Summary of Mg surface modification techniques.
\r\n\t
",isbn:"978-1-83968-571-2",printIsbn:"978-1-83968-570-5",pdfIsbn:"978-1-83968-599-6",doi:null,price:0,priceEur:0,priceUsd:0,slug:null,numberOfPages:0,isOpenForSubmission:!0,hash:"dd81bc60e806fddc63d1ae22da1c779a",bookSignature:"Dr. Sebahattin Demirkan and Dr. Irem Demirkan",publishedDate:null,coverURL:"https://cdn.intechopen.com/books/images_new/10818.jpg",keywords:"Decision Making, Blockchain, Accounting, Earnings Management, Strategic Alliances, Innovation, Performance, Corporate Governance, Accounting Quality, Digital Assets, Internationalization, MNCs",numberOfDownloads:null,numberOfWosCitations:0,numberOfCrossrefCitations:null,numberOfDimensionsCitations:null,numberOfTotalCitations:null,isAvailableForWebshopOrdering:!0,dateEndFirstStepPublish:"January 28th 2021",dateEndSecondStepPublish:"February 25th 2021",dateEndThirdStepPublish:"April 26th 2021",dateEndFourthStepPublish:"July 15th 2021",dateEndFifthStepPublish:"September 13th 2021",remainingDaysToSecondStep:"6 hours",secondStepPassed:!1,currentStepOfPublishingProcess:2,editedByType:null,kuFlag:!1,biosketch:"Academician in the area of accounting who believes in the impact of interdisciplinary research. Dr. Sebahattin Demirkan's research interests are in the areas of financial accounting, capital markets, auditing, corporate governance, strategic alliances, taxation, CSR, and data analytics.",coeditorOneBiosketch:"Researcher of strategic management, corporate entrepreneurship, and international business; specific interests include innovation, the ambidexterity framework, inter-organizational relationships, and networks. Experienced in teaching graduate and undergraduate courses in strategy, entrepreneurship, and international business and management areas.",coeditorTwoBiosketch:null,coeditorThreeBiosketch:null,coeditorFourBiosketch:null,coeditorFiveBiosketch:null,editors:[{id:"336397",title:"Dr.",name:"Sebahattin",middleName:null,surname:"Demirkan",slug:"sebahattin-demirkan",fullName:"Sebahattin Demirkan",profilePictureURL:"https://mts.intechopen.com/storage/users/336397/images/system/336397.jpg",biography:"Dr. Sebahattin Demirkan is a Professor of Accounting. He earned his Ph.D. in Accounting/Management Science at Jindal School of Management of the University of Texas at Dallas where he got his MS in Accounting, MSA Supply Chain, and MBA degrees. He got his BA in Economics and Management at the Faculty of Economics and Administrative Sciences at Bogazici University, Istanbul. He worked at Koc Holding, a private venture capital firm, and the University of California, Berkeley during and after his education at Bogazici University. His research interests are in the areas of financial accounting, capital markets, auditing, corporate governance, strategic alliances, taxation, CSR, and data analytics. Dr. Sebahattin Demirkan has published articles in Contemporary Accounting Research, JAPP, JAAF, TEM, Journal of Management, and other top academic journals. He teaches several different classes in both undergraduate and graduate levels in Accounting and Analytics programs. He is a treasurer and vice president of the TASSA, board member of the BURCIN and member of the American Accounting Association.",institutionString:"Manhattan College",position:null,outsideEditionCount:0,totalCites:0,totalAuthoredChapters:"0",totalChapterViews:"0",totalEditedBooks:"0",institution:{name:"Manhattan College",institutionURL:null,country:{name:"United States of America"}}}],coeditorOne:{id:"342242",title:"Dr.",name:"Irem",middleName:null,surname:"Demirkan",slug:"irem-demirkan",fullName:"Irem Demirkan",profilePictureURL:"https://s3.us-east-1.amazonaws.com/intech-files/0033Y000033HrA8QAK/Profile_Picture_1606729803873",biography:"Dr. Irem Demirkan earned her Ph.D. in International Management Studies and M.S. in Administrative Studies at Jindal School of Management at the University of Texas at Dallas, USA. She got her BA in Economics at the Faculty of Economics and Administrative Sciences at Bogazici University, Istanbul, Turkey. She worked in the finance and textile industries before joining to academia. Dr. Demirkan has published research in the areas of strategic management and corporate entrepreneurship in journals such as the Journal of Management, Journal of Business Research, Management Science, European Journal of Innovation and Management, IEEE Transactions on Engineering Management, among others. 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Whether that be identifying an exceptional author and proposing an editorship collaboration, or contacting researchers who would like the opportunity to work with IntechOpen, I establish and help manage author and editor acquisition and contact."}},relatedBooks:[{type:"book",id:"1591",title:"Infrared Spectroscopy",subtitle:"Materials Science, Engineering and Technology",isOpenForSubmission:!1,hash:"99b4b7b71a8caeb693ed762b40b017f4",slug:"infrared-spectroscopy-materials-science-engineering-and-technology",bookSignature:"Theophile 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"}},{type:"book",id:"3092",title:"Anopheles mosquitoes",subtitle:"New insights into malaria vectors",isOpenForSubmission:!1,hash:"c9e622485316d5e296288bf24d2b0d64",slug:"anopheles-mosquitoes-new-insights-into-malaria-vectors",bookSignature:"Sylvie Manguin",coverURL:"https://cdn.intechopen.com/books/images_new/3092.jpg",editedByType:"Edited by",editors:[{id:"50017",title:"Prof.",name:"Sylvie",surname:"Manguin",slug:"sylvie-manguin",fullName:"Sylvie Manguin"}],productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"3161",title:"Frontiers in Guided Wave Optics and Optoelectronics",subtitle:null,isOpenForSubmission:!1,hash:"deb44e9c99f82bbce1083abea743146c",slug:"frontiers-in-guided-wave-optics-and-optoelectronics",bookSignature:"Bishnu Pal",coverURL:"https://cdn.intechopen.com/books/images_new/3161.jpg",editedByType:"Edited by",editors:[{id:"4782",title:"Prof.",name:"Bishnu",surname:"Pal",slug:"bishnu-pal",fullName:"Bishnu Pal"}],productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"72",title:"Ionic Liquids",subtitle:"Theory, Properties, New Approaches",isOpenForSubmission:!1,hash:"d94ffa3cfa10505e3b1d676d46fcd3f5",slug:"ionic-liquids-theory-properties-new-approaches",bookSignature:"Alexander Kokorin",coverURL:"https://cdn.intechopen.com/books/images_new/72.jpg",editedByType:"Edited by",editors:[{id:"19816",title:"Prof.",name:"Alexander",surname:"Kokorin",slug:"alexander-kokorin",fullName:"Alexander Kokorin"}],productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"1373",title:"Ionic Liquids",subtitle:"Applications and Perspectives",isOpenForSubmission:!1,hash:"5e9ae5ae9167cde4b344e499a792c41c",slug:"ionic-liquids-applications-and-perspectives",bookSignature:"Alexander Kokorin",coverURL:"https://cdn.intechopen.com/books/images_new/1373.jpg",editedByType:"Edited by",editors:[{id:"19816",title:"Prof.",name:"Alexander",surname:"Kokorin",slug:"alexander-kokorin",fullName:"Alexander Kokorin"}],productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"57",title:"Physics and Applications of Graphene",subtitle:"Experiments",isOpenForSubmission:!1,hash:"0e6622a71cf4f02f45bfdd5691e1189a",slug:"physics-and-applications-of-graphene-experiments",bookSignature:"Sergey Mikhailov",coverURL:"https://cdn.intechopen.com/books/images_new/57.jpg",editedByType:"Edited by",editors:[{id:"16042",title:"Dr.",name:"Sergey",surname:"Mikhailov",slug:"sergey-mikhailov",fullName:"Sergey Mikhailov"}],productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"371",title:"Abiotic Stress in Plants",subtitle:"Mechanisms and Adaptations",isOpenForSubmission:!1,hash:"588466f487e307619849d72389178a74",slug:"abiotic-stress-in-plants-mechanisms-and-adaptations",bookSignature:"Arun Shanker and B. Venkateswarlu",coverURL:"https://cdn.intechopen.com/books/images_new/371.jpg",editedByType:"Edited by",editors:[{id:"58592",title:"Dr.",name:"Arun",surname:"Shanker",slug:"arun-shanker",fullName:"Arun Shanker"}],productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"878",title:"Phytochemicals",subtitle:"A Global Perspective of Their Role in Nutrition and Health",isOpenForSubmission:!1,hash:"ec77671f63975ef2d16192897deb6835",slug:"phytochemicals-a-global-perspective-of-their-role-in-nutrition-and-health",bookSignature:"Venketeshwer Rao",coverURL:"https://cdn.intechopen.com/books/images_new/878.jpg",editedByType:"Edited by",editors:[{id:"82663",title:"Dr.",name:"Venketeshwer",surname:"Rao",slug:"venketeshwer-rao",fullName:"Venketeshwer Rao"}],productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"4816",title:"Face Recognition",subtitle:null,isOpenForSubmission:!1,hash:"146063b5359146b7718ea86bad47c8eb",slug:"face_recognition",bookSignature:"Kresimir Delac and Mislav Grgic",coverURL:"https://cdn.intechopen.com/books/images_new/4816.jpg",editedByType:"Edited by",editors:[{id:"528",title:"Dr.",name:"Kresimir",surname:"Delac",slug:"kresimir-delac",fullName:"Kresimir Delac"}],productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"3621",title:"Silver Nanoparticles",subtitle:null,isOpenForSubmission:!1,hash:null,slug:"silver-nanoparticles",bookSignature:"David Pozo Perez",coverURL:"https://cdn.intechopen.com/books/images_new/3621.jpg",editedByType:"Edited by",editors:[{id:"6667",title:"Dr.",name:"David",surname:"Pozo",slug:"david-pozo",fullName:"David Pozo"}],productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}}]},chapter:{item:{type:"chapter",id:"53740",title:"Surface Modification of Magnesium and its Alloys Using Anodization for Orthopedic Implant Application",doi:"10.5772/66341",slug:"surface-modification-of-magnesium-and-its-alloys-using-anodization-for-orthopedic-implant-applicatio",body:'\nBiodegradable metallic implant material has received considerable attention in biomedical field such as blood vessels or orthopedic application as load-bearing implant [1, 2]. Mg is suitable for implant application in human body, for example, Mg stent, bone fixation screw, microclips in laryngeal microsurgery, bone fixation and wound-closing devices, as shown in \nFigure 1\n. Mg has many appealing properties such as light weight, high strength-to-weight ratio, good castability and osteoconductivity [3]. However, Mg has limitations mainly due to its high surface chemical reactivity resulting in high degradation rate [4]. The poor corrosion resistance of Mg limits its clinical applications, as hydrogen evaluation is one of the corrosion products that increase alkalinity of the surrounded media and causing inflammation of the surrounding tissues due to the formation of gas pockets [5, 6]. The high degradation rate may eventually hinder the bone formation and hamper the long-term success of the implants and decrease its bioactivity as well as loss its mechanical properties [7]. Mg-based implants exhibited rough surfaces as well as shallow pits and small cavities after one day of implantation, which formed during the on-going corrosion process to form cracks until the implant totally dissolves [8]. The high purity of Mg finds to corrode uniformly in vivo [9]. Biodegradable metals (BMs) are typically degraded through the corrosion process when exposed to a corrosive media. For example, when BMs especially Mg implanted in human body, the corrosion/degradation process generated electrochemically in different reactions of metals with an electrolyte and produced metal oxides and hydroxides [7]. Moreover, hydrogen gas evaluation is a combined corrosion product; these reactions could be represented in the following chemical equations at anodic dissolution of Mg and the cathodic reaction [10].
\nDifferent applications of Mg-based implant material: (a) cardiovascular Mg stents, (b) MAGNEZIX screw, (c) microclip for laryngeal microsurgery (pure magnesium), (d) biodegradable orthopedic implants and (e) wound-closing devices (WZ21) [11].
Surface modification is considered one of the most useful and effective methods to control the initial degradation of Mg and its alloys [12]. \nTable 1\n summarizes the previous research on Mg coating with different applied techniques and chemical composition. Among these techniques, anodization is a widely and traditional process for metal surface modification to improve the physiochemical properties of metals [13]. A suitable electrolyte of anodization for the specific application of Mg is one of the essential requirements when it employed. For example, hydroxyapatite (HA) is a bioactive ceramic material which widely used in bone application [14]. HA could be engineered to mimic the three-dimensional inorganic component of the bone which is composed of 65% of bone. The structure of HA could provide the space and area necessary for vascularization and tissue regeneration. In this chapter, HA coating with different nanostructures (nanoplates/nanospheres) by means of anodization is discussed with the associated mechanical integrity, biodegradability and biocompatibility. Formation of nanoplates could promote the osseointegration and eliminate the mismatching of the implant material. Accordingly, using stimulated body fluid (SBF) finds to form apatite film on the surface of Mg in a short duration.
\nSubstrate | \nExperimental and coating type | \nReference | \n
---|---|---|
Mg-Zn-Ca alloy | \nFabrication of hydroxyapatite nanorod on MAO coating to increase bioactivity and improve the biodegradation behavior | \n[15] | \n
Mg-1.0Ca alloys | \nSodium phytate (Na12Phy) used as an electrolyte with anodic coatings fabricated in an organic phosphate containing solution on the Mg-1.0Ca alloys. In order to achieve a proper degradation rate, acceptable biocompatibility and good antibacterial ability | \n[16] | \n
AZ31B | \nDifferent electrolytes such as KOH, Na2SiO3 and Na2B4O7 were used for pulsed DC micro-arc oxidation (MAO) process | \n[17] | \n
Mg-Zn-Ca | \nA porous bioceramic containing tricalcium phosphate in (TCP) coating was prepared by (MAO) at different voltages | \n[18] | \n
Pure Mg | \nAnodic oxide coatings were prepared using 0.3 M NaOH + 15 g/l ZrO2 and 3 M NaOH + 15 g/l ZrO2\n | \n[19] | \n
AZ31B | \nA chemical conversion film on magnesium alloys is proposed based on the interaction of a deep eutectic solvent (DES) with the substrate | \n[20] | \n
Mg-3Zn | \nA nanostructured hydroxyapatite (HA) coating was grown on through the electrophoretic deposition (EPD) technique | \n[21] | \n
ZK61 | \nMAO coating film with low crystallinity is composed of MgO, Mg2SiO4 and Mg2Si2O6 employed | \n[22] | \n
AZ31 | \nA dopamine-induced hydroxyapatite coating was successfully developed on the AZ31 alloy | \n[23] | \n
AZ31 | \nUse of a microwave-assisted coating technology to improve the in vitro corrosion resistance and biocompatibility of AZ31Mg alloy | \n[24] | \n
Pure Mg | \nA simple strontium phosphate (SrP) conversion coating process was developed to protect magnesium (Mg) from the initial degradation postimplantation | \n[25] | \n
AZ31 | \nA Si-doped calcium phosphate coating was achieved via pulse ED on the AZ31 alloy. A novel dual-layer structure was observed with a porous lamellar-like and outer block-like apatite layer | \n[26] | \n
Summary of Mg surface modification techniques.
Anodization is an electrochemical process that converts the metal surface into a decorative, durable, corrosion-resistant and anodic oxide finish [27]. The coating thicknesses can range from 5 to 200 μm. Typically, anodic oxide layers grow depending on the process time and applied voltages [28], leading to a direct dependence of the oxide thickness on the applied voltage as shown in \nFigure 2\n. For metals and alloys with barrier-type anodic oxide films, blocking electron conduction under anodic polarization an anodization can be carried out at high voltages in aqueous solution [29]. Therefore, thick oxides that can be grown on the conductive oxide layers on the metal surface by means of anodization are limited to the applied voltage. The applied voltage is lower than that at which water can dissociated with evaluation of oxygen, whereas, above that potential water tends to decompose rather than thickening of the oxide layer. For example, Mg has potential and conductivity; therefore, the resulting potential while anodization applied depends mainly on the electrolyte composition [29]. The incorporation of electrolyte materials with growing oxide/hydroxide layers can form an oxide layers that have higher blocking efficiency toward the corrosive ions. Therefore, thick and compact film is a challenge for Mg interface anodization treatment, however, obtaining a low Pilling-Bedworth ratio for the formed anodized film [30]. This could cause an internal stresses on the generated anodic film and subsequently crack defects [31]. The degree of porosity and oxide layer quality could be enhanced by anodization parameter adjustment. These parameters include electrolyte composition, anodization voltage, current and time [32]. Anodization performed in different baths, for example an alkaline electrolyte is based on potassium hydroxide, phosphate, fluoride, or silicate-containing baths. Electrolyte composition plays a critical role not only to enable anodization at high voltage but also to reduce Mg dissolution during the process [33]. There are various methods and techniques with a wide range of patents to produce such anodic films [34]. In addition to anodization approaches which are mainly used to thicken the native oxide/hydroxide films on metal surfaces, dedicated anodization approaches have been explored to obtain nanoporous oxide layers. Therefore, the appropriate electrolyte composition leads to competition between Mg dissolution during anodization and anodic oxide film growth. Thus, optimized parameters based on the electrochemical process self-organized growth of nanoporous or nanotubular oxide layers could performed; however, it is still at early stage for Mg and its alloys [35, 36]. \nTable 2\n summarizes different Mg alloy anodization/PEO on different electrolytes with the resulted film thickness and chemical composition and the mainly electrochemical corrosion parameters (E\ncorr and i\ncorr).
\nSubstrate | \nElectrolyte | \nThickness | \nLayer composition | \n\ni\ncorr\n | \n\nE\ncorr\n | \nRefs. | \n
---|---|---|---|---|---|---|
AZ91D | \nPEO in NaOH + (NaPO3)6 + Ca(H2PO2)2 solution | \n3–5 μm | \nMg, Al, P and Ca and little crystallized MgO | \nX | \nX | \n[40] | \n
AZ91D | \nPEO in Na2SiO3 + (NaPO3)6 + Ca(H2PO2)2 solution | \n8–10 μm | \nMg, Al, Si, P and Ca, crystallized Mg2SiO4 and MgO | \nX | \nX | \n[2] | \n
AM50 | \nPEO in CaOH2 + Na3PO4 solution in different mass ratios | \nin the range of 20–70 μm | \nMgO, Mg3(PO4)2, amorphous Ca-phases, CaH(PO4)2, CaO2\n | \nX | \nX | \n[41] | \n
AZ91 | \nNaOH | \n1–2 μm | \nMgO and Mg | \nX | \nX | \n[42] | \n
ZK60 | \n100 g/l NaOH + 20 g/l Na2B4O7⋅10H2O + 50 g/l C6H5Na3O7⋅2H2O + 60 g/l Na2SiO3⋅9H2O | \n10–60 μm | \nMgO and Mg2SiO4\n | \n1.829 × 10−2 (mA/cm2) | \n−1.46 | \n[43] | \n
AZ31 | \nSBF solution | \n5–25 μm | \nMgO, Mg and amorphous apatite | \n103 ˷ to 0.9 μA/cm2\n | \n−1.39 to −1.45 | \n[44] | \n
AZ31 | \n(ZrO2-NPs) dispersed in SBF | \nX | \nMg, MgO, ZrO2, and Mg2Zr5O12\n | \n−1.46 to −1.38 | \n2.796 to 1.9 | \n[45] | \n
AZ31 | \n(SBF solution + HA) then hydrothermal in 5 M NaOH at 60°C for 2 h | \nX | \nMg, MgO, CaO and HA | \n7.6 to 1025 nA/cm2\n | \n1.52 to1.31 | \n[46] | \n
Anodization of Mg alloys in different electrolyte.
Illustrative diagram shows the mechanism of anodization technique. Mg acts as an anode where it converted to Mg2+ then reacts with O2− to form MgO in the presence of OH ions, Mg(OH)2 formation on the metal surface and hydrogen formed the surrounding cathode part.
The design of surface morphology structure of biodegradable implant is an important factor since the interconnection of biomaterial interface with surrounding tissues is important for implant engagement and cell attachment [37]. In bone implant, nanoplate and nanosphere structure of HA coating as a biomimetic films are considered for the Mg coating interface, which is characterized by mimicking that of bone [38]. \nFigure 3a\n shows the nanoplates formation on the surface of AZ31 Mg alloy by the anodization method in SBF solution at 50 V and 30 mA with a process time of 10 min followed by the hydrothermal process in NaOH solution at 60°C for 2 h. However, adding HA powder to SBF solution resulted in nanosphere structure. Natural bone consists of HA nanocrystals in a plate-like shape with a length of 30–200 nm and a thickness of 2–7 nm [39]. As a result, designing HA films with the specific orientation and morphology is an important approach to improve Mg biological properties such as bioactivity and mimic that on natural bone. Furthermore, such nanoplates can promote the porosity of the implant interface, as a result avoid a mechanical mismatch between the hosts and implant interface, stress shield effect can be eliminated by altering surface porosity.
\nSurface morphology of anodized AZ31 Mg alloy in different electrolytes followed by the hydrothermal process in NaOH at 60°C for 2 h and SBF for 2 days at 37°C: (a) SBF as an electrolyte resulted in nanoplates morphology and (b) (SBF-10 g/l HA) resulted in nanospheres structure.
The chemical composition of HA coating finds is composed of Mg, MgO, HA and CaO peaks as shown in XRD peaks in \nFigure 4a\n. Furthermore, FT-IR spectra can indicate the outer HA film formation as shown in \nFigure 4b\n. The bands at a wave number of around 530 cm−1 is assigned to \n
XRD patterns and FTIR spectra of the surface treatment samples, two anodized samples in (i) SBF solution and (ii) SBF/HA solution followed by the hydrothermal process according to reference [46].
Mechanical tuning is one of the most effective factors for biodegradable Mg implant in load-bearing application and stent application [48]. Basically, implant materials act as a mechanical support during the healing process thereafter degrade and loss their mechanical properties. Because of that the chemical and mechanical stabilities of implant materials during the healing period are critically important. While implants are exposed to human body fluid, it often experiences considerable loadings and, thus, may undergo environmentally assisted cracking (stress corrosion cracking (SCC) and corrosion fatigue). \nFigure 5\n shows the mechanical behavior of Mg implant in vivo for 12 weeks of implantation and the resulted tensile strength. The coated samples with HA indicated a higher mechanical stability than uncoated samples. The degradation volumes of the bare and HA-coated Mg specimens after 6 and 12 weeks of implantation find that the coated samples have lower degradation with addition protection (\nFigure 5b\n). The concept of the mechanically tuned with degradation rate during tissue regeneration is illustrated in \nFigure 5c\n. In period of 1–7 days, inflammation process, hematoma formation with atypical inflammatory response, occurs. Next stage repairs, hematoma, granulation tissue, connective tissue, cartilage, mineralization, woven bone, continuous for 3–6 months depend on the fracture position and type. In the final stage, remodeling, woven bone is replaced by cortical bone and the medullary cavity is restored, which persists for several years.
\n(a) In vivo evaluation of the mechanical and degradation properties of Mg coated with calcium orthophosphate coatings. (a) Optical images of the HA coated and bare samples at different implantation times at top and tensile strength of the HA-coated Mg samples comparing to the bare one after interval time. Reprinted from reference [49]. (b) Degradation volumes of the bare and HA-coated Mg specimens after 6 and 12 weeks of implantation [50]. (c) The schematic diagram of degradation behavior and the change of mechanical integrity of BM implants during the bone healing process [51].
Electrochemical polarization is an efficient technique used to evaluate metal corrosion potential in a short duration. Metals are commonly performed using electrochemical corrosion tests in SBF solution (pH = 7.4) at 37°C to mimic that of human blood plasma. The experimental setup consisted of three conventional electrodes within a cell, which named as working electrode, a saturated calomel electrode (SCE), or Ag/AgCl as a reference electrode, third is counter electrode such as a platinum wire. The experiment is conducted and monitored the current density as a function of the free open-circuit potential using the potentiostat of an electrochemical device. Initially, the samples are exposed to the solution for 10–20 min, a scan rate (mV/s) of the potentiodynamic polarization test is main parameter when test was performed. Corrosion current density (i\ncorr) could be estimated from the linear fit and Tafel extrapolation to the cathodic and anodic parts of the polarization curves. Thereafter, the corrosion rate can be calculated based on Faraday\'s laws.
\nwhere Ew is the equivalent weight of the corroding species in grams and ρ is the density of the corroding material in g/cm3.
\nFaraday’s laws assume a uniform corrosion in terms of the penetration, here the corrosion current (i\ncorr) is an effective factor in the corrosion rate and therefore the resulted value does not typically indicate an absolute corrosion rate for Mg. However, it represents indication of the corrosion distortion, which occurs at a selected point in time, in terms of current density. It is seldom in Mg and its alloys to degrade uniformly. The corrosion rate expressed with a current density value is highly accurate and can be considered to have the highest resolution of all methods. Current could be originating from a different local site on the surface and the method is considered as short-term and destructive one. In addition to the potentiodynamic polarization technique, it is essential to understand the mechanism of the corrosion rate; it may not suitable as a good indication for long-term corrosion rates. Electrochemical impedance spectroscopy (EIS) is a powerful tool that is used to evaluate a different application such as biosensors and conductivity as well as corrosion resistance of different metals using the frequency response of AC polarization [52]. EIS conducted with a range of low magnitude polarizing voltages that cycle from a peak anodic to peak cathodic voltage spanning a spectra of voltage frequencies. However, the technique has different components to understand the corrosion resistance of a metal surface. The components such as capacitance and resistance are obtained for each frequency and can then be used to explain a number of phenomena and properties of the metal surface. A determination of the corrosion rate is possible when the EIS determined polarization resistance (R\npolar) parameter at the zero frequency limit is used. The R\npolar is inversely proportional to the current density (j\ncorr) as described by the Stern-Geary relationship [44].
\nwhere βa and βc are the anodic and cathodic Tafel slopes, respectively.
\nA corrosion rate can be determined by transferring the current using EIS, the primary function of performing EIS on Mg and its alloys in an electrolytic solution is the identification and quantification of the formation behavior of corrosion layers which produced by the corrosion process. However, EIS results have some limitations as it can be affected by the Mg dissolution at low frequencies and therefore the chosen equivalent circuit. As a result, to employ EIS properly, a deep understanding of the corrosion processes takes place through the process and the best model. \nFigure 6a\n shows the potentiodynamic polarization curve of bare samples and anodized ones in SBF and SBF/ZrO2 NPs as an electrolyte with the resulted potential and current density. Moreover, EIS results in terms of Nyquist plot and bode diagrams are shown in \nFigure 6b and c\n. Both techniques find corrosion resistance in anodized samples comparing to the bare samples.
\nCorrosion evaluation of AZ31 Mg and anodizing samples in the SBF and SBF/ZrO2 NP electrolyte. Test was performed in SBF solution at 37°C under a three-electrode system where Mg samples act as an electrode, platinum as a counter electrode and Ag/AgCl as a reference electrode. (a) Potentidynamic polarization curves, (b) Nyquist plot and (c) Bode plot diagrams.
In this technique, an in vitro degradation rate in terms of mass loss is evaluated, before sample sterilization the initial weight of the samples was recorded. Then samples were immersed in a ratio of 30:1 volume to a weight ratio of SBF solution or Dulbecco\'s modified eagle\'s medium (DMEM) supplemented with 10% fetal bovine serum (FBS) for 168 h (1 week) under cell culture conditions (37°C, 20% O2, 5% CO2 and 95% humidity). The immersion medium recommended to be changed every 2–3 days to mimic the semistatic immersion test and to avoid saturation effects according to the standard ASTM-G31-72. After immersion time, the formed corrosion products were removed by treating the corroded disc with chromic acid (200 g/L Cr2O3 + 10 g/L AgNO3) at least for 20 min at room temperature [52].
\nFinally, the degradation rate (DR) was calculated in mm/year using the equation [53]:
\nwhere Δg is the change in weight by grams, A is the exposed surface area of the sample in cm2, t is the immersion time in hours and ρ is the density in g/cm3.
\nWhile exposure of the Mg substrate to aqueous solution generates H2 and OH− ions along the process of its degradation reaction with the medium, because of that the fluid pH value tends to be increase around the Mg surface. However, the instability of Mg occurs at pH values less than 11, a soluble compound formation with most inorganic ions would inhibits the formation of passive films of magnesium hydroxide in the biological environment. Moreover, the released Mg ions are another factor to indicate the dissolution of Mg in the aqueous solution process according to Eq. (8).
\nThere are various corrosion types during the Mg degradation process, including uniform corrosion [54, 55], localized corrosion [54, 55], flow-induced corrosion [55], erosion corrosion [56], galvanic corrosion [57], stress corrosion [58], atmospheric corrosion, hydrogen cracking [59] and intergranular corrosion [60]. It is worth noting that localized corrosion is always a source of stent fracture. In order to evaluate the biodegradability of Mg implant, a comparison between the anodized and the bare samples under in vitro conditions using SBF solution at 37°C up to 30 days usually is carried out. The corrosion rate based on mass loss and Mg2+ ions release rate and pH value is shown in \nFigure 7a–c\n. Both two factors can indicate the biodegradability behavior of Mg and its alloys. The tendency of bare samples to corrosion is significantly different from that of the anodized samples which has more corrosion resistance. The formation of Mg(OH)2 as a corrosion product in bare samples generated once exposed to aqueous solution. Similarly, more Mg ions release from bare samples resulted due to the high degradation and high sensitive surface of Mg. When Mg exposed to a corrosive medium similar to the human plasma corrosion product such as Mg(OH)2 and hydrogen release effect on the pH value of the surrounded solution. In short immersion time, Mg interface exhibits different features especially when treated with the CaP apatite film [61]. For example, when the surface rich with labile ions of CaP it was found to form HA nanoplates, as shown in \nFigure 7e\n; however, a pours structure formed in the case of apatite film enriched with ZrO2 NPs, as shown in \nFigure 7f\n. In contrast, the bare sample exhibits cracks and corrosion occurrence. These results can be attributed to the formation of a porous layer of nanoparticles/plates/rods of the CaP compounds with corrosion products such as magnesium hydroxide and calcium magnesium phosphate.
\nImmersion test results are showing, (a) corrosion rate, (b) Mg ions released and (c) pH value. FESEM images show the surface morphology of (d) bare sample (e) anodized sample in SBF (f) anodized samples in SBF/ZrO2 NPs after 3 days of immersions in SBF solution at 37°C.
Magnesium sample employed to the anodization technique is more stable in aqueous solutions and corrosive media due to the formation of a thin ceramic layer on the Mg interface. Therefore, Mg biodegradability can be controlled and delayed. An illustrative diagram in \nFigure 8b\n illustrates a corrosion mechanism before and after anodization treatment in 0.9 NaCl solution. First, the corrosive solution reacts with the substrate interface and starts to corrode and induce cracks and pitting corrosion. Upon increasing the exposure time, anodized film penetrated and the solution reached substrate surface. Thereafter, both the Mg(OH)2 and MgO by means of Cl− ions penetration are converted and degraded according to the chemical equation:
\nSchematic illustration of the degradation process: (a) bare AZ31 Mg alloy and (b) anodized samples.
Instantaneously, the corrosive solution contact substrate surface Mg+2 ions released and hydrogen gas evaluation occurs. As a result, Mg (OH)2 will deposit and react with Cl− ions to form MgCl2 leading to corrosion occurrence according to the chemical equation:
\nThe pitting corrosion on the metal surface is due to chloride ions; therefore, the main concept of anodization film is to block Cl− ions and retard corrosion occurrence on the Mg surface [6].
\nBiomaterials must be designed to be biocompatible; however, the majority of biomaterials community has failed to understand the biocompatibility paradigm [62].
\nBasically, biocompatibility is a characteristic and a complex characteristic at a system and not a material. There are different effects of materials in biological systems as, tissue processing involved in wound healing, the endothelium in contact with intravascular implant devices and the stem cells in bioreactors, the target cells in gene therapy, emphasize that there is no material with complete biocompatibility characteristics [63]. In biodegradable implant such as Mg, bare substrates without any surface modifications show few round shapes of cells on its surface. These attributed to many factors which mainly show corrosion behavior with combined hydrogen gas and induce toxicity to surround tissues. Moreover, surface tribology has additional effect, for example, a rough surface has more cell attachment comparing to smooth one in nanoscale, which behaves as accommodation for cells [64]. In addition, biomimetic nanostructure on the implant surface can enhance biocompatibility and cell proliferation. The Mg substrate that employed to surface modifications using the anodization/hydrothermal process with nanoplate structure shows flat and well-spread features among the nanoplates, as shown in \nFigure 9\n. Cell proliferation of the extraction of HA nanoplates on the Mg alloy surface finds higher cell proliferation. This can conclude that cells can modify their morphology to match the surface topography as shown in the inset images in \nFigure 9\n. These findings indicate that how surface modification can influence surface bioactivity and cell adhesion to the implant interface. Implant surface adheres with the cells and eliminates the mismatch between the surface of the biomaterials and the connected tissue [65]. Extraction of anodized layers shows more cell viability and proliferation as shows in \nFigure 10\n using confocal microscopic images comparing to the bare substrates extraction.
\nFigure shows the cell viability of anodized/hydrothermal treated Mg samples using cck-8 (a). FESEM images show cell attachment after 5 days (b); naked (c); (SBF-HA) (d); and (SBF-HA)/HT samples. Cell proliferation is presented in means ± STD (n = 4) based on ANOVA one-way test (*indicates p < 0.05).
Microscopic florescent images for the live/dead cells of the (a) negative control, (b) positive control, (c) bare sample, (d) AZ31 Mg alloy anodized in SBF at 37°C.
Magnesium and its alloys are exhibit biodegradable in physiological media as well as its stiffness close to bone. In addition characteristics of Mg such as low weight, high specific strength and good biocompatibility bring a revolution in medical field toward new generation of biomaterials. However, the high degradation is accompanied by the hydrogen gas effect on the healing of the surrounded tissues. During its healing period, Mg implants lose their mechanical integrity before the bone heals due to the high degradation process. To overcome these limitations, different methods and techniques have been proposed to control the degradation rate of Mg to acceptable levels. Anodization as one of the surface modification techniques finds to increase the surface bioactivity and control degradation rate. In bone substitute Mg acts as a mechanical support during the healing process; moreover, the presence of apatite film on the surface of implant materials can enhance osseointegration of the defected bone. Furthermore, more research studies are devoted to Mg to be used in the future as implant materials in clinical application.
\nThis chapter was supported by grant from the Basic Science Research Program through the National Research Foundation of Korea (NRF) by Ministry of Education, Science and Technology (Project no. 2016R1A2A2A07005160) and (Project no NRF-2015R1C1A1A02036404).
\nHeavy metals are those elements which have density greater than 5 g cm−3 [1]. Some heavy metals namely, cobalt (Co), copper (Cu), molybdenum (Mo), manganese (Mn), nickel (Ni) iron (Fe), and zinc (Zn) are considered to be essential for plants. These heavy metal elements directly impact on plant growth, development, senescence and energy producing processes and other physiological process due to their high reactivity. The concentration of heavy metals in soil after the admissible limits is toxic to plants either provoke oxidative stress through free radicals or crumbling up the functions of enzymes by replacing metals and nutrients which are essential [2, 3]. Cell metabolism changes by the affect of heavy metals at first reduce the plant growth. However, toxicity of metals depends on various stage of their growth stage [4]. Maksymiec and Baszynski [5] reported that dicotyledonous plants like various beans and Medicago sativa were more resistant to heavy metals at the early growth stage [6]. So, the heavy metals toxicity on the plant physiology and metabolism are much more noticeable. Among the heavy metals, chromium and cadmium are of special concern due to their potential toxicity on plants even at low concentrations [7, 8, 9]. The various types of chromium toxicity in plant had described by [10], and the inhibition of enzymatic activity by vaeious types mutagenesis had also be described. The visible symptoms are reduction in growth, leaf chlorosis, stunting, and yield reduction [7, 11]. [12] has explain that Cadmium (Cd) is particularly is one of the most dangerous pollutant due to its high level of toxicity and much solubility in water. [13, 14], have reported that in some plant species Cd interacts with the absorption of metal nutrients such as Fe, Zn, Cu and Mn, in addition to inducing a process named as peroxidation and breakdown of chlorophyll in plants, resulting in an enhanced production of reactive oxygen species (ROS) [15]. According to [16], Cadmium also inhibits the uptake of elements such as K, Ca, Mg, Fe because it uses the same transmembrane carriers. Cadmium acquisition in plants may also cause serious health hazard to human beings through food chain; however, it causes an extra risk to the children by direct ingestion of Cd-contaminated soil [17].
Heavy metals remain in environment in various forms like colloidal, ionic, particulate and dissolved phases. The soluble forms of heavy metal elements are remain in environment as ionised or unionized organometallic chelates. According to [18], the metal concentrations of soil ranges from low to 100,000 mg kg−1 which depends on the location, area and the types of metals. [19], studied that among chemical elements, Cr is considered to be the seventh most abundant elements on earth and constitutes 0.1 to 0.3 mg kg − 1 of the crystal rocks. According to McGrath [20], In alloys and 15 percent in chemical industrial processes, mainly leather tanning, pigments, electroplating and wood preservation, about 60–70 percent of the total world production of Cr is used. Chromium has many oxidation states ranging from Cr2− to Cr6 +; however, in a number of compounds, valences of I, II, IV and V have been shown to exist [21]. Cr (VI) is, however, considered the most toxic form of chromium and is also generally associated with oxygen as chromate (CrO42−) or dichromate (CrO42−) and dichromate (Cr2O72−) oxyanions. [22], observed that Cr (III) is less mobile and less toxic and is mainly bound to organic matter in soil and aquatic environments. According to [23], Cr present mostly in the form of Cr (III) in soil, and mineral environment. [24], has described that Cr and Fe (OH)3 is a solid phase of Cr(III) having even lower solubility than Cr(OH)3. Consequently, within the soil add up to solvent Cr(III) remains inside the allowable limits for drinking water for a wide extend of pH (4–12) due to precipitation of Cr(OH)3, Fe(OH)3[25, 26], moreover, major source of Cd is the parental fabric. Anthropogenic exercises have too been improved the sum of Cd in soil [27]. Overwhelming metals are regularly show at exceptionally moo concentrations in freshwaters [28], but the release of fluid squander from a wide assortment of businesses such as electroplating, metal wrapping up, calfskin tanning, chrome planning, generation of batteries, phosphate fertilizers, shades, stabilizers, and amalgams has solid impact in sea-going situations [29, 30, 31]. Cadmium pollution is also happened from rubber when car tires run over streets, and after a rain, the Cd is washed into sewage disposal systems and collected in the slush.
Heavy metals are enter in environment are transported by water and air, also deposited in soil and sediments where they could be immobilized [32]. However, the bonding process of metals may take considerably long time. At the starting of the official handle the bio accessible division of metal components in soil is tall, but diminishes continuously in due course of time [33]. Metal dissolvability and bioavailability to plant is basically affected by the chemical properties of soil such as, soil pH, stacking rate, cation trade capacity, soil surface, redox potential, clay substance and natural matter [34, 35, 36]. For the most part, higher the slime or natural matter and soil pH, the metals will be relentlessly bound to soil with longer time and will be less organically accessible to the plants. Soil temperature is additionally an vital calculate for varieties in metal amassing by crops [37]. The bioavailability of metals is make greater in soil through several means, the secretion of phytosiderophores into the rhizosphere to chelate and solubilise metals that are soil bound [38]. Acidification of the rhizosphere and exudation of carboxylates are deliberated potential means to enhancing metal consumption.
Heavy metals are taken through root cells of the vegetation after their mobilization inside the soil, and their improvement inside the soil relies upon in the main upon: (i) dissemination of steel additives alongside the attention attitude which has formed because of take-up of factors and ultimately inanition of the aspect inside the root region; (ii) interferences through roots, in which soil extent is uprooted through root extent after developing (iii) move of steel additives from enormous soil association down the water capacity slope [39]. Cell divider acts as a particle exchanger of relatively moo partiality and moo selectivity in which metals are first of all bound. From the mobileular divider, the shipping frameworks and intracellular high-affinity authoritative locations intercede and power the take-up of those metals over the plasma layer. A stable using power for the take-up of steel additives thru auxiliary transporters is made because of the layer capacity, that is bad at the indoors of the plasma movie and can exceed −200 mV in root epiderm. This is examined both in soil culture and in solution culture for Cd which might probably be due to low concentration of heavy metals per unit of absorption area [40, 41]. Both non-essential and essential metals are also preoccupied through leaves. Within the shape of gases, they input via thestomata withinside the leaves, while in ionic shape metals specifically input via theleaf cuticle [39, 42]. Hg in gaseous shape istaken up through stomata [43] and its uptake is recommended to bebetter in C3 than C4 flora [44]. The uptake of metals takes place viaectodesmata, non-plasmatic “channels” at a excessive level whichare much less dense elements of the cuticular layer which are located fundamental withinside theepidermal mobileular wall or cuticular membrane machine among shield cells andsubsidiary cells. Furthermore, the cuticle overlaying shield cells are oftenspecific to it overlaying everyday epidermal mobileular [39]. Most of the metallic factors are insoluble that won’t capin an edge toflow freely withinside the vascular machine of flora and, as a result typically shapesulphate, phosphate or carbonate precipitates immobilizing them inextracellular booths i.e. apoplastic and intracellular compartment i.e. symplastic [45]. In the apoplastic pathway solute and also the water debris diffuse via mobileular membrane, consequently the pathway stays unregulated. The mobileularwall of the endodermal mobileular layer acts as an impediment for apoplastic diffusioninto the vascular machine. Generally, prior to the access of metallic ions withinside thexylem, solutes must be haunted through root symplasm [46]. Ifmetals are obsessed through the premise symplasm, their similarly motion from root tothe xylem is specifically ruled through 3 processes, including: (i) metallicsequestration arise into the premise symplasm, (ii) symplastic shipping ariseinto the stele, and (iii) launch of metals arise into the xylem. The ionshipping into the xylem is often occured through membrane shipping proteins. Metal factors which are not wished through the flora successfully compete thecritical heavy metals for his or her shipping the usage of the equal transmembranecarriers. Cr(III) uptake through the plant is specifically a passive process, whilst Cr(VI) shipping is mediated through sulphate carrier [47]. Inhibitors like, sodium azide and di nitrophenol inhibits the uptake of Cr(VI) through barley seedlings however this is not happened just in case of Cr(III) [47]. In keeping with [48], Group VI anions like SO4−2 additionally inhibit the uptake of chromateswhile Ca2+ stimulates its shipping. This inhibition of chromate shipping is passed thanks to the aggressive inhibitiondue to the chemical similarity, whilst inspired shipping of Cr(VI) because of Ca is attributed to its critical position in flora for the receive and shipping of metallic factors [26, 49].
According to Kumar et al. [50], many plants species show an unusual capability to absorbe heavy metals through root system and accumulate of these heavy metals in their parts. Zayed and Terry [26] said that it seems a common tendency of all plant species to maintain Cr in their roots, but with quantitative differences. It is found that for the translocation of Cr to the plant tip, leafy vegetables such as spinach, turnip leaves that tend to acquire Fe appear to be the most effective [51]. While those leafy vegetables such as lettuce were considerably less effective for translocating Cr to their leaves, cabbage which accumulated relatively low Fe levels in their leaves. Zayed and Terry [26] have reported that some plant species attain substantially higher root or shoot concentration ratio than other species. However, a ‘Soil–Plant Barrier’ well protects the food chain from heavy metal toxicity, implying that, due to one or more of the following processes, heavy metal levels in edible plant tissues are reduced to safe levels for animals and humans: (i) prevention of metal element uptake due to soil insolubility, (ii) prevention of metal element translocation by making them immobile in roots, or (iii) prevention of metal element translocation for animals and humans to the permissible level [52]. Within plant tissues, some elements such as B, Mo, Cd, Mn, Se, and Zn are readily absorbed and translocated, while others such as Al, Ag, Cr, Fe, Hg, and Pb are less mobile because of their strong binding to soil components or root cell walls. However, at certain concentrations, all of these elements are mobilised, even against a concentration gradient, within the transport system of the plant. Kinetic data show, for instance, that essential Cu2 +, Ni2 + and Zn2 + and non-essential Cd2 + compete for their transport with the same transmembrane carrier [53]. As is the case of phytosiderophore such as Fe-transport in graminaceous species, metal chelate complexes can be transported by plasma membrane [54]. Among the most important parameter the most influencing factor of heavy metal accumulation in plants is soil pH [55, 56, 57, 58]. At higher soil pH, metal elements in soil solution decrease their bioavailability, and at lower soil pH metalelements in soil solution increase their bioavailability to plants [59].
Heavy metals mitigate the growth and development of the plant [60, 61]. The plant parts which are associated with the heavy metals polluted soils normally the roots express rapid and sensorial changes in their growth and development [62]. It is well observed that the very significant effects of a number of metals (Cd, Al, Cu, Fe, Ni, Pb, Hg, Cr, Zn,) on the growth of above ground plant parts vary [63]. Through the formation of free radicals and reactive oxygen species (ROS), heavy metals mainly affect plant growth, which causes constant oxidative damage by decreasing important cellular components. [64, 65]. For example, rice seedlings irradiated to Cd or Ni [66] and runner bean plants treated with Cd and Cu have shown an increase in carbohydrate content and a decrease in photosynthesis process, resulting in growth inhibition [67]. Similarly, in cucumber plants, Cu limits K uptake by leaf and inhibits the photosynthesis via sugar acquisition resulting into the inhibition of cell expansion [68]. Limped leaves, growth inhibition, progressive chlorosis in certain leaves and leaf sheaths and browned root systems, especially the root tips, are the symptoms of Cd toxicity in rice plants [7, 69]. Moreover, plant growth has also been retarded in maize (Zea mays) Cd [70, 71]. Some phenotypic abnormalities such as stunted growth, less branching and less fruiting are also shown by tomato plants irrigated with polluted water. However, acquisition of heavy metals is much more appears in stems, roots, and leaves as compared to fruits [72].
Seed germination is the breaking of seed dormancy which is inhibited by heavy metals. Germination of seeds and growth of seedling may sensitive towards environmental conditions [59]. So as per [73], the performance of germination, breaking of seed dormancy and seedlings growth rates are therefore often used to assess the abilities of plant tolerance to metal elementsIn comparison to control, higher concentrations such as 1 μM, 5 μM and 10 μM of heavy metals such as Cu, Zn, Mg and Na significantly inhibit seed germination and early growth of rice, barley, wheat and maize seedlings [74]. The ability of a seed to germinate in a moderate containing any metal element like Cr would be a direct indication of its level of tolerance to this metal, but seed germination is the first physiological process affected by toxic elements [73]. At 200 μM of Cr treatment, the seed germination of Echinochloa colona is decreased to 25 percent [75], and high levels (500 ppm) of Cr (VI) in soil decreased Phaseolus vulgaris germination by up to 48 percent [76]. Jain et al. [77] observed reductions in sugarcane bud germination of up to 35 per cent and 60 per cent at 20 and 80 ppm Cr application, respectively. In another study by Peralta et al. [73], at 40 ppm Cr (VI) treatment, Medicago sativacv germination was reduced to 23 percent.
Among the plant parts, roots are firstly come into contact with toxic elements and they usually absorbed more metals by root hair through absorbption process but shoots are not that [78, 79, 80]. The inhibition or retard of root elongation appears to be the first visible effect of metal toxicity. Elongations of root are reduced by the inhibition of cell division, the decrease of cell expansion, decrease of cell size in the elongation zone [81]. So the first visible effect of metal toxicity is the inhibition of root elongation, the root length can be used as most important tolerance index [82, 83, 84, 85]. Medicago sativa plants grown in solid media watered with 20 mg L−1 of Cr (VI) in another [73] study, the ratio of Cr in shoots to Cr in roots was approximately 43 percent. This is an indication that in the roots, 50 percent of the absorbed Cr is held. The response of roots to heavy metals in both herbaceous plant species and trees has been extensively studied. [86, 87, 88, 89]. After the work of numerous researchers [86, 87, 89, 90]. The main morphological and structural effects of metal root toxicity can be summarised as: (i) decrease in root elongation, (ii) decrease in biomass, (iii) decrease in vessel diameter, (iv) damage to tip, (v) collapse of root hair or decrease in number of roots, (vi) increase or decrease in lateral root formation, (vii) enhancement of suberification, (viii) enhancement of lignifications, (ix) translocation process become hampered. The research work of [91], revealed that Cr affects the root length than the other parts of plant as compared to other heavy metals. Mokgalaka-Matlala et al. [92], have observed that when increasing concentrations of As (V) and As (III) in Prosopis juliflora, the root elongation decreased significantly. It is reported that when Cr has applied on Salix viminalisis then the root length is affected more than by Cd and Pb [91]. In fact, the inhibition effect of Cr on the growth of the Salix alba root is similar to that of Hg and stronger than that of Cd and Pb, whereas the root length of Ni decreased less than Cr [93, 94]. In Salix viminalisis, the order of metal toxicity to the new root rimordial was reported to be Cd > Cr > Pb [91].
The heavy metal elements highly affect the plant height as well as shoot growth [95]. Cr transport to the various part of the plant have a direct impact on cellular metabolism as a result shoots contributing affected so plant height ultimately reduces [61]. It is observed that reduction of 11, 22 and 41% respectively compared to control in oat plants at 2, 10 and 25 ppm of Cr content in nutrient solutions in sand cultures [96]. Joseph et al. [97] observed a similar reduction in the height of Curcumas sativus, Lactuca sativa and Panicum miliaceum due to Cr (VI). Shoot growth in Medicago sativa is inhibited by Cr (III) [98]. In a glasshouse experiment after 32 and 96 days, Sharma and Sharma [99] noted a significant decrease in the height of Triticum aestivum when sown in sand with 0.5 μM sodium dichromate. A significant reduction in height of Sinapsis albaat a level of 200 or 400 mg kg−1 of Cr in soil along with N, P, K and S fertilizers was reported by Hanus and Tomas [100]. Very recently, it is found that a reduction in stem height at various concentrations (10, 20, 40 and 80 ppm) of Cd and Cr have been reported in Dalbergia sisso seedlings compared to the control [101].
The heavy metal elements severely affect the leaf height as well as leaf growth. Metal elements like Cd induce morphological changes such as drying of older leaves, wilt, and chlorosis and necrosis of younger leaves. Datura innoxia, D. metel, plants grown in a contaminated environment with Cr(VI) exhibited toxic symptoms at 0.1 mM to 0.2 mM of Cr(VI) in the form of leaf fall and wilting of leaves at 0.4 to 0.5 mM Cr(VI) in soil [97, 102]. A similar reduction in the height of Curcumas sativus, Lactuca sativa and Panicum miliaceum due to Cr(VI) was observed (1995). In Medicago sativa, shoot growth is inhibited by Cr(III) [98]. Sharma and Sharma [99] noted a significant drop in the height of Triticum aestivum when sown in sand with 0.5 μM sodium dichromate in a glasshouse experiment after 32 and 96 days [103]. In Zea mays, Acacia holosericeaOryza sativa, and Leucaena leucocephala plants treated with tannery effluent of varying concentrations, leaf dry weight and leaf area slowly decreases [104]. The effect of Cr(III) and Cr(VI) on the Spinacia oleracea plant was found in a study. Singh [105] reported that Cr applied to soil at a rate of 60 mg kg−1 and higher levels decreased the size of the leaves, causing leaf foliage, leaf tips or margins to burn, and slowed the rate of leaf growth.
The physiological process of the plant is severely affected by heavy metal elements. In reaction to heavy metal stress, plants show morphological, physiological, biochemical and metabolic changes which are thought to be adaptive responses [106]. Cd not only inhibits growth, for example, but also changes different physiological and biochemical features such as water balance, nutrient uptake, photosynthesis, breathing, mineral, nutrition and ion uptake, translocation, plant hormone [107, 108, 109] and Photosynthetic electron transport around PS I and PS II photosystems [110, 111, 112]. Likewise, Cr inhibits electron transport, decreases CO2 fixation, malformation of chloroplast [113, 114, 115], decreases water potential, increases transpiration rate, decreases diffusive resistance, and causes a reduction intercalary meristem [116].
The photosynthetic mechanism is significantly impacted by the heavy metal elements. The photosynthetic apparatus tends to be very susceptible to the toxicity of heavy metals, which directly or indirectly affect the photosynthetic process by inhibiting the enzyme activities of the Calvin cycle and CO2 deficiency in the plant body due to stomatal closure [59, 117, 118]. Cr has a well-cited detrimental effect on the photosynthic process in terrestrial plants. The influence of Cr on the PS I was more conspicuous than on the PS II operation in isolated chloroplasts of Pisumsativum plant [119] according to different reports. Photo inhibition in the leaves of Lolium perenne due to the influence of 250 μM Cr on the primary photochemistry of PS II, according to the Vernay et al. [120] report and A decrease in the overall photochemical efficiency of plant PS II at 500 μM of Cr was noted. Shanker et al. [61] argued that Cr triggered oxidative stress in plants because, due to the loss of molecular oxygen, Cr improves alternate sinks for the electrons. The ultimate influence of Cr ions on photosynthesis and conversion of excitation energy will be attributed to Cr-induced anomalies such as thylakoid expansion and reduction in the amount of grana in the ultrastructure of the chloroplast [121]. The impact of Cr on photosynthesis in higher plants is widely known [122, 123], it is not well known to what degree Cr induces photosynthesis inhibition either because of ultra-structure chloroplast malformation and the influence of Cr on the Calvin cycle enzymes or because of electron transport inhibition [116]. Krupa and Baszynski explained in 1995 that some theories applied to all photosynthesis pathways of heavy metal toxicity and introduced a list of primary photosynthetic carbon reduction enzymes that inhibited mainly cereal and legume crops in heavy metal treated plants. The 40 percent inhibition of whole plant photosynthesis in 52-day-old Pisum sativum seedlings at 0.1 mM Cr(VI) was further increased to 65 and 95 percent after 76 and 89 days of growth respectively [119]. A potential explanation of Cr mediated reduction rate of photosynthetic is a malformation of the chloroplast ultra structure and inhibition or returdation of electron transport processes due to Cr and a diversion of electrons from the electron donation side of PS-I to Cr (VI). It is likely that, as demonstrated by the low photosynthetic rate of the Cr stressed plants, electrons generated by the photo chemical process are not generally used for carbon fixation. According to [124, 125, 126], bioaccumulation of Cr and its toxicity to photosynthetic pigments in various crops and trees has been investigated. [127]; has extensively studied the effect of Cr present in tannery effluent sludge which directly get into chloroplast pigment content in Vigna radiata and reported that irrespective of Cr concentration, chlorophyll a, chlorophyll b, chlorophyll d and total chlorophyll decreased in 6 days old seedlings as compared to control. Chatterjee and Chatterjee [128] have reported that a dramatic decrease in chlorophylls a, b and d in leaves was recorded in Brassica oleracea grown in distilled sand with full nutrition with control and Co, Cr and Cu at 0.5 mM each. The stress order was Co > Cu > Cr. Conversely, a broad analysis on the tolerance of Cr and Ni in Echinochloa colona found that in terms of survival under elevated Cr concentration, the chlorophyll content was high in resistant calluses [129]. Chromium (VI) at 1 and 2 mg L−1 significantly decreased chlorophylls a, b and d and carotenoid concentrations in Salvinia minima [130]. The decrease in the chlorophyll a/b ratio brought about by Cr indicates that Cr toxicity possibly reduces the size of the peripheral part of the antenna complex [114]. It has been hypothesized that the decrease in chlorophyll b due to Cr could be due to the destabilization and degradation of the proteins of the peripheral part [61]. The interaction of heavy metals with the functional SH groups of proteins according to Van Assche and Clijsters [131, 132] is a possible mechanism of action for heavy metals.
Every physiological process is directly linked to water’s chemical potential. Water’s chemical potential is a quantitative expression of water-related energy. In plant growth regulation, water can be considered as the most important factor because it affects all growth processes directly or indirectly [133]. Plants grown in contaminated heavy metal soils often suffer from drought stress due primarily to poor physicochemical properties of the soil and shallow root system; researchers are interested in investigations on plant water relation under heavy metal stress. According to Barcelo et al. [134], Selection of drought resistance species can be considered to be an important trait in phytoremediation of soils polluted with heavy metals. The heavy metal stress can induce stress in plants through a series of events leading to decreased water loss like enhanced water conservation, decrease in number and size of leaves, decrease in root hair, malformation of parenchymatous cells stomatal size, number and diameter of xylem vessels, increased stomatal resistance, enhancement of leaf rolling and leaf abscission, higher degree of root suberization [90]. It has been suggested that through various mechanisms operating on the apoplastic and/or the symplastic pathway, heavy metals may influence root hydraulic conductivity. Reduced cell expansion can occur in the growth medium at relatively low concentrations without damaging the integrity of the cells. In bean plants, for instance, leaf expansion growth was inhibited after 48 h in bean plants exposed to 3 uM Cd. The most significant higher toxic effect of Cr (VI) is to degenerate the stomatal conductance that could damage the cells and membranes of stomatal guard cells. In this way, the relationship between water and many plant species has been affected.
Complex processes has used by plants to adjust their metabolism to rapidly changing environment. These processes include transduction, transcription, perception, and transmission of stress stimuli [135, 136, 137]. During stressing conditions plants adopt various process likes mechanisms of resistance and tolerance, later involves the immobilization of a metal in roots and in cell walls [138]. The plants adopt a series of mechanisms to avoid heavy metal toxicity which include: (i) Through auto oxidation and Fenton reaction plant produce reactive oxygen, (ii) blocking of main functional group, and (iii) from biomolecules displacement of metal ions, [139]. Plants are capable of growing in polluted soils because; (i) plants avoid metal absorption by aerial components or sustain low metal concentrations over a wide range of metal concentrations in soil by trapping metals in their roots [140]; (ii) plants deliberately absorb metals in their epidermal tissues due to the development of metal binding chelators (iii) they storing metals in non-sensitive parts by alter metal compartmentalisation pattern that is called metal indicators, and (iv) by the process of hyperaccumulators i.e. they can accumulate metals at much higher levels than soil in their aerial components [141, 142]. The processes used for hyperaccumulation are still unclear. Plants that can accumulate either As, Cu, Cr, Ni, Pb, or Co > 1000 mg kg−1 or zinc >10,000 mg kg−1 in their shot dry matter ([141, 143, 144, 145]; Baker and Reeves 2000) or Mo > 1500 mg kg−1 [146] are the standard for classifying plants as hyperaccumulators. (ii) Plants that absorb metals 10–500 times higher than average amounts in shoots [147], (iii) plants that accumulate metal components more in shoots than in roots [141]. Very few higher plant species have adaptations that enable them to live and replicate with Zn, Cu, Pb, Cd, Ni, and As highly polluted soils. [148, 149]. The tree roots of these plants can deliberately forage towards less polluted soil areas [150] and can “rest and wait” for optimal growth conditions even with highly reduced growth [151].
For the biological, biochemical and physiological functions of plants, various types of heavy metal elements are very important, including protein biosynthesis, lipids, nucleic acids, growth substances, hormones, chlorophyll and secondary metabolism synthesis, stress tolerance, morphological, structural and functional integrity of different membranes and other cellular compounds. These metal components, however, become poisonous in nature, above allowable limits, depending on the types of plants and the nature of the metal. Metal toxicity can inhibit the transport chain of electrons, reduce CO2 fixation, decrease the production of biomass, and cause chloroplast malformation. It can also affect plant growth by generating free radicals and ROS and other substances, which, by decreasing important cellular components, pose a threat to continuous oxidative damage. In addition, heavy metal stress can induce many events in plants leading to decrease in number and size of leaves, enhancement of leaf rolling and leaf abscission, leave erosion, changes in stomatal size, guard cell size, and stomatal resistance, and higher degree of root ligninization, suberization. Symptoms that are visible in plant by the affect of heavy metal toxicity include drying of older leaves, chlorosis, and necrosis of young leaves, stunting, wilting, canker, colour changes, blotch wrinkling and yield reduction. However, plants use complex processes (perception, transduction, and transmission of stress stimuli) and several non enzymatic and enzymatic mechanisms such as CAT, SOD, POD, and APX that activate the cell for their metabolism to heavy metal stress.
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