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Original article
Comparison of stromal corneal nerves between normal and keratoconus patients using confocal microscopy
Análisis de nervios estromales en pacientes con queratocono
M. Ramírez Fernández
Corresponding author
, E. Hernández Quintela, R. Naranjo Tackman
Servicio de Córnea y Cirugía Refractiva, Asociación Para Evitar la Ceguera en México (APEC), Hospital Luis Sánchez Bulnes, Universidad Nacional Autónoma de México, Mexico City, Mexico
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therefore&#44; the study was more limited&#46;<a class="elsevierStyleCrossRefs" href="#bib0005"><span class="elsevierStyleSup">1&#44;7&#44;8</span></a></p><p id="par0010" class="elsevierStylePara elsevierViewall">Although corneal stroma nerves are not as abundant as those in the subepithelial nerve plexus&#44; upon usual eye examination by slit lamp&#44; due to their size&#44; corneal stroma nerves are the only ones that can be seen&#59; for years they have been described as more apparent and thicker in keratoconus patients than in normal patients or those without this condition<a class="elsevierStyleCrossRef" href="#bib0045"><span class="elsevierStyleSup">9</span></a>&#59; it has even been suggested that corneal stroma nerves are linked to keratoconus progression&#46;<a class="elsevierStyleCrossRef" href="#bib0050"><span class="elsevierStyleSup">10</span></a> This study aims to take images and thus be able to compare as many nerves in the corneal stroma&#44; between those from normal subjects and those from keratoconus patients&#44; using confocal microscopy&#46;</p></span><span id="sec0010" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0070">Subjects&#44; material and methods</span><p id="par0015" class="elsevierStylePara elsevierViewall">All people involved signed the consent form&#59; the study was divided into two groups&#58; group A&#44; control group where 140 eyes of 70 normal subjects without any ocular or systemic disease were studied&#59; and group B where 122 eyes of 87 patients diagnosed with keratoconus using topography &#40;Bausch &#38; Lomb Surgical&#44; Orbtek Inc&#46;&#44; Salt Lake City&#44; UT&#44; USA&#41; in stages <span class="elsevierStyleSmallCaps">ii</span> and <span class="elsevierStyleSmallCaps">iii</span> based on the Amsler-Krumeich&#44; classification&#44; which has been used in multiple keratoconus studies&#46;<a class="elsevierStyleCrossRefs" href="#bib0055"><span class="elsevierStyleSup">11&#8211;14</span></a></p><p id="par0020" class="elsevierStylePara elsevierViewall">Confocal microscopy&#58; At the corneal imaging unit of the Cornea and Refractive Surgery Department of the hospital of the Association to Prevent Blindness in Mexico&#44; after topical anesthesia of cornea with tetracaine hydrochloride 5&#46;0<span class="elsevierStyleHsp" style=""></span>mg per ml &#40;Ponti Ofteno&#44; Laboratorios Sophia&#44; S&#46;A&#46; Guadalajara&#44; Mexico&#41;&#44; all study subjects in both groups underwent a central scan of the total cornea thickness using Confoscan 4 confocal microscope &#40;Fortune Technologies&#44; Vigonza&#44; Italy&#41;&#46; Each confocal microscopy test rendered scanned images in JPEG format&#44; consisting of 2 consecutive scans of total central cornea thickness depth&#59; this scan is equivalent to the endothelium and epithelium and back to endothelium imaging scan&#44; i&#46;e&#46;&#44; from posterior to anterior and back to posterior&#44; to allow movement in the <span class="elsevierStyleItalic">Z</span> axis of central cornea thickness&#46; A Z-Ring Scan &#40;Confoscan&#44; Fortune Technologies&#44; Italy&#41; was used&#59; this device maintains contact with the cornea surface to obtain reliable thickness measurements without anteroposterior eyeball movement&#46;</p><p id="par0025" class="elsevierStylePara elsevierViewall">An average of 350 images per scan were obtained&#59; they were 340<span class="elsevierStyleHsp" style=""></span>&#956;m<span class="elsevierStyleHsp" style=""></span>&#215;<span class="elsevierStyleHsp" style=""></span>255<span class="elsevierStyleHsp" style=""></span>&#956;m at axes <span class="elsevierStyleItalic"><span class="elsevierStyleSmallCaps">X</span></span>&#44; <span class="elsevierStyleItalic">Y</span>&#59; they are automatically saved to a computer hard disk for further analysis using Navis v&#46; 3&#46;5&#46;0 microscopic image analysis software &#40;NIDEK&#44; Multi-Instrument Diagnostic System&#44; Japan&#41;&#46;</p><p id="par0030" class="elsevierStylePara elsevierViewall">Each image from every scan obtained by confocal cornea microscopy was checked by searching for those with pictures of nerves in stroma&#46; Only nerves in focus and with sharp edges were assessed and measured&#46; Stroma nerve morphology and thickness were analyzed&#59; they were measured using Navis v&#46; 3&#46;5&#46;0 microscopic image analysis software &#40;NIDEK&#44; Multi-Instrument Diagnostic System&#44; Japan&#41;&#46; Nerve thickness was obtained from the average of the thickest and narrowest portion of each nerve tested &#40;<a class="elsevierStyleCrossRefs" href="#fig0005">Figs&#46; 1 and 2</a>&#41;&#46; Nerves with bifurcations &#40;<a class="elsevierStyleCrossRef" href="#fig0015">Fig&#46; 3</a>&#41; were not measured&#46; Only corneal stroma nerves were analyzed&#46; Subepithelial nerve plexus nerves were not measured&#46;</p><elsevierMultimedia ident="fig0005"></elsevierMultimedia><elsevierMultimedia ident="fig0010"></elsevierMultimedia><elsevierMultimedia ident="fig0015"></elsevierMultimedia></span><span id="sec0015" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0075">Results</span><p id="par0035" class="elsevierStylePara elsevierViewall">All confocal and central cornea thickness microscopy scans were integral&#44; given that epithelium&#44; subepithelial nerve plexus&#44; endothelium&#44; and corneal stroma were visible with keratocyte nuclei&#44; starting with the first image beneath the subepithelial nerve plexus&#44; up to the image above the corneal endothelium&#46;</p><p id="par0040" class="elsevierStylePara elsevierViewall">A total of 243 stroma nerves in focus and with well-defined edges were obtained for analysis from all confocal microscopy scan images&#59; 127 were from group A &#40;subjects without pathology&#41;&#44; and 116 from group B &#40;keratoconus patients&#41;&#46; All stroma nerves were observed in cornea confocal microscopy as well-defined linear and highly reflective structures&#44; surrounded by corneal stroma keratocytes&#44; the latter with moderate reflection &#40;<a class="elsevierStyleCrossRefs" href="#fig0005">Figs&#46; 1 and 2</a>&#41;&#46; Corneal stroma nerves were arranged in an oblique direction in 100&#37; of all images assessed&#44; also in all cases with coarse and narrow morphology in some portions &#40;<a class="elsevierStyleCrossRefs" href="#fig0005">Figs&#46; 1 and 2</a>&#41;&#46;</p><p id="par0045" class="elsevierStylePara elsevierViewall">Mean stroma nerve thickness in group A&#44; i&#46;e&#46; patients without disease &#40;<a class="elsevierStyleCrossRef" href="#fig0005">Fig&#46; 1</a>&#41; was 5&#46;7<span class="elsevierStyleHsp" style=""></span>&#177;<span class="elsevierStyleHsp" style=""></span>1&#46;7 ranging from 3&#46;3 to 10&#46;4<span class="elsevierStyleHsp" style=""></span>&#956;&#44; mean stroma nerves thickness in group B&#44; keratoconus patients &#40;<a class="elsevierStyleCrossRef" href="#fig0010">Fig&#46; 2</a>&#41; was 7&#46;2<span class="elsevierStyleHsp" style=""></span>&#177;<span class="elsevierStyleHsp" style=""></span>1&#46;9 ranging from 3&#46;5 to 12&#46;0<span class="elsevierStyleHsp" style=""></span>&#956;m&#46; The difference in the thickness of corneal stroma nerves between group A and group B was statistically significant &#40;<span class="elsevierStyleItalic">p</span><span class="elsevierStyleHsp" style=""></span>&#60;<span class="elsevierStyleHsp" style=""></span>0&#46;05&#44; unpaired <span class="elsevierStyleItalic">t</span> test&#41; &#40;<a class="elsevierStyleCrossRef" href="#fig0020">Fig&#46; 4</a>&#41;&#46;</p><elsevierMultimedia ident="fig0020"></elsevierMultimedia></span><span id="sec0020" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0080">Discussion</span><p id="par0050" class="elsevierStylePara elsevierViewall">Under normal conditions&#44; various structures such as corneal surface epithelium&#44; basal epithelial cells&#44; nerve plexus subepithelial&#44; cores stromal keratocytes and endothelial cells may be assessed by confocal microscopy&#46;<a class="elsevierStyleCrossRefs" href="#bib0005"><span class="elsevierStyleSup">1&#44;7</span></a> Stroma nerves are no exception&#46; In this study&#44; they show as highly reflective linear structures with thick and thin portions&#44; as already reported in the literature&#44;<a class="elsevierStyleCrossRefs" href="#bib0075"><span class="elsevierStyleSup">15&#44;16</span></a> surrounded by keratinocytes nuclei with moderate reflection&#46;<a class="elsevierStyleCrossRefs" href="#bib0005"><span class="elsevierStyleSup">1&#44;17</span></a></p><p id="par0055" class="elsevierStylePara elsevierViewall">Most studies of corneal nerves using confocal microscopy have focused mainly on subepithelial nerve plexus because it is across the cornea&#44; beneath its epithelium&#44; which makes it very easy to take images of the plexus&#46;<a class="elsevierStyleCrossRefs" href="#bib0015"><span class="elsevierStyleSup">3&#44;4&#44;18</span></a> Therefore&#44; there have been studies of the subepithelial nerve plexus under normal conditions<a class="elsevierStyleCrossRefs" href="#bib0015"><span class="elsevierStyleSup">3&#44;19</span></a> &#40;or changes the nerve plexus undergoes resulting from surgical procedures such as corneal transplantation<a class="elsevierStyleCrossRef" href="#bib0010"><span class="elsevierStyleSup">2</span></a> or refractive surgery<a class="elsevierStyleCrossRefs" href="#bib0020"><span class="elsevierStyleSup">4&#44;20&#44;21</span></a>&#41; and diseases such as dry eye&#46;<a class="elsevierStyleCrossRef" href="#bib0080"><span class="elsevierStyleSup">16</span></a> Subepithelial nerve plexus studies have been conducted on keratoconus patients&#44; including linking them to corneal sensitivity&#46;<a class="elsevierStyleCrossRefs" href="#bib0025"><span class="elsevierStyleSup">5&#44;6</span></a> Furthermore&#44; it is harder to find corneal stroma nerves&#44; since it is not a plexus&#44;<a class="elsevierStyleCrossRef" href="#bib0110"><span class="elsevierStyleSup">22</span></a> and given that images by confocal microscopy are 340<span class="elsevierStyleHsp" style=""></span>&#956;m<span class="elsevierStyleHsp" style=""></span>&#215;<span class="elsevierStyleHsp" style=""></span>255<span class="elsevierStyleHsp" style=""></span>&#956;m in the axes <span class="elsevierStyleItalic"><span class="elsevierStyleSmallCaps">X</span></span>&#44; <span class="elsevierStyleItalic">Y</span> &#40;Confoscan 4 Fortune Technologies&#44; Vigonza&#44; Italy&#41;&#44; it has to match the path of a corneal stroma nerve with the tiny image taken in the scan&#46; This study focused on corneal stroma nerves&#46;</p><p id="par0060" class="elsevierStylePara elsevierViewall">Keratoconus is a disease in which the cornea undergoes a usually progressive ectasia&#44; which may appear in childhood or later in life and often leads to the need for a corneal transplant to correct the problem&#46;<a class="elsevierStyleCrossRef" href="#bib0115"><span class="elsevierStyleSup">23</span></a> Early diagnosis of keratoconus today is of utmost importance due to the rise of refractive surgery&#44; since it is a major contraindications for it&#46;<a class="elsevierStyleCrossRef" href="#bib0120"><span class="elsevierStyleSup">24</span></a> Development and evolution of corneal topography equipment have allowed the diagnosis of keratoconus at increasingly early stages&#46;<a class="elsevierStyleCrossRefs" href="#bib0125"><span class="elsevierStyleSup">25&#8211;30</span></a> However&#44; for years it has been shown that there are subtle details such as observing corneal stroma nerves&#44; most evident in keratoconus patients undergoing ophthalmic examination by slit lamp&#44; which could prove incipient keratoconus<a class="elsevierStyleCrossRef" href="#bib0045"><span class="elsevierStyleSup">9</span></a> and thus support diagnosis in cases only suspected via corneal topography&#46; There is even speculation on the role corneal nerves may have on keratoconus progression&#46;<a class="elsevierStyleCrossRef" href="#bib0050"><span class="elsevierStyleSup">10</span></a> Even Al-Alqaba et al&#46;<a class="elsevierStyleCrossRef" href="#bib0155"><span class="elsevierStyleSup">31</span></a> reported a case series of 14 corneal buttons disease in keratoconus patients&#44; comparing them to 6 corneal buttons in patients without this condition&#44; where the corneal stroma nerves were thicker&#46; This study was performed to compare the thickness of corneal stroma nerves by confocal microscopy in vivo and in a larger group&#46;</p><p id="par0065" class="elsevierStylePara elsevierViewall">Studies have been conducted using confocal microscopy in corneal stroma keratoconus patients&#44; mainly focused on changes in density and reflection of stromal keratocytes&#44; due to their easy capture by confocal microscopy since keratinocytes are found across stroma thickness&#46;<a class="elsevierStyleCrossRefs" href="#bib0160"><span class="elsevierStyleSup">32&#44;33</span></a> This study aimed to compare morphological characteristics and thickness of corneal stroma nerves using confocal microscopy&#44; between normal subjects and patients diagnosed with keratoconus&#46; We found no morphological differences between the two groups&#59; both showed corneal stroma nerves and well-defined highly reflective linear structures&#44; surrounded by keratinocytes as shown in confocal microscopy images of corneal stroma&#59; they were arranged in an oblique direction in 100&#37; of all images assessed&#44; with various degrees of tortuosity of the thick and narrow portions in both groups&#46; We did find a statistically significant difference regarding thickness of corneal stroma nerves&#59; it was thicker in patients diagnosed with keratoconus&#44; which underpins the classical idea that corneal stroma nerves are more apparent in ophthalmic examination by slit lamp in keratoconus patients&#46;<a class="elsevierStyleCrossRef" href="#bib0045"><span class="elsevierStyleSup">9</span></a></p><p id="par0070" class="elsevierStylePara elsevierViewall">Corneal stroma nerves thickness has been studied in other diseases where they are known to be abnormally thick&#44; such as the Mocan et al&#46; study&#44;<a class="elsevierStyleCrossRef" href="#bib0085"><span class="elsevierStyleSup">17</span></a> in which mean corneal stroma thickness in diabetic patients was 8&#46;99<span class="elsevierStyleHsp" style=""></span>&#177;<span class="elsevierStyleHsp" style=""></span>2&#46;32<span class="elsevierStyleHsp" style=""></span>&#956;m&#46; In that study&#44; 35 eyes were assessed in the group of patients with diabetes and 24 in the healthy subject control group&#59; this study assessed 122 in the keratoconics group&#44; and 140 in the control group without disease&#44; and&#44; even so&#44; the latter shows corneal stroma nerve thickness of 5&#46;7<span class="elsevierStyleHsp" style=""></span>&#177;<span class="elsevierStyleHsp" style=""></span>1&#46;7<span class="elsevierStyleHsp" style=""></span>&#956;m&#44; similar to the control group reported by Mocan et al&#46; of 5&#46;69<span class="elsevierStyleHsp" style=""></span>&#177;<span class="elsevierStyleHsp" style=""></span>2&#46;32<span class="elsevierStyleHsp" style=""></span>&#956;m&#46;<a class="elsevierStyleCrossRef" href="#bib0085"><span class="elsevierStyleSup">17</span></a></p><p id="par0075" class="elsevierStylePara elsevierViewall">In this study&#44; stromal nerve with bifurcations were not measured&#44; to avoid overestimating their thickness or altering the actual mean thickness of measured nerves&#46;</p><p id="par0080" class="elsevierStylePara elsevierViewall">Only patients with a diagnosis of keratoconus in stages <span class="elsevierStyleSmallCaps">ii</span> and <span class="elsevierStyleSmallCaps">iii</span> based on the Amsler-Krumeich classification were included because stage <span class="elsevierStyleSmallCaps">iv</span> patients already have scarring that limits the taking and acquisition of quality corneal stroma images using confocal microscopy&#46; Likewise&#44; for stage <span class="elsevierStyleSmallCaps">i</span> patients&#44; keratoconus diagnosis may be doubtful&#44; and bias the study by measuring nerves in non-keratoconus patients&#44; as we needed to be certain to measure corneal stroma nerves in patients with unquestionable keratoconus diagnosis&#46;</p><p id="par0085" class="elsevierStylePara elsevierViewall">Thanks to confocal microscopy it is possible to evaluate corneal stroma nerves&#59; although they are more difficult to capture in confocal microscopy images than subepithelial nerve plexus&#44; a higher number of patients will yield enough images for analysis and measurement&#46;</p><p id="par0090" class="elsevierStylePara elsevierViewall">This study can confirm that keratoconus patients have corneal stroma nerves thicker than those of normal subjects&#44; and the aforementioned notion that&#44; under ophthalmologic examination with slit lamp&#44; corneal stroma nerves are more evident in keratoconus patients than in those without the disease&#46;<a class="elsevierStyleCrossRef" href="#bib0045"><span class="elsevierStyleSup">9</span></a> The latter may provide additional data on clinical suspicion of keratoconus&#46;</p></span><span id="sec0025" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0085">Funding</span><p id="par0095" class="elsevierStylePara elsevierViewall">APEC&#44; <span class="elsevierStyleSmallCaps">I</span>&#46;A&#46;P&#46;&#44; Mexico City&#44; Mexico&#46;</p></span><span id="sec0030" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0090">Conflicts of interest</span><p id="par0100" class="elsevierStylePara elsevierViewall">The authors declare that they have no conflicts of interest&#46;</p></span></span>"
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            0 => "Resumen"
            1 => "Objetivo"
            2 => "M&#233;todos"
            3 => "Resultados"
            4 => "Conclusiones"
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          "identificador" => "xpalclavsec352984"
          "titulo" => "Palabras clave"
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          "titulo" => "Introduction"
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          "titulo" => "Subjects&#44; material and methods"
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        6 => array:2 [
          "identificador" => "sec0015"
          "titulo" => "Results"
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          "titulo" => "Discussion"
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        8 => array:2 [
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          "titulo" => "Funding"
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          "identificador" => "sec0030"
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    "fechaRecibido" => "2013-07-24"
    "fechaAceptado" => "2014-02-24"
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          "clase" => "keyword"
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          "palabras" => array:5 [
            0 => "Cornea"
            1 => "Stromal"
            2 => "Keratoconus"
            3 => "Nerves"
            4 => "Confocal"
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          "clase" => "keyword"
          "titulo" => "Palabras clave"
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          "palabras" => array:5 [
            0 => "C&#243;rnea"
            1 => "Estroma"
            2 => "Queratocono"
            3 => "Nervios"
            4 => "Confocal"
          ]
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    "resumen" => array:2 [
      "en" => array:2 [
        "titulo" => "Abstract"
        "resumen" => "<span class="elsevierStyleSectionTitle" id="sect0010">Objective</span><p id="spar0025" class="elsevierStyleSimplePara elsevierViewall">To evaluate the differences in stromal corneal nerves between normal patients and keratoconus patients&#46;</p> <span class="elsevierStyleSectionTitle" id="sect0015">Material and methods</span><p id="spar0030" class="elsevierStyleSimplePara elsevierViewall">A total of 140 eyes of 70 normal patients &#40;group A&#41; and 122 eyes of 87 keratoconus patients &#40;group B&#41; were examined with the confocal microscope&#44; with a central scan of the total corneal thickness being taken&#46; The morphology and thickness of the corneal stromal nerves were evaluated by using the Navis v&#46; 3&#46;5&#46;0 software&#46; Nerve thickness was obtained from the mean between the widest and the narrowest portions of each stromal nerve&#46;</p> <span class="elsevierStyleSectionTitle" id="sect0020">Results</span><p id="spar0035" class="elsevierStyleSimplePara elsevierViewall">Corneal stromal nerves were observed as irregular linear hyper-reflective structures with wide and narrow portions in all cases&#46; Mean corneal stromal nerves thickness in group A was 5&#46;7<span class="elsevierStyleHsp" style=""></span>&#177;<span class="elsevierStyleHsp" style=""></span>1&#46;7 &#40;range from 3&#46;3 to 10&#46;4<span class="elsevierStyleHsp" style=""></span>&#956;m&#41;&#44; mean corneal stromal nerves thickness in group B was 7&#46;2<span class="elsevierStyleHsp" style=""></span>&#177;<span class="elsevierStyleHsp" style=""></span>1&#46;9 &#40;range from 3&#46;5 to 12&#46;0<span class="elsevierStyleHsp" style=""></span>&#956;m&#41;&#46; There was a statistical significant difference &#40;<span class="elsevierStyleItalic">p</span><span class="elsevierStyleHsp" style=""></span>&#60;<span class="elsevierStyleHsp" style=""></span>&#46;05&#41; in stromal corneal nerves thickness between group A and group B&#46;</p> <span class="elsevierStyleSectionTitle" id="sect0025">Conclusion</span><p id="spar0040" class="elsevierStyleSimplePara elsevierViewall">Stromal corneal nerves morphology was similar in both groups&#44; but stromal nerves were thicker in keratoconus patients&#46;</p>"
      ]
      "es" => array:2 [
        "titulo" => "Resumen"
        "resumen" => "<span class="elsevierStyleSectionTitle" id="sect0035">Objetivo</span><p id="spar0005" class="elsevierStyleSimplePara elsevierViewall">Evaluar las diferencias de los nervios del estroma de la c&#243;rnea entre sujetos normales y pacientes con queratocono&#46;</p> <span class="elsevierStyleSectionTitle" id="sect0040">M&#233;todos</span><p id="spar0010" class="elsevierStyleSimplePara elsevierViewall">Un total de 140 ojos de 70 sujetos normales &#40;grupo A&#41; y 122 ojos de 87 pacientes con queratocono &#40;grupo B&#41;&#44; fueron evaluados con el microscopio confocal&#44; realizando un rastreo central del espesor total de la c&#243;rnea&#46; La morfolog&#237;a y el espesor de los nervios fueron evaluados utilizando el programa Navis v&#46; 3&#46;5&#46;0&#46; El espesor de los nervios se obtuvo del promedio de la porci&#243;n m&#225;s delgada y la m&#225;s gruesa de cada nervio&#46;</p> <span class="elsevierStyleSectionTitle" id="sect0045">Resultados</span><p id="spar0015" class="elsevierStyleSimplePara elsevierViewall">Los nervios del estroma se observaron como estructuras lineales de alta reflexi&#243;n e irregulares&#44; con porciones gruesas y angostas en todos los casos&#46; El promedio del espesor de los nervios en el grupo A fue de 5&#44;7<span class="elsevierStyleHsp" style=""></span>&#177;<span class="elsevierStyleHsp" style=""></span>1&#44;7 &#40;rango de 3&#44;3 a 10&#44;4<span class="elsevierStyleHsp" style=""></span>&#956;&#41;&#44; en el grupo B fue de 7&#44;2<span class="elsevierStyleHsp" style=""></span>&#177;<span class="elsevierStyleHsp" style=""></span>1&#44;9 &#40;rango de 3&#44;5 a 12&#44;0<span class="elsevierStyleHsp" style=""></span>&#956;&#41;&#46; La diferencia en el espesor de los nervios entre el grupo A y el grupo B fue estad&#237;sticamente significativa &#40;p<span class="elsevierStyleHsp" style=""></span>&#60;<span class="elsevierStyleHsp" style=""></span>0&#44;05&#41;&#46;</p> <span class="elsevierStyleSectionTitle" id="sect0050">Conclusiones</span><p id="spar0020" class="elsevierStyleSimplePara elsevierViewall">La morfolog&#237;a de los nervios del estroma de la c&#243;rnea fue similar en ambos grupos&#59; el espesor de estos fue mayor en los pacientes con queratocono&#46;</p>"
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        "nota" => "<p class="elsevierStyleNotepara" id="npar0005">Please cite this article as&#58; Ram&#237;rez Fern&#225;ndez M&#44; Hern&#225;ndez Quintela E&#44; Naranjo Tackman R&#46; An&#225;lisis de nervios estromales en pacientes con queratocono&#46; Arch Soc Esp Oftalmol&#46; 2014&#59;89&#58;308&#8211;312&#46;</p>"
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          "en" => "<p id="spar0045" class="elsevierStyleSimplePara elsevierViewall">Corneal stroma confocal microscopy image 340<span class="elsevierStyleHsp" style=""></span>&#956;m<span class="elsevierStyleHsp" style=""></span>&#215;<span class="elsevierStyleHsp" style=""></span>255<span class="elsevierStyleHsp" style=""></span>&#956;m&#46; Corneal stroma nerve of group A is shown&#44; seen as a linear&#44; highly reflective structure with thick and narrow portions &#40;indicated by arrows&#41; surrounded by keratinocytes&#46;</p>"
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          "en" => "<p id="spar0050" class="elsevierStyleSimplePara elsevierViewall">Corneal stroma confocal microscopy image of 340<span class="elsevierStyleHsp" style=""></span>&#956;m<span class="elsevierStyleHsp" style=""></span>&#215;<span class="elsevierStyleHsp" style=""></span>255<span class="elsevierStyleHsp" style=""></span>&#956;m&#46; Corneal stroma nerve of group B is shown&#44; seen as a linear&#44; highly reflective structure with thick and narrow portions &#40;indicated by arrows&#41; surrounded by keratinocytes&#46;</p>"
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          "en" => "<p id="spar0055" class="elsevierStyleSimplePara elsevierViewall">Corneal stroma confocal microscopy image of 340<span class="elsevierStyleHsp" style=""></span>&#956;m<span class="elsevierStyleHsp" style=""></span>&#215;<span class="elsevierStyleHsp" style=""></span>255<span class="elsevierStyleHsp" style=""></span>&#956;m&#46; Corneal stroma nerve bifurcation of group A is shown &#40;indicated by arrows&#41; surrounded by keratocytes&#46;</p>"
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          "en" => "<p id="spar0060" class="elsevierStyleSimplePara elsevierViewall">Mean<span class="elsevierStyleHsp" style=""></span>&#177;<span class="elsevierStyleHsp" style=""></span>standard deviation of corneal stroma nerve thickness of group A&#44; normal subjects&#44; and group B&#44; keratoconus patients&#46; Difference in thickness between the two groups was statistically significant &#40;<span class="elsevierStyleItalic">p</span><span class="elsevierStyleHsp" style=""></span>&#60;<span class="elsevierStyleHsp" style=""></span>0&#46;05&#44; unpaired <span class="elsevierStyleItalic">t</span> test&#41;&#46;</p>"
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      "titulo" => "References"
      "seccion" => array:1 [
        0 => array:2 [
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            0 => array:3 [
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              "etiqueta" => "1"
              "referencia" => array:1 [
                0 => array:2 [
                  "contribucion" => array:1 [
                    0 => array:2 [
                      "titulo" => "Microscopia confocal de la c&#243;rnea"
                      "autores" => array:1 [
                        0 => array:2 [
                          "etal" => false
                          "autores" => array:4 [
                            0 => "V&#46; Sanchez-Huerta"
                            1 => "M&#46; Ramirez"
                            2 => "E&#46; Hernandez-Quintela"
                            3 => "R&#46; Naranjo Tackman"
                          ]
                        ]
                      ]
                    ]
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