Biomedical subjects

Shigeru Kinoshita

Publications and source records attributed to Shigeru Kinoshita.

At least 19 recordsLinked to original sources

PubMed2026

Crosstalk mediators implicated in the Stevens-Johnson Syndrome through gene regulatory network analysis.

Stevens-Johnson syndrome (SJS) is a rare and severe mucocutaneous disorder often triggered by medications or infections. Our previous research identified that four key genes, Ikzf1, Ptger3, Mavs, and Tlr3 are involved in SJS susceptibility and the conjunctival epithelial innate immune response, demonstrating their role in regulating interferon-stimulated genes. However, the interplay among these regulatory factors remains unclear. This study aimed to elucidate the crosstalk mechanisms between the pathways regulated by these four genes in conjunctival epithelial cells. We constructed a comprehensive gene regulatory network using transcriptomic data from murine conjunctival epithelial cells under 16 distinct conditions, including polyI:C stimulation across wild-type, knockout, and transgenic backgrounds for the key genes. A targeted network analysis systematically identified numerous candidate genes mediating the crosstalk between the regulatory pathways initiated by Ikzf1, Ptger3, Mavs, and Tlr3. The identified candidates suggest the involvement of diverse signaling pathways previously unlinked to SJS pathology. Our findings suggest that the pathogenesis of SJS may arise not from the dysfunction of isolated genes but from the disruption of a balance maintained by intricate pathway crosstalk.

Animals↗

PubMed2026

Descemet Stripping Only in Fuchs Endothelial Corneal Dystrophy: Results of a Randomized Clinical Trial of Topical Ripasudil and Directions for Future Innovation.

PURPOSE: To review history of Descemet stripping only (DSO) in Fuchs endothelial corneal dystrophy, describe the results of a clinical trial of topical ripasudil after DSO (K-321-201 study), and discuss future directions. METHODS: A 1-year, phase 2, randomized, placebo-controlled multicenter clinical trial of two doses of K-321 (ripasudil) administered for 12 weeks after DSO surgery in Fuchs endothelial corneal dystrophy was performed. The primary endpoint, central corneal endothelial cell density (ECD) at 12 weeks after surgery, was determined by an independent reading center that was masked to study group assignment. Duration of corneal edema, need for medical or surgical rescue therapy, corneal thickness, and central ECD throughout the entire study period were also examined. Adverse events and exploratory endpoints were collected. RESULTS: Sixty-five subjects were enrolled (21 in the QID group, 22 in the BID, and in the placebo group). Over 95% of subjects completed the trial. The QID group had a higher central ECD 12 weeks after DSO than the placebo group (531 ± 312 cells/mm2 vs 228 ± 298 cells/mm2, P = .0065). Corneal edema cleared in 17/21 (81.0%) of the QID group at 12 weeks, compared with 2/22 (9.1%) of the placebo group (P < .0001). Rescue was required in 2/21 (9.5%) subjects in the QID group and 6/22 (27.3%) subjects in the placebo group (P = .0092). Adverse events were mild and did not lead to discontinuation of treatment. CONCLUSIONS: Topical K-321 given QID improves DSO outcomes, as demonstrated by a higher ECD, more rapid resolution of corneal edema, and reduced failure rate. The medication was well-tolerated.

Humans↗

PubMed2007

Polyclonality of Staphylococcus epidermidis residing on the healthy ocular surface.

Staphylococcus epidermidis is part of the normal bacterial flora on the ocular surface. The chromosomal DNA of bacterial isolates obtained from the conjunctival sac, upper and lower lid margins, and upper and lower Meibomian glands of healthy volunteers was subjected to SmaI digestion and PFGE to study the genetic diversity of the organisms. Multiple colonies were also examined of S. epidermidis derived from the conjunctival sac of the same subjects. Lastly, commensal bacteria were harvested from the ocular surfaces of four healthy subjects once a month for 6 months, and the genetic background of the S. epidermidis isolates was analysed. It was found that bacterial strains not only from different subjects but also from multiple ocular surface sites of the same subject exhibited different PFGE patterns. In five of 42 subjects multiple colonies of S. epidermidis were isolated from the conjunctival sac; three harboured multiple colonies with different PFGE patterns, and two manifested multiple colonies with identical PFGE patterns. S. epidermidis isolated from the conjunctival sac of the same subjects over a 6-month period exhibited varying PFGE patterns. The data demonstrate the polyclonality of S. epidermidis on the healthy ocular surface.

Conjunctiva↗

PubMed2006

Identification of human oral keratinocyte stem/progenitor cells by neurotrophin receptor p75 and the role of neurotrophin/p75 signaling.

This study was undertaken to determine whether human oral keratinocyte stem cells characteristically express higher levels of the low-affinity neurotrophin receptor p75 and to elucidate the function of p75 in oral keratinocytes. Examination of their expression patterns and cell-cycling status in vivo showed that p75 was exclusively expressed in the basal cell layer of both the tips of the papillae and the deep rete ridges. These immunostaining patterns suggest a cluster organization; most p75(+) cells did not actively cycle in vivo. Cell sorting showed that cells in the p75(+) subset were smaller and possessed higher in vitro proliferative capacity and clonal growth potential than the p75(-) subset. Clonal analysis revealed that holoclone-type (stem cell compartment), meroclone-type (intermediate compartment), and paraclone-type (transient amplifying cell compartment) cells, previously identified in skin and the ocular surface, were present in human oral mucosal epithelium. Holoclone-type cells showed stronger p75 expression at both the mRNA and protein level than did meroclone- and paraclone-type cells. Among the several neurotrophins, nerve growth factor (NGF) and neurotrophin-3 stimulated p75(+) oral keratinocyte cell proliferation, and only NGF protected them from apoptosis. Our in vivo and in vitro findings indicate that p75 is a potential marker of oral keratinocyte stem/progenitor cells and that some neurotrophin/p75 signaling affects cell growth and survival.

Apoptosis↗

PubMed2006

Identification of resident and inflammatory bone marrow derived cells in the sclera by bone marrow and haematopoietic stem cell transplantation.

AIMS: To characterise bone marrow derived cells in the sclera under normal and inflammatory conditions, we examined their differentiation after transplantation from two different sources, bone marrow and haematopoietic stem cells (HSC). METHODS: Bone marrow and HSC from green fluorescent protein (GFP) transgenic mice were transplanted into irradiated wild-type mice. At 1 month after transplantation, mice were sacrificed and their sclera examined by histology, immunohistochemistry (CD11b, CD11c, CD45), and transmission and scanning electron microscopy. To investigate bone marrow derived cell recruitment under inflammatory conditions, experimental autoimmune uveitis (EAU) was induced in transplanted mice. RESULTS: GFP positive cells were distributed in the entire sclera and comprised 22.4 (2.8)% (bone marrow) and 28.4 (10.9)% (HSC) of the total cells in the limbal zone and 18.1 (6.7)% (bone marrow) and 26.3 (3.4)% (HSC) in the peripapillary zone. Immunohistochemistry showed that GFP (+) CD11c (+), GFP (+) CD11b (+) cells migrated in the sclera after bone marrow and HSC transplantation. Transmission and scanning electron microscopy revealed antigen presenting cells among the scleral fibroblasts. In EAU mice, vast infiltration of GFP (+) cells developed into the sclera. CONCLUSION: We have provided direct and novel evidence for the migration of bone marrow and HSC cells into the sclera differentiating into macrophages and dendritic cells. Vast infiltration of bone marrow and HSC cells was found to be part of the inflammatory process in EAU.

Animals↗

PubMed2006

Ocular surface reconstruction with combination of cultivated autologous oral mucosal epithelial transplantation and penetrating keratoplasty.

PURPOSE: To report an assessment of the two-step surgical combination of cultivated autologous oral mucosal epithelial transplantation (COMET) and penetrating keratoplasty (PKP) used to treat patients with severe limbal deficiency disorders, and to investigate the keratin expression patterns of transplanted surviving oral mucosal epithelium. DESIGN: Observational case series. METHODS: Two patients with Stevens-Johnson syndrome and chemical eye injury were treated by COMET followed, approximately six months later, by a PKP triple procedure. In the course of a mean follow-up period of 22.5 months, their clinical outcomes and the efficacy of this two-step surgical procedure were assessed. In addition, the keratin expression in corneal buttons excised during PKP were immunohistochemically examined to characterize the oral mucosal epithelium that survived ectopically on the cornea. In vivo laser confocal microscopy was used to investigate the structure of the epithelium on the corneal grafts. RESULTS: The ocular surfaces were successfully reconstructed with cultivated autologous oral mucosal epithelial sheets and PKP. No clinical complications, such as persistent epithelial defects, rejections, or recurrence of cicatrization, were encountered. Postoperative best-corrected visual acuity was 20/125 in one patient and 20/100 in the other. The surviving oral mucosal epithelium, distinguished by its fluorescence pattern, consisted of an irregular, nonkeratinized, stratified epithelium without goblet cells. Immunohistochemical study demonstrated that K3, but not K12, was expressed in the transplanted cultivated oral mucosal epithelium that was similar to oral mucosal tissue. In vivo, the epithelial structure and cell density in the basal cell layer of the corneal grafts were similar to normal cornea. CONCLUSIONS: This study presents a two-step surgical approach to treat severely scarred ocular surfaces by means of a combination of COMET and PKP. Clinical outcomes suggest that this treatment may be beneficial for the maintenance of the reconstructed ocular surface by providing oral mucosal epithelium around the corneal graft.

Aged↗

PubMed2006

Double-mirror goniolens with dual viewing system for goniosurgery.

PURPOSE: To compare the performance of the double-mirror goniolens with dual viewing system in goniosurgery, goniosynechialysis, and trabeculotomy ab externo with that of the conventional Swan-Jacob gonioprism (SJG). DESIGN: Prospective interventional case series. METHODS: We treated 10 patients (10 eyes) with medically uncontrollable chronic angle-closure glaucoma by goniosynechialysis and another 10 eyes with open-angle glaucoma by trabeculotomy ab externo (TLO). The 20 patients were divided randomly into two equal groups and underwent operation with the double-mirror goniolens or SJG instrument. RESULTS: There were no intraoperative complications, irrespective of the procedure or instrument that was used. Patients in the SJG group were required to tilt the head at a >30-degree angle to facilitate observation and manipulation of the quadrants. On the other hand, all structures could be observed and manipulated without head-tilting in the double-mirror goniolens group. CONCLUSION: The double-mirror goniolens with dual viewing system is more suitable than the conventional SJG for goniosurgery.

Glaucoma, Angle-Closure↗

PubMed2006

A comparison between cultivated and conventional limbal stem cell transplantation for Stevens-Johnson syndrome.

PURPOSE: To compare the resolution of inflammation and long-term results of cultivated and conventional limbal stem cell transplantation (LSCT) in a patient with Stevens-Johnson syndrome (SJS). DESIGN: Interventional case report. METHODS: A 32-year-old man with SJS and bilateral total limbal stem cell deficiency underwent cultivated LSCT in the right eye, followed by conventional LSCT in the left eye three weeks later. The postoperative medication included dexamethasone 0.1% and ofloxacin 0.3% eyedrops and a tapering dose of systemic corticosteroid, cyclosporine, and cyclophosphamide. Tear samples were collected and analyzed for interleukin (IL) 8 levels. RESULTS: Complete corneal epithelialization was achieved 48 hours after cultivated LSCT, compared with three weeks after conventional LSCT. Ocular inflammation and IL-8 levels decreased more rapidly in the eye with cultivated LSCT. Four years after surgery, more severe corneal scarring and opacification were noted in the conventional LSCT eye. CONCLUSIONS: Cultivated LSCT resulted in a better clinical result and vision, with less stromal scarring compared with conventional LSCT.

Adult↗

PubMed2006

Functional visual acuity in Stevens-Johnson syndrome.

PURPOSE: To evaluate the correlation of functional visual acuity (FVA) measurement with ocular surface findings in patients with Stevens-Johnson syndrome (SJS). DESIGN: Prospective comparative study. METHODS: Sixty-nine eyes of 38 patients with chronic SJS assessed at the Tokyo Dental College, Tokyo Medical Center, and the Kyoto Prefectural University of Medicine, Department of Ophthalmology, Kyoto, Japan, were studied. Twenty eyes of 10 normal subjects and 40 eyes of 20 patients with Sjögren syndrome (SS) were also studied. Conventional Landolt visual acuity (VA) and FVA examinations and slit-lamp examinations were performed. FVA was measured continuously by the FVA measurement system during a 30-second blink-free period in one eye. The visual maintenance ratio (VMR) was calculated as follows: VMR = [(2.7 - FVA)/(2.7 - baseline VA)], where logarithm of minimal angle of resolution values of FVA were entered into the formula and 2.7 represented the lowest visual acuity in this series. Slit-lamp examinations, Schirmer test, and fluorescein vital stainings were also performed in all subjects. RESULTS: VMR was markedly lower in patients with SJS compared with patients with SS and controls. FVA values showed a relation with the presence of corneal opacity and vascularization. CONCLUSIONS: The FVA measurement system is not only a useful tool in the evaluation of dynamic VA changes, but also reflects the ocular surface clinical findings in SJS.

Adult↗

PubMed2006

Transplantation of autologous serum-derived cultivated corneal epithelial equivalents for the treatment of severe ocular surface disease.

PURPOSE: To evaluate the use of autologous serum (AS)-derived cultivated corneal epithelial transplantation for the treatment of severe ocular surface disease. DESIGN: Retrospective noncomparative case series. PARTICIPANTS: Nine eyes from 9 patients with total limbal stem cell deficiency were studied. These consisted of 2 eyes with Stevens-Johnson syndrome, 1 with chemical injury, 1 with ocular cicatricial pemphigoid, 1 with Salzmann corneal dystrophy, 1 with aniridia, 1 with graft-versus-host disease, and 2 with idiopathic ocular surface disease. METHODS: Autologous serum obtained from patients was used for cultivating corneal epithelial cells on an amniotic membrane substrate. These AS-derived corneal epithelial equivalents were compared with those derived from fetal bovine serum (FBS)-supplemented medium. At the time of surgery, complete removal of the corneal pannus and conjunctiva up to 3 mm from the limbus was performed. Allogeneic (7 cases) and autologous (2 cases) AS-derived cultivated corneal epithelial equivalents were transplanted onto the ocular surface. Postoperative follow-up included serial slit-lamp examinations with fluorescein staining, as well as photographic documentation. MAIN OUTCOME MEASURES: Ocular surface reconstruction with corneal epithelialization, graft integrity, visual acuity, and postoperative complications. RESULTS: The corneal epithelial sheets cultivated in AS- and FBS-supplemented media were morphologically similar, and demonstrated the normal expression of tissue-specific keratins and junctional specialization assembly proteins. After transplantation, complete corneal epithelialization was achieved within 2 to 5 days. All eyes demonstrated an improvement in visual acuity by > or =2 lines. During the follow-up period of 14.6+/-4.36 months, the corneal surface of all patients remained stable and transparent, without significant complications. CONCLUSIONS: Transplantation of AS-derived cultivated corneal epithelial equivalents was shown to be a feasible method of treating patients with severe ocular surface disease. The use of AS is of clinical importance in the development of autologous xenobiotic-free bioengineered ocular surface equivalents for clinical transplantation.

Adult↗

PubMed2006

Human corneal epithelial cells respond to ocular-pathogenic, but not to nonpathogenic-flagellin.

In this study, we investigated the expression of TLR5 in human corneal epithelial cells (CEC), and the functional outcome of TLR5 triggering by flagellins of pathogenic- and nonpathogenic bacteria. Flagellins derived from Pseudomonas aeruginosa, Salmonella typhimurium, Serratia marcescense or Bacillus subtilis were used. The TLR5 protein and TLR5 specific mRNA expression was evident on human CEC. In human corneal epithelium tissues, TLR5 protein was detected at the basal and wing cells of the tissues. Ocular pathogenic bacteria, namely P. aeruginosa and S. marcescense, derived flagellin induced the significantly increased level of gene activation and IL-6 and IL-8 production. In contrast, ocular nonpathogenic S. typhimurium- and B. subtilis-derived flagellin induced neither the gene activation nor the increased production of IL-6 and IL-8 in human CEC. Human CEC would respond only to flagellin derived of ocular pathogenic bacteria, but not to those derived of ocular nonpathogenic bacteria, to generate pro-inflammatory cytokines.

Bacterial Proteins↗

PubMed2006

Neural conversion of ES cells by an inductive activity on human amniotic membrane matrix.

Here we report a human-derived material with potent inductive activity that selectively converts ES cells into neural tissues. Both mouse and human ES cells efficiently differentiate into neural precursors when cultured on the matrix components of the human amniotic membrane in serum-free medium [amniotic membrane matrix-based ES cell differentiation (AMED)]. AMED-induced neural tissues have regional characteristics (brainstem) similar to those induced by coculture with mouse PA6 stromal cells [a common method called stromal cell-derived inducing activity (SDIA) culture]. Like the SDIA culture, the AMED system is applicable to the in vitro generation of various CNS tissues, including dopaminergic neurons, motor neurons, and retinal pigment epithelium. In contrast to the SDIA method, which uses animal cells, the AMED culture uses a noncellular inductive material derived from an easily available human tissue; therefore, AMED should provide a more suitable and versatile system for generating a variety of neural tissues for clinical applications.

Amnion↗

PubMed2006

Different expression of angiogenesis-related factors between human cultivated corneal and oral epithelial sheets.

We developed a cultivated oral mucosal epithelial sheet (COE) transplantation system to address severe human ocular surface disorders. Unlike the cultivated corneal epithelial sheet (CCE), the COE induces mild superficial peripheral neovascularization although central clarity is maintained. To evaluate the characteristic differences between CCE and COE regarding to angiogenesis, we examined the expression of angiogenesis-related factors in CCE and COE. Using samples of CCE and COE, we immunohistochemically determined protein expression of the angiogenesis related factors: Thrombospondin-1 (TSP-1), pigment epithelium derived factor (PEDF), endostatin, angiostatin, vascular endothelial growth factor (VEGF), Fms-like tyrosine kinase 1 (Flt-1), kinase insert domain receptor (KDR), and basic fibroblast growth factor (bFGF). We used Western blot analysis to confirm the factors that were immunohistochemically different in CCE and COE. The immunohistochemical staining intensity of TSP-1 was higher in CCE than COE and by Western blot analysis the expression of TSP-1 was significantly higher in CCE than COE (P<0.05). PEDF and endostatin stained moderately stronger in CCE than COE. Immunohistochemically there was no obvious difference between CCE and COE with respect to angiostatin, VEGF, Flt-1, KDR, and bFGF. In comparison with CCE, COE showed decreased expression of anti-angiogenic factors particularly TSP-1. This different expression may relate to the superficial peripheral neovascularization encountered after COE transplantation.

Angiogenesis Inducing Agents↗

PubMed2006

Comparison of intact and denuded amniotic membrane as a substrate for cell-suspension culture of human limbal epithelial cells.

BACKGROUND: We have previously developed a limbal epithelial culture system using a cell-suspension method on denuded amniotic membrane (AM). However, other workers reported that intact AM is advantageous for limbal epithelial culture in that it preserves stem cell characteristics. In this study, we cultivated human limbal epithelial cell-suspensions on both intact and denuded AM and compared the morphology and adhesion of the limbal epithelial cells on these two substrates. METHODS: Human limbal epithelial cells were dissociated from donor eyes using dispase and gentle pipetting and then seeded onto intact and denuded AM as cell suspension. Limbal epithelial cells on AM were co-cultured with a MMC-treated 3T3 fibroblast feeder layer and epithelial differentiation was promoted by air lifting. Cultures were examined by light, scanning and transmission electron microscopy and differences in cellular attachments and intercellular spacing were quantified. Basement membrane complexes were examined by indirect immunofluorescence. RESULTS: Limbal cells grown on denuded AM were well stratified and differentiated. Cells were well attached to each other and to the basement membrane. In contrast, limbal cells cultured on intact AM failed to stratify and in places formed a monolayer. The culture on denuded AM had significantly (P<0.001) more desmosomal junctions as well as significantly (P<0.001) more junctional attachments to the carrier than the intact culture. In addition, the intercellular spaces between cells cultivated on denuded AM were significantly (P<0.001) smaller than those between cells grown on the intact substrate. In cultures on both denuded and intact AM, the basement membrane zone displayed a positive staining for collagen VII, integrins alpha-6 and beta-4 and laminin 5. CONCLUSIONS: We successfully cultivated well-stratified and -differentiated limbal cells on denuded AM, while on the intact AM limbal cells failed to stratify and in places formed only a monolayer of cells. The limbal cells cultivated on denuded AM were well attached to the AM stroma and were morphologically superior to the limbal epithelium cultivated on intact AM. We conclude that for purposes of transplantation of differentiated epithelial sheets, denuded AM is probably the more practical carrier for human limbal epithelial cell cultures when using our cell-suspension culture system.

3T3 Cells↗

PubMed2006

Neurensin-1 expression in the mouse retina during postnatal development and in the cultured retinal neurons.

Neurensin-1/Neuro-p24 (previously named Neuro-p24) is a neuron-specific membrane protein that is localized particularly in neurites. Neurensin-1 is considered to play an essential role in neurite extension during nervous development, regeneration and plasticity. To understand what role Neurensin-1 plays in retinal differentiation, we examined Neurensin-1 distribution and gene expression pattern in the postnatally developing retina of the mouse, because the retina is an excellent model for nervous development. In the postnatal day (PD) 1 retina, intense Neurensin-1 immunoreactivity was found in the optic nerve fiber layer. Faint staining was seen in the ganglion cells, presumptive amacrine and horizontal cells. As the postnatal development proceeded, the optic fibers became more intensely stained in addition to other parts of the retina such as the ganglion cells, inner plexiform layer and horizontal cells. In PD 10 retinas, the horizontal cell processes showed a prominently stained configuration. As the retina developed further to attain maturity, the staining in the retina became less pronounced, although the optic nerves remained positively stained. The distribution of Neurensin-1 mRNA was consistent with these results and confirmed that the ganglion, amacrine and horizontal cells actively synthesize Neurensin-1 in the developing retina. In the retinal cell culture from newborn mice, two types of neural cells were stained for Neurensin-1, one of which showed long processes and appeared presumptive ganglion cells. These results suggest that Neurensin-1 plays a role in the fiber extension of the retinal neurons, as has been observed in other central nervous tissues, and indicate that the developing retina is a suitable experimental model for the analysis of Neurensin function, both in vivo and in vitro.

Animals↗

PubMed2006

Autologous serum-derived cultivated oral epithelial transplants for severe ocular surface disease.

OBJECTIVE: To evaluate the use of autologous serum (AS)-derived cultivated oral epithelial transplants for the treatment of severe ocular surface disease. METHODS: We used AS from 10 patients with severe ocular surface disease and total limbal stem cell deficiency to develop autologous cultivated oral epithelial equivalents. These were compared with epithelial equivalents derived from conventional fetal bovine serum-supplemented medium. Surgery involved removal of the corneal pannus and surrounding diseased tissue and transplantation of the AS-derived epithelial equivalents. The oral equivalents were analyzed by review of histologic and immunohistochemical findings. RESULTS: Oral epithelial sheets cultivated in AS- and fetal bovine serum-supplemented media were similar in morphology, and both formed basement membrane assembly proteins important for maintaining graft integrity. Complete corneal epithelialization was achieved within 2 to 5 days postoperatively. The ocular surface remained stable without major complications in all eyes during a mean +/- SD follow-up of 12.6 +/- 3.9 months. The visual acuity improved by more than 2 lines in 9 of 10 eyes, with transplanted oral epithelium surviving up to 19 months. CONCLUSION: The successful use of an AS-derived oral epithelial equivalent to treat severe ocular surface disease represents an important advance in the pursuit of completely autologous xenobiotic-free bioengineered ocular equivalents for clinical transplantation.

Adult↗

PubMed2006

The persistence of transplanted amniotic membrane in corneal stroma.

PURPOSE: To characterize the long-term incorporation of transplanted amniotic membrane into corneal stroma. DESIGN: Experimental study. METHODS: Rabbit amniotic membrane, stained with a fluorescent dye (DTAF), was implanted unilaterally into the corneal stroma of four adult rabbits. Corneas were examined clinically and by transmission electron microscopy and fluorescent microscopy at 1, 3, 7, and 10 months after surgery. RESULTS: Visibility of the transplanted amniotic membrane, in situ, on slit-lamp examination decreased over time. However, fluorescent and electron microscopy clearly demonstrated that the amniotic membrane remained structurally unchanged and intact within the corneal stroma up to 10 months after implantation. CONCLUSIONS: Amniotic membrane allografts persist intact within an intracorneal space for many months postoperatively and are not quickly broken down or dissolved by the host tissue.

Amnion↗

PubMed2006

Midterm results on ocular surface reconstruction using cultivated autologous oral mucosal epithelial transplantation.

PURPOSE: To perform a midterm assessment of the integrity and reproducibility of cultivated autologous oral mucosal epithelial sheets, and to evaluate the clinical efficacy of their transplantation in ocular surface. DESIGN: Observational case series. METHODS: Cultivated autologous oral mucosal epithelial sheets were created using amniotic membrane and buccal mucosal epithelium from 12 patients with Stevens-Johnson syndrome, chemical and thermal injury, pseudo-ocular cicatricial pemphigoid, and idiopathic ocular surface disorder. They were transplanted onto 15 eyes from these patients who were then followed up for a mean of 20 months; with the longest follow-up being 34 months. We assessed their clinical outcomes with special reference to neovascularization. RESULTS: Cultivated autologous oral mucosal epithelial sheets could be generated from all patients. On the second postoperative day, 14 of 15 sheets transplanted demonstrated total re-epithelialization on the cornea. During the follow-up, the ocular surface was stable and transparent without any major complications in 10 of 15 eyes (67%), and the transplanted epithelium survived for at least 34 months. There were five eyes (33%) with small but long-standing epithelial defects, three of these healed spontaneously, and two (13%) required reoperation. In 10 eyes, postoperative visual acuity was improved by more than 2 lines. All eyes manifested some peripheral corneal vascularization. CONCLUSIONS: We established a successful tissue-engineering technique to generate cultivated autologous oral mucosal epithelial sheets and succeeded in reconstructing the ocular surface. We suggest that this surgical modality may be both safe and useful, especially in younger patients with the most severe ocular surface disorders.

Adolescent↗