[Show abstract][Hide abstract] ABSTRACT: Objectives:
Present study examines the relationship between the estimated risk of developing type 2 diabetes (T2D) and health-related quality of life (HRQoL). We quantify the association between Finnish Diabetes Risk Score (FINDRISC) and HRQoL, and examine the potential use of FINDRISC as tool to evaluate HRQoL indirectly.
We conducted a cross-sectional study comprising 707 Finnish people without a diagnosis of T2D between the ages of 51 and 75 years. The risk of developing T2D was assessed using the validated and widely used FINDRISC (range 0-26 points), and quality of life was measured using two preference-based HRQoL instruments (15D and SF-6D) and one health profile instrument (SF-36). Effects of the individual FINDRISC items and demographic and clinical characteristics, such as co-morbidities, on HRQoL were studied using multivariable Tobit regression models.
Low HRQoL was significantly and directly associated with the estimated risk of developing T2D. An approximate 4-5 point change in FINDRISC score was observed to be associated with clinically noticeable changes in the preference-based instrument HRQoL index scores. The association between HRQoL and the risk of developing T2D was also observed for most dimensions of HRQoL in all applied HRQoL instruments. Overall, old age, lack of physical activity, obesity, and history of high blood glucose were the FINDRISC factors most prominently associated with lower HRQoL.
The findings may help the health care professionals to substantiate the possible improvement in glucose metabolism and HRQoL potentially achieved by lifestyle changes, and better convince people at high risk of T2D to take action towards healthier lifestyle habits. FINDRISC may also provide an accurate proxy for HRQoL, and thus by estimating the risk of T2D with the FINDRISC, information about patients' HRQoL may also be obtained indirectly, when it is not feasible to use HRQoL instruments.
[Show abstract][Hide abstract] ABSTRACT: In this study, we investigated the suitability of ultrathin and porous polyimide (PI) membrane as a carrier for subretinal transplantation of human embryonic stem cell (hESC) -derived retinal pigment epithelial (RPE) cells in rabbits. The in vivo effects of hESC-RPE cells were analyzed by subretinal suspension injection into Royal College of Surgeons (RCS) rats. Rat eyes were analyzed with electroretinography (ERG) and histology. After analyzing the surface and permeability properties of PI, subretinal PI membrane transplantations with and without hESC-RPE were performed in rabbits. The rabbits were followed for three months and eyes analyzed with fundus photography, ERG, optical coherence tomography (OCT), and histology. Animals were immunosuppressed with cyclosporine the entire follow-up time. In dystrophic RCS rats, ERG and outer nuclear layer (ONL) thickness showed some rescue after hESC-RPE injection. Cells positive for human antigen were found in clusters under the retina 41 days post-injection but not anymore after 105 days. In rabbits, OCT showed good placement of the PI. However, there was loss of pigmentation on the hESC-RPE-PI over time. In the eyes with PI alone, no obvious signs of inflammation or retinal atrophy were observed. In the presence of hESC-RPE, mononuclear cell infiltration and retinal atrophy were observed around the membranes. The porous ultrathin PI membrane was well-tolerated in the subretinal space and is a promising scaffold for RPE transplantation. However, the rejection of the transplanted cells seems to be a major problem and the given immunosuppression was insufficient for reduction of xenograft induced inflammation.
[Show abstract][Hide abstract] ABSTRACT: Corneal epithelium is renewed by limbal epithelial stem cells (LESCs), a type of tissue-specific stem cells located in the limbal palisades of Vogt at the corneo-scleral junction. Acute trauma or inflammatory disorders of the ocular surface can destroy these stem cells, leading to limbal stem cell deficiency (LSCD) - a painful and vision-threatening condition. Treating these disorders is often challenging and complex, especially in bilateral cases with extensive damage. Human pluripotent stem cells (hPSCs) provide new opportunities for corneal reconstruction using cell-based therapy. Here, we investigated the use of hPSC-derived LESC-like cells on bioengineered collagen matrices in serum-free conditions, aiming for clinical applications to reconstruct the corneal epithelium and partially replace the damaged stroma. Differentiation of hPSCs towards LESC-like cells was directed using small-molecule induction followed by maturation in corneal epithelium culture medium. After four to five weeks of culture, differentiated cells were seeded onto bioengineered matrices fabricated as transparent membranes of uniform thickness, using medical-grade porcine collagen type I and a hybrid cross-linking technology. The bioengineered matrices were fully transparent, with high water content and swelling capacity, and parallel lamellar microstructure. Cell proliferation of hPSC-LESCs was significantly higher on bioengineered matrices than on collagen-coated control wells after two weeks of culture, and LESC markers p63 and cytokeratin 15, along with proliferation marker Ki67 were expressed even after 30 days in culture. Overall, hPSC-LESCs retained their capacity to self-renew and proliferate, but were also able to terminally differentiate upon stimulation, as suggested by protein expression of cytokeratins 3 and 12. We propose the use of bioengineered collagen matrices as carriers for the clinically-relevant hPSC-derived LESC-like cells, as a novel tissue engineering approach for corneal reconstruction.
No preview · Article · Nov 2015 · Experimental Eye Research
[Show abstract][Hide abstract] ABSTRACT: Focusing on seasonality, gender, age, and suicide methods a Finnish nation-wide cohort-based study was carried out to compare suicide data between sighted, visually-impaired (WHO impairment level I-II, i.e., visual acuity >0.05, but <0.3) and blind (WHO impairment level III-V, i.e., visual acuity <0.05) victims. Standardized mortality ratios (SMR) of age- and gender-matched populations from official 1982-2011 national registers were used. Group differences in categorical variables were assessed with Pearson's Chi-square or Fisher's Exact test and in continuous variables with Mann-Whitney U-test. Seasonality was assessed by Chi-square for multinomials; ratio of observed to expected number of suicides was calculated with 95% confidence level. Hanging, poisoning, drowning, but rarely shooting or jumping from high places, were preferred suicide methods of the blind. Mortality was significantly increased in the visually impaired (SMR = 1.3; 95% CI 1.07-1.61), but in gender-stratified analyses the increase only affected males (1.34; 95% CI = 1.06-1.70) and not females (1.24; 95% CI 0.82-1.88). Age-stratified analyses identified blind males of working age rather than older men (as in the general population) as a high risk group that requires particular attention. The statistically significant spring suicide peak in blind subjects mirrors that of sighted victims and its possible cause in the blind is discussed.
[Show abstract][Hide abstract] ABSTRACT: Limbal epithelial stem cells (LESCs) are tissue-specific stem cells responsible for renewing the corneal epithelium. Acute trauma or chronic disease affecting LESCs may disrupt corneal epithelial renewal, causing vision threatening and painful ocular surface disorders, collectively referred to as LESC deficiency (LESCD). These disorders cannot be treated with traditional corneal transplantation and therefore alternative cell sources for successful cell-based therapy are needed. LESCs derived from human pluripotent stem cells (hPSCs) are a prospective source for ocular surface reconstruction, yet critical evaluation of these cells is crucial before considering clinical applications. In order to quantitatively evaluate hPSC-derived LESCs, we compared protein expression in native human corneal cells to that in hPSC-derived LESCs using isobaric tag for relative and absolute quantitation (iTRAQ) technology. We identified 860 unique proteins present in all samples, including proteins involved in cell cycling, proliferation, differentiation and apoptosis, various LESC niche components, and limbal and corneal epithelial markers. Protein expression profiles were nearly identical in LESCs derived from two different hPSC lines, indicating that the differentiation protocol is reproducible, yielding homogeneous cell populations. Their protein expression profile suggests that hPSC-derived LESCs are similar to the human ocular surface epithelial cells, and possess LESC-like characteristics.
Full-text · Article · Oct 2015 · Scientific Reports
[Show abstract][Hide abstract] ABSTRACT: Introduction
The efficacy, safety and tolerability of the preservative-free (PF) fixed combination (FC) of tafluprost 0.0015% and timolol 0.5% (once daily) were compared to those of the individual components (PF tafluprost 0.0015% once daily and PF timolol 0.5% twice daily) in patients with open-angle glaucoma or ocular hypertension inadequately controlled on prior timolol or prostaglandin monotherapy for 6 months.
A stratified, double-masked, randomized, multicenter phase III study was conducted. A total of 189 prior timolol users were randomized within the timolol stratum (TS) to receive either FC (n = 95) or timolol 0.5% (TIM; n = 94). Furthermore, a total of 375 prior prostaglandin analog (PGA) users were randomized within the prostaglandin stratum (PS) to receive either FC (n = 188) or tafluprost 0.0015% (TAF; n = 187). To be eligible for participation in the study, the patients were required to have an intraocular pressure (IOP) of ≥22 mmHg when on timolol (TIM) or of ≥20 mmHg when on PGA in either treated eye at the screening and end-of-run-in visits. In addition to these, the study included visits at baseline, 2 and 6 weeks, 3 and 6 months and at a post-study visit. IOP was measured at 8 a.m., 10 a.m., 4 p.m., and 8 p.m.
In the TS, a significant reduction from baseline IOP was seen with FC and TIM throughout the study. Average diurnal IOP change from baseline at month 3 was −8.55 mmHg (32%) for FC and −7.35 mmHg (28%) for TIM. The model-based treatment difference (FC–TIM) was −0.885 mmHg [95% confidence interval (CI) −1.745 to −0.024; p = 0.044] demonstrating the superiority of FC over TIM. In the PS, a significant reduction in IOP was seen with both FC and TAF throughout the study. The average diurnal IOP change from baseline at month 3 was −8.61 mmHg (33%) for FC and −7.23 mmHg (28%) for TAF. The model-based treatment difference (FC–TAF) was −1.516 mmHg (95% CI −2.044 to −0.988; p
Full-text · Article · Dec 2014 · Advances in Therapy
[Show abstract][Hide abstract] ABSTRACT: PurposeChronic conjunctival inflammation, caused by various reasons, for example long-term use of topical drugs and/or their preservatives, affects the outcome of glaucoma surgery by interfering with wound healing. Matrix metalloproteinases (MMPs) remodel extracellular matrix (ECM) and are involved in the wound healing process. This study was designed to evaluate the conjunctival expression of MMPs and their tissue inhibitors (TIMPs) in the normal eye, primary open-angle glaucoma (POAG) and exfoliation glaucoma (ExG) and whether there is an association between staining intensities and deep sclerectomy outcome.Methods
Immunohistochemical procedures were performed on conjunctival samples which were obtained from POAG (n = 11) and ExG (n = 14) patients as well as normal (n = 7) subjects. Antibodies against MMPs (MMP-1, -2, -3 and -9) and TIMPs (TIMP-1, -2 and -3) were used.ResultsIn conjunctival stroma, expression levels of MMP-2 (p = 0.047), MMP-3 (p = 0.009), MMP-9 (p < 0.001), TIMP-1 (p = 0.003), TIMP-2 (p < 0.001) and TIMP-3 (p < 0.001) in ExG and MMP-9 (p = 0.008), TIMP-2 (p = 0.02) and TIMP-3 (p = 0.002) in POAG were significantly increased compared to control. We further found correlations between expression of MMP-1 and MMP-3 and the length of pilocarpine treatment.Conclusion
The expression of MMPs and TIMPs is increased in the conjunctiva of POAG and ExG patients having a long history of topical antiglaucoma drops. Antiglaucoma agents and/or their preservatives alter the remodelling balance of ECM in conjunctiva of POAG and ExG eyes. The balance between MMPs and TIMPs may play a crucial role in the conjunctival wound healing process and the outcome of glaucoma surgery.
No preview · Article · Oct 2014 · Acta ophthalmologica
[Show abstract][Hide abstract] ABSTRACT: Human induced pluripotent stem cells (hiPSCs) offer unique opportunities for developing novel cell-based therapies and disease modeling. In this study, we developed a directed differentiation method for hiPSCs toward corneal epithelial progenitor cells capable of terminal differentiation toward mature corneal epithelial-like cells. In order to improve the efficiency and reproducibility of our method, we replicated signaling cues active during ocular surface ectoderm development with the help of two small-molecule inhibitors in combination with basic fibroblast growth factor (bFGF) in serum-free and feeder-free conditions. First, small-molecule induction downregulated the expression of pluripotency markers while upregulating several transcription factors essential for normal eye development. Second, protein expression of the corneal epithelial progenitor marker p63 was greatly enhanced, with up to 95% of cells being p63 positive after 5 weeks of differentiation. Third, corneal epithelial-like cells were obtained upon further maturation.
Full-text · Article · Feb 2014 · Stem Cell Reports
[Show abstract][Hide abstract] ABSTRACT: To examine the general health status and health-related quality of life (HRQOL) along the diabetes continuum of middle-aged and older Finns, and to determine the glucose metabolism stage by which the HRQOL has decreased noticeably.
The cross-sectional sample consisted of 920 persons aged 51-75 from the general population in a single municipality in a rural area of Eastern Finland. Data were adjusted for age, sex, smoking history, current alcohol consumption, employment and marital status. The HRQOL and health status were evaluated using two preference-based HRQOL instruments, 15D and SF-6D, and one health profile instrument, 36-Item Short Form Health Survey (SF-36).
Individuals with impaired glucose tolerance (IGT) and type 2 diabetes had noticeably low mean SF-6D, 15D and general health status. The decrease in overall HRQOL was mainly due to a decline in the physical dimensions of HRQOL. The adjusted odds ratios (95 % CI) for having noticeably low HRQOL on SF-6D, 15D and general health dimension of SF-36 associated with IGT were 1.95 (1.18-3.25), 1.35 (0.84-2.18) and 2.00 (1.21-3.29), respectively.
The progression along the diabetes continuum was significantly associated with a decrease in HRQOL and health status. Furthermore, the data indicate that when a person is detected to have IGT, the HRQOL and general health status have already diminished noticeably. The prevailing evidence suggests that detection and intervention before a patient develops IGT is essential in order to minimize the loss of quality of life and quality-adjusted life years.
No preview · Article · Feb 2014 · Quality of Life Research
[Show abstract][Hide abstract] ABSTRACT: To present the outcomes of laser-assisted in situ keratomileusis (LASIK) operations performed with the new three-dimensional, transportable FEMTO LDV Z6 I femtosecond laser (Ziemer Ophthalmic Systems, Port, Switzerland) and the Allegretto Wave Concerto 500 Hz excimer laser (Wavelight AG, Erlangen, Germany) in terms of accuracy, reproducibility and safety of flap creation.
This is a retrospective study of 309 consecutive eyes of 160 patients treated with the FEMTO LDV Z6 I for corneal flap creation. The target flap thickness was 90 μm. The size of the suction ring varied from 9.5 to 10.0 mm and target flap diameter from 9.3 to 9.6 mm, respectively. The target hinge length was 4.0 mm.
The FEMTO LDV Z6 I produced the 90-μm targeted flaps very consistently (mean 90.1 ± 2.7 μm, range 78-100). Mean flap diameter with the 9.3-mm target flap diameter was 9.3 ± 0.1 mm (range 9.0-9.6) and with the 9.6-mm target flap diameter 9.6 ± 0.1 mm (range 9.0-9.8). Mean hinge length was 3.9 ± 0.1 mm (range 3.3-4.2). Minor complications were reported in 15 (5%) eyes, but none of them prevented refractive laser treatment. The most common complications were bubbles in the conjunctiva (n = 7, 2%) and an opaque bubble layer inside the flap margin (n = 6, 2%). None of the eyes lost two Snellen lines of corrected distance visual acuity during 1-month follow-up.
In the hands of an experienced surgeon, the transfer from the Classic FEMTO LDV to Z6 I was a safe and straight forward process yielding accurate and reproducible flaps.
No preview · Article · Dec 2013 · Acta ophthalmologica
[Show abstract][Hide abstract] ABSTRACT: Extracellular matrix (ECM) interactions play a vital role in cell morphology, migration, proliferation and differentiation of cells. We investigated the role of ECM proteins to the structure and function of human embryonic stem cell derived retinal pigment epithelial (hESC-RPE) cells during their differentiation and maturation from hESCs into RPE cells in adherent differentiation cultures on several human ECM proteins found in native human Bruch's membrane namely collagen I, collagen IV, laminin, fibronectin, vitronectin, as well as on commercial substrates of xeno-free CELLstartTM and MatrigelTM. Cell pigmentation, expression of RPE specific proteins, fine structure as well as the production of basal lamina by hESC-RPE on different protein coatings were evaluated after 140 days of differentiation. The integrity of hESC-RPE epithelium and barrier properties on different coatings were investigated by measuring transepithelial resistance. All coatings supported the differentiation of hESC-RPE cells as demonstrated by early onset of cell pigmentation and further maturation to RPE monolayers after enrichment. Mature RPE phenotype was verified by RPE specific gene and protein expression, correct epithelial polarization and phagocytic activity. Significant differences were found in the degree of RPE cell pigmentation and tightness of epithelial barrier between different coatings. Furthermore, the thickness of self-assembled basal lamina and secretion of the key ECM proteins found in the basement membrane of the native RPE varied between hESC-RPE cultured on compared protein coatings. In conclusion, this study shows that the cell culture substrate has a major effect on the structure and basal lamina production during the differentiation and maturation of hESC-RPE potentially influencing the success of cell integrations and survival after cell transplantation.
No preview · Article · Sep 2013 · Tissue Engineering Part A
[Show abstract][Hide abstract] ABSTRACT: To detect by immunohistochemical means the changes in bleb capsules at the cellular level between functioning and non-functioning glaucoma drainage implants (GDIs).
Three samples each of functioning (1 Baerveldt and 2 Molteno implants) and non-functioning filtration blebs (1 Molteno and 2 Ahmed implants) were studied. A non-functioning bleb was defined as an intra-ocular pressure (IOP) >21 mmHg or a <20% reduction in IOP from baseline on three consecutive follow-up visits with maximal tolerated medication. The capsules were obtained between 6 and 108 months after GDI insertion for medical reasons only. Primary antibodies were used to stain fibronectin, tenascin, laminin, collagen IV and smooth muscle actin (SMA). The samples were graded on the basis of the intensity and quantity of immunohistochemical staining into four categories as follows: no staining or a mild, moderate or marked staining.
The non-functioning blebs expressed more fibronectin, tenascin and SMA through the whole capsule wall than the functioning blebs. In the functioning blebs, tenascin was found mainly in the inner layer of the capsule. More type IV collagen and laminin were also found in the non-functioning bleb capsules than in the functioning blebs. No difference was found between the bleb capsules irrespective of whether they had been perfused with aqueous humour immediately after surgery (Ahmed) or after a delay (Molteno, Baerveldt).
Accumulation of extracellular matrix components and activated fibroblasts in the bleb capsules of non-functioning GDI indicates the presence of an active wound healing process, suggesting a possible reduction in filtration through the bleb wall.
No preview · Article · Sep 2013 · Acta ophthalmologica
[Show abstract][Hide abstract] ABSTRACT: The aim of this study was to characterize the ocular morphology of low-density lipoprotein receptor-deficient apolipoprotein B-100-only mice, where overexpression of insulin-like growth factor II (IGF-II) has been shown to induce glucose intolerance and increase atherosclerotic lesion progression and calcification.
Fifteen-month-old mice were examined on a normal chow diet and after 3 months of a high-fat Western diet. IGF-II-negative LDLR(-/-)ApoB(100/100) littermates and C57Bl/6J mice served as controls. In vivo color images of the fundi were obtained, and eyes were processed either for retinal flat mounts for assessment of neovascularization or for paraffin-embedded samples for immunohistochemical analyses.
IGF-II overexpression and the resulting prediabetic phenotype did not induce microvascular damage when assessed in fundus photographs and retinal whole mounts, and the number of capillaries in IGF-II/LDLR(-/-)ApoB(100/100) mice was not significantly different from LDLR(-/-)ApoB(100/100) mice. However, morphology of the inner nuclear, outer plexiform, and outer nuclear layers was altered in the IGF-II/LDLR(-/-)ApoB(100/100) mice. Moreover, photoreceptor atrophy and thinning of the outer nuclear layer were present. Caspase-3 staining was positive in the photoreceptor inner segment. In addition, retinas of the IGF-II/LDLR(-/-)ApoB(100/100) mice displayed reduced rhodopsin positivity, consistent with the decreased number of photoreceptor cells.
This study reports a novel form of retinopathy with photoreceptor atrophy and abundant changes in retinal morphology in a mouse model of prediabetes and atherosclerosis.
[Show abstract][Hide abstract] ABSTRACT: We address the performance evaluation practices for developing medical image analysis methods, in particular, how to establish and share databases of medical images with verified ground truth and solid evaluation protocols. Such databases support the development of better algorithms, execution of profound method comparisons, and, consequently, technology transfer from research laboratories to clinical practice. For this purpose, we propose a framework consisting of reusable methods and tools for the laborious task of constructing a benchmark database. We provide a software tool for medical image annotation helping to collect class label, spatial span, and expert's confidence on lesions and a method to appropriately combine the manual segmentations from multiple experts. The tool and all necessary functionality for method evaluation are provided as public software packages. As a case study, we utilized the framework and tools to establish the DiaRetDB1 V2.1 database for benchmarking diabetic retinopathy detection algorithms. The database contains a set of retinal images, ground truth based on information from multiple experts, and a baseline algorithm for the detection of retinopathy lesions.
Preview · Article · Jun 2013 · Computational and Mathematical Methods in Medicine
[Show abstract][Hide abstract] ABSTRACT: Purpose:
Aquaporins (AQPs), a family of transmembrane water channel proteins, are essential for allowing passive water transport through retinal pigmented epithelial (RPE) cells. Even though human native RPE cells and immortalized human RPEs have been shown to express AQPs, the expression of AQPs during the differentiation in stem cell-derived RPE remains to be elucidated.
In human embryonic (hESCs) and induced pluripotent stem cells (hiPSCs)-derived RPE cells, the expression of several AQPs was determined by quantitative real-time PCR and the localization of AQP1 was assessed with confocal microscopy. The functionality of AQP water channels was determined by cell volume assay in hESC-derived RPE cells.
AQP1, AQP3, AQP4, AQP5, AQP6, AQP7, AQP10, AQP11, and AQP12 were expressed in hESC- and hiPSC-derived RPE cells. Furthermore, the expression of AQP1 and AQP11 genes were significantly upregulated during the maturation of both hESC and iPSC into RPE. Confocal microscopy shows the expression of AQP1 at the apical plasma membrane of polarized cobblestone hESC- and hiPSC-derived RPE cells. Lastly, aquaporin inhibitors significantly reduced AQP functionality in hESC-RPE cells.
hESC-RPE and hiPSC-RPE cells express several AQP genes, which are functional in mature hESC-derived RPE cells. The localization of AQP1 on the apical plasma membrane in mature RPE cells derived from both hESC and hiPSC suggests its functionality. These data propose that hESC- and hiPSC-derived RPE cells, grown and differentiated under serum-free conditions, resemble their native counterpart in the human eye.
No preview · Article · May 2013 · Investigative ophthalmology & visual science
[Show abstract][Hide abstract] ABSTRACT: Purpose:
The aim of the study is to examine the health-related quality-of-life (HRQOL) impact of the nocturnal awakenings and the duration of the sleep in the Finnish middle-aged and older population.
Cross-sectional sample consisted of 823 community-dwelling persons aged 55-75 living in a single municipality in a rural area of Eastern Finland. Frequency of the nocturnal awakenings was dichotomized as reporting "frequent," if the participant reported subjectively awakening "often" or "very often," and "infrequent" if the participant reported awakening "sometimes" or less frequently. HRQOL was measured with a preference-based HRQOL-index instrument, 15D. Analyses were adjusted for gender, BMI, morbidities, depression, employment and marital status, current smoking and drinking, exercise, recommendation to exercise from a health care professional, and subjective opinion about own exercise habits.
Frequent nocturnal awakenings had statistically and clinically significant negative impact on HRQOL, the mean (SE) adjusted marginal HRQOL impact being -0.0416 (0.006). More than 10 and less than 6.5 h of daily sleep were associated with higher probability of having low HRQOL, adjusted odd ratios (95 % CI) being 2.65 (1.11-6.33) and 2.65 (1.55-4.52), respectively. However, the changes in daily sleep duration did not have noticeable influence on the significance or magnitude of the negative HRQOL impact of the frequent nocturnal awakenings.
Nocturnal awakenings displayed a strong independent association with decreased HRQOL. The findings suggest that both clinicians and researchers should pay closer attention to nocturnal awakenings and other sleep problems in order to find ways to improve the quality of life in individuals with such conditions.
No preview · Article · Apr 2013 · Quality of Life Research
[Show abstract][Hide abstract] ABSTRACT: We address performance evaluation practices for developing medical image analysis methods, and contribute to the practice to establish and to share databases of medical images with verified ground truth and solid evaluation protocols. This helps to develop better algorithms, to perform profound method comparisons, including the state-of-the-art methods, and consequently, supports technology transfer from research laboratories to clinical practice. For this purpose, we propose a framework consisting of reusable methods and tools for the laborious task of constructing a benchmark database. We provide a medical image annotation software tool which helps to collect and store ground truth for retinopathy lesions from experts, including the fusion of spatial annotations from several experts. The tool and all necessary functionality for method evaluation are provided as a public software package. For demonstration purposes, we utilise the framework and tools to establish the DiaRetDB1 V2.1 database for benchmarking diabetic retinopathy detection algorithms. The database contains a set of retinal images, ground truth from several experts, and a strawman algorithm for the detection of retinopathy lesions.
[Show abstract][Hide abstract] ABSTRACT: Purpose:
Ocular surface reconstruction with cultivated oral mucosal epithelial transplantation technique is a viable treatment option for severe ocular surface injuries and diseases with limbal stem cell deficiency. Currently, this technique is based on utilization of xenogenic, allogenic or undefined components such as murine 3T3 feeders, serum and amniotic membrane. In this study, we aimed to find a more defined culture method to generate stratified human oral mucosal epithelium.
In this study, we have examined the formation of stratified cell sheets from human oral mucosal epithelial cells under serum-free culture environment both in the absence and presence of fibroblast-conditioned culture medium and elevated epidermal growth factor (EGF) concentration.
In all examined culture conditions, the cultivated oral epithelial cells formed a stratified tissue, which was positive for keratins K3/12, K4 and K13. The tissue-engineered oral epithelia also expressed proliferation and progenitor markers Ki67 and p63 in the basal layer of the cell sheets, suggesting that the epithelia still had regenerative capacity. The cultures presented expression of tight junction proteins ZO-1 and occludin and high transepithelial electrical resistance values.
In this culture method, we have been able to produce stratified cell sheets successfully without serum, conditioning of the medium or increased EGF concentration. We provide a novel protocol to produce tight multi-layered epithelium with proliferative potential, which can be easily adapted for cultivated oral mucosal epithelial transplantation.
No preview · Article · Sep 2012 · Acta ophthalmologica