For the treatment of bilateral limbal stem cell deficiency (LSCD), cell therapy with transplantation of cultivated oral mucosa epithelial cells (COMET) is a promising alternative. Although not yet established, current protocols on the cultivation of oral mucosal epithelial cell (OMECs) sheets are based mainly on substrates and xenobiotic additives that may lead to variable outcomes and undesirable immune responses by the patient. The aim of this study was to characterize OMECs cultivated in xenobiotic-free media (XF) seeded on fibrin gel, in comparison to conventional complex (COM) medium. Oral mucosal biopsies were retrieved from 31 donors. After cultivation in COM or XF medium, OMECs were compared based on growth kinetics, morphology, cell size and viability. Using immunofluorescence and gene expression analyses, the degree of stemness, proliferation and differentiation was evaluated in OMEC cultures. Our findings showed that although OMECs showed a similar morphology and viability, and comparable growth kinetics, immunofluorescence revealed the preservation of stemness (p63 + p40 positivity in cells ≤11 μm) and proliferation in both COM and XF. Gene expression analyses showed that keratin (K)13 and K15 expression levels were significantly higher in XF (adj. p < 0.001), but otherwise COM and XF-treated OMECs had comparable transcriptional profiles in a panel of stemness, proliferation and differentiation genes. These results demonstrate the feasibility of culturing OMECs on fibrin gel without xenogeneic additives, while maintaining their undifferentiated state and preserving stemness. In conclusion, both in terms of results and methodology, the procedures presented here are suitable for implementation in clinical practice.
- MeSH
- buněčná diferenciace MeSH
- buněčné kultury * MeSH
- deficit limbálních kmenových buněk MeSH
- dospělí MeSH
- epitelové buňky * metabolismus účinky léků MeSH
- fibrin * MeSH
- gely MeSH
- kmenové buňky * metabolismus cytologie MeSH
- kultivační média MeSH
- kultivované buňky MeSH
- lidé středního věku MeSH
- lidé MeSH
- limbus corneae * cytologie patologie metabolismus MeSH
- nemoci rohovky patologie farmakoterapie metabolismus MeSH
- proliferace buněk účinky léků MeSH
- rohovkový epitel metabolismus cytologie účinky léků patologie MeSH
- senioři MeSH
- transplantace kmenových buněk metody MeSH
- ústní sliznice * cytologie MeSH
- viabilita buněk MeSH
- xenobiotika farmakologie MeSH
- Check Tag
- dospělí MeSH
- lidé středního věku MeSH
- lidé MeSH
- mužské pohlaví MeSH
- senioři MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
[Scleral Grafts in Ophthalmic Surgery. A Review]
Cíle: Shrnout historii i současné trendy ve využití sklérálních štěpů v oftalmologii. Materiál a metody: Provedli jsme analýzu literatury prostřednictvím databází MEDLINE a Cochrane Library. Hledaná hesla byla “sclera”, “graft”, “surgery”. Výsledkem vyhledávání bylo 1596 článků, z nichž jsme 192 vyhodnotili jako relevantní. Relevantní články byly seřazeny chronologicky a dle způsobu využití sklerálních štěpu, což umožnilo vypracování přehledového článku. Výsledky: Skléra se v oftalmologii rutinně používá od padesátých let dvacátého století, a to v mnoha různých indikacích. Některé z nich se časem staly prakticky obsoletními (například využití při operačním řešení amoce sítnice), ale velká část nachází uplatnění dodnes (zejména využití v glaukomové či okuloplastické chirurgii, případně jako záplata při defektu skléry nebo rohovky). Závěr: I přesto, že je v současné době alogenní skléra oproti jiným produktům tkáňového bankovnictví v oftalmologii využívána spíše méně často a okruh jejích indikací se částečně zúžil, zůstává vzhledem ke své dostupnosti a vlastnostem užitečným a perspektivním materiálem.
Aim: To summarize the history and current trends in the use of scleral grafts in ophthalmology. Materials and methods: We conducted a review of the literature through the MEDLINE and Cochrane Library databases. The search terms were "sclera", "graft", and "surgery". The search resulted in 1596 articles, of which we evaluated 192 as relevant. The relevant articles were sorted chronologically and according to the method of using scleral grafts, which enabled the development of a review article. Results: The sclera has been routinely used in ophthalmology since the 1950s in many different indications. Some of these indications have become practically obsolete over time (for example, use in the surgical management of retinal detachment), but a large number still find application today (especially use in glaucoma or oculoplastic surgery, or as a patch for a defect in the sclera or cornea). Conclusion: Even though allogeneic sclera is currently used less frequently in ophthalmology compared to other tissue banking products and the range of its indications has partially narrowed, it remains a useful material due to its availability and properties.
- MeSH
- alografty * klasifikace MeSH
- dějiny 20. století MeSH
- lidé MeSH
- oftalmologické chirurgické výkony dějiny klasifikace metody MeSH
- přehledová literatura jako téma MeSH
- skléra * chirurgie MeSH
- skleroplastika dějiny klasifikace metody MeSH
- transplantáty dějiny klasifikace MeSH
- vitreoretinální chirurgie dějiny klasifikace metody MeSH
- Check Tag
- dějiny 20. století MeSH
- lidé MeSH
- Publikační typ
- přehledy MeSH
Bowman's layer is an acellular corneal structure, which is considered to be a specially modified anterior stroma. It is presumed, that it forms as a result of ongoing epithelial-stromal interactions and no clear physiological purpose has been proven. Despite this fact, Bowman's layer has found its place in corneal transplantation. It has been performed for over a decade, mainly in treatment of advanced keratoconus with multiple modifications. Transplantation of Bowman's layer can be expected to become a widely used surgical procedure in the treatment of many corneal pathologies involving fragmentation and destruction of Bowman's layer. This article aims to summarize information available on its structure, possible function, and transplantation. A thorough literature search was performed in the PubMed database and Google Scholar using keywords: Bowman's layer, structure, function, preparation and corneal transplantation. All the relevant sources were used, which represent 77 peer-reviewed articles with information corcerning the topic of this article.
- MeSH
- Bowmanova membrána patologie MeSH
- keratokonus chirurgie terapie MeSH
- lidé MeSH
- nemoci rohovky chirurgie terapie MeSH
- transplantace rohovky * MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- přehledy MeSH
Glaucoma is a significant cause of blindness worldwide, and its treatment remains challenging. The disease progressively leads to damage to the optic disc and thus loss of visual acuity and visual field. High intraocular pressure (IOP) is a common risk factor. There are three major methods to treat this disease: topical, laser, and surgical. None of these are completely satisfactory; therefore, alternatives using new biomaterials are being sought. Since biomaterial engineering has experienced significant growth in recent decades, its products are gradually being introduced to various branches of medicine, with the exception of ophthalmology. Biomaterials, such as glaucoma drainage implants, have been successfully used to treat glaucoma. There is significant ongoing research on biomaterials as drug delivery systems that could overcome the disadvantages of topical glaucoma treatment, such as poor intraocular penetration or frequent drug administration. This article summarizes the use of novel biomaterials for glaucoma treatment presented in the literature. The literature search was based on articles published in English on PubMed.gov, Cochranelibrary.com, and Scopus.com between 2018 and 2023 using the following term "biomaterials in glaucoma." A total of 103 published articles, including twenty-two reviews, were included. Fifty-nine articles were excluded on the basis of their titles and abstracts.
- Publikační typ
- časopisecké články MeSH
- přehledy MeSH
PRCIS: Deep sclerectomy (DS) with fibrin adhesive can constitute a safe alternative to the classic procedure using sutures, providing nonallergenic, nontoxic, and secure adhesion with no sign of aqueous humor outflow obstruction postoperatively. OBJECTIVE: To evaluate short and medium-term postoperative results of DS with a fibrin sealant. PATIENTS AND METHODS: This prospective, noncomparative, interventional case series involves 12 eyes of 12 patients with uncontrolled open angle glaucoma who underwent DS with Esnoper (Clip or V2000) implant between February 2021 and March 2022. A novel method of wound closure (sclera, Tenon fascia, and conjunctiva) employing fibrin glue was used instead of classic sutures. Surgical outcomes assessed include: intraocular pressure and glaucoma therapy reduction, best-corrected visual acuity changes, and number of complications registered peri and postoperatively. All measurements were performed preoperatively, as well as at 1 day, at 1 and 2 weeks, and at 1, 2, 3, 6, 9, and 12 months after surgery. RESULTS: The mean intraocular pressure decreased from 24.0 ± 9.1 mm Hg to 13.8 ± 6.3 mm Hg at 1 year postoperatively ( P < 0.001). Kaplan-Meier survival analysis revealed complete and qualified success rates of 83.3% and 91.7%. The mean glaucoma therapy decreased from 3.2 ± 1.1 to 0.8 ± 1.3 drugs 12 months after surgery ( P < 0.001). Nd:YAG goniopunture was performed in 2 eyes at 1 and 12 months postoperatively. No significant best-corrected visual acuity changes were registered. Perioperatively, we noted a trabeculo-descemet microperforation in 1 eye, transient hypotony in 5 eyes, and mild hyphema in 2 eyes. CONCLUSIONS: Fibrin adhesive provided an effective closure in sutureless DS in the patients included in our study. This modification of classical DS may simplify the surgical technique, ensure secure wound adaptation, optimize healing, and lower the risk of inflammation and fibrosis postoperatively.
- MeSH
- bezstehová chirurgie * metody MeSH
- drenážní implantáty glaukomové MeSH
- fibrinová tkáňová adheziva * terapeutické užití MeSH
- glaukom s otevřeným úhlem * chirurgie patofyziologie MeSH
- lidé středního věku MeSH
- lidé MeSH
- následné studie MeSH
- nitrooční tlak * fyziologie MeSH
- prospektivní studie MeSH
- senioři nad 80 let MeSH
- senioři MeSH
- skléra * chirurgie MeSH
- sklerostomie * metody MeSH
- tkáňová adheziva * terapeutické užití MeSH
- tonometrie oční MeSH
- výsledek terapie MeSH
- zraková ostrost * fyziologie MeSH
- Check Tag
- lidé středního věku MeSH
- lidé MeSH
- mužské pohlaví MeSH
- senioři nad 80 let MeSH
- senioři MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
The cornea is one of the most commonly transplanted tissues worldwide. It is used to restore vision when severe visual impairment or blindness occurs in patients with corneal diseases or after trauma. Due to the global shortage of healthy donor corneas, decellularized corneal tissue has significant potential as an alternative to corneal transplantation. It preserves the native and biological ultrastructure of the cornea and, therefore, represents the most promising scaffold. This article discusses different methods of corneal decellularization based on the current literature. We searched PubMed.gov for articles from January 2009 to December 2023 using the following keywords: corneal decellularization, decellularization methods, and corneal transplantation. Although several methods of decellularization of corneal tissue have been reported, a universal standardised protocol of corneal decellularization has not yet been introduced. In general, a combination of decellularization methods has been used for efficient decellularization while preserving the optimal properties of the corneal tissue.
- Publikační typ
- časopisecké články MeSH
- přehledy MeSH
PURPOSE: To evaluate the safety and efficacy of using corneal stromal lenticules (CSLs) obtained during refractive surgery Refractive Lenticule Extraction (ReLEx) with the Small Incision Lenticule Extraction (SMILE) procedure for the treatment of corneal ulcers. METHODS: This retrospective study included 12 eyes of 12 patients, 7 men and 5 women with varying degrees of corneal ulcer. The mean age was 64 ± 18 (range 34 to 95 years). The monitoring included corrected distance visual acuity (CDVA), slit-lamp biomicroscopy examination, a Seidel test, stability of the graft and anterior segment optical coherence tomography (AS-OCT) inspection. Patients were closely monitored for possible postoperative complications for at least 6 months. RESULTS: In 7/12 (58%) eyes, the corneal ulcer was successfully sealed with CSL and amniotic membrane (AM) without the need for any additional surgical intervention. In 3 eyes, penetrating keratoplasty (PK) was needed in addition to CSL transplantation and in 2 eyes the scleral patch was used to fully seal after CSL transplantation. During the follow-up period no signs of rejection or infection were detected in any patient. CONCLUSION: The use of CSLs from ReLEx SMILE may be considered as an alternative method for the treatment of corneal ulcers before a more extensive and definitive solution - PK - is used. Our preliminary findings suggest that properly performed CSL transplantation using cryopreserved lenticules is a safe and effective method to temporarily cover the corneal partial-thickness defect or even perforation.
- MeSH
- dospělí MeSH
- lidé středního věku MeSH
- lidé MeSH
- retrospektivní studie MeSH
- senioři nad 80 let MeSH
- senioři MeSH
- stroma rohovky chirurgie MeSH
- transplantace rohovky * metody MeSH
- vřed rohovky * chirurgie MeSH
- vřed chirurgie MeSH
- Check Tag
- dospělí MeSH
- lidé středního věku MeSH
- lidé MeSH
- mužské pohlaví MeSH
- senioři nad 80 let MeSH
- senioři MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
PURPOSE: The aim of our presentation is to introduce future eye bank product - corneal stromal lenticule from living donors, which can be used for allotransplantation. METHODS: ReLEx (refractive lenticule extraction) SMILE (small incision lenticule extraction) is a common approach in laser eye surgery. It is minimally invasive and flap-free procedure. During this procedure part of corneal stroma (lenticule) is created by femtosecond laser and consequently removed through small incision. The lenticule is basically waste material of the ReLEx SMILE procedure. In the International Eye Bank of Prague, we decided to establish new protocol for lenticule withdrawal, storage and release for transplantation. RESULTS: All donors signed an informed consent, and their serum was tested for the presence of infectious diseases. After ReLEx SMILE procedure the lenticule was stored in container with cryopreservation solution and frozen in the eye bank using the same protocol for frozen amniotic membrane. After 6 months in -80°C tissues were defrosted and examined histologically, using conventional light histology staining and electron microscopy. CONCLUSION: We believe, that lenticule from living donor is a safe and effective tissue, that can be used for many indications and in particular situations represents good alternative to whole donor cornea and amniotic membrane.
- MeSH
- lidé MeSH
- oční banky * MeSH
- rohovka MeSH
- stroma rohovky * chirurgie MeSH
- usmívání se MeSH
- žijící dárci MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
PURPOSE: The goal of our study is to find an optimal approach to the preparation and preservation of corneal stromal tissue. We want to compare different methods of corneal stromal tissue creation and storage to optimize the efficacy of this process under the conditions of an eye bank. After we find the most suitable method to create a safe high quality product, we want to prove the possibility of using a single donor cornea for more than one patient. We would also like to verify the feasibility of making more corneal lenticules after the removal of a corneal endothelium for DMEK transplantation. METHODS: We provided morphological (histology, scanning electron microscope) and microbiological analysis in order to compare different methods of corneal lenticule and corneal stromal lamellae preparation and preservation. We also tested the surgical handling of the tissue to secure a safe manipulation of the tissue for clinical use. We compared two methods of corneal lenticule preparation: microkeratome dissection and femtosecond laser. As methods of preservation, we tested hypothermia, cryopreservation at -80 degrees Celsius in DMSO (dimethyl sulfoxide) and storage at room temperature with glycerol. Some intrastromal lenticules and lamellae in each group were previously irradiated with gamma radiation of 25 kGy (KiloGray). RESULTS: Corneal stromal lamellae prepared with a microkeratome have a smoother cut - side surface compared to lamellae prepared with a femtosecond laser. Femtosecond laser preparation caused more irregularities on the surface and we detected more conglomerates of the fibrils, while lamellae made with microkeratome had more sparse network. Using femtosecond laser, we were able to make more than five lenticules from a single donor cornea. Gamma irradiation led to damage of collagen fibrils in corneal stroma and a loss of their regular arrangement. Corneal tissue stored in glycerol showed collagen fibril aggregates and empty spaces between fibrils caused by dehydration. Cryopreserved tissue without previous gamma irradiation showed the most regular structure of the fibrils comparable to storage in hypothermia. CONCLUSION: Our results suggest that formation of a corneal lenticule lamellae by microkeratome results in smoother corneal lenticules, while being much cheaper than formation by femtosecond laser. Gamma irradiation of 25 kGy caused damage of the collagen fibres as well as their network arrangement, which correlated with loss of transparency and stiffer structure. These changes impair possible surgical utilisation of gamma irradiated corneas. Storage in glycerol at room temperature and cryopreservation had similar outcomes and we believe that both methods are appropriate and safe for further clinical use .
- MeSH
- dimethylsulfoxid MeSH
- glycerol * MeSH
- hypotermie * MeSH
- kolagen MeSH
- lidé MeSH
- rohovka chirurgie MeSH
- stroma rohovky chirurgie MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
PURPOSE: To evaluate changes in corneal refractive parameters after implantation of a stromal lenticule of different thickness. We assume that the refractive outcome depends on the optical power of the used lenticule. METHODS: We conducted an ex-vivo non-human study on 33 normotonic porcine eyeballs divided into two groups, for 4D and 8D human lenticule implantation. Corneal stromal lenticules were obtained as a by-product from a laser procedure ReLEx SMILE. We evaluated corneal refractive parameters measured on Oculus Pentacam© device before and immediately after the intrastromal lenticule implantation. RESULTS: There was no statistically significant difference in corneal refractive parameters between the eyeball groups before lenticule implantation. In both groups, the intrastromal implantation in the depth of 300um led to a significant increase of central corneal pachymetry and corneal anterior steepening. In the 4D group the average central corneal pachymetry increased from 903 ± 124.59 to 1230 ± 148.99 (p = 0.0022) and in 8D group from 733.35 ± 69.60 to 1109 ± 161.64 (p = 0.0008). Induced changes in other studied parameters were not statistically significant, Kmax changed from 45.57 ± 2.78 to 72.07 ± 16.83 (p = 0.0094) and Km front from 40.72 ± 1.60 to 48.87 ± 5.83 (p = 0.0037) in 4D group and in the 8D group average Kmax increased from 42.22 ± 1.54 to 62.95 ± 12.67 (p = 0.0001) and K2 front 40.46 ± 1.64 to 51.51 ± 9.63 (p = 0.0037). There were no significant differences in refractive changes between the 4D and 8D groups after lenticule implantation. CONCLUSION: Intrastromal corneal lenticule implantation induces changes in corneal refractive parameters. In both groups, the implantation induced a significant increase of an anterior corneal steepening without any significant influence on posterior corneal flattening. Corneal lenticule implantation did not lead to any significant change of corneal astigmatism. However, in order to have more precise data for future clinical applications we need to continue with the experiments and verify the results on human corneas.
- MeSH
- laserová chirurgie rohovky * metody MeSH
- lasery excimerové terapeutické užití MeSH
- lidé MeSH
- nemoci rohovky * MeSH
- prasata MeSH
- refrakce oka MeSH
- rohovka MeSH
- rohovková topografie MeSH
- stroma rohovky chirurgie MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH