised primarily of cholesteryl esters (CE), minor amounts of triglycerides (TG) and some free unesterified cholesterol (FC). The core is surrounded by an amphipathic outer shell. This shell is composed of a phospholipid (PL) monolayer containing the larger part (>2/3) of the FC molecules and one single copy of apo-B100, which is one of the largest known monomeric glycoproteins [13]. Figure 1 provides an overview on characteristic properties of LDL together with a schematic presentation of an LDL particle. Since molecular interactions between different kinds of lipids have turned out to be highly complex, it is almost impossible to separate the surface and core regions exactly from each other. Accordingly, in some recent reports an additional hydrophobic interfacial layer composed of phospholipid acyl chains, FC, some CE molecules and hydrophobic protein domains is defined. This description takes account for the interplay between neutral core lipids and the surface layer [14].
Molecular organisation of LDL. LDL particles are isolated from human plasma within a defined density range. Their particle size varies between 20 to 25 nm. LDLs are built up by a hydrophobic lipid core of cholesterylester (CE) and triglyceride (TG) molecules, which make up more than 40% of particle mass surrounded by a phospholipid (PL) monolayer corresponding to about 20% of particle mass. Varying amounts of free cholesterol (FC) are incorporated in the shell and the core regions. One single copy of apo-B100 (550 kDa) is embedded in the surface monolayer, partially penetrating the core and covering about 40 to 60% of the surface area. The carbohydrate moieties are distributed along the protein chain and are surface exposed. The N-terminal end of apo-B100 (about 26% of total) is hydrophilic and shows a high homology to lamprey lipovitellin. The C-terminal end was shown to be located close to the N-terminus.
Since LDL particles are highly heterogeneous, especially with respect to the chemical composition of the core lipids, the actual size of LDL particles varies between 20 to 25 nm, with an average particle diameter of about 22 nm. This intrinsic heterogeneity allows a subdivision of LDLs into distinct highly homogeneous LDL subspecies, which are identified on the basis of their hydrated densities, which normally lies between the extremes of d, 1.019 and 1.063 g ml-1 [15]. These subspecies also differ in their physico-chemical characteristics, receptor binding affinity [16], susceptibility to oxidative modifications [17,18], and in their atherogenic behaviour. Following these lines, it is important to consider LDL as a flexible construct, which needs to respond to changing environmental conditions during lipid exchange. Hence, during particle remodelling, apo-B100 and the surface PL molecules have to rearrange to compensate for changes in the surface area and surface pressure [6]. It is known, that apo-B100 predominantly resides on the surface of LDL and displaces PL molecules, concomitantly changing the diffusion and order parameter of lipids as shown in a recent near atomistic simulation study [19]. Based on simple geometrical considerations taking into account the surface PL monolayer (about 700 PL molecules) with an average area per lipid of 0.65 nm2 and an LDL particle diameter of 22 nm, large parts of the surface layer must be covered by the protein to avoid unfavourable hydrophobic contacts. In support of these considerations, a loose surface packing of PL molecules was derived from molecular dynamics simulations [19]. This low surface pressure enables hydrophobic amino acid regions of apo-B100 to penetrate into the interfacial regions, predominantly formed by the acyl chains of PLs. Consequently, apo-B100 might interact more readily with the neutral core lipids, and indeed it was shown that some of the CE molecules align along the β-sheet structures of apo-B100 [20], thereby driving CE molecules to the surface, where they become part of the interfacial layer. Particularly noteworthy is the fact that the lipids within the interfacial layer are not homogeneously distributed but form local microenvironments [14]. More precisely, two nanodomains were identified, one rich on sphingomyelin and FC, the other one rich in phosphatidylcholine and poor in FC. The latter was shown to be associated more closely with apo-B100 [21]. Even though, one has to keep in mind that these domains are not static or confined in size and number and co-determine the intrinsic dynamics of LDL. Based on these types of findings, it seems reasonable to suggest that variations in the molecular organisation of lipid/apo-B100 impact the structure of LDL, and have to be considered to act as physiological determinants of LDL function.
Our present understanding of the structure of LDL particles has emerged from the concerted application of different physico-chemical techniques with early ground-breaking findings derived from neutron- or X-ray small angle scattering data [22-25] complemented by results from negative staining electron micoscopic (e.m.) [26,27] and spectroscopic techniques [28,29]. For comprehensive reviews on different biophysical studies applied on LDL species see refs. [30,31]. In recent years structural investigations using cryo-e.m. reconstruction techniques have become prevalent and with time 3-dimensional models with improved resolution were presented [32-37]. While in earlier studies LDLs are described as quasi-spherical particles, later studies presented a new view of the overall particle structure displaying an oblate elliptical particle shape. Moreover, recent 3D-images show convincing data that LDL can be considered as discoidal-shaped particle with two flat surfaces on opposite sides. In this model, apo B100 encircles LDL at the edge of the particle, while the PL monolayer is rather located at the flat surfaces which are parallel to the CE layers in the core [36,37]. To get a better impression of what LDL looks like in a structure map obtained by 3D-reconstruction from cryo-e.m, we show some images in Figure 2 revealing the surface density distribution on LDL. It has to be stated that this model strictly holds true for LDL particles with the core lipids being in a frozen liquid-crystalline state.
Density distribution at the surface of LDL. The 3D-density map derived from cryo e.m. images by reconstitution reveals the oblate overall particle shape of LDL shown in gray. The overlaid high density regions represent the backbone of apo-B100, colored in orange. The belt surrounds the particle to form an enclosed circle. The second group of high density regions (green) contours the rims and complements the backbone enclosing lower-density regions. The high density regions on the sidewall (yellow) are structures extending from the backbone. A knob-like protrusion is visible at the pointed end (indicated by triangles in the right and top views). The 3D-map is turned 90° in each frame. Reprinted with permission from ref. [
Despite of compositional heterogeneity, LDL particles share one common feature: the CE molecules in the core undergo a structural transition from an ordered liquid-crystalline phase to a fluid oil-like state as function of temperature and chemical composition [38]. More precisely, the actual transition temperature, which is close to body temperature, is inversely correlated to the content of triglycerides within the lipid core [22,39]. Based on these characteristics, several models for CE packing have been suggested including a spherical concentric layer model derived primarily from X-ray and neutron scattering data [40,41]. More recently, the concept of a flat lamellar structure came up. This model is derived from single-particle reconstructions from cryo-e.m. images of LDL in vitreous ice [32,34]. An ordered three-layer internal lamellar structure with a distance of about 3.6. nm between the single lamellae was reported [32], in agreement with repeat distances derived from X-ray scattering patterns for LDL below the transition temperature. While these images were observed for LDL particles being in the liquid crystalline phase before snap-freezing, diverse results were reported for LDL particles frozen from a state above the phase transition temperature [42,43]. One plausible explanation for these discrepancies might be that the melting rate of the core lipids proceeds extremely fast. It has been shown that the physical state of core lipids changes within milliseconds [44]. This fast kinetics has caused experimental difficulties for long time, however, a recent experimental approach by speeding up freezing allowed to trap the lipids in the molten state [45]. The authors report on a co-existing phase of layered and broken shells for LDL particles, which are shock-frozen in a state above the phase transition. This is the first time to visualize the nucleation process of CEs within LDL. Most interesting, the images indicate intermediate states between the order/disorder phase transition. Figure 3 shows the dynamic model of the core CE packing during the phase transition and gives a comparison of the internal features of reconstructed 3D-volumes of LDL.
Schematic picture of the dynamic model of LDL core lipid packing during the phase transition. Comparision of the internal features of the reconstructed 3D-volume of LDL snap-frozen from below (22°C) and above (53°C) the phase transition temperature (Tm). Samples prepared from 22°C show a layered organisation while samples prepared from 53°C reveal a disorded shell like structure, which is concentric to the surface. Note, the overall shape of LDL has also changed slightly. The lower panel shows a hypothetical model for the core lipid packing depicting the dynamic process of the core lipid phase transition upon cooling from isotropic to layered passing through an intermediate state. Modified with permission from ref. [
In summary, it seems reasonable to argue that both the overall shape and core lipid packing of LDL particles are highly sensitive to changes in temperature and lipid composition. Indeed, this newly proposed patch nucleation behavior permits the temporary formation of local molecular microenvironments as suggested previously by our group in terms of trigylceride segregation [46]. In the next paragraph we will address some interesting questions in support of above hypotheses.
Does a lipid microphase separation occur in LDL particles as a function of the relative core content of CE and TG ?
As already mentioned, the transition temperature correlates with the lipid composition, however, a discontinuity in the concentration dependence was observed [46]. A break in the concentration dependence of a transition temperature in a mixed lipid system constitutes an index for the existence of a phase separation at the break point. In isolated triglyceride - cholesteryl ester systems no indication of a phase separation at similar compositions was found [39,47]. It appears therefore, that structural constraints within the LDL particle determine this effect. Experimental data provide evidence that at low TG content (below 12%) the TG molecules separate into distinct hydrophobic nanoenvironments while the CEs form a smectic liquid crystalline layer. With increasing TG content the thermal stability of the CE layer is decreased by intermixing with TG [46]. This hypothesis implies that the TG-rich fluid nanodomains can serve as a reservoir for lipophilic minor constituents, such as vitamins (tocopherol, carotenoids etc.) below the phase transition. The local concentration of these antioxidants and hence their efficiency in scavenging lipophilic free radicals is higher than if they were dissolved in the bulk volume of total apolar lipids. At the same conditions the CE molecules are strongly immobilized and the intracellular degradation of LDL is decelerated [48], equally the activity of lipid transfer proteins is diminished [49,50]. Based on these considerations it is tempting to speculate that circulating LDL, as a consequence of the variation in blood temperature, periodically undergoes a thermal transition resulting in a transient increase in the local core concentration of minor constituents [46]. Here, it should be emphasized that a periodic redistribution of lipophilic solutes, and also for example of drugs, into the confined LDL core volume could represent an attractive approach to the modulation of biochemical reactions, which would not occur at sufficient rates under the normal conditions of relative concentration. Studies along these lines could indeed verify the long missing physiological role of the thermal LDL transition.
Can LDL structure follow quasi-isothermal changes in blood temperature during its circulation, or does it remain adiabatically metastable in the molten-lipid state?
In order to provide evidence to answer this question we have applied time resolved X-ray scattering experiments using a high flux synchrotron generated X-ray beam. Thus, we have been able to trigger the thermal transition in either direction (heating and cooling) simultaneously monitoring associated structural changes in sub-second time intervals. With our special instrumental setup we managed to evaluate the kinetics of core-transition by T-jump and T-drop experiments [44]. We found that the melting transition proceeds faster than 10 milliseconds indicating that thermal-induced lipid reorganisation takes place at the time scale of blood circulation. As the velocity of blood-flow can be as low as 0.3 mm/s in peripheral blood capillaries the residence time for LDL particles in cooler regions of the body can be several seconds. Consequently, LDL can easily follow periodic temperature changes during blood circulation and assist the redistribution of lipophilic constituents within its core nanodomains forming fluid defect zones. For biomedicine, this strengthens the hypothesis that the core lipids of LDL not only act as passive chemical substrates in metabolism, but that their physical state within the LDL nanoparticles has the potential to control their metabolic fate in normal and atherosclerotic cholesterol transport.
Does the core lipid transition have a physiological meaning ?
Despite its occurrence conspicuously close to blood temperature and the variation of the transition temperature of LDL among different subjects, no clear evidence for a physiological or patho-biochemical role of this transition has so far been found. It is now generally accepted in literature that the rearrangement of the core lipids also affects the overall structure and shape of the LDL particle. Morphological changes in turn can impact receptor-binding activity as well as the action of lipid hydrolyzing enzymes. Equally, the susceptibility of LDL particles to oxidative modifications and lipid peroxidation might be correlated to temperature [18]. As oxidized LDL play a crucial role in the pathogenesis of atherosclerosis, any contribution to the comprehension of antioxidant efficiency may be of therapeutic potential [2,51], further pointing to the physiological relevance of the lipid core organisation. However, this vital question still remains unanswered.
As already indicated above, the physicochemical state of the core lipids is intimately related to the structure and dynamics of the particle surface, which consists of about 700 phospholipid molecules and one single copy of apo-B100. Apo-B100 is a huge glycoprotein and its polypeptide chain consists of 4536 amino acid residues with an estimated molecular mass of about 550 kDa for the glycosilated form [52,53]. Apo-B100 is a single chain protein with a total contour length of about 70 nm [54] and can be viewed as a highly flexible molecular string composed of single domains [20]. Five consecutive domains were identified based on secondary structure elements representing the main conformational motifs of apo-B100. The single domains are connected by flexible interdomain linker regions, which allow relative movements of domains to each other. The feasibility of such motions was shown in a low resolution model of detergent solubilized apo-B100, which was derived from small angle neutron scattering data [55]. In this model, compact rigid domains are visible being connected by flexible interdomain linkages, which possess a substantial degree of freedom in their spatial orientation. A hypothetical spatial arrangement of the apo-B100 molecule on a spherical LDL particle was created after assigning the secondary structure elements, which were deduced from a secondary structure prediction, to the surface of apo-B100 (Figure 4). Likewise, the averaged surface shape of the 3D-model would allow for variations in the thickness of the apo-B100 molecule by about 1 nm. Such variations are most likely required to compensate for changes in the surface area upon lipid exchange and particle shrinking during endogeneous lipoprotein conversion from very low density lipoprotein (VLDL) to LDL.
Reconstituted low resolution model of lipid-free apo-B100 derived from small angle neutron scattering data. Apo-B100 shows an elongated arch-like morphology indicating single domains and mobile less defined linker regions. A hypothetical model of a spherical LDL particle after superposition of the structural model of apo-B100 is shown (adapted from ref. [
Concerning the topology of apo-B100 on the surface of LDL the most detailed information is obtained from cryo-e.m. images (see also Figure 2). Chatterton et al. were among the first to visualize apo-B100 as string circumventing LDL, and to report on mapped epitopes of apo-B100 distributed over one hemisphere of the LDL particle [56,57]. Recent single particle 3D-reconstructions from immuno cryo e.m. images delineated a more accurate picture of apo-B100 revealing a looped topology of the protein backbone with distinct epitopes identified along the protein chain. According to this model, epitopes in the LDL receptor binding domain are located on one side of LDL, whereas epitopes located in the N-terminal and C-terminal domains are in close vicinity to each other on the opposite side of LDL [36]. In addition, a prominent protrusion is visible in the images at the pointed end of the particle. A similar knob-like region was apparent in the low resolution model of lipid-free apo-B100 shown in Figure 4. This protrusion most probable represents the non-lipid associated globular N-terminal domain of apo-B100, which shows a high homology to lamprey lipovitellin [58]. Except for the N-terminal domain, little is known about the molecular organisation of the structural motifs, whose amphipathic nature determine lipid association. However, to evaluate lipid-protein interactions physical parameter like interfacial elasticity or molecular dynamics have to be considered. In this context, it was suggested that the hydrophobic β-sheet domains of apo-B100 act as elastic lipid anchors, whereas the amphipathic α-helical domains respond rapidly to changes in surface pressure [59,60]. In any case, it can be assumed that alterations in the adsorption and penetration depth of apo-B100 in the phospholipid monolayer and in the lipid core are accompanied by structural rearrangements of the domains and changes in the orientation of the domains relative to each other. In the course of such elastic motions, intramolecular rearrangements are likely to alter the overall hydrophobicity and surface activity of single protein domains. These modifications not only affect lipid-protein interaction, but are equally important for molecular and cellular recognition of apo-B100.
LDL particles are formed in the circulation by lipolytic conversion of TG-rich VLDL particles. This enzyme mediated endogenous transformation is accompanied by an extensive shrinking in particle size from about 50-80 nm for VLDL to ~20 nm for LDL. In the course of remodelling, apo-B100 remains bound to its nanocarrier stabilizing the lipid assembly by maintaining structural integrity. To accomplish this, apoB100 has to become more condensed or relaxed depending on the lipid packing density. Likewise, this dynamic process is modulated by the molecular mobility of the surrounding microenvironment. To test for this hypothesis we have recorded temperature dependent molecular motions in VLDL and LDL particles using elastic incoherent neutron scattering [61]. With this technique, motions in the nano- to picosecond time scale can be recorded. The calculated dynamic force constants are a direct measure for the resilience of the particles. The results show that at physiological temperatures VLDL particles are very soft, elastic and mobile as compared to LDL, which is more rigid (see Figure 5). This observation supports the notion that apo-B100 in VLDL is loosely packed at the interface covering a large surface area with low interfacial surface tension [59]. During particle conversion from VLDL to LDL, however, the relative number of surface molecules increases and a higher molecular packing density leads to a compression of the lipid anchored protein regions and an overall stiffening of the LDL particle [60].
Molecular motions in LDL and VLDL. Elastic temperature scans are recorded with elastic incoherent neutron scattering. The mean square thermal fluctuations (<u2>) are shown as function of temperature. The molecular resiliences are derived from the slopes in the curves. It is seen that VLDL has an increased motion at elevated temperatures compared to LDL. Parts of this figure are reproduced, with permission, from ref. [
To conclude, the intrinsic conformational flexibility and elasticity of apo-B100 containing lipoprotein particles is most likely critical for specific affinities of lipoproteins to receptors, antibodies or enzymes. Moreover, it would seem that the susceptibility of lipoproteins to oxidative modifications and hence their atherogenicity is influenced by their dynamic nature.
In the search for new and improved therapeutics, the field of nanomedicine dealing with functionalized nanoparticles for molecular imaging and therapy is rapidly emerging. Nanoparticles offer new opportunities to transfer active substances directly to the diseased site in the body. By additional surface coatings or functionalizations, the properties of nanoparticles can be tuned to specific needs. Within the last two decades, a variety of artificial nanoparticles have been designed for targeted delivery of drugs or contrast agents. Many of these nanoconstructs are developed for cancer therapy taking advantage of the
leaky vasculature of tumours. Apart from tumour targeting, increasing efforts are devoted to the treatment and imaging of atherosclerotic plaques (for a recent review see ref. [62]). Over time, a broad and versatile nanoparticle platform was created in which liposomes and biodegradable polymers have turned out to be the most promising candidates. It is important to mention that several nanomedicine products have already been established on the market and numerous products are successfully applied in clinical trials [63]. However, inherent problems of nanoparticles are biocompatibility and low stability in vivo, since most nanoparticles become rapidly cleared by the reticuloendothelial system. In contrast to artificial systems, lipoproteins are naturally occurring nanoparticles evading recognition by the body´s immune system. Hence, lipoproteins are excellent candidates with attractive properties to be considered as molecular transporters. A great advantage of LDL over other nanoparticles is the fact that LDL particles stay in circulation for several days, and are not cleared immediately by the mononuclear phagocyte system of the liver and spleen. The average lifetime of an LDL particle is 2-3 days and this time span is about three times longer as reported for long-circulating liposomes, currently applied in chemotherapy [64]. It was recognized that certain tumor cells overexpress LDL receptor, however, the targeting specificity is limited as the LDL receptor is ubiquitously expressed throughout the body, most prominent in the liver. However, using apo-B100 as inherent targeting sequence the enhanced circulation times in blood enable drug-loaded LDL particles to bind to specific receptors exposed on the surface of e.g. tumor or atherosclerotic plaque. Once recognized by the receptor, the functionalized LDL particles become internalized, accumulate in the tissue and exert an enhanced effect (reviewed by [65]). The intrinsic targeting properties of LDL to atherosclerotic plaques are already utilized for early diagnosis and detection of atherosclerotic lesions by different imaging modalities (for reviews see refs. [66,67]). However, to modify lipoprotein particles for medical purposes, care has to be taken not to compromise essential biophysical and structural features of LDL with the goal to preserve the biological activity. In general, there are several possibilities to create multi-functionalized lipoprotein particles. Some representative examples are shown in the scheme in Figure 7. One possibility is to load hydrophobic drugs (e.g. chemotherapeutics, antibiotics, vitamins, signal emitting molecules or small nanocrystals) in the lipophilic inner core of LDL. This can be accomplished by different techniques including lyophilisation, solvent evaporation and reconstitution procedures [68,69]. However, LDL particles can not be reconstituted so easily and remote drug/contrast agent loading into native lipoprotein particles is still a tedious approach currently not being standardized. Amphiphilic substances (drugs or marker molecules) or fatty acid modified chelator complexes can be incorporated in the PL monolayer [70,71]. This has successfully been done in numerous biophysical studies and for diagnostic purposes. Finally, the surface of LDL can be modified by protein labeling. This is done by covalent attachment of substances to the lysine and cysteine amino acid residues of apo-B100. Such substances include fluorophores, radionuclides or metal ions for molecular imaging [65]. Alternatively, targeting sequences (e.g. folic acid) can be coupled to apo-B100 with the purpose to reroute LDL to alternate receptors, which, in case of folate, are more specifically expressed in tumor cells [72].
Scheme giving some examples of how LDL particles can be modified to act as natural endogenous nanoparticles for targeted drug delivery or multifunctional molecular imaging.
To construct lipoprotein mimetic particles, also referred to as lipoprotein related particles, artificial lipoprotein particles have to be reassembled from individual lipid and protein entities. This approach was highly successful for high density lipoproteins using apo-AI mimetic peptide sequences [73]. For LDL, this approach was not pursued yet and will be much more complicated considering the complex dynamic nature of apo-B100.
Over the last few years, a promising nanoparticle platform was established, which exploits the endogenous properties of natural lipoproteins being non-toxic, non-immunogenic and biodegradable. Although this platform still offers vast potential for improvements, first promising results in enhanced multimodal imaging of tumors and atherosclerotic plaques are achieved giving hope that further endeavors to combine diagnostics and personalized therapeutics will also be successful.
The intrinsic flexibility and dynamics of LDL lipids and protein in conjunction with the inherent compositional heterogeneity of LDL particles has hitherto hampered successful structure determinations at atomic level. Recent technological developments, however, allowed to restore characteristic structural features of individual LDL particles at low resolution. In particular, using cryo e.m. 3D-reconstruction techniques several groups have succeeded in imaging morphological and topological details of LDL to a resolution limit of approximately 2 nm [34-36]. Now, new concepts will be needed to make further progress in the development of high resolution models of LDL. One promising way is to put stronger emphasis on protein crystallography in combination with computational modelling and molecular dynamics simulations. X-ray crystallography apprears to be a hopeless pursuit with heterogeneous and flexible particles like LDL. Nevertheless, our earlier attempts of crystallisation have been partially successful [74]. Additional efforts, however, have to be focussed on the stabilization of apo-B100 in a more rigid state, perhaps by co-crystallisation with monoclonal antibodies. An alternative way ahead would be to work with lipid-free apo-B100 stabilized by detergent-mimetic systems, e.g. amphipathic designer peptides, or to proceed with truncated fragments of apo-B100.
At present there is still a deficit in our knowledge concerning the molecular lipid trafficking mechanisms of LDL. To know the atomic structure of LDL, in particular of apo-B100, may well contribute to a better understanding of biologic aspects of cardiovascular diseases, especially with respect to future strategies towards rational pharmaceutical interventions.
AcknowledgementThis manuscript is based in part upon work supported by the Austrian Science Fund under grant number P-20455.
There is no other substitute for concrete that can be supplemented with an alternative because of its intrinsic qualities like get into any shape, quality and ability to consume locally available fine and coarse materials along with its strength, resistance to fire, with little support [1]. The use of concrete and its consumption is on par with wans and the activity of construction is set to have been emitting 8% of CO2 [2]. Even though the substitute for concrete is not identified the area of development becomes the centre stage for identifying substitute materials. The contribution of nano technology has made big strides and effect on various aspects of science. Nano silica is preferably utilized in numerous approaches and is easily available as well.
The research and investigation in nano particles and their utilization in cement and mortar invariably becomes popular [3]. The properties of cement in both hardened as well as in fresh phase are majorly laid focus on. The density of concrete and cement can be improvised with the inclusion of nano as well as micro NS which have the tendency of acting as a material of filler, which results in up gradation of its quality [4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17]. Nano silica with separated molecule impact got an extraordinary pozzolonic property than other pozzolonic materials on comparison because of the nano size of particles and greater silica contents, accordingly both impacts are important in making concrete [4, 6]. Expansion of Nano silica to solidify concrete and cement mortar then resulted in improvement of sharp and long haul considers and over utilisation of nano silica results in decrease in quality. Nano silica greatly improved qualities of cement and concrete in fresh and solidified stage and in durability in different environmental conditions [10, 11, 18, 19]. The nano particles are centered and a considerable research is carried out.
Therefore, the present research includes the impact and durability of 40 nm silica particles in cement mortar, durability existence under different environments both physical and chemical are taken up for examination [20, 21, 22, 23]. For acquiring an accurate output, instruments of Scanning Electron Microscope (SEM) and X-ray diffraction (XRD) were resorted to. In this research, cement to sand proposition was 1: 3 and water to cement (concrete plus NS) proposition was set at 0.4.
In the set forth work area, the proportion of 1:3 is considered for cement-sand and 0.4 for water-cement. Also, nanosilica is incorporated along with cement starting from zero percentage and with every sample, an increase of 2% is followed until an overall 14% is achieved. Every sample is treated individually with starting with M0 and concluding with M7.
Sticking to the particulars of Jo B W [24], the blending of NS with cement mortar is done for a minute maintaining 285 rpm. Prior to the blending process, the water is treated with NS and this was put into the rotational blender which runs at 140 rpm. This was done for 30 seconds so as to solidify the mixture and addition of a fine total was done. Adding to the process, a super plasticizer was annexed to the mix when the blender was running at a speed of 285 rpm. Now, a resting phase of 90 seconds is allocated to the blender and it resumes working for a minute with the same speed before rest. Finally, the mix was set out in a hardened figure.
IS of code 5513-1976 was used for detecting the time taken for setting.
The part 6 of 4031-1998 was considered to find the compressive quality of mortar.
Determining the flexural standards using the BS section 188 of 1881 of the year 1983 was done.
AR graded magnesium sulphate and hydrochloric acid, which are synthetic in nature, were used in the research. The ACI of 318-99 were followed and the compound grouping was done. An exemplar of the fixations projects over a span of 120 days for 1%, 240 days for 2%, 360 days for 3% and 480 days for 4%, provided the recharge of the fixations was for each 4-month tenure.
The blended samples of M0 as well as M5 were tested upon for the temperature check at 900 °C in a suppress heater. Also, the blended sample out-righting 28 days were dried for a span of 6 hours at normal temperature and then they were transferred to a muffle furnace. An altering is done with temperatures where each temperature has got three samples assessed for a period of two and half hours. The altering temperatures start from 100 °C and every alteration has an add-on of 100 °C up to 600 °C. Finally, the samples were put out to cool down at a temperature of 30 °C i.e., room temperature (Tables 1–5).
Calcium oxide | 62.5 |
Silicon dioxide | 20.5 |
Aluminium oxide | 6.1 |
Ferrous oxide | 3.1 |
Magnesium oxide | 1.6 |
Sulphur trioxide | 1.5 |
Oxides of potassium and sodium | 1 |
Constitutes of cement.
Compressive Strength (days) 3 7 28 | 27 MPa (Min) 37 MPa (Min) 53 MPa (Min) | 35.91 MPa 46.72 MPa 62.90 MPa |
Relative Density | — | 3.10 |
Setting Span Initial Final | 30 min 600 min | 155 min 267 min |
Soundness (Le- Chatlier Expansion) | 10 mm (Max) | 0.85 mm |
Specific Surface Area (m2/Kg) | 226 | 314 |
Cement attributes.
Passing through Sieve of 2 mm | 100 |
Retained over Sieve of 90 m | 100 |
1 mm < Size of the particle | 33.33 |
500 m < 1 mm > Size of the particle | 33.33 |
500 m > Size of the particle | 33.33 |
Ennore sand attributes.
Relative density | 1.33 |
Surface area | 50.5 |
Particle size nm | 40 |
SiO2 | 99.7% |
Loss on Ignition | 0.3% |
Attributes of nanosilica.
power of hydrogen, ph | 6.5- 8.5 | 7.2 | 7.5 |
Total Dissolved Solids (mg/L) | 2000 | 4.6 | 11.0 |
Organic Solids (mg/L) | 200 | 1.1 | 6.0 |
Inorganic Solids (mg/L) | 3000 | 6.2 | 17.0 |
Alkalinity (mg/L) | 250 | 3.6 | 11.0 |
Acidity (mg/L) | 50 | 0 | 5.0 |
Sulphates (mg/L) | 400 | 3.2 | 12.0 |
Chlorides | 500(RCC) | 1.5 | 9.0 |
Attributes of water.
Figure 1 reveals the corollary on setting time due to NS and uncovers the fact of lowering the hardening time with increment in NS quantity. The initial and final setting time for M0 was 155 min and 266 min, M1 was 153 min and 263 min, M2 was 148 min and 260 min, M3 was 143 min and 255 min, M4 was 137 min and 245 min, M5 was 131 min and 240 min, M6 was 126 min and 234 min, M7 was 123 min and 227 min. The cycle was advanced because of NS has a vast surface region; subsequently, hydration measure turns out to be quick.
Efficacy of nanosilica over setting span.
A rise in the strength upon inclusion of nanosilica up to 10% was portrayed in Figure 2. With further addition of NS, decrement in strength can be observed from the diagram. The finest of all samples is the M5 sample comprised of 90% cement and 10% NS. For 3 days, the increment strength observed was 17.8 MPa, 7 days resulted 21.58 MPa, 28 days directed a strength of 21.18 MPa, 90 days showcased 21.50 MPa, 180 days witnessed 21.32 MPa and finally, 365 days exhibited a strength of 20.66 MPa. From the analysis, it is truely clear that M6 blended with 88% of cement and the remaining with NS has seen a decrease in compressive strength whereas M5 figured the best strength notable.
Efficacy of nanosilica over strength attribute (compression).
The analysis was done for a couple of samples having 0% NS and 10% NS i.e., for M0 and M5 by X-ray diffraction, XRD. The samples were of OPC type and were hydrated for a span of 28 days. Figure 3 shows that hydroxide of calcium showed up at 18° whereas the calcium silicate hydrate showed up at 26° individually for both the samples. Concentration and strength of hydroxide of calcium was predominantly high and that of silicate hydroxides of calcium was less when compared to OPC of 90% and nano silica of 10% test. The variation of strength for the sample of M0 and the sample of M5 was due to the reaction of oxides of silica with hydroxides of calcium (finished results of cement hydrate).
Depiction of M0 & M5 samples XRD (hydrated for 28 days).
Figures 4 and 5 demonstrate the results for the sample M0 and also for the sample M5 which were hydrated for 28 days. The diagram of Scanning Electron Microscope (SEM) portrays the hydroxides of calcium’s needle like structure along with the silicate hydroxide of calcium which was spread widely. The calcium hydroxide’s long needle like structures in Figure 4 can be collated with Figure 5. Also, a high in silicate hydroxide of calcium’s content can be seen in Figure 5 when compared to Figure 4. The major difference in the structure of hydroxides of calcium and quantity of silicate hydroxide of calcium was due to the presence of oxides of silica in NS. These react with hydroxides of calcium in the cement hydrate and thereby, causing the differences.
Depiction of M0’s SEM diagram (28 days of hydration).
Depiction of M5’s SEM diagram (28 days of hydration).
The NS impact on flexural strength can be identified from the Figure 6. It was evident that cement with less than 10% NS exhibited expand in flexural strength by 10% whereas cement with NS in more than 10% showcased decline in the strength. The samples were identified upon M5 blend with a higher expansion rate with time. It was seen that 2 MPa was recorded for 3 days, 2.2 MPa for 7 days, 2.5 MPa for 28 days, 2.4 MPa for 90 days, 2.4 MPa for 180 days and finally 2.6 MPa for 365 days. From the analysis, we can notice that samples has shown a decrease in strength with NS percentage more than 10 but the value was higher when compared with the strength of sample with zero percentage of NS. The major concerns that led the expansion of the strength characteristics were the size of the particles and content of oxides of silica. An extra quantity of silicate hydroxide of calcium was witnessed due to the action of hydroxides of calcium and oxides of silica. Furthering this, the NS acts as a framework’s filler material of cement mortar.
Efficacy of nanosilica upon strength (Flexural).
Figures 7 and 8 render the effect of Magnesium Sulphate (MgSO4) over compressive strength of the samples. If one observes in Figure 7, by and large, cement mortar strength quality increased irrespective of age, the equivalent can be observed in M0 examples of no MgSO4 fixation., Compressive strength qualities decreased as for time and focus with less than 4% fixation, and 4% centralization of MgSO4 has resulted in the greatest quality decrease.
Corollary of MgSO4 over Sample M0.
Corollary of MgSO4 over Sample M5.
M5 blend examples equivalents the above and were provided in Figure 8. Concerning the concentration and age of MgSO4, M0 blend examples get altogether result in quality decrease compared to M5 blend examples Also the M0’s compressive strength for 4% was noted to be 57.30 MPa for 120 days, 53.55 MPa for 240 days, 48.50 MPa for 260 days and finally 44.16 MPa for 480 days. In the similar context, the strength values of M5 were 78.26 MPa for 120 days, 74.1 MPa for 240 days, 67.0 MPa for 260 days and finally 64.0 MPa for 480 days. The contrast of strength characteristic between M0 and M5 was 21, 20.4, 20.3 and 19.7 MPa respectively.
The blended samples of OPC’s M0 and M5 were hydrated for 28 days and the samples were kept in water with 4% of MgSO4 for a span of 360 days. The results of the sample M0 and the sample M5 were represented as charts in the Figure 9 where magnesium silicate hydrate was presented at 48°; at 18°, the hydroxide of calcium turned up and at 33°, magnesium’s hydroxide was presented and finally the silica hydroxide of calcium was shown at 26°. It was noted that the hydroxide of calcium exhibited a high force of ascent and, additionally, the lower power ascent of silicate hydroxide of calcium of M0 diversified from M5. The contrast happened because of nanosilica quality. Also, hydroxide and silicate hydrate of magnesium were directed because of the reaction of MgSO4 with hydroxide and silicate hydroxide of calcium. The quality of silicate hydrate of magnesium declined due to the non-binding nature, but M5 exhibits a finer standard when collated with M0 due to the inclusion of nanosilica.
Depicting M0 & M5 samples XRD (Sample placed for 360 days in water with 4% MgSO4).
The blended samples of OPC’s M0 and M5 were hydrated for 28 days and the samples were kept in water with 4% of MgSO4 for a span of 360 days. The results were represented as Scanning Electron Microscope diagrams in the Figures 9 and 10 where hydroxide of calcium, silicate hydroxide of calcium, hydroxide of magnesium and silicate hydrate of magnesium were presented. Calcium hydroxide’s needle structure was indicated in the Figure 10 and further, it was made in contrast with the Figure 11 whereas the content of calcium silicate hydroxide is high when compared to Figure 10. In further addition, magnesium’s silicate hydrate and hydroxide witnessed a fine and more elegant outlook when collated with Figure 11. The inclusion of NS shows diversified nature of OPC. Also, placing the sample in water mixed with MgSO4 adds up to the work.
Depicting M0’s SEM diagram (Sample placed for 360 days in water with 4% MgSO4).
Depicting M5’s SEM diagram (Sample placed for 360 days in water with 4% MgSO4).
Figures 12 and 13 render the effect of Hydrochloric Acid (HCl) over compressive strength of the samples. By and large, concrete mortar quality expanded irrespective of age, the equivalent can be seen at sample M0 which was placed in zero HCl fixations. It was also seen that a decline in compressive quality with concentration and time in Figure 12 when M0 blend examples kept in less than 4% convergence of HCl and examples were given for HCl centralization with 4% has the greatest quality decrease.
HCl Corollary over Sample M0.
HCl Corollary over Sample M5.
The equivalent to above can likewise be seen in M5 blend examples which appear in Figure 13. Yet, M0 blend examples get noteworthy quality decrease contrasted with that of M5 blend examples. M0’s and M5’s compressive quality for a focus of 4% was is 55.35 MPa and 77.33 MPa for 120 days, 49.00 MPa and 72.30 MPa for 240 days, 42.60 MPa and 65.55 MPa for 260 days and 37.31 MPa and 56.30 MPa for 480 days. The differentiation in strength at 4% fixation for 120 days was 21.3 MPa, 240 days was 23.25 MPa, 260 days was 22.82 MPa and for 480 days was 18.96 MPa.
The blended samples of OPC’s M0 and M5 were hydrated for 28 days and the samples were kept in water with 4% of HCl for a span of 360 days. The results were represented in the Figure 14 where hydroxide of calcium, silicate hydroxide of calcium, chlorides of calcium, Friedel’s mixes of salt of M0 sample and M5 sample were presented individually at 18°, 26°, 33°, 37° and finally at 48.5°. It was notified that the ascent power of calcium hydroxide is high whereas the force ascent of silicate hydroxide of calcium is more for M5 than M0. The observed variation in ascent of M0 sample and M5 sample was majorly due to the expansion taking place in cement mortar due to the inclusion of NS. Adding to it, the reaction of hydroxides of calcium with HCl and chlorides of calcium with C3A frames the chlorides of calcium and Friedel’s mixes of salt. Silicate hydroxide of calcium got destabilized due to its reaction with HCl resulting a decline in the quality of strength but the performance of M5 was way too good when collated to that of M0 because of essence of NS in cement mortar.
Depicting M0 & M5 samples XRD (Sample placed for 360 days in water with 4% HCl).
The blended samples of OPC’s M0 and M5 were hydrated for 28 days and the samples were kept in water with 4% of HCl for a span of 360 days. The results were represented as Scanning Electron Microscope diagrams in the Figures 15 and 16 where hydroxides of calcium, silicate hydroxides of calcium, chlorides of calcium were shown. A significant improvement in case of above mentioned compounds can be noted in the Figure 15 when contrasted with the Figure 16. The diagrams of Scanning Electron Microscope test uphold the investigation of X-ray Diffraction.
Depicting M0’s SEM diagram (Sample placed for 360 days in water with 4% HCl).
Depicting M5’s SEM diagram (Sample placed for 360 days in water with 4% HCl).
Temperature’s corollary on the blend samples of M0 and M5 were shown in the Figure 17. It was evidently clear that an increase in temperature of M0 and M5 samples results the compressive strength to lower. The strength values noted for M0 sample and M5 sample at various temperature conditions are 62.93 MPa and 84.12 MPa at 27 °C, 51.90 MPa and 73 MPa at 400 °C, 28.30 MPa and 48 MPa at 600 °C, 15.40 MPa and 28.41 MPa at 800 °C. The M0 sample and M5 sample exhibits decrease in compressive strength quality because of between layer water, losing free water and synthetically reinforced water. Furthermore warm extension of cement mortar and was unique. The quality decreases in case of compressive strength for sample of M0 and sample of M5 was critical over a temperature of 400 °C. But, more compressive quality was exhibited by M0 blend examples than M5 blend examples.
Corollary of temperature.
The strength qualities (compression and flexural) expanded by and large due to the NS of 40 nm in case of M5 blend. Cement’s extraordinary pozzolonic property and its property as a good material for filling of NS instigated the rise in strength with cement and brilliant filling material.
NS has quickened setting measure because of its enormous surface area.
M5 mix examples have given much better execution in solidness in different environmental conditions than that of M0 blend examples.
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I am also a member of the team in charge for the supervision of Ph.D. students in the fields of development of silicon based planar waveguide sensor devices, study of inelastic electron tunnelling in planar tunnelling nanostructures for sensing applications and development of organotellurium(IV) compounds for semiconductor applications. I am a specialist in data analysis techniques and nanosurface structure. I have served as the editor for many books, been a member of the editorial board in science journals, have published many papers and hold many patents.",institutionString:null,institution:{name:"Sheffield Hallam University",country:{name:"United Kingdom"}}},{id:"54525",title:"Prof.",name:"Abdul Latif",middleName:null,surname:"Ahmad",slug:"abdul-latif-ahmad",fullName:"Abdul Latif Ahmad",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:null},{id:"20567",title:"Prof.",name:"Ado",middleName:null,surname:"Jorio",slug:"ado-jorio",fullName:"Ado Jorio",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:{name:"Universidade Federal de Minas Gerais",country:{name:"Brazil"}}},{id:"47940",title:"Dr.",name:"Alberto",middleName:null,surname:"Mantovani",slug:"alberto-mantovani",fullName:"Alberto Mantovani",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:null},{id:"12392",title:"Mr.",name:"Alex",middleName:null,surname:"Lazinica",slug:"alex-lazinica",fullName:"Alex Lazinica",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/12392/images/7282_n.png",biography:"Alex Lazinica is the founder and CEO of IntechOpen. After obtaining a Master's degree in Mechanical Engineering, he continued his PhD studies in Robotics at the Vienna University of Technology. Here he worked as a robotic researcher with the university's Intelligent Manufacturing Systems Group as well as a guest researcher at various European universities, including the Swiss Federal Institute of Technology Lausanne (EPFL). During this time he published more than 20 scientific papers, gave presentations, served as a reviewer for major robotic journals and conferences and most importantly he co-founded and built the International Journal of Advanced Robotic Systems- world's first Open Access journal in the field of robotics. Starting this journal was a pivotal point in his career, since it was a pathway to founding IntechOpen - Open Access publisher focused on addressing academic researchers needs. Alex is a personification of IntechOpen key values being trusted, open and entrepreneurial. 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