\r\n\tThis book aims to explore the issues around the rheology of polymers, with an emphasis on biopolymers as well as the modification of polymers using reactive extrusion.
",isbn:null,printIsbn:"979-953-307-X-X",pdfIsbn:null,doi:null,price:0,priceEur:0,priceUsd:0,slug:null,numberOfPages:0,isOpenForSubmission:!1,hash:"5bc21841d2b87388ad498bc09910944b",bookSignature:"Dr. Casparus Johannes Verbeek and Dr. Velram Mohan",publishedDate:null,coverURL:"https://cdn.intechopen.com/books/images_new/8880.jpg",keywords:"Extrusion, Injection Moulding, Thermoplastics, Natural Polymers, Biomass, Polymer Modification, Polymer Blends, Compatibilization, Processing Challenges, Reactive Compounding, Screw Design, Process Design",numberOfDownloads:null,numberOfWosCitations:0,numberOfCrossrefCitations:0,numberOfDimensionsCitations:0,numberOfTotalCitations:0,isAvailableForWebshopOrdering:!0,dateEndFirstStepPublish:"September 6th 2019",dateEndSecondStepPublish:"September 27th 2019",dateEndThirdStepPublish:"November 26th 2019",dateEndFourthStepPublish:"February 14th 2020",dateEndFifthStepPublish:"April 14th 2020",remainingDaysToSecondStep:"a year",secondStepPassed:!0,currentStepOfPublishingProcess:5,editedByType:null,kuFlag:!1,biosketch:null,coeditorOneBiosketch:null,coeditorTwoBiosketch:null,coeditorThreeBiosketch:null,coeditorFourBiosketch:null,coeditorFiveBiosketch:null,editors:[{id:"102391",title:"Dr.",name:"Casparus",middleName:"Johannes",surname:"Verbeek",slug:"casparus-verbeek",fullName:"Casparus Verbeek",profilePictureURL:"https://mts.intechopen.com/storage/users/102391/images/system/102391.jpeg",biography:"Dr Verbeek is a Chemical Engineer, currently an associate professor at the School of Engineering at the University of Waikato and is also the R&D manager for Aduro Biopolymers. 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1. Introduction
As one of the remarkable products of evolution, lactation is a very dynamic and complex process. The process of lactation involves the development of the mammary gland (MG) and the synthesis and secretion of milk. The lactation process is affected by many factors, including genetics, epigenetics, non-genetics and environmental factors. The knowledge of lactation regulation is not only important for improvement of milk production and quality but also provides a model for basic cellular processes (proliferation, differentiation, survival and death) [1], which may have important implications for productivity (milk yield) and disease status (e.g. breast cancer, mastitis, etc.). The endocrine regulation and physiological processes as well as the signalling pathways involved in these processes are fairly understood [1, 2]. Facilitated by the release of the whole genome sequences of cattle, sheep and goat [3–6] as well as availability of single nucleotide polymorphism (SNP) genotyping chips [7–11], the genetic mechanisms of ruminant lactation have been extensively explored (Figure 1). As a consequence, many quantitative trait loci (QTL) and genetic markers for lactation-related traits (for instance, milk yield, milk components, lactation persistency, etc.) have been detected and catalogued in the animal QTL database (http://www.animalgenome.org/cgi-bin/QTLdb/index).
Figure 1.
Growing research by year in the field of cattle genomics and transcriptomics (including non-coding RNA) from January 2000 to August 2016.
Transcriptomics research by both microarray and RNA sequencing methods has allowed for a better understanding of the genes and regulatory networks of complex traits in animals [12], such as the biosynthesis of major milk components (reviewed in Refs. [13, 14]). Emerging studies now suggest that non-coding RNAs (ncRNAs) are key regulators of mammary gland development and lactation processes [15–17]. The results from the ENCODE (ENCyclopedia of DNA elements) project [18, 19] indicate that only a small portion of the genome, about 1.5%, codes for proteins while most of the genome is transcribed into non-coding regulatory elements or ncRNA. This indicates that ncRNAs play significant regulatory roles in complex animal traits. A similar project to functionally annotate regulatory elements in animal genomes (FAANG project, www.faang.org) started in 2014 [20] and will generate data that will foster understanding of how the genome is read and translated into complex animal traits of economic importance. Indeed, the recent explosion of data on the regulatory functions of ncRNAs proves their importance in the regulation of multiple/major biological processes impacting development, differentiation and metabolism. This chapter explores recent developments on the expression, regulation and functions of ncRNAs, in particular microRNA (miRNA) and long non-coding RNA (lncRNA), in ruminant (cattle, sheep and goat) mammary gland development and the lactation process, as well as illustrate our own studies on the roles of ncRNAs in these processes.
2. Non-coding RNAs: biosynthesis and classification
Non-coding RNAs are transcribed RNA molecules that are not translated into proteins. They play a remarkable variety of biological functions by engaging target transcripts through sequence-specific interactions. They regulate many biological processes, including gene expression (transcription, RNA processing and translation), protect genomes from foreign nucleic acids and can guide DNA synthesis or genome rearrangement [21]. In general, ncRNAs are classified according to size or function. According to size, ncRNAs are classified as (1) small or short ncRNA: <200 nucleotides in their mature forms (e.g. miRNA, PIWI-interacting RNA [piRNA], small nuclear RNA [snRNA], small nucleolar RNA [snoRNA] and endogenous small interfering RNA [siRNA]) and (2) long ncRNA: >200 nucleotides long (e.g. lncRNA). According to function, ncRNAs are classified as (1) housekeeping or translation-related ncRNAs: they are constitutively expressed and crucial for normal cellular function and viability and include tRNA, rRNA and snoRNA and (2) regulatory ncRNAs and include miRNA, lncRNA, siRNA and piRNA [22, 23]. The biogenesis of these various types of ncRNAs has been discussed extensively [23–26]. This chapter focuses particularly on the involvement of miRNA and lncRNA in ruminant mammary gland development and lactation.
2.1. MicroRNAs
MiRNAs are an abundant class of short ncRNAs of about 22 nucleotides long. They regulate a variety of cellular processes through post-transcriptional repression of gene expression. MiRNAs consequently control the activities of about 60% of all protein-coding genes and participate in the regulation of almost every cellular process investigated in mammals [25]. Mature miRNAs are generated from a series of biochemical events beginning in the nucleus and culminating in the cytoplasm [24, 27, 28]. Briefly, these events occur in several main steps as follows: (1) nuclear processing of primary miRNA transcripts (pri-miRNAs) into precursor miRNAs (pre-miRNAs) by the DiGeorge Syndrome Critical Region Gene 8 (DGCR8)/Drosha complex, (2) cytoplasmic processing of pre-miRNAs into imperfectly paired miRNA duplexes by dicer, and (3) preferential incorporation of one strand (the ‘guide’ miRNA strand) onto the RNA-induced silencing complex (RISC) [25]. Most miRNA genes located in introns of protein-coding genes share the promoter of the host gene [29]. MiRNAs often have multiple transcription start sites and regulate gene expression through inhibition of translation initiation or elongation, co-translational protein degradation and premature termination of translation [25, 30].
Since the discovery of the first miRNA, lin-4, in 1993 [31] and aided by deep sequencing technologies and developments in bioinformatics processing of deep sequence data, thousands of miRNAs have been detected in humans, mouse, farm animal species and plants and deposited in the miRNA data base (Table 1). Due to the crucial regulatory roles of miRNAs in many biological processes across species, they are being considered as candidate biomarkers of various human diseases, such as autoimmune [32], metabolic [33] and cardiovascular diseases [34], and various types of cancers [35–37].
Species
MiRNA
lncRNA
Precursor
Mature
Transcripts
Genes
Cattle
808
793
22,386
23,696
Sheep
106
153
–
Goat
267
436
–
Pig
382
411
–
Chicken
740
994
13,085
9681
Human
1881
2588
141,353
90,062
Mouse
1193
1915
117,405
79,940
Table 1.
Number of detected miRNAs and lncRNAs in farm animal species, mouse and human*.
Data source: MiRBase release 21 (http://www.mirbase.org/[38], and NONCODE database (www.noncode.org, Noncode 2016 [39]).
2.2. Long non-coding RNAs
Long non-coding RNAs are a diverse collection of non-coding RNAs with emerging regulatory roles in many biological processes in every branch of life [26, 40–42]. LncRNA transcripts are >200 nucleotides long and constitute the largest portion of the mammalian non-coding RNA transcriptome [40]. LncRNA closely resembles mRNA than other classes of ncRNA in terms of their biogenesis pathways and form. Most lncRNAs are transcribed by the activities of RNA polymerase II, have a 5′ terminal methylguanosine cap and are often spliced and polyadenylated [41]. Some non-polyadenylated lncRNAs arise through alternative pathways probably expressed from RNA polymerase III promoters [43, 44] or arise during splicing and small nucleolar RNA production [45]. Furthermore, some lncRNAs are regulated in different ways at different stages of their biogenesis, maturation and decay [26]. Thousands of genes encoding lncRNAs have been identified in mammalian genomes (including livestock species), birds and plants studied so far and deposited in the NONECODE database (www.nonecode.org [39], Table 1).
3. MicroRNA in mammary gland development and lactation biology
3.1. Occurrence of microRNA in ruminant mammary gland and in milk
The regulatory roles of miRNAs in livestock species have emerged and are growing quickly [46, 47]. The most recent release of miRBase (release 21, http://www.mirbase.org/, [38]) contains 793 mature miRNAs for cattle, 436 for goat and 153 for sheep [38] (Table 1). However, with the increase in the application of RNA sequencing in expression profiling of miRNAs in different livestock species, the number of novel livestock miRNAs is expected to increase.
3.1.1. Cattle
The profiles of miRNAs in bovine MG tissue or milk have been investigated using different approaches, such as microarray [48, 49], genome sequencing [4] and RNA sequencing [50–57]. A total of 496 miRNA genes were identified following sequencing of the cattle genome of which 135 were novel [4].The expression profiles of miRNAs in MG tissues and cells facilitate discovery of novel miRNAs and also identification of candidate miRNAs for different cell types, lactation stages, periods, disease response and so on. Before the release of the bovine genome sequence, Gu et al. [49] pioneered miRNA discovery in the bovine MG by cloning and sequencing small RNAs from MG tissue followed by identification of 59 distinct bovine miRNAs. Using next-generation sequencing techniques, Chen et al. [58] identified 230 and 213 known miRNAs in cow colostrum and mature milk, respectively. The authors also observed that 108 and 8 miRNAs were upregulated and downregulated, respectively, in colostrum compared to mature milk [58]. Using microarray, Izumi et al. [59] identified 100 and 53 known miRNAs in colostrum and mature milk, respectively. Using Solexa sequencing method, Li et al. [60] reported 884 unique miRNAs sequences in the bovine MG (283 known, 505 novel and 96 conserved miRNAs). Le Guillou et al. [61] identify 167 novel miRNAs in the bovine MG, many of which were also detected in mouse MG. Analysing three milk fractions (fat, whey and cells) and mammary gland tissues, we reported 210, 200 and 249 known and 33, 31 and 36 novel miRNAs in milk fat, whey and cells, respectively, and 321 known and 176 novel miRNAs in mammary gland tissues [62]. Deep sequencing the milk fat across the lactation curve, we also identified a total of 475 known and 238 novel miRNAs [63].
3.1.2. Goat
A total of 487 miRNAs were identified when the goat genome was sequenced and the largest miRNA clusters were found on chromosome 21 [6]. Using the Illumina-Solexa high-throughput sequencing technology to analyse goat MG tissues during early lactation, Ji et al. [64] reported 131 novel and 300 conserved miRNAs. Using the same method (Illumina-Solexa sequencing), Li et al. [65] reported 346 conserved and 95 novel miRNAs in goat MG tissues from dry off and peak lactation does.
3.1.3. Sheep
Most miRNAs identified in sheep come from tissues other than the MG. For example, Caiment et al. [66] identified 747 miRNAs from the skeletal muscle through deep sequencing, whereas McBride et al. [67] reported 212 miRNAs from sheep ovarian follicles and corpus lutea at various reproductive stages. In the MG, Galio et al. [68] showed the presence of three known miRNAs including miR-21, miR-205 and miR-200 family in pregnant and lactating sheep.
3.2. MicroRNA function in ruminant mammary gland and milk synthesis
3.2.1. Expression patterns of microRNAs in lactation stages
3.2.1.1. Temporal and spatial expression of microRNAs
Indication of involvement of miRNAs in MG functions was gained through observation of differences in type and expression levels of miRNAs between lactation stages, under different nutritional regimes and presence of disease pathogens. Li et al. [50] identified 56 miRNAs that were significantly differentially expressed between lactation and non-lactation periods. Similarly, Wang et al. [48] detected 12 downregulated miRNAs (miR-10a, miR-15b, miR-16, miR-21, miR-33b, miR-145, miR-146b, miR-155, miR-181a, miR-205, miR-221 and miR-223) in the dry period (30 days prepartum) compared to early lactation period (7 days postpartum) and one upregulated miRNA (miR-31) in early lactation compared to the dry period. Previously, we examined miRNA expression pattern during a lactation cycle to explore it regulatory mechanisms during lactation using milk fat as input tissue for sampling [63]. In a previous investigation, we have shown that milk fat miRNA transcriptome closely resemble the miRNome of MG tissue [62]. We collected samples at the lactogenesis (LAC) (day 1 and 7), galactopoiesis (GAL) (day 30, 70, 130, 170 and 230) and involution (INV) (day 290 and when milk production dropped to 5 kg/day) stages from nine cows for deep sequencing [63]. We observed that 15 miRNAs (miR-30a-5p, miR-30d, miR-21-5p, miR-26a, miR-148a, let-7a-5p, let-7b, let-7f, let-7g, miR-99a-5p, miR-191, miR-200a, miR-200c, miR-186, miR-92a) were highly expressed across lactation stages [63]. MiR-148a and miR-26a were the most abundantly expressed accounting for more than 10% of the read counts in each stage of lactation. We also performed a differential expression (DE) analysis and detected miR-29b/miR-363 and miR-874/miR-6254 as important mediators of transition signals from LAC to GAL and from GAL to INV stages, respectively [63]. Furthermore, DE analysis indicated various patterns of miRNA expression across the lactation curve. For instance, some miRNAs were highly expressed during early lactation (lactogenesis) followed by decreased expression at later stages, whereas others were slightly expressed during early lactation but showed increased expression during mid-lactation and decreased expression during late lactation and vice versa [63] (Figure 2).
Figure 2.
Differential miRNA expression patterns during a bovine lactation curve. (a) Fold change values of six miRNAs whose expression patterns changed significantly during each lactation switch and (b) box plots of their normalized read count values by lactation day. 1LAC: lactogenesis; GAL: galactopoiesis; INV: involution; 2D: downregulated and U: upregulated.
The temporal expression pattern of miRNAs has been reported in other ruminant species. For example, Galio et al. [68] reported a change in the expression pattern of miR-21, miR-205 and miR-200 family in MG tissues from pregnant and lactating sheep. From the early, middle and late stages of pregnancy and during lactation, the expression of miR-21 and miR-25 decreased, whereas miR-200 family (miR-200a, miR-200b, miR-200c, miR-141 and miR-429) showed increased expression [68]. Similarly, investigating the expression pattern of miRNAs during early and peak lactation and dry period, Li et al. [65] identified 15 differentially expressed miRNAs when comparing peak lactation and dry period including three significantly highly expressed miRNAs (miR-2887, miR-451 and miR-2478) during peak lactation and 12 significantly highly expressed miRNAs (miR-199b, miR-128, miR-25, miR-145, miR-98, miR-222, miR-181b, miR-199a-3p, miR-93, miR-221, let-7b and let-7c) during the dry period.
3.2.1.2. MicroRNAs synergistically regulate lactation control mechanisms
A wealth of evidence indicates that several miRNAs can work together to regulate target genes in the same or different biological pathways [69, 70]. We have successfully characterized a group of highly interacting miRNAs (modules) using a weighted co-expression network analysis [71] and correlated important miRNA modules to milk yield and milk components [72]. We identified three consensus (BLUE [62 miRNAs], TURQUOISE [133 miRNAs] and BROWN [59 miRNAs]) modules and the GREY module reserved for unclassified genes, throughout lactation stages (Figure 3). Based on module trait relationship, we were able to determine important modules (with absolute correlation >0.6) for milk components at each lactation stage. The BROWN and BLUE modules were highly related to protein and somatic cell count, respectively, in early lactation, the BLUE module to somatic cells in middle lactation and the BLUE module to urea and lactose in late lactation stage. We also found the most important component or hub miRNAs, which potentially coordinated miRNA synergetic mechanisms in their respective modules. MiR-149-5b and miR-874 were hub miRNAs in the BLUE module for milk somatic cells at early and middle lactation, respectively, whereas miR-330 was the hub miRNA in the BLUE module for milk urea and lactose at late lactation (Figure 3). Three miRNAs (mir-149-5b, miR-874 and miR-30) in the BLUE module play important roles in cell cycle [73–77], so it could be expected that these miRNAs regulate secretion of somatic cells in milk from MG.
Figure 3.
Important consensus modules and their hub miRNAs for milk component traits in different lactation periods. (a) Dynamic cut tree (dendrogram) based on topological overlap distance in gene expression profile; (b) module trait relationship in early, middle and late lactation and (c) hub miRNAs in the modules. GREY colour is for genes that do not belong to a specific module.
3.2.2. Networks and pathways regulated by microRNAs during a lactation cycle
Through their target genes, miRNAs have been shown to control signal transduction in different species [78]. MiRNA roles in important pathways such as transforming growth factor beta (TGF-β), prolactin and protein kinase signalling in MG development and lactation have been reviewed by several authors [79–83]. MiRNA regulation of three important signalling pathways (NOTCH, PTEN and HIPPO) in MG and breast cancer cells was recently reviewed [15]. Important miRNAs regulating these pathways include mir-34, mir-29, mir-146, mir-199 and mir-200 families for NOTCH signalling pathway, miR-21 and miR-155 for PTEN signalling pathway and miR-934 for HIPPO pathway. In Canadian Holstein cows, we performed the enrichment of differentially expressed miRNA target genes to signalling pathways and noted that relevant signalling pathways for transition between lactation stages are involved in apoptosis (PTEN and SAPK/JNK), intracellular signalling (protein kinase A, TGF-β and ERK5), cell cycle regulation (STAT3), cytokines (prolactin), hormone and growth factors (growth hormone and glucocorticoid receptor). PTEN is an important target gene for miR-29b in the regulation of mammary gland development [84]. PTEN signalling is crucial for the activities of prolactin autocrine [85]. The initiation of lactation is known to require induction of autocrine prolactin, and the level of this autocrine is known to be endogenously regulated by the signal of PTEN-PI3K-AKT pathway [85]. Figure 4 is an illustration of some miRNAs that target genes in relevant signalling pathways during lactation [63]. Pathways, such as PTEN and growth hormone signalling, have been identified as important for regulatory mechanisms during lactation [85, 86].
Figure 4.
Illustration of miRNA-gene-pathway networks obtained from dynamic differentially expressed miRNAs during a bovine lactation curve. The outer layer shows miRNAs (blue arrow heads), which targets at least two genes (white dots) in significantly enriched pathways (red dots).
3.2.3. Functional validation of microRNA target genes
Since in vivo experiments for functional validation of MG miRNAs are not feasible, such studies have mostly relied on the use of knock-out/mimics and MG-specific cell types. Using bovine mammary epithelial cells (BMEC), miR-15a was shown to regulate growth hormone receptor, viability of BMEC and the expression of casein genes [86]. MiR-486 regulation of lactation by targeting the PTEN gene in cow MGs has been demonstrated [87]. Bian et al. [88] recently reported that epigenetic regulation of miR‐29s affects the lactation activity of BMEC. MiR-181a was shown to regulate the biosynthesis of bovine milk fat through targeting acyl-CoA synthetase long-chain family member 1 (ACSL1) [89]. MiR-103 was reported to control milk fat accumulation in goat MG during lactation [90]. Moreover, miR-27a was shown to suppress triglyceride accumulation as well as altered gene expression associated with fat metabolism in dairy goat mammary epithelial cells (GMEC) [91]. In another study, miR-135a was reported to target and regulate prolactin receptor (PRLR) gene in GMEC [92]. Inhibition of the expression of miR-145 in GMEC was shown to increase methylation levels of fatty acid synthase (FASN), stearoyl-CoA desaturase 1 (SCD1), peroxisome proliferator-activated receptor gamma (PPARG) and sterol regulatory element binding transcription factor 1 (SREBF1) [93]. MiR-24 control of triacylglycerol synthesis in goat mammary epithelial cells by targeting FASN gene has been demonstrated [94]. The ability of miR‐145 to regulate lipogenesis in GMEC through targeting insulin-induced gene 1 (INSIG1) and epigenetic regulation of lipid‐related genes has been demonstrated [93]. MiR-143 was shown to inhibit proliferation as well as induce apoptosis of GMEC [95]. MiR130b regulation of PPARγ coactivator-1α suppressed fat metabolism in GMEC [96]. In non-ruminant species, many miRNAs, including let-7 family members, mir-17/92, miR-30b, miR-93, miR-99a and miR-b, miR-101a, miR-126-3p, miR-138, miR-146b, miR-200 family members, mir-203, miR-205, miR-206, miR-210, miR-212/132, miR-221 and miR-424/50, have been reported to play roles in mammary gland development and disease [15]. Some miRNAs with functionally validated targets are summarized in Table 2.
MicroRNAs with functionally validated target genes using ruminant mammary gland cells.
3.3. Nutritional modulation of microRNA expression and function
The miRNA expression profile in response to dietary treatments has been studied in adipose tissues of lambs and cattle and bovine mammary gland tissues [56, 100–102]. A change in diet that interferes with energy balance has been shown to change miRNA expression pattern in cow liver [103]. Wang et al. [104] fed cows with high- and low-quality forage diets (corn stover and rice straw) and showed that miR-125b, miR-141, miR-181a, miR-221 and miR-15b changed their expression patterns across different tissues including MG. We have examined the expression pattern of miRNAs following MG adaptation to dietary supplementation with 5% linseed oil or 5% safflower oil using miRNA sequencing and identified seven differentially regulated miRNAs, including six upregulated (miR-199c, miR-199a-3p, miR-98, miR-378, miR-148b and miR-21-5p) and one downregulated (miR-200a) by both linseed and safflower oil. The target genes of these seven miRNAs have functions related to gene expression and general cellular metabolism and are enriched in four pathways of lipid metabolism (3-phosphoinositide biosynthesis, 3-phosphoinositide degradation, D-myo-inisitol-5-phosphate metabolism and the superpathway of inositol phosphate compounds) [51]. The largest number of target genes (39) were associated with two functions (synthesis of lipid and concentration of lipid) related with lipogenesis. In goat, Mobuchon et al. [105] detected 30 miRNAs with expression patterns potentially modulated by food deprivation (14 and 16 were upregulated and downregulated, respectively). Among them, miR-204-5p and miR-223-3p were most remarkably affected by food deprivation and potentially played roles in the nutritional regulation of gene expression in the MG.
3.4. MicroRNA functions in mammary gland health
MiRNAs have been shown to play roles in bovine infection and immunity in a wide range of tissues [54, 106–113]. For mammary gland, Naeem et al. [114] studied the expression of 14 miRNAs (miR-10a, miR-15b, miR-16a, miR-17, miR-21, miR-31, miR-145, miR-146a, miR-146b, miR-155, miR-181a, miR-205, miR-221 and miR-223) in MG tissue challenged with Streptococcus uberis and identified three downregulated miRNAs (miR-181a, miR-16 and miR-31) and one upregulated miRNA (miR-223) in infected versus healthy tissue. Lawless et al. [107] showed that 21 miRNAs were differentially expressed upon Streptococcus uberis infection of bovine primary epithelial cells. Using BMEC, Jin et al. [108] reported a differential expression of nine miRNAs (miR-184, miR-24-3p, miR-148, miR-486, let-7a-5p, miR-2339, miR-499, miR-23a and miR-99b) upon challenge with heat inactivated Escherichia coli and Staphylococcus aureus bacteria. Hou et al. [115] identified three upregulated miRNAs (miR-296, miR-2430 and miR-671) and one downregulated miRNA (miR-2318) in mastitis affected compared with healthy mammary gland tissues. Li et al. [111] sequenced RNA isolated from S. aureus-induced mastitis and control cows and identified 77 miRNAs with significant expression differences between the two groups. Li et al. [116] showed that miR-23 might be an important immune miRNA through its target mastitis candidate gene, high mobility group box 1 (HMGB1).
3.5. MicroRNA function in milk recipients
Recent evidence suggesting that milk-derived miRNAs may have potential regulatory roles in modulating the immune system or metabolic processes of milk recipients still remain controversial [117–124]. Currently, there are two hypotheses about miRNA function in infants/offspring: the first proposes that milk miRNAs exert physiological regulatory functions after transferring to offspring, and the second assumes that miRNAs do not have any function but merely provide nutrition. According to Zhang et al. [117], the rice-derived miRNA, miR-168a, can bind to the mRNA of human/mouse low-density lipoprotein receptor adapter protein 1 (LDLRAP1) and inhibit its expression in the liver, and consequently decrease LDL removal from mouse plasma. Baier et al. [118] reported that miR-29b-3p and miR-200c-3p could be absorbed by humans in biologically meaningful amounts, which could affect related gene expression in peripheral blood mononuclear cells while Izumi et al. [125] confirmed that whey exosomes containing miRNAs and mRNA could be absorbed by human macrophages. These results opened a new aspect of the nutritional control of metabolism [119]. However, other studies have not succeeded to validate the hypothesis that milk miRNAs exert physiological regulatory functions after transferring to offspring [126–129]. For instance, Auerbach et al. [129] observed that drinking bovine milk increased circulating levels of miRNAs (miR-29b-3p and miR-200c-3p) but found no evidence that they significantly altered miRNA signals after milk ingestion. These authors concluded that milk miRNAs likely serve as a source of nutrition but not as post-transcriptional regulators in recipients.
4. Long non-coding RNA in mammary gland development and lactation biology
4.1. Prolife and expression of long non-coding RNAs
A limited number of studies have examined the occurrence and potential functions of lncRNAs in ruminant livestock species [130–132]. A pioneer study screened reconstructed transcript assemblies of bovine-specific expressed sequence tags and identified 449 putative lncRNAs located in 405 intergenic regions [130]. Following this initial study, Weikard et al. [131] used RNA sequencing technique and identified 4848 potential lncRNAs, which were predominantly intergenic (4365) in bovine skin. In another study, Billerey et al. [132] characterized 584 lncRNAs in bovine muscle in addition to significant correlated expression between 2083 pairs of lncRNA/protein encoding genes. Koufariotis et al. [133] characterized the lncRNA repertoire across 18 bovine tissues including the mammary gland and reported 9778 transcripts. Ibeagha-Awemu et al. [134] studied the lncRNA profile of the bovine mammary gland by RNA sequencing and identified 4227 lncRNAs (338 known and 3889 novel). In goats, Zhan et al. [135] sequenced libraries from developing longissimus dorsi fetal (45, 60 and 105 days of gestation) and postnatal (3 days after birth) muscles and identified 3981 lncRNA transcripts corresponding to 2739 lncRNA genes. Ren et al. [136] identified 1336 specific lncRNAs in fetal skin of Youzhou dark goat (dark skin) and Yudong white goat (white skin). Similarly, Chao et al. [137] in a study with aim to identify and classify new transcripts in Dorper and small-tail Han sheep muscle transcriptomes predicted with high confidence 1520 transcripts to be lncRNAs.
4.2. Function of long non-coding RNAs
While the regulatory roles of lncRNAs have been associated with several human disease conditions including tumourigenesis, cardiac development, aging and immune system development [138–143], little information exist on livestock species. Our previous study on bovine mammary gland identified 26 lncRNAs that were significantly differentially regulated in response to a diet rich in α-linolenic acid thus suggesting potential regulatory roles of lncRNAs in fatty acid synthesis and lipid metabolism [134]. In a study with goat fetal muscle tissues at different stages of development, Zhan et al. [135] identified 577 significantly differentially expressed lncRNA transcripts thus suggesting roles in muscle development.
5. Genome editing technology and non-coding RNA
Genome engineering has been considered as the next genomic revolution [144], and it is expected to significantly improve livestock production by precision genome editing [145–147] favouring markers associated with improved productivity, reproduction and health status. The history of genome editing in livestock has been extensively reviewed [145, 148–150]. The advent of engineered endonucleases (EENs), including zinc finger nucleases (ZFNs) [151], transcription activator-like effector nucleases (TALENs) [152] and clustered regularly interspaced short palindromic repeats (CRISPR/Cas9) [153]), allows to cut a specific position in DNA sequence and then use endogenous cellular pathways to direct DNA repair to introduce specified alterations to the DNA sequence. Genome-editing approaches have been successfully used in different livestock species, such as pig [154, 155], goat [156], cattle [157] and sheep [158]. In dairy cows, these technologies have been used to manipulate the genome so that they produce specific milk types, such as milk that causes less allergic problems (e.g. milk with less β-lactoglobulin protein) [159, 160]. These genome-editing tools also helped to improve mammary gland health by generating mastitis-resistant cattle [161, 162]. From an animal breeding perspective, a simulation study showed that genomic prediction combined with genome editing could be of benefit [163]. A total of 10,000 additive loci were simulated and shown to contribute to the variation in selected traits and benefits could be achieved with only 20 of those loci being edited in each selected sire [163]. Similar to other genome sequences, miRNA gene sequences within mammalian genomes can be easily edited with high efficacy and precision [144]. Targeted miRNA editing will enable revelation of the complex regulatory circuits governed by miRNAs and realization, in the long term, of their full diagnostic and therapeutic potentials. For instance, Chen et al. [164] successfully used TALEN to disrupt the function of miR-21 in cancerous cells. A transgenic calf engineered to express miRNA-4 and miR-6 showed an absence of β-lactoglobulin and a concurrent increase in casein proteins in milk [165].
6. Conclusion and remarks
Up to now, it is well known that the mammalian genome encodes thousands of ncRNAs and these ncRNAs play important roles in many processes related to MG development, health and disease as well as roles in milk secretion and lactation processes. Regarding animal breeding, several ncRNAs target specific processes and their target genes could be important biomarkers for specific traits of interest. Therefore, the application of ncRNA to improve mammary gland health and milk production as well as enhance milk quality is very promising. However, the first step is a better understanding of ncRNA function in MG development and lactation. In fact, the MG is a complex tissue and lactation is a complicated process, but what we known about the regulatory networks underling MG function and the lactation process is very limited. For instance, through RNA sequencing, many novel ncRNAs have been detected in the MG but knowledge of their actual functions remains elusive. Therefore, integrated ‘omics’ approaches (genomics, transcriptomics, epigenomics and proteomics) should be used to identify and explore the potential roles of ncRNAs in mammary gland development and lactation biology. Moreover, a miRNA can target hundreds of genes thus making it difficult, costly and labour-intensive to functionally validate each miRNA gene target. Thus, integrative approaches such as combination of miRNA and mRNA expression in the same sample will refine computational predictions and increase our understanding of miRNA function and its application.
Acknowledgments
We acknowledge financial support from Agriculture and Agri-Food Canada.
\n',keywords:"non-coding RNA, microRNA, long non-coding RNA, mammary gland, lactation, genome editing, signalling pathways",chapterPDFUrl:"https://cdn.intechopen.com/pdfs/53907.pdf",chapterXML:"https://mts.intechopen.com/source/xml/53907.xml",downloadPdfUrl:"/chapter/pdf-download/53907",previewPdfUrl:"/chapter/pdf-preview/53907",totalDownloads:1436,totalViews:281,totalCrossrefCites:3,totalDimensionsCites:8,hasAltmetrics:0,dateSubmitted:"May 10th 2016",dateReviewed:"December 8th 2016",datePrePublished:null,datePublished:"May 10th 2017",dateFinished:null,readingETA:"0",abstract:"The ruminant mammary gland (MG) is an important organ charged with the production of milk for young and human nourishment. Many factors influence MG productivity, including nutrition, genetics, breed, epigenetics (including non-coding RNA [ncRNA]), disease pathogens and other environmental factors. In recent years, increasing research is beginning to determine the role of non-coding RNA in MG functions. Non-coding RNAs (small interfering RNA [siRNA], microRNA [miRNA], PIWI-interacting RNA [piRNA], small nucleolar RNA [snoRNA] and long non-coding RNA [lncRNA]) are a class of untranslated RNA molecules that function to regulate gene expression, associated biochemical pathways and cellular functions and are involved in many biological processes. This chapter presents a review of the current state of knowledge on the role of ncRNAs (particularly miRNAs and lncRNAs) in the MG and lactation processes, lactation signalling pathways, lipid metabolism, MG health of ruminants as well as miRNA roles in milk recipients. Finally, the potential application of new genome editing technology for ncRNA studies in MG development, the lactation process and milk components is presented.",reviewType:"peer-reviewed",bibtexUrl:"/chapter/bibtex/53907",risUrl:"/chapter/ris/53907",book:{slug:"current-topics-in-lactation"},signatures:"Duy N. Do and Eveline M. Ibeagha-Awemu",authors:[{id:"191336",title:"Dr.",name:"Eveline",middleName:null,surname:"Ibeagha-Awemu",fullName:"Eveline Ibeagha-Awemu",slug:"eveline-ibeagha-awemu",email:"eveline.ibeagha-awemu@agr.gc.ca",position:null,institution:{name:"Agriculture and Agriculture-Food Canada",institutionURL:null,country:{name:"Canada"}}},{id:"191339",title:"Dr.",name:"Duy",middleName:null,surname:"Do",fullName:"Duy Do",slug:"duy-do",email:"Do.DuyNgoc@AGR.GC.CA",position:null,institution:null}],sections:[{id:"sec_1",title:"1. Introduction",level:"1"},{id:"sec_2",title:"2. Non-coding RNAs: biosynthesis and classification",level:"1"},{id:"sec_2_2",title:"2.1. MicroRNAs",level:"2"},{id:"sec_3_2",title:"2.2. Long non-coding RNAs",level:"2"},{id:"sec_5",title:"3. MicroRNA in mammary gland development and lactation biology",level:"1"},{id:"sec_5_2",title:"3.1. Occurrence of microRNA in ruminant mammary gland and in milk",level:"2"},{id:"sec_5_3",title:"3.1.1. Cattle",level:"3"},{id:"sec_6_3",title:"3.1.2. Goat",level:"3"},{id:"sec_7_3",title:"3.1.3. Sheep",level:"3"},{id:"sec_9_2",title:"3.2. MicroRNA function in ruminant mammary gland and milk synthesis",level:"2"},{id:"sec_9_3",title:"3.2.1. Expression patterns of microRNAs in lactation stages",level:"3"},{id:"sec_9_4",title:"3.2.1.1. Temporal and spatial expression of microRNAs",level:"4"},{id:"sec_10_4",title:"3.2.1.2. MicroRNAs synergistically regulate lactation control mechanisms",level:"4"},{id:"sec_12_3",title:"3.2.2. Networks and pathways regulated by microRNAs during a lactation cycle",level:"3"},{id:"sec_13_3",title:"Table 2.",level:"3"},{id:"sec_15_2",title:"3.3. Nutritional modulation of microRNA expression and function",level:"2"},{id:"sec_16_2",title:"3.4. MicroRNA functions in mammary gland health",level:"2"},{id:"sec_17_2",title:"3.5. MicroRNA function in milk recipients",level:"2"},{id:"sec_19",title:"4. Long non-coding RNA in mammary gland development and lactation biology",level:"1"},{id:"sec_19_2",title:"4.1. 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Agriculture and Agri-Food Canada, Sherbrooke Research and Development Centre, Sherbrooke, Quebec, Canada
Department of Animal Science, McGill University, Ste-Anne-De-Bellevue, Quebec, Canada
'},{corresp:"yes",contributorFullName:"Eveline M. Ibeagha-Awemu",address:"Eveline.ibeagha-awemu@agr.gc.ca",affiliation:'
Agriculture and Agri-Food Canada, Sherbrooke Research and Development Centre, Sherbrooke, Quebec, Canada
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\n
1. Introduction
\n
In spite of deep inspiration, nasal tip maintains its general shape through different mechanisms. Major mechanisms include medialis and lateralis cartilages, the fibrous union between both crus medialis to the caudal septum and the ligaments that bind the caudal aspect of the superior lateral cartilages to the cephalic aspect of the inferior lateral cartilages. Secondary mechanisms encompass the cartilaginous septal dorsum, interdomal ligaments, the membranous septum, the nasal spine, the adherence between skin soft tissues and alar cartilages as well as the lateral alar walls (Figure 1).
\n
Figure 1.
A representation of the main cartilages from a basal perspective: 1. domus; 2. septal angle; 3. crus lateralis; 4. crus intermedia; 5. soft triangle; 6. crus medialis; 7. rima alaris; 8. ‘feet’ of the crura medialis; 9. nostrils; 10. caudal septum; 11. lobe of the nasal ala; 12. anterior nasal spine; and 13. nasolabial ridge.
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Normal alar contour is defined by well-defined alar margins and extension from the tip lobe to the alar lobe. The inferior lateral cartilages act as a dynamic spring that can resist small traumatisms, providing some elasticity to the nasal tip to prevent a collapse of the nasal alae during inspiration.
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Positioning of the alar cartilages is a fundamental element when planning a rhinoplasty. An overenthusiastic resection may cause their weakness and inability to perform their sustaining role.
\n
An inadequate positioning of the alar cartilages entails an instable Anderson tripod with alar pinching. This may be a consequence of the resection of ligaments and the septal angle in order to achieve a reduction in nasal tip projection in open rhinoplasty procedures.
\n
Less frequently, inborn asymmetries of the alar cartilages produce unbalance and rotation of the nasal tip. Moreover, some patients may suffer from a cephalad rotation of the alar cartilages; they may orientate their main axis toward the inner cantus without giving adequate support to the external valve.
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\n
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2. Historical notes
\n
Alar and perialar rim grafts are rods or little splints made of cartilage. They are placed under the caudal margin of alar cartilages. They may be applied for primary and secondary rhinoplasty. Alar rim grafts were heralded as early as the 1950s by Fomon [1, 2] and Denecke [3] for the correction of alar base in cleft lip nose. Composite paranasal grafts on the nasal mucosa (instead of under) were an interesting, easier variation proposed by Farrior [4]. However, these techniques had a moderate predicament for the treatment of stenoses of the vestibule [5, 6]. In a parallel evolution, cartilage struts under the anterior half of the alae of a pinched nose tip were popularised, while the cartilage of the auricular concha became the donor site of choice for nasal procedures [7].
\n
With greater emphasis focused on correcting the collapse of the internal valve, this approach by alar rim grafts was a sleeper. It was not until recent times when some surgeons [8, 9, 10] pointed to its potential role in aesthetic cases and added some technical refinements (Figure 2).
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Figure 2.
Testing of the general elasticity and passive collapse of the nasal tip and alae.
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Alar rim grafts may be used for treatment and prevention of disorders of the nasal tip outline. They have been advised for the treatment of alar deformities. These deformities may stem not only from malposition or congenital hypoplasia of inferior lateral cartilages but also from a loss of continuity or a weakening of crus lateralis as a result of previous surgeries [11]. In rare cases, scar tissue may cause local synechiae that easily resolve with local section [12] (Table 1).
\n
\n
\n\n
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Congenital asymmetry
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Dynamic alar collapse
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Alar flare (without functional impairment)
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Primary retraction or notching
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Secondary (surgical or traumatic) retraction as in pinched tip
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Malposition of the lateral cartilages (upwards/downwards, bulbous tip, square tip)
\n
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Congenital microrrhinia (all nasal dimensions affected as seen in foetal alcoholic syndrome)
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\n\n
Table 1.
Main indications for alar rim grafts.
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3. Pre-operative assessment
\n
The position and dimension of the nasal alae must be assessed from frontal, lateral and basal points of view. As a rule of thumb, the distance between each inner cantus of the eyelids is roughly equivalent to the width of nasal base in frontal view. An ideal nasal base is schematised as an equilateral triangle. However, there is considerable interethnic variability. In shallow, general speaking, patients with East Asian and African ancestry may present a wider base [13]. The alar contour displays an oval nostril, and the alar fringe follows a smooth curvature with an inferior concavity. According to the distances between columella-nose axis and alar rim-nose axis, we may consider four different alterations of the alar position [14, 15] (Figure 3A and B):
alar retraction—an elevation of the inferior concavity of the arch;
hanging alar rim;
retracted columella; and
hanging columella—the inner mucosal lining of the medial aspect of each narine is conspicuous.
\n
Figure 3.
(A) Anatomical variations and primary alterations of alar contour include (from left to right) pinched lobes, undulating alar cartilages, visible margins, bulbous tip and parallel orientation of both alar cartilages. (B) Some features that may involve alar rim grafts as a treatment include (from left to right) lack of sustentation of alar rim, high positioned soft triangle, bulbous tip and square tip.
\n
Functional primary disturbances may be the main motivation for a rhinoplasty (though they are also seen as an undesired side effect of a previous operation). These disturbances may cause difficulties in breathing, altered olfactory function, bleeding and frequent infection. Pre-operative rhinoscopy in order to exclude upper functional conditions (septal deviation, hypertrophic cornets, collapse of the upper internal valve and polyps) must be always carried out. A hanging tip is assessed by pinching the skin of the nasal dorsum. The collapse of the upper internal valve is sometimes evident after mild finger traction on the maxillary ascending apophysis. A cotton tip moisturised in adrenaline produces vasoconstriction and reduction in the size of hypertrophied cornets. A collapse of the lower valve may be corrected by gently opening the tweezers inside the air passage.
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\n
\n
4. Surgical technique
\n
Inspection of the nasal external valve constitutes an unavoidable step toward the end of any rhinoplasty procedure. Whenever we have performed a reduction of the projection of the nasal tip, we shall get an alar excess. Scar lines or excessive resection may entail a narrowing of the air passage. We use these grafts for open and closed rhinoplasties. The graft consists of a rod of septal or auricular cartilage (Figure 4A,B and 5) that we lay as reinforcement inside a pocket along the alar margin. We perform an incision (less than 5 mm) on the hairy area of alar vestibulum (scarcely two-or-three millimetres away from the narine marge) by means of a number 11 blade. We use blunt scissors to create a double pocket backwards to the alar lobe and frontwards to the soft triangle (Figure 6). Some authors propose infiltration with 1% lidocaine with epinephrine in the skin caudal to the marginal incision [12]. In the so-created tunnels, we lay our rod of cartilage (from 1 × 6 mm up to 2 × 12 mm), from incision to back in the posterior tunnel and from incision to front in the anterior tunnel. We perform a single stitch in reabsorbable polydioxanone (PDS® 5/0). We must crush the anterior shaft with the tissue forceps in order to prevent it from exteriorizing through the skin. In any case, the graft should never extend anterior to the nasal tip [16, 17]. It must never widen the tip or be palpable [18]. We usually perform this step at the end of the procedure. We can check the final texture of the nasal tip and alae by pressing with fingertips or tissue forceps. Alar rim grafts allow us to caudally displace the alar rim margin up to 2 mm but this small gain reveals itself crucial in many cases. Whenever the alar rim graft is an isolated procedure, a safe, less traumatic way to incise the skin maybe achieved by using an ophthalmic slit blade [19].
\n
Figure 4.
(A) A sketch of the positioned alar rim graft in a frontal view. (B) The positioned alar rim graft in a lateral view.
\n
Figure 5.
Intraoperatory appreciation of the required length of grafts.
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Figure 6.
Incision, blunt dissection and insertion of an alar rim graft.
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\n
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5. Variations
\n
When performing an open rhinoplasty, we use the same pre-rimal incision. Thus, we can check the symmetry of graft positioning. When performing a closed rhinoplasty, we place the grafts through a marginal incision (1 mm from the alar rim) and we extend the pocket frontwards to the soft triangle and backwards to the caudal end of the ala. We first lay the graft inside the posterior pocket and by careful sliding, we position its crushed edge inside the anterior pocket.
\n
An alternative method involves conchal cartilage extension grafts fixed to the caudal margins of the lateral crura as described by Jang et al. [20]. This hybrid method focuses on correcting anterior contraction of the alar rim as seen in East Asian patients with nostril exposure. Alar vestibular skin is dissected at the end of an open approach for augmentation rhinoplasty. Conchal cartilage grafts are fashioned in a semilunar shape (13 mm × 6 mm) and sutured to the caudal margins of each crus lateralis.
\n
Articulated alar rim grafts [21, 22] stand as an interesting concept. In this widespread variation, the anterior margin of each alar rim graft is sutured to the tip complex instead of just being freely sited in a pocket. Emphasis is mainly made to stabilise the nasal tip.
\n
A peculiar variation [23] elevates a 2–3 mm flap from the caudal portion of the crus lateralis, pulls it caudally and extends it with a cartilage graft. This extension of the alar rim flap is placed along the alar rim for support.
\n
Selected cases of external nasal valve collapse as an isolated condition have been treated by a microinvasive technique that creates the pocket from the cutaneous, facial aspect of the posterior margin of the ala [24].
\n
Needless to say, versatile surgeons should bear in mind alternative donor sites as part of their armamentarium [25]. A posterior incision is the less conspicuous choice when taking conchal grafts Adequate semicompressive dressings and anaesthetic infiltration of the margins of the skin (for instance with bupivacaine or ropivacaine) would minimise haematoma and post-operative pain in this donor area.
\n
\n
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6. Complications
\n
As with any surgical procedures, patients should be informed about potential problems as transient inflammation, haemorrhage, haematoma, seroma, adherences, conspicuous scarring, keloids and pigmentation alterations. The same goes for undesirable infectious conditions as chondritis, osteitis, myositis and abscess. More specific conditions are paraesthesia, loss of temperature sensation and partial resorption of the cartilaginous graft. Jarring right-left asymmetry of the grafts may entail pyramid deviation.
\n
Local necrosis and extrusion of the graft are very rarely seen. They may be the result of local traumatism or inadequate dressing as well as previous ischaemic features as seen in chain smoking (Figures 7, 8, 9, 10).
\n
Figure 7.
(A) Broad, bilobed nasal tip in a 27-year-old female patient. Slight deviation of dorsum and retracted columella. (B) Surgical planning for the patient in Figure 5A. This patient underwent lipectomy (yellow), cephalic resection of alar cartilages (red), caudal extension graft (from septum), submucous section of triangle area, medial and lateral fracture (spiked line), septal cartilage graft over tip region, septal cartilage graft inside alar rim. (C) and (D) Post-operative result of patient in (A).
\n
Figure 8.
(A) A 32-year-old female patient suffering for traumatic deviation. Hanging tip, acute nasolabial angle, broad tip and unbalance between lobe and alae. (B) Surgical planning for the patient in Figure 6A. A trans-columellar incision was extended along pre-rimal areas. The procedure included a partial septoplasty (red), tension-discharging incisions on the left side, dorsal expansion grafts, a columellar stick graft, cephalic resection of alar cartilages, several interdomal, intradomal and intraalar stitches, as well as septal cartilage graft to the left cartilaginous wall, a suspension-rotation suture of the crus medialis, predomal grafts (light blue), a medial and lateral fracture (interrupted line), a batten graft to the radix, pre-rimal grafts and a resection with rotation of the skin and mucosa of the alae. (C) and (D) Comparison between pre-operative and post-operative result of patient in (A).
\n
Figure 9.
(A) A 35-year-old male patient with supratip deformation after secondary rhinoplasty. He showed a thick skin, a saddle (broad and flat) dorsum as well as an asymmetric, pinched tip, other features include an osseous dehiscence on the left side and malpositioned, undulating alae. (B) Surgical planning for patient in (A). (C) and (D) Comparison between pre-operative and post-operative result of patient in (A).
\n
Figure 10.
(A) Secondary rhinoplasty in a 29-year-old female patient that showed deviation, irregularities of her dorsum, upper valve collapse, pinched tip, light alar retraction, hanging columella and maxillary hypotrophy. (B) Surgical planning for the patient in Figure 8A included an osteocartilaginous septoplasty with removal of previous graft and fibrotic tissue (red star). The procedure involved reconstruction of the left alar cartilage (blue star), expansor grafts for dorsum, batten alar graft on the right lateral wall, a left alar graft, a predomal graft, a fixation suture between crus medialis and caudal septum, pre-rimal grafts, a lateral fracture (spiked line), partial resection of the mucosa of the membranous septum, an expansion mesh of polypropylene on the maxilla and partial resection of the skin and mucosa of the alae. (C) and (D) Comparison between pre-operative and post-operative result of patient in (A).
\n
\n
\n
7. Conclusion
\n
These grafts are useful to prevent an alar retraction and post-operative shifts on those patients that show primary alterations of alar outline. They provide support and steadiness for the alar rim by creating a structure that counteracts the forces of scar contraction [26, 27, 28, 29]. Whenever we use them we shall prevent descent (or rotation) of caudal margin of alar cartilages and a trilobulate, pinched nose. At the same time, we enhance a correct functioning of the external valve and prevent its collapse.
\n
These grafts are also very useful for the treatment of pinched, nasal tips with a very long crus intermedium, i.e. lack of sustentation of skin in the soft triangle. We may use them as a complement for domal sutures and alar grafts in order to compensate the modifications that these procedures induce on the sustaining tip structures; they favour projection and balance of the nasal tip while preserving the function of external valve. They allow us to provide triangularity to the nasal base, by re-establishing the Anderson tripod, as well as to preserve the pyramidal shape of nasal tip. This will help to achieve a natural allure of the nasal tip contour.
\n
\n\n',keywords:"graft, alar, nose, cartilage, rhinoplasty",chapterPDFUrl:"https://cdn.intechopen.com/pdfs/66026.pdf",chapterXML:"https://mts.intechopen.com/source/xml/66026.xml",downloadPdfUrl:"/chapter/pdf-download/66026",previewPdfUrl:"/chapter/pdf-preview/66026",totalDownloads:465,totalViews:0,totalCrossrefCites:0,dateSubmitted:"November 21st 2018",dateReviewed:"January 29th 2019",datePrePublished:"March 26th 2019",datePublished:"December 4th 2019",dateFinished:null,readingETA:"0",abstract:"Alar rim grafts date back to the 1950s for the correction of alar base in cleft lip nose. Cartilage struts under the anterior half of the alae of a pinched nose tip were popularised and the cartilage of the auricular concha became the donor site of choice for nasal procedures. Recently, some surgeons pointed to its potential role in aesthetic cases and added some technical refinements. These grafts are used for open and closed rhinoplasties. They usually consist of a rod of septal or auricular cartilage that we lay as reinforcement inside a pocket along the alar margin. Indications include the following: congenital or traumatic asymmetry, dynamic alar collapse, alar flare, primary retraction or notching, secondary (surgical or traumatic) retraction and malposition of the lateral cartilages (upwards or downwards). Harvesting and implanting techniques as well as the possible drawbacks are discussed.",reviewType:"peer-reviewed",bibtexUrl:"/chapter/bibtex/66026",risUrl:"/chapter/ris/66026",signatures:"Pedro S. Arquero, Wenceslao M. Calonge, Daniel P. Espinoza and Diana Oesch",book:{id:"8616",title:"Contemporary Rhinoplasty",subtitle:null,fullTitle:"Contemporary Rhinoplasty",slug:"contemporary-rhinoplasty",publishedDate:"December 4th 2019",bookSignature:"Sebastian Torres Farr",coverURL:"https://cdn.intechopen.com/books/images_new/8616.jpg",licenceType:"CC BY 3.0",editedByType:"Edited by",editors:[{id:"177686",title:"M.D.",name:"Sebastian",middleName:null,surname:"Torres Farr",slug:"sebastian-torres-farr",fullName:"Sebastian Torres Farr"}],productType:{id:"1",title:"Edited Volume",chapterContentType:"chapter",authoredCaption:"Edited by"}},authors:[{id:"202013",title:"M.D.",name:"Wenceslao M",middleName:null,surname:"Calonge",fullName:"Wenceslao M Calonge",slug:"wenceslao-m-calonge",email:"wcalonge@yahoo.es",position:null,institution:null},{id:"286266",title:"Dr.",name:"Daniel",middleName:null,surname:"Espinoza Kauer",fullName:"Daniel Espinoza Kauer",slug:"daniel-espinoza-kauer",email:"daneskau@hotmail.com",position:null,institution:null},{id:"286267",title:"Dr.",name:"Diana",middleName:null,surname:"Oesch Ortiz",fullName:"Diana Oesch Ortiz",slug:"diana-oesch-ortiz",email:"dianaoesch@gmail.com",position:null,institution:null},{id:"292634",title:"Dr.",name:"Pedro S.",middleName:null,surname:"Arquero",fullName:"Pedro S. Arquero",slug:"pedro-s.-arquero",email:"arquero@clinicaarquero.com",position:null,institution:null}],sections:[{id:"sec_1",title:"1. Introduction",level:"1"},{id:"sec_2",title:"2. Historical notes",level:"1"},{id:"sec_3",title:"3. Pre-operative assessment",level:"1"},{id:"sec_4",title:"4. Surgical technique",level:"1"},{id:"sec_5",title:"5. Variations",level:"1"},{id:"sec_6",title:"6. Complications",level:"1"},{id:"sec_7",title:"7. Conclusion",level:"1"}],chapterReferences:[{id:"B1",body:'Fomon S, Bell JW, Berger EL, Goldman IB, Neivert H, Schattner A. Management of deformities of lower cartilaginous vault. A.M.A. Archives of Otolaryngology. 1951;54(5):467-472'},{id:"B2",body:'Fomon S, Bell JW, Lubart J, Schattner A, Syracuse VR. Rhinoplastic problems in the lower cartilaginous vault. Archives of Otolaryngology. 1964;79:512-521'},{id:"B3",body:'Denecke HJ, Meyer R. Plastische Operationen an Kopf und Hals. In: Korrigierende und Rekonstruktive Nasenplastik. Berlin: Springer; 1964'},{id:"B4",body:'Farrior RT. The problem of the unilateral cleft lip nose. A composite operation for revisión of the secondary deformity. Laryngoscope. 1962;72:289-352'},{id:"B5",body:'Janeke JB, Wright WK. Studies on the support of the nasal tip. Archives of Otolaryngology. 1971;93(5):458-464'},{id:"B6",body:'Vecchione TR. Reconstruction of the ala and nostril sill using proximal composite grafts. Annals of Plastic Surgery. 1980;5(2):148-150'},{id:"B7",body:'Orticoechea M. A new method for total reconstruction of the nose: The ears as donor areas. Clinics in Plastic Surgery. 1981;8(3):481-505'},{id:"B8",body:'Troell RJ, Powell N, Riley RW, Li KK. Evaluation of a new procedure for nasal alar rim and valve collapse: Nasal alar rim reconstruction. Otolaryngology and Head and Neck Surgery. 2000;122(2):204-211'},{id:"B9",body:'Rohrich RJ, Raniere J Jr, Ha RY. The alar contour graft: Correction and prevention of alar rim deformities in rhinoplasty. Plastic and Reconstructive Surgery. 2002;109(7):2495-2505'},{id:"B10",body:'Toriumi DM, Checcone MA. New concepts in nasal tip contouring. Facial Plastic Surgery Clinics of North America. 2009;17(1):55-90'},{id:"B11",body:'Daniel RK, Palhazi P, Gerbault O, Kosins AM. Rhinoplasty: The lateral crura-alar ring. Aesthetic Surgery Journal. 2014;34(4):526-537'},{id:"B12",body:'Alexander AJ, Shah AR, Constantinides MS. Alar retraction: Etiology, treatment, and prevention. JAMA Facial Plastic Surgery. 2013;15(4):268-274'},{id:"B13",body:'Tas S, Colakoglu S, Lee BT. Nasal base retraction: A treatment algorithm. Aesthetic Surgery Journal. 2017;37(6):640-653'},{id:"B14",body:'Ellenbogen R, Bazell G. Nostrilplasty: Raising, lowering, widening, and symmetry correction of the alar rim. Aesthetic Surgery Journal. 2002;22(3):227-237'},{id:"B15",body:'Unger JG, Roostaeian J, Small KH, Pezeshk RA, Lee MR, Harris R, et al. Alar contour grafts in rhinoplasty: A safe and reproducible way to refine alar contour aesthetics. Plastic and Reconstructive Surgery. 2016;137(1):52-61'},{id:"B16",body:'Gruber RP, Fox P, Peled A, Belek KA. Grafting the alar rim: Application as anatomical graft. Plastic and Reconstructive Surgery. 2014;134(6):880e-887e'},{id:"B17",body:'Toriumi DM, Josen J, Weinberger M, Tardy ME Jr. Use of alar batten grafts for correction of nasal valve collapse. Archives of Otolaryngology—Head & Neck Surgery. 1997;123:802-808'},{id:"B18",body:'Guyuron B, Bigdeli Y, Sajjadian A. Dynamics of the alar rim graft. Plastic and Reconstructive Surgery. 2015;135(4):981-986'},{id:"B19",body:'Li YK, Greensmith A. Facilitated alar rim graft placement with an ophthalmic slit blade. Plastic and Reconstructive Surgery. Global Open. 2018;6(4):e1721'},{id:"B20",body:'Jang YJ, Kim SM, Lew DH, Song SY. Simple correction of alar retraction by conchal cartilage extension grafts. Archives of Plastic Surgery. 2016;43(6):564-569 [Epub Nov 18, 2016]'},{id:"B21",body:'Goodrich JL, Wong BJ. Optimizing the soft tissue triangle, alar margin furrow, and alar ridge aesthetics: Analysis and use of the articulate alar rim graft. Facial Plastic Surgery. 2016;32(6):646-655'},{id:"B22",body:'Ballin AC, Kim H, Chance E, Davis RE. The articulated alar rim graft: Reengineering the conventional alar rim graft for improved contour and support. Facial Plastic Surgery. 2016;32(4):384-397'},{id:"B23",body:'Kemaloğlu CA, Altıparmak M. The alar rim flap: A novel technique to manage malpositioned lateral crura. Aesthetic Surgery Journal. 2015;35(8):920-926'},{id:"B24",body:'Deroee AF, Younes AA, Friedman O. External nasal valve collapse repair: The limited alar-facial stab approach. The Laryngoscope. 2011;121(3):474-479'},{id:"B25",body:'Field LM. Nasal alar rim reconstruction utilizing the crus of the helix, with several alternatives for donor site closure. The Journal of Dermatologic Surgery and Oncology. 1986;12(3):253-258'},{id:"B26",body:'Losquadro WD, Bared A, Toriumi DM. Correction of the retracted alar base. Facial Plastic Surgery. 2012;28(2):218-224'},{id:"B27",body:'Boahene KD, Hilger PA. Alar rim grafting in rhinoplasty: Indications, technique, and outcomes. Archives of Facial Plastic Surgery. 2009;11(5):285-289'},{id:"B28",body:'Kalan A, Kenyon GS, Seemungal TA. Treatment of external nasal valve (alar rim) collapse with an alar strut. The Journal of Laryngology and Otology. 2001;115(10):788-791'},{id:"B29",body:'Cárdenas-Camarena L, Guerrero MT. Use of cartilaginous autografts in nasal surgery: 8 years of experience. Plastic and Reconstructive Surgery. 1999;103(3):1003-1014'}],footnotes:[],contributors:[{corresp:null,contributorFullName:"Pedro S. Arquero",address:null,affiliation:'
Clinica Arquero, Spain
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Altmetric and Dimensions from Digital Science
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