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Dive into the research topics where M. Carmen Martínez-García is active.

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Featured researches published by M. Carmen Martínez-García.


Biomedical Optics Express | 2011

Femtosecond infrared intrastromal ablation and backscattering-mode adaptive-optics multiphoton microscopy in chicken corneas

Emilio J. Gualda; Javier R. Vázquez de Aldana; M. Carmen Martínez-García; Pablo Moreno; Juan Hernández-Toro; Luis Roso; Pablo Artal; Juan M. Bueno

The performance of femtosecond (fs) laser intrastromal ablation was evaluated with backscattering-mode adaptive-optics multiphoton microscopy in ex vivo chicken corneas. The pulse energy of the fs source used for ablation was set to generate two different ablation patterns within the corneal stroma at a certain depth. Intrastromal patterns were imaged with a custom adaptive-optics multiphoton microscope to determine the accuracy of the procedure and verify the outcomes. This study demonstrates the potential of using fs pulses as surgical and monitoring techniques to systematically investigate intratissue ablation. Further refinement of the experimental system by combining both functions into a single fs laser system would be the basis to establish new techniques capable of monitoring corneal surgery without labeling in real-time. Since the backscattering configuration has also been optimized, future in vivo implementations would also be of interest in clinical environments involving corneal ablation procedures.


Investigative Ophthalmology & Visual Science | 2017

Biomechanical Changes After In Vivo Collagen Cross-Linking With Rose Bengal–Green Light and Riboflavin-UVA

Nandor Bekesi; Patricia Gallego-Muñoz; Lucía Ibares-Frías; Pablo Pérez-Merino; M. Carmen Martínez-García; Irene E. Kochevar; Susana Marcos

Purpose To compare corneal biomechanical properties after in vivo and ex vivo cross-linking (CXL) using rose bengal-green light (RGX) or riboflavin-UVA (UVX). Methods Corneas of 30 rabbits were treated in vivo by the two CXL modalities monolaterally (Group 1) or bilaterally (Group 2). Rabbits in Group 1 were euthanized 1 month after treatments and in Group 2 two months after treatment. Ex vivo CXL was also performed. Eyes were measured by Scheimpflug air puff corneal deformation imaging (Corvis ST) under constant IOP. Corneal deformation parameters were assessed. Inherent corneal biomechanical properties were estimated using inverse finite element modeling. Results Peak to peak distance decreased 16% 2 months after RGX, and 4% and 20% 1 and 2 months after UVX, respectively. The equivalent Youngs modulus (Eeq) increased relative to the control during the post treatment period for both RGX and UVX. The Eeq increased by factors of 3.4 (RGX) and 1.7 (UVX) 1 month and by factors of 10.7 (RGX) and 7.3 (UVX) 2 months after treatment. However, the Eeq values for ex vivo CXL were much greater than produced in vivo. The ex vivo Eeq was greater than the 1-month in vivo values by factors of 8.1 (RGX) and 9.1 (UVX) and compared with 2 month by factors of 2.5 (RGX) and 2.1 (UVX). Conclusions These results indicate that corneal stiffness increases after CXL, and further increases as a function of time after both RGX and UVX. Also, while biomechanical properties determined after ex vivo CXL are indicative of corneal stiffening, they may not provide entirely accurate information about the responses to CXL in vivo.


Optometry and Vision Science | 2010

Corneal light transmission and roughness after refractive surgery.

Pablo Pérez-Merino; M. Carmen Martínez-García; Santiago Mar-Sardaña; Alfonso Pérez-Escudero; Tomas Blanco-Mezquita; Agustín Mayo-Iscar; Jesús Merayo-Lloves

Purpose. To determine the relation between the corneal light transmission measurements and the epithelial surface properties in hen corneas after different refractive surgery techniques photorefractive keratectomy, laser in situ keratomileusis, and laser-assisted subepithelial keratomileusis, and a group with only epithelial corneal removal (deepithelialization). Methods. Five groups of hen corneas with different treatments and a control group were analyzed at 30 days. Direct transmittance and corneal light scattering were measured by a scatterometer developed by our group. Quantitative and systematic measurements of external and internal roughness and epithelium thickness were assessed using standard techniques developed for quantitative analysis of microphotographs of the corneal epithelium. Results. Data analysis revealed that the roughness in the epithelial surface was associated with the corneal light transmission. The direct transmittance of light showed a significant correlation with the epithelial roughness in the control (r = −0.99, p < 0.05) and photorefractive keratectomy (r = −0.99, p < 0.05) groups. However, there was no relation between the epithelial thickness and the corneal light transmission measurements. Conclusions. The experimental results suggested that the roughness of the epithelial surfaces is related to the light transmission in the cornea.


Cytokine | 2017

Effects of TGFβ1, PDGF-BB, and bFGF, on human corneal fibroblasts proliferation and differentiation during stromal repair

Patricia Gallego-Muñoz; Lucía Ibares-Frías; María Cruz Valsero‐Blanco; Roberto Cantalapiedra-Rodriguez; Jesus Merayo-Lloves; M. Carmen Martínez-García

HighlightsTGF&bgr;1 induces slow and constant proliferation and myofibroblast differentiation, reducing cellular motility during closure.PDGF‐BB induces a quick proliferation and high cellular motility with a low level of myofibroblast differentiation during closure.bFGF accelerates wound closure by increasing cell proliferation inhibiting myofibroblast differentiation. Abstract In an effort to improve the regenerative nature of corneal repair, this study reports the use of an in vitro human corneal fibroblasts (HCFs) wound model after treatment with three of the main growth factors (GFs) involved in corneal healing: transforming growth factor beta 1 (TGF&bgr;1), platelet‐derived growth factor BB‐isoform (PDGF‐BB), and basic fibroblast growth factor (bFGF) in order to delve in cell proliferation and differentiation processes. HCFs were mechanically wounded. The individual effect of TGF&bgr;1, PDGF‐BB, and bFGF on cell proliferation and differentiation during the repair process was studied at different time points until wound closure. Wound dimensions and morphological changes were evaluated by microscopy. Cell proliferation and myofibroblast differentiation were analyzed by immunofluorescence cytochemistry. Changes in cell morphology were apparent at Day 4. PDGF‐BB‐ and bFGF‐treated cells had fibroblast‐like morphology. TGF&bgr;1 stimulated proliferation in the wound edge and surrounding area, induced myofibroblast differentiation and inhibited cellular migration. PDGF‐BB induced rapid wound closure due to proliferation, high motility, and late myofibroblast differentiation. The time course of closure induced by bFGF was similar to that for PDGF‐BB, but was mostly due to proliferation in the wound area, and inhibited myofibroblast differentiation. Each of the GFs induced increases in responses promoting stromal repair differently. This study provides insight regarding how to optimize the outcome of stromal repair following corneal injury.


Experimental Eye Research | 2019

Nidogen-2: Location and expression during corneal wound healing

Patricia Gallego-Muñoz; Elvira Lorenzo-Martín; Itziar Fernández; Cristina Herrero-Pérez; M. Carmen Martínez-García

ABSTRACT Nidogen‐2 is a basement membrane (BM) glycoprotein that could be a key to understanding why defects in BM regeneration occur after severe trauma to the cornea. We monitored the location and expression of nidogen‐2 during corneal repair after alkali burn in rabbits. In rabbits that received both general and ocular topical anaesthesia, the central cornea of the left eye was burned by placing an 8‐mm diameter filter paper soaked in 0.5 N NaOH for 60 s. Right corneas were used as controls. The eyes were evaluated at 2, 7, 15, and 30 days after burning and analysed by immunohistochemistry for nidogen‐2 and &agr;‐smooth muscle actin, a myofibroblast marker. Nidogen‐2 mRNA expression levels were determined by quantitative real‐time polymerase chain reaction. In control corneas, nidogen‐2‐positive cells were in all epithelial layers, the endothelium, and the anterior and posterior stromal regions. At Day 2 after the alkali burn, the wound area epithelium and the peripheral epithelium were made up of only 1 to 2 cell layers, all of them nidogen‐2 positive. At Day 7 in the wound area, the epithelium consisted of two cell layers, and the basally located cells were mostly nidogen‐2 positive. The greatest change was observed at Day 30. At this time, the ulcer prevalence in the alkali‐burned corneas was approximately 50% and the central epithelial defects remained. In unepithelialized corneas, frequent epithelial detachments were present, in which almost of the epithelial cells were nidogen‐2 negative. The injured stroma was repopulated by activated stromal cells that synthesized nidogen‐2. The nidogen‐2 was retained in the newly secreted, but disordered, matrix produced mainly by the myofibroblasts localized in the stroma at 7, 15, and 30 days after burning. Thus, even though nidogen‐2 was present, it was unable to contribute to the effective regeneration of the BM. HighlightsCorneal stromal cells produce nidogen‐2 after alkali burn.Corneal nidogen‐2 expression levels are upregulated during corneal repair.During corneal repair, the epithelial and stromal nidogen‐2 location changes.


Journal of Tissue Engineering and Regenerative Medicine | 2018

Human corneal fibroblast migration and extracellular matrix synthesis during stromal repair: Role played by platelet-derived growth factor-BB, basic fibroblast growth factor, and transforming growth factor-β1: HCFs migration and ECM synthesis during stromal repair: GF effects

Patricia Gallego-Muñoz; Lucía Ibares-Frías; José A. Garrote; María Cruz Valsero‐Blanco; Roberto Cantalapiedra-Rodriguez; Jesus Merayo-Lloves; M. Carmen Martínez-García

The development of treatments that modulate corneal wound healing to avoid fibrosis during tissue repair is important for the restoration of corneal transparency after an injury. To date, few studies have studied the influence of growth factors (GFs) on human corneal fibroblast (HCF) expression of extracellular matrix (ECM) proteins such as collagen types I and III, proteoglycans such as perlecan, or proteins implicated in cellular migration such as α5β1‐integrin and syndecan‐4. Using in vitro HCFs, a mechanical wound model was developed to study the influence of the GFs basic fibroblast GF (bFGF), platelet‐derived GF (PDGF‐BB) and transforming GF‐β1 (TGFβ1) on ECM protein production and cellular migration. Our results show that mechanical wounding provokes the autocrine release of bFGF and TGFβ1 at different time points during the wound closure. The HCF response to PDGF‐BB was a rapid closure due to fast cellular migration associated with a high focal adhesion replacement and a high expression of collagen and proteoglycans, producing nonfibrotic healing. bFGF stimulated nonfibrotic ECM production and limited the migration process. Finally, TGFβ1 induced expression of the fibrotic markers collagen type III and α5β1 integrin, and it inhibited cellular migration due to the formation of focal adhesions with a low turnover rate. The novel in vitro HCF mechanical wound model can be used to understand the role played by GFs in human corneal repair. The model can also be used to test the effects of different treatments aimed at improving the healing process. Copyright


Investigative Ophthalmology & Visual Science | 2018

Rose Bengal and Green Light Versus Riboflavin–UVA Cross-Linking: Corneal Wound Repair Response

Elvira Lorenzo-Martín; Patricia Gallego-Muñoz; Lucía Ibares-Frías; Susana Marcos; Pablo Pérez-Merino; Itziar Fernández; Irene E. Kochevar; M. Carmen Martínez-García

Purpose To study corneal wound healing after two cross-linking techniques using either rose bengal and green light (RGX) or the conventional treatment using riboflavin and UVA radiation (UVX). Methods Corneas of New Zealand rabbits were monolaterally treated with UVX (21 eyes) or RGX (25 eyes). Treatments involved corneal de-epithelialization (8-mm diameter), soaking with photosensitizer (0.1% riboflavin in 20% dextran for 30 minutes for UVX; 0.1% rose bengal for 2 minutes for RGX), and light irradiation (370 nm, 3 mW/cm2, 30 minutes for UVX; 532 nm, 0.25 W/cm2, 7 minutes for RGX). Contralateral eyes were used as controls. Clinical follow-up included fluorescein staining, haze measurement, and pachymetry. Healing events analyzed after euthanasia at 2, 30, and 60 days included cell death (TUNEL assay), cell proliferation (BrdU [bromodeoxyuridine] immunofluorescence), and differentiation to myofibroblasts (α-SMA [alpha smooth muscle actin] immunohistochemistry). Results Re-epithelialization and pachymetries were similar after RGX and UVX. The haze from day 1 to 15 was greater after UVX. Cell death was deeper after UVX, being localized in the anterior and middle stroma, and was superficial (anterior third) after RGX. Cell proliferation appeared after 2 days and was localized in the middle and posterior stroma in the UVX group but was superficial in the RGX group. After 60 days the number of stromal cells had not returned to the control number in either group. Conclusions The deeper and longer-lasting cell damage caused by UVX compared to RGX may underlie the slower cell repopulation after UVX and other differences in healing. Shallower damage and a shorter treatment time suggest that RGX may be appropriate for stiffening thin corneas.


Journal of Tissue Engineering and Regenerative Medicine | 2017

Human corneal fibroblast migration and extracellular matrix synthesis during stromal repair: Role played by platelet‐derived growth factor‐BB, basic fibroblast growth factor, and transforming growth factor‐β1

Patricia Gallego-Muñoz; Lucía Ibares-Frías; José A. Garrote; María Cruz Valsero‐Blanco; Roberto Cantalapiedra-Rodriguez; Jesus Merayo-Lloves; M. Carmen Martínez-García

The development of treatments that modulate corneal wound healing to avoid fibrosis during tissue repair is important for the restoration of corneal transparency after an injury. To date, few studies have studied the influence of growth factors (GFs) on human corneal fibroblast (HCF) expression of extracellular matrix (ECM) proteins such as collagen types I and III, proteoglycans such as perlecan, or proteins implicated in cellular migration such as α5β1‐integrin and syndecan‐4. Using in vitro HCFs, a mechanical wound model was developed to study the influence of the GFs basic fibroblast GF (bFGF), platelet‐derived GF (PDGF‐BB) and transforming GF‐β1 (TGFβ1) on ECM protein production and cellular migration. Our results show that mechanical wounding provokes the autocrine release of bFGF and TGFβ1 at different time points during the wound closure. The HCF response to PDGF‐BB was a rapid closure due to fast cellular migration associated with a high focal adhesion replacement and a high expression of collagen and proteoglycans, producing nonfibrotic healing. bFGF stimulated nonfibrotic ECM production and limited the migration process. Finally, TGFβ1 induced expression of the fibrotic markers collagen type III and α5β1 integrin, and it inhibited cellular migration due to the formation of focal adhesions with a low turnover rate. The novel in vitro HCF mechanical wound model can be used to understand the role played by GFs in human corneal repair. The model can also be used to test the effects of different treatments aimed at improving the healing process. Copyright


Journal of Tissue Engineering and Regenerative Medicine | 2016

Human corneal fibroblast migration and ECM synthesis during stromal repair: Role played by PDGF-BB, bFGF, and TGFβ1

Patricia Gallego-Muñoz; Lucía Ibares-Frías; José A. Garrote; María Cruz Valsero‐Blanco; Roberto Cantalapiedra-Rodriguez; Jesus Merayo-Lloves; M. Carmen Martínez-García

The development of treatments that modulate corneal wound healing to avoid fibrosis during tissue repair is important for the restoration of corneal transparency after an injury. To date, few studies have studied the influence of growth factors (GFs) on human corneal fibroblast (HCF) expression of extracellular matrix (ECM) proteins such as collagen types I and III, proteoglycans such as perlecan, or proteins implicated in cellular migration such as α5β1‐integrin and syndecan‐4. Using in vitro HCFs, a mechanical wound model was developed to study the influence of the GFs basic fibroblast GF (bFGF), platelet‐derived GF (PDGF‐BB) and transforming GF‐β1 (TGFβ1) on ECM protein production and cellular migration. Our results show that mechanical wounding provokes the autocrine release of bFGF and TGFβ1 at different time points during the wound closure. The HCF response to PDGF‐BB was a rapid closure due to fast cellular migration associated with a high focal adhesion replacement and a high expression of collagen and proteoglycans, producing nonfibrotic healing. bFGF stimulated nonfibrotic ECM production and limited the migration process. Finally, TGFβ1 induced expression of the fibrotic markers collagen type III and α5β1 integrin, and it inhibited cellular migration due to the formation of focal adhesions with a low turnover rate. The novel in vitro HCF mechanical wound model can be used to understand the role played by GFs in human corneal repair. The model can also be used to test the effects of different treatments aimed at improving the healing process. Copyright


Investigative Ophthalmology & Visual Science | 2017

Corneal Wound Repair After Rose Bengal and Green Light Crosslinking: Clinical and Histologic Study

Patricia Gallego-Muñoz; Lucía Ibares-Frías; Elvira Lorenzo; Susana Marcos; Pablo Pérez-Merino; Nandor Bekesi; Irene E. Kochevar; M. Carmen Martínez-García

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Pablo Pérez-Merino

Spanish National Research Council

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Susana Marcos

Spanish National Research Council

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