Tsutomu Inatomi

Bio: Tsutomu Inatomi is an academic researcher from Kyoto Prefectural University of Medicine. The author has contributed to research in topics: Transplantation & Cornea. The author has an hindex of 40, co-authored 111 publications receiving 6175 citations. Previous affiliations of Tsutomu Inatomi include Kyoto Prefectural University.

Growth factor mRNA and protein in preserved human amniotic membrane.

Abstract: Purpose. To investigate the expression of growth factor mRNA and the level of growth factor protein in preserved human amniotic membrane (AM). Methods. RT-PCR was used to examine the expression of ...

Transplantation of cultivated autologous oral mucosal epithelial cells in patients with severe ocular surface disorders

Abstract: Background/aims: To determine outcomes of transplants of cultivated autologous oral epithelial cells in patients with severe ocular surface disorders. Methods: The eyes (n = 6) of four patients with Stevens-Johnson syndrome (three eyes) or chemical burns (three eyes) were studied. Autologous oral epithelial cells, grown for 2–3 weeks on a denuded amniotic membrane carrier in the presence of 3T3 fibroblasts, were air lifted. The resultant sheet was transplanted onto the damaged eye, and acceptance of the sheet by the corneal surface was confirmed 48 hours after surgery. The success of ocular surface reconstruction, graft survival, changes in visual acuity, and postoperative complications were assessed and the quality of the cultivated oral epithelial sheet was evaluated histologically. Results: At 48 hours after transplant, the entire corneal surface of all six eyes was free of epithelial defects indicating complete survival of the transplanted oral epithelium. Visual acuity was improved in all eyes. During follow up (mean 13.8 (SD 2.9) months), the corneal surface remained stable, although all eyes manifested mild peripheral neovascularisation. Conclusions: Autologous oral epithelial cells grown on denuded amniotic membrane can be transplanted to treat severe ocular surface disorders.

Injection of Cultured Cells with a ROCK Inhibitor for Bullous Keratopathy.

Abstract: Background Corneal endothelial cell (CEC) disorders, such as Fuchs’s endothelial corneal dystrophy, induce abnormal corneal hydration and result in corneal haziness and vision loss known as bullous keratopathy. We investigated whether injection of cultured human CECs supplemented with a rho-associated protein kinase (ROCK) inhibitor into the anterior chamber could increase CEC density. Methods We performed an uncontrolled, single-group study involving 11 persons who had received a diagnosis of bullous keratopathy and had no detectable CECs. Human CECs were cultured from a donor cornea; a total of 1×106 passaged cells were supplemented with a ROCK inhibitor (final volume, 300 μl) and injected into the anterior chamber of the eye that was selected for treatment. After the procedure, patients were placed in a prone position for 3 hours. The primary outcome was restoration of corneal transparency, with a CEC density of more than 500 cells per square millimeter at the central cornea at 24 weeks after ...

Cultivation of Corneal Epithelial Cells on Intact and Denuded Human Amniotic Membrane

Abstract: PURPOSE. Surgery to reconstruct the ocular surface is greatly facilitated by the use of amniotic membrane, either as a biologic drape or, more recently, as a substrate for the transplantation of cultivated corneal epithelial cells. This study was designed to compare the usefulness of intact and denuded human amniotic membranes as a substrate for corneal epithelial cell culture. METHODS. Small (3-mm-diameter) biopsy specimens of superficial cornea including epithelium were excised from the central and limbal regions in rabbits. They were cultured on human amniotic membrane with or without amniotic epithelial cells and examined by light, scanning electron, and transmission electron microscopy. RESULTS. Cellular outgrowth from the central explants (n = 10) after 14 days in culture measured 1.82 ± 2.62 mm2 on intact amniotic membrane and 131.83 ± 28.31 mm2 on denuded amniotic membrane. In contrast, outgrowths from the limbal explants (n = 10) at the same time measured 4.58 ± 4.56 and 505.39 ± 134.20 mm2 on intact and denuded amniotic membranes, respectively. The leading edges of the outgrowths on intact amniotic membrane were much less uniform than those on denuded amniotic membrane, and, in the former, corneal epithelial cells appeared to migrate over the top of amniotic epithelial cells. Limbal cells cultivated on denuded amniotic membrane formed a nicely stratified layer that adhered well to the underlying amniotic membrane. CONCLUSIONS. Denuded amniotic membrane appears to be an excellent substrate for the cultivation of corneal epithelial cells, with a view to transplantation.

Investigative Ophthalmology and Visual Science

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Corneal reconstruction with tissue-engineered cell sheets composed of autologous oral mucosal epithelium.

Abstract: Background Ocular trauma or disease may lead to severe corneal opacification and, consequently, severe loss of vision as a result of complete loss of corneal epithelial stem cells. Transplantation of autologous corneal stem-cell sources is an alternative to allograft transplantation and does not require immunosuppression, but it is not possible in many cases in which bilateral disease produces total corneal stem-cell deficiency in both eyes. We studied the use of autologous oral mucosal epithelial cells as a source of cells for the reconstruction of the corneal surface. Methods We harvested 3-by-3-mm specimens of oral mucosal tissue from four patients with bilateral total corneal stem-cell deficiencies. Tissue-engineered epithelial-cell sheets were fabricated ex vivo by culturing harvested cells for two weeks on temperature-responsive cell-culture surfaces with 3T3 feeder cells that had been treated with mitomycin C. After conjunctival fibrovascular tissue had been surgically removed from the ocular surfa...

Grading of corneal and conjunctival staining in the context of other dry eye tests.

Abstract: PurposeTo describe the Oxford Scheme for grading ocular surface staining in dry eye and to discuss optimization of stain detection using various dyes and filters. Also, to propose a sequence of testing for dry eye diagnosis.MethodsThe grading of corneal and conjunctival staining is described

Concise review: isolation and characterization of cells from human term placenta: outcome of the first international Workshop on Placenta Derived Stem Cells.

Abstract: Placental tissue draws great interest as a source of cells for regenerative medicine because of the phenotypic plasticity of many of the cell types isolated from this tissue. Furthermore, placenta, which is involved in maintaining fetal tolerance, contains cells that display immunomodulatory properties. These two features could prove useful for future cell therapy-based clinical applications. Placental tissue is readily available and easily procured without invasive procedures, and its use does not elicit ethical debate. Numerous reports describing stem cells from different parts of the placenta, using nearly as numerous isolation and characterization procedures, have been published. Considering the complexity of the placenta, an urgent need exists to define, as clearly as possible, the region of origin and methods of isolation of cells derived from this tissue. On March 23-24, 2007, the first international Workshop on Placenta Derived Stem Cells was held in Brescia, Italy. Most of the research published in this area focuses on mesenchymal stromal cells isolated from various parts of the placenta or epithelial cells isolated from amniotic membrane. The aim of this review is to summarize and provide the state of the art of research in this field, addressing aspects such as cell isolation protocols and characteristics of these cells, as well as providing preliminary indications of the possibilities for use of these cells in future clinical applications.

TFOS DEWS II pathophysiology report

Abstract: The TFOS DEWS II Pathophysiology Subcommittee reviewed the mechanisms involved in the initiation and perpetuation of dry eye disease. Its central mechanism is evaporative water loss leading to hyperosmolar tissue damage. Research in human disease and in animal models has shown that this, either directly or by inducing inflammation, causes a loss of both epithelial and goblet cells. The consequent decrease in surface wettability leads to early tear film breakup and amplifies hyperosmolarity via a Vicious Circle. Pain in dry eye is caused by tear hyperosmolarity, loss of lubrication, inflammatory mediators and neurosensory factors, while visual symptoms arise from tear and ocular surface irregularity. Increased friction targets damage to the lids and ocular surface, resulting in characteristic punctate epithelial keratitis, superior limbic keratoconjunctivitis, filamentary keratitis, lid parallel conjunctival folds, and lid wiper epitheliopathy. Hybrid dry eye disease, with features of both aqueous deficiency and increased evaporation, is common and efforts should be made to determine the relative contribution of each form to the total picture. To this end, practical methods are needed to measure tear evaporation in the clinic, and similarly, methods are needed to measure osmolarity at the tissue level across the ocular surface, to better determine the severity of dry eye. Areas for future research include the role of genetic mechanisms in non-Sjogren syndrome dry eye, the targeting of the terminal duct in meibomian gland disease and the influence of gaze dynamics and the closed eye state on tear stability and ocular surface inflammation.