Arresting proliferation improves the cell identity of corneal endothelial cells in the New Zealand rabbit.

Purpose: Corneal endothelium engineering aims to reduce the tissue shortage for corneal grafts. We investigated the impact of mitogenic and resting culture systems on the identity of corneal endothelial cells (CECs) for tissue engineering purposes. Methods: Rabbit CECs were cultured in growth factor-supplemented media (MitoM) until confluence. At the first passage, the CECs were divided into two populations: P1 remained cultured in MitoM, and P2 was cultured in a basal medium (RestM) for another passage. Morphologic changes in the CECs were analyzed, and RNA was isolated for transcriptome analysis. Quantitative PCR and immunocytochemistry validation of selected differentially expressed markers were performed. Results: The CECs in MitoM showed fibroblastic morphology, whereas the CECs in RestM exhibited polygonal morphology. Circularity analysis showed similar values in human (0.75±0.056), rabbit basal (before cultured; 0.77±0.063), and CECs in RestM (0.73±0.09), while MitoM showed lower circularities (0.41±0.19). Genes related to collagen type IV and the extracellular matrix, along with the adult CEC markers ATP1A1, ATP1B1, COL8A2, GPC4, and TJP1, were highly expressed in RestM. Conversely, the IL-6, F3, and ITGB3 genes and the non-adult CEC markers CD44, CNTN3, and CD166 were more expressed in MitoM. Overall, from the transcriptome, we identified 832 differentially expressed probes. A functional analysis of the 308 human annotated differentially expressed genes revealed around 13 functional clusters related to important biological terms, such as extracellular matrix, collagen type 4, immune responses, cell proliferation, and wound healing. Quantitative PCR and immunocytochemistry confirmed the overexpression of ATP1A1, TJP1, and GPC4 in CECs in RestM. Conclusions: The addition of a stabilization step during CEC culture improves the cells' morphology and molecular identity, which agrees with transcriptome data. This suggests that stabilization is useful for studying the plasticity of the corneal endothelium's morphology, and stabilization is proposed as a necessary step in corneal endothelium engineering.

HEMATOLOGIC AND BIOCHEMICAL PARAMETERS OF KELP GULLS (LARUS DOMINICANUS) CAPTURED IN THE CITY OF TALCAHUANO, CHILE.

In order to provide tools for future health-based monitoring programs, we developed reference intervals for hematology and plasma biochemistry and partitioned data for sex and season (winter and autumn). Ninety-one physically healthy kelp gulls (Larus dominicanus) were sampled in the city of Talcahuano, Chile, during winter (July-September) of 2007, autumn (April-June) and winter of 2008, and autumn of 2009. After blood sampling, the kelp gulls were euthanized by cervical dislocation. Packed cell volume (PCV), hemoglobin, total plasma protein (TPP), and complete blood count were performed. Alanine amino transferase, gamma-glutamyl transpeptidase, lactate dehydrogenase, alkaline phosphatase (ALP), creatinine, urea, calcium, phosphorus, and uric acid were analyzed. No significant differences were found between sexes (P>0.05). When a comparison was made between the two seasons, the values of PCV, TPP, white blood cell, heterophils, eosinophils, and monocytes were significantly higher in winter than in autumn (P<0.05) while and urea and uric acid concentrations were higher in autumn. Heterophils were the predominant circulating leukocyte for all birds. There was no significant difference in body condition between autumn and winter nor between sexes. Body condition showed a significant relationship with TPP and basophil concentration and ALP activity.

A comparative wordlist for investigating distant relations among languages in Lowland South America.

The history of the language families in Lowland South America remains an understudied area of historical linguistics. Panoan and Tacanan, two language families from this area, have frequently been proposed to descend from the same ancestor. Despite ample evidence in favor of this hypothesis, not all scholars accept it as proven beyond doubt. We compiled a new lexical questionnaire with 501 basic concepts to investigate the genetic relation between Panoan and Tacanan languages. The dataset includes data from twelve Panoan, five Tacanan, and four other languages which have previously been suggested to be related to Pano-Tacanan. Through the transparent annotation of grammatical morphemes and partial cognates, our dataset provides the basis for testing language relationships both qualitatively and quantitatively. The data is not only relevant for the investigation of the ancestry of Panoan and Tacanan languages. Reflecting the state of the art in computer-assisted approaches for historical language comparison, it can serve as a role model for linguistic studies in other areas of the world.

Public Awareness on Oral Cancer: A Population- Based Study in Asturias.

OBJECTIVE: Through awareness campaigns, we can change the patient's abilities to detect oral cancer at an early stage and their ability to seek help. To focus these campaigns, we need to know the level of knowledge of the population and its interest in learning about this disease. The aim of this study was to assess the level of oral cancer awareness in Asturias and the interest of the population in learning about this pathology. METHODS: A representative community-based survey was carried out online using Google Forms®. Responses were transferred to a Microsoft Excel and analysed using the R-program. The relationship between two qualitative variables was studied using Pearson's Chi-square test or Fisher's test. Univariate and multivariate logistic regression models were used to determine which factors are associated with knowledge of oral cancer. RESULT: We found that those having over 50 years and being health professionals are more likely to know about the existence of oral cancer. Almost 85.1% of participants mentioned tobacco as a risk factor, only 39.8% identified alcohol. The ulcer was the most frequently recognized alarm sign (70.6%). The primary care physician was chosen as the first option for consultation by the 56.5% of the sample. Only 12.4% of the participants reported knowing how to self-examine their mouth. The number of views of a video of how-to self-inspection oral cavity displayed at the end of the questionnaire increased in a 39.38% during our study period. CONCLUSION: This survey showed a worrying lack of awareness and knowledge about oral cancer among the population of Asturias, especially among those under 50 years old. The interest shown in increasing their knowledge, give us hope in the success of future awareness campaigns.

Grammars Across Time Analyzed (GATA): a dataset of 52 languages.

Grammars Across Time Analyzed (GATA) is a resource capturing two snapshots of the grammatical structure of a diverse range of languages separated in time, aimed at furthering research on historical linguistics, language evolution, and cultural change. GATA comprises grammatical information on 52 diverse languages across all continents, featuring morphological, syntactic, and phonological information based on published grammars of the same language at two different time points. Here we introduce the coding scheme and design features of GATA, and we describe some salient patterns related to language change and the coverage of grammatical descriptions over time.

Increased conformity offers diminishing returns for reducing total knee replacement wear.

Wear remains a significant problem limiting the lifespan of total knee replacements (TKRs). Though increased conformity between TKR components has the potential to decrease wear, the optimal amount and planes of conformity have not been investigated. Furthermore, differing conformities in the medial and lateral compartments may provide designers the opportunity to address both wear and kinematic design goals simultaneously. This study used a computational model of a Stanmore knee simulator machine and a previously validated wear model to investigate this issue for simulated gait. TKR geometries with different amounts and planes of conformity on the medial and lateral sides were created and tested in two phases. The first phase utilized a wide range of sagittal and coronal conformity combinations to blanket a physically realistic design space. The second phase performed a focused investigation of the conformity conditions from the first phase to which predicted wear volume was sensitive. For the first phase, sagittal but not coronal conformity was found to have a significant effect on predicted wear volume. For the second phase, increased sagittal conformity was found to decrease predicted wear volume in a nonlinear fashion, with reductions gradually diminishing as conformity increased. These results suggest that TKR geometric design efforts aimed at minimizing wear should focus on sagittal rather than coronal conformity and that at least moderate sagittal conformity is desirable in both compartments.

Parasites of the Neotropic cormorant Nannopterum (Phalacrocorax) brasilianus (Aves, Phalacrocoracidae) in Chile.

The Neotropic cormorant Nannopterum (Phalacrocorax) brasilianus (Suliformes: Phalacrocoracidae) is widely distributed in Central and South America. In Chile, information about parasites for this species is limited to helminths and nematodes, and little is known about other parasite groups. This study documents the parasitic fauna present in 80 Neotropic cormorants' carcasses collected from 2001 to 2008 in Antofagasta, Biobío, and Ñuble regions. Birds were externally inspected for ectoparasites and necropsies were performed to examine digestive and respiratory organs in search of endoparasites. Ectoparasites collected were cleared and mounted for identification under a microscope. Fecal samples were also evaluated to determine the presence of protozoan parasites employing a flotation technique. A total of 44 (42.5%) of birds were infested with at least one ectoparasite species, while 77 (96.25%) were carrying endoparasites. No protozoan forms were found after examination. Most prevalent endoparasite species found were Contracaecum rudolphii s. l. (72/80, 90%), followed by Pectinopygus gyroceras (33/80, 41.25%), and Profilicollis altmani (26/80, 32.5%). This is the first report of P. altmani, Baruscapillaria carbonis, Avioserpens sp., Cyathostoma (Cyathostoma) phenisci, and Eidmaniella pelucida in the Neotropic cormorant. These findings also expand the distributional range of Andracantha phalacrocoracis, Paradilepis caballeroi, Hysteromorpha triloba, and P. gyroceras to Chile.

External and gastrointestinal parasites of the Franklin's Gull, Leucophaeus pipixcan (Charadriiformes: Laridae), in Talcahuano, central Chile.

Parasitological studies of the Franklin's gull, Leucophaeus pipixcan, are scarce, and knowledge about its endoparasites is quite limited. In order to describe its parasitic community, a total of 60 Franklin's gulls were captured in the coastal area in central Chile, using modified Bal-chatri traps. Ectoparasites were collected from all 60 live individuals through inspection of their plumage, while 30 were examined for endoparasites by standard parasitological necropsy. The prevalence of ectoparasites was 78.3%, including the feather mite Zachvatkinia larica (43.3%) and four species of lice: Actornithophilus piceus lari (15.0%), Austromenopon transversum (6.7%), Quadraceps punctatus (10.0%) and Saemundssonia lari (46.7%). Some 25 of 30 (83.3%) of birds necropsied were parasitized with the following helminths: Aporchis sp. (6.7%), Tetrabothrius cylindraceus (56.7%), Cyclophyllidea gen. sp. (3.3%), Profilicollis altmani (56.7%), Eucoleus contortus (10.0%), Cosmocephalus obvelatus (13.3%), Paracuaria adunca (10.0%), Stegophorus sp. (3.3%) and Tetrameres skrjabini (3.3%). To our knowledge, with the exception of P. altmani, these helminths are reported for first time in the Franklin's gull.

Parasites of the Shiny Cowbird, Molothrus bonariensis, and the Austral Blackbird, Curaeus curaeus, (Passeriformes: Icteridae) in Chile.

Comparative studies of parasites in sympatric bird species have been generally scarce. Parasitic infection/transmission can be spread in a number of ways that suggests possible direct and indirect, horizontal transmission between avian hosts. In order to determine whether two sympatric icterids from Central and Southern Chile share their parasite fauna (ecto- and endoparasites), we examined parasites of 27 Shiny Cowbirds, Molothrus bonariensis, and 28 Austral Blackbirds, Curaeus curaeus, including individuals captured in the wild and carcasses. We found that Shiny Cowbirds were infected with the chewing lice Brueelia bonariensis, Philopterus sp. 1, the feather mites Amerodectes molothrus, Proctophyllodes spp. (species 1 and 2), and the helminths Mediorhynchus papillosus, Plagiorhynchus sp., Dispharynx nasuta and Tetrameres paucispina, while Austral Blackbirds had the chewing lice Myrsidea sp., Philopterus sp. 2, the feather mites Proctophyllodes sp. 3, Amerodectes sp., and three helminths: Anonchotaenia sp., Capillaria sp. and M. papillosus. The flea Dasypsyllus (Neornipsyllus) cteniopus was found only on the Austral Blackbird. The only parasite species shared by both icterids was the acanthocephalan M. papillosus, possibly due to their feeding on the same intermediate insect hosts. With the exception of B. bonariensis and Philopterus sp. 1 found on the Shiny Cowbird, all species reported in this study represent new parasite-host associations and new records of parasite diversity in Chile.

Reprimo as a potential biomarker for early detection in gastric cancer.

PURPOSE: Gastric cancer is a curable disease if diagnosed at early stage. However, most cases are diagnosed at advanced stage because of the lack of screening programs. Therefore, the identification of plasma biomarkers for early detection is necessary. EXPERIMENTAL DESIGN: To search for these biomarkers, we evaluated the DNA methylation patterns of 24 genes by Methylation-specific PCR in primary tissues from 32 retrospectively collected gastric cancer cases (testing group). Correlation between methylation and gene expression was evaluated in the MKN-45 cell line after treatment with 5-aza-2'-deoxycytidine. The most frequently hypermethylated genes were next evaluated in primary tissues and plasma samples from 43 prospectively collected gastric cancer cases as well as plasma samples from 31 asymptomatic age- and gender-matched controls (validation group). RESULTS: In the testing group, 11 genes were hypermethylated in at least 50% of cases (APC, SHP1, E-cadherin, ER, Reprimo, SEMA3B, 3OST2, p14, p15, DAPK, and p16). Eight genes (BRCA1, p73, RARbeta, hMLH1, RIZI, RUNX3, MGMT, and TIMP3) were statistically associated with a particular variant of gastric cancer, the signet-ring cell type (P = 0.03). Seven genes (APC, SHP1, E-cadherin, ER, Reprimo, SEMA3B, and 3OST2) were next evaluated in the validation group. We confirm the high frequency of methylation in primary tumors for all seven genes. However, only APC and Reprimo were frequently methylated in pair plasma samples. In asymptomatic controls, only Reprimo was infrequently methylated in comparison with plasma from gastric cancer cases (P < 0.001). CONCLUSION: Our results identified specific methylation profile associated to signet-ring cell-type histology and aberrant hypermethylation of Reprimo as a potential biomarker for early detection of gastric cancer.

Gastrointestinal and external parasites of the Variable hawk Geranoaetus polyosoma (Accipitriformes: Accipitridae) in Chile.

Information about parasites associated with diurnal raptors from Chile is scarce. Between 2006 and 2017, a total of 15 specimens of the Variable hawk, Geranoaetus polyosoma (Quoy & Gaimard, 1824) were collected, 14 of them from different localities in the Biobío region and one specimen from the Valparaíso region. An external examination of the plumage was made to collect ectoparasites, and necropsies were performed, focusing primarily on the gastrointestinal tract. Chewing lice (Phthiraptera) were found on five (33.3%) of the birds corresponding to three species: 97 specimens of Degeeriella fulva (Giebel, 1874), six specimens of Colpocephalum turbinatum Denny, 1842 and nine belonging to an unidentified species of the genus Craspedorrhynchus Kéler, 1938. Endoparasites found in three (20%) of the birds included round worms (Nematoda) of the genus Procyrnea Chabaud, 1958, and spiny-headed worms (Acanthocephala) of the genus Centrorhynchus Lühe, 1911. The species Colpocephalum turbinatum and the genera: Craspedorrhynchus sp., Procyrnea sp. and Centrorhynchus sp. are new records for the Variable hawk.

External and intestinal parasites of the Austral thrush Turdus falcklandii (Aves, Turdidae) in central Chile.

A total of thirty Austral thrushes Turdus falcklandii Quoy & Gaimard, 1824 (Turdidae) carcasses were brought to the Departamento de Ciencia Animal, Facultad de Ciencias Veterinarias, Universidad de Concepción, to be examined for ecto- and endoparasites. Ectoparasites were found on 20% (6/30) of the thrushes and belonged to species Brueelia magellanica Cichino, 1986 (Phthiraptera), Menacanthus eurysternus Burmeister, 1838 (Phthiraptera) and Tyrannidectes falcklandicus Mironov & González-Acuña, 2011 (Acari). Endoparasites were isolated from 26.6% (8/30) of the birds and identified as Lueheia inscripta Westrumb, 1821 (Acanthocephala), Plagiorhynchus cylindraceus Goeze, 1782 (Acanthocephala), Wardium sp. sensu Mayhew, 1925 (Cestoda), Dilepis undula (Cestoda) Schrank, 1788, and Zonorchis sp. (sensu Travassos, 1944) (Trematoda). To our knowledge, all endoparasites collected in this study are new records in T. falcklandii and expand their distributional range to Chile.

Gastrointestinal and ectoparasites of plumbeous rail, Pardirallus sanguinolentus (Aves: Rallidae) in Central Chile.

With the aim to identify the parasite fauna of plumbeous rail, Pardirallus sanguinolentus (Aves: Rallidae) in Chile, 26 carcasses were parasitologically necropsied. The present study revealed the presence of 14 species of parasites (inverse Simpson index = 4.64; evenness = 0.332), including ectoparasites: feather mites: Analloptes megnini , Grallobia sp., Grallolichus sp., Megniniella sp., and Metanalges sp.; the feather lice Pseudomenopon meinertzhageni, Rallicola andinus, and Fulicoffula sp.; and six species of gastrointestinal helminths: Heterakis psophiae, Porrocaecum ardeae, Tetrameres sp., Capillaria sp., Diorchis sp., and Plagiorhynchus sp. The relatively high parasite richness that was found could be attributed to the highly favorable conditions of wetlands for parasite development. All parasites found, except feather lice, are new records for plumbeous rail. A checklist of parasites for plumbeous rail is presented.

Gastrointestinal and external parasitism in the Magellanic Horned Owl Bubo magellanicus (Strigiformes: Strigidae) in Chile.

To describe the parasitic community of the Magellanic Horned Owl, Bubo magellanicus (Aves, Strigiformes), 19 carcasses from central Chile were analyzed. Ectoparasites were collected through plumage inspection, while endoparasites were collected through traditional techniques of parasitological necropsy. Sixteen owls were infected with at least one species of ectoparasite (84.21%) or endoparasite (31.58%). Eleven of 19 birds (57.89%) harbored feather mites of the three species Pandalura cirrata (42.11%), Glaucalges attenuatus (47.37%), and Kramerella sp. (10.53%), whereas 16 individuals (84.21%) harbored the chewing louse Strigiphilus chilensis. Only six birds (31.58%) were infected with helminths; the nematodes Capillaria tenuissima (26.32%) and Dispharynx nasuta (5.26%); the acanthocephalan Centrorhynchus spinosus (5.26%); and the trematode Neodiplostomum sp. (5.26%). Apart from S . chilensis, all parasites comprised new records for B . magellanicus.

Primary explant culture and collagen I substrate enhances corneal endothelial cell morphology.

OBJECTIVES: Corneal endothelial cell (CEC) isolation and harvest aim to produce engineered grafts to solve donor corneal tissue shortage. To yield high amounts of CEC maintaining morphological and molecular characteristics, several isolation and culture conditions are reported. Here, we combined direct explant culture, with three different coating conditions and a two-step media approach to compare confluence efficiency, morphology, and specific molecular markers expression. DATA DESCRIPTION: Confluence was reached after 2 weeks in the three coating conditions (Matrigel, collagen I, and in uncoated plates) using a two-step approach (proliferative medium without pituitary extract, followed by stabilizer basal medium). Na/K-ATPase and GPC4 markers were detected by immunocytochemistry while GPC4, CD200, and TJP1 by RT-PCR in the three CEC coating culture conditions. CEC in proliferative medium showed spindle morphology in the three conditions. Polygonal morphology was seen in CEC cultures using basal medium under uncoated and collagen I coated plates. CEC cultured in Matrigel-coated plates remained with spindle morphology in basal medium.

External and gastrointestinal parasites of the rufous-collared sparrow Zonotrichia capensis (Passeriformes, Emberizidae) in Chile.

A total of 277 rufous-collared sparrows, Zonotrichia capensis Müller, 1776 (Emberizidae), were examined for external parasites. The birds were captured using mist nets in seven locations in northern and central Chile. Additionally, seven carcasses from central Chile (the Biobío region) were necropsied to evaluate the presence of endoparasite infection. Ectoparasites were found on 35.8% (99/277) of the examined birds and they were represented by the following arthropods: feather mites Amerodectes zonotrichiae Mironov and González-Acuña, 2014 (Analgoidea: Proctophyllodidae), Proctophyllodes polyxenus Atyeo and Braasch, 1966 (Analgoidea: Proctophyllodidae), and Trouessartia capensis Berla, 1959 (Analgoidea: Trouessartiidae); a louse Philopterus sp. (Phthiraptera: Ischnocera); and ticks Amblyomma tigrinum Koch, 1844 (Acari: Ixodidae) and Ixodes auritulus Neumann, 1904 (Acari: Ixodidae). Two of the seven necropsied carcasses were infected with the acanthocephalan Mediorhynchus papillosus Van Cleave, 1916 (Gigantorhynchida: Gigantorhynchidae). To our knowledge, this study reports P. polyxenus, Philopterus sp., A. tigrinum, and M. papillosus for the first time for Z. capensis and expands the distributional range for T. capensis to Chile.

Parasites of the Southern silvery grebe Podiceps occipitalis (Aves, Podicipedidae) in Chile.

A total of 97 southern silvery grebes (Podiceps occipitalis), which died as the result of an oil spill on the coast of central Chile, were examined for ecto- and endoparasites. Two lice species including Aquanirmus rollandii (Philopteridae) and Pseudomenopon dolium (Menoponidae) were found from 6.2% (6/97) of birds. In 91.7% (89/97) of cases, grebes were infected with some kind of helminths. Three species of gastrointestinal helminths were detected: Eucoleus contortus (Nematoda), Profilicollis bullocki (Acanthocephala), and Confluaria sp. (Cestoda). In addition, Pelecitus fulicaeatrae (Nematoda) was removed from the tibiotarsal-tarsometatarsal articulation in 13.4% (13/97) of the specimens examined. To our knowledge, these are the first records of A. rollandii, E. contortus, and Confluaria sp. as parasites of P. occipitalis. In addition, these findings expand the distributional range of A. rollandii, E. contortus, P. fulicaeatrae, and Confluaria sp. to Chile.

Silk Fibroin Films for Corneal Endothelial Regeneration: Transplant in a Rabbit Descemet Membrane Endothelial Keratoplasty.

Purpose: Develop a silk fibroin (SF)-based artificial endothelial graft for its use in a rabbit Descemet membrane endothelial keratoplasty (DMEK). Methods: Human and rabbit artificial corneal endothelial grafts were developed through the culture of human and rabbit corneal endothelial cells (CECs) on SF films. Rabbit artificial SF endothelial grafts were transplanted in a DMEK surgery into a rabbit in vivo model. Results: SF artificial endothelial grafts showed the characteristic endothelial markers: zonula occludens (ZO-1) and Na+/K+ ATPase. In a rabbit model of DMEK surgery, SF artificial endothelial graft restored the corneal transparency and thickness at 6 week of follow-up. Anterior segment optical coherence tomography revealed the SF graft as a fully integrated component in the corneal tissue, displaying a similar corneal thickness and endothelial cell count when compared with its healthy contralateral cornea. Histologic analysis showed that the SF artificial endothelial graft was attached and integrated on the surface of the corneal stroma without a significant inflammatory reaction, and rabbit CECs consisted in a monolayer that showed their characteristic markers ZO-1 and Na+/K+ ATPase, suggesting proper intercellular junctions and cellular pump function. Conclusions: We have developed SF films with biological properties that supported the growth of rabbit and human CECs, which showed normal morphology and characteristic markers; and with mechanical properties that allowed its use in a DMEK surgery, proving its in vivo functionality in a rabbit model of endothelial dysfunction.

Ectoparasites of the black-chinned siskin Spinus barbatus (Passeriformes: Fringillidae) in Chile.

Despite being a bird with a broad and extensive distribution in Chile, the black-chinned siskin, Spinus barbatus Molina, 1782 is not well studied in relation to its parasites. This paper aims to describe the ectoparasite fauna of S. barbatus in central and southern Chile. A total of 125 individuals caught with mist nets were examined alive; a total of 22 parasites were found dead and were exposed to parasit autopsy. The extracted parasites were preserved in 70% alcohol for subsequent mounting and identification. Ectoparasites were found in 56 black-chinned siskins (38%); 48 of them (33%) had 870 mites - 680 feather mites (Astigmata: Analgoidea) were identified as Proctophyllodes spini, 167 as Knemidokoptes jamaicensis, 19 as Strelkoviacarus critesi, and one as Analges passerinus. Moreover, three mites were chiggers belonging to the tribe Schoengastiini (Prostigmata: Trombiculidae). In 21 birds (14%), 54 lice were found, 21 of which were identified as Philopterus roehreri, 18 as Myrsidea serini, and 15 as Ricinus carolynae. Endoparasites were not found in the necropsied individuals. All of the parasites that were found represent new records for Chile, and they also serve as new records of host-parasite associations for S. barbatus.

Human Bone Derived Collagen for the Development of an Artificial Corneal Endothelial Graft. In Vivo Results in a Rabbit Model.

Corneal keratoplasty (penetrating or lamellar) using cadaveric human tissue, is nowadays the main treatment for corneal endotelial dysfunctions. However, there is a worldwide shortage of donor corneas available for transplantation and about 53% of the world's population have no access to corneal transplantation. Generating a complete cornea by tissue engineering is still a tough goal, but an endothelial lamellar graft might be an easier task. In this study, we developed a tissue engineered corneal endothelium by culturing human corneal endothelial cells on a human purified type I collagen membrane. Human corneal endothelial cells were cultured from corneal rims after corneal penetrating keratoplasty and type I collagen was isolated from remnant cancellous bone chips. Isolated type I collagen was analyzed by western blot, liquid chromatography -mass spectrometry and quantified using the exponentially modified protein abundance index. Later on, collagen solution was casted at room temperature obtaining an optically transparent and mechanically manageable membrane that supports the growth of human and rabbit corneal endothelial cells which expressed characteristic markers of corneal endothelium: zonula ocluddens-1 and Na+/K+ ATPase. To evaluate the therapeutic efficiency of our artificial endothelial grafts, human purified type I collagen membranes cultured with rabbit corneal endothelial cells were transplanted in New Zealand white rabbits that were kept under a minimal immunosuppression regimen. Transplanted corneas maintained transparency for as long as 6 weeks without obvious edema or immune rejection and maintaining the same endothelial markers that in a healthy cornea. In conclusion, it is possible to develop an artificial human corneal endothelial graft using remnant tissues that are not employed in transplant procedures. This artificial endothelial graft can restore the integrality of corneal endothelium in an experimental model of endothelial dysfunction. This strategy could supply extra endothelial tissue and compensate the deficit of cadaveric grafts for corneal endothelial transplantation.