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PubMed2003

Ocular surface reconstruction using cultivated mucosal epithelial stem cells.

PURPOSE: To investigate the possibility of using cultivated oral epithelial cells in ocular surface reconstruction. METHODS: An ocular surface injury was created in adult albino rabbits by a lamellar keratectomy. Oral mucosal biopsy specimens taken from both adult albino rabbits and human volunteers were cultivated for 2-3 weeks on a denuded amniotic membrane (AM) carrier. The cultivated epithelium was examined by histologic and immunohistochemical analysis. At 3-4 weeks after the ocular surface injury, the rabbit conjunctivalized corneal surfaces were surgically reconstructed by transplanting both the rabbit and human cultivated oral epithelial cells on the AM carrier. RESULTS: Both the rabbit and human cultivated oral epithelial sheets had 5 or 6 layers of stratified, well-differentiated cells. Histologic examination revealed that the cultivated epithelial cells were similar in appearance to those of in vivo normal corneal epithelium. Immunohistochemistry confirmed the presence of the keratin pair 4 and 13 and keratin 3 in the cultivated oral epithelial cells. Corneas that were grafted with rabbit and human cultivated oral epithelial cells on an AM carrier were clear and were epithelialized 10 and 2 days after surgery, respectively. CONCLUSIONS: We have generated confluent cultures of both rabbit and human oral epithelial cells on AM expanded ex vivo from biopsy-derived oral mucosal tissues. We have successfully carried out xeno- and autologous transplantation of these cultivated oral epithelial cells onto the ocular surfaces of keratectomized rabbit eyes. We believe that xeno- and autologous transplantation of cultivated oral epithelium is a feasible method for ocular surface reconstruction.

Animals↗
PubMed1998

Cellular origin of mucins of the ocular surface tear film.

In summary, we have demonstrated that the ocular surface epithelia express at least three mucin genes. We suggest that the gel-forming mucin MUC5AC is a major mucin forming the mucus gel of the tear film. We further suggest that MUC1 facilitates the spread of the MUC5-containing mucus on the ocular surface and, along with the mucus gel, prevents cell and debris adhesion to the ocular surface. The function of MUC4 at the ocular surface remains to be elucidated.

Animals↗
PubMed1999

Cultured corneal epithelia for ocular surface disease.

PURPOSE: To evaluate the potential efficacy for autologous and allogeneic expanded corneal epithelial cell transplants derived from harvested limbal corneal epithelial stem cells cultured in vitro for the management of ocular surface disease. METHODS: Human Subjects. Of the 19 human subjects included, 18 (20 procedures) underwent in vitro cultured corneal epithelial cell transplants using various carriers for the epithelial cells to determine the most efficacious approach. Sixteen patients (18 procedures on 17 eyes) received autologous transplants, and 2 patients (1 procedure each) received allogeneic sibling grafts. The presumed corneal epithelial stem cells from 1 patient did not grow in vitro. The carriers for the expanded corneal epithelial cells included corneal stroma, type 1 collagen (Vitrogen), soft contact lenses, collagen shields, and amniotic membrane for the autologous grafts and only amniotic membrane for the allogeneic sibling grafts. Histologic confirmation was reviewed on selected donor grafts. Amniotic membrane as carrier. Further studies were made to determine whether amniotic membrane might be the best carrier for the expanding corneal epithelial cells. Seventeen different combinations of tryspinization, sonication, scraping, and washing were studied to find the simplest, most effective method for removing the amniotic epithelium while still preserving the histologic appearance of the basement membrane of the amnion. Presumed corneal epithelial stem cells were harvested and expanded in vitro and applied to the amniotic membrane to create a composite graft. Thus, the composite graft consisted of the amniotic membrane from which the original epithelium had been removed without significant histologic damage to the basement membrane, and the expanded corneal epithelial stem cells, which had been applied to and had successfully adhered to the denuded amniotic membrane. Animal model. Twelve rabbits had the ocular surface of 1 eye damaged in a standard manner with direct removal of the presumed limbal stem cells, corneal epithelium, and related epithelium, followed by the application of n-heptanol for 60 seconds. After 6 weeks, all damaged eyes were epithelialized and vascularized. Two such treated eyes were harvested without further treatment, to be used for histologic study as damaged controls. The remaining 10 rabbits received composite grafts (consisting of amniotic membrane with expanded allogeneic rabbit corneal epithelial cell transplants) applied to the ocular surface in a standard manner followed by the application of a contact lens. At 16 days following transplantation, 5 of the rabbits were sacrificed and the corneal rims were removed for histologic study. At 28 days, the remaining rabbits were sacrificed and the previously damaged eyes were harvested for histologic and immunohistochemical study. RESULTS: Human subjects. Of the 19 total patients admitted to the study, the presumed corneal epithelial stem cells of 1 patient did not grow in vitro. Of the remaining 18 patients (20 procedures, 19 eyes), 3 patients had unsuccessful results (3 autologous procedures), 1 patient had a partially successful procedure (allogeneic procedure), and 1 patient had a procedure with an undetermined result at present (allogeneic procedure). One unsuccessful patient had entropion/trichiasis and mechanically removed the graft and eventually went into phthisis. The other 2 unsuccessful patients suffered presumed loss of autologous donor epithelium and recurrence of the ocular surface disease (pterygium). The partially successful patient receiving an allogeneic transplant had infectious keratitis delay of his re-epithelialization; he has only minimal visual improvement but has re-epithelialized. The patient receiving the second allogeneic graft lost his donor epithelium at day 4. Additional donor epithelium was reapplied, but the result is undetermined at present. Amniotic membrane as carrier. The in vitro preparation of the amniotic membrane with corneal epithelial stem cell graft overlay was successful. Histology documented removal of the amniotic epithelium and reapplication of corneal epithelial cells. Animal model. The 2 rabbits that had no reparative surgery following standard ocular surface injury had histology and immunopathology consistent with incomplete corneal epithelial stem cell failure with vascularization and scarring of the ocular surface. Light microscopy and immunohistologic staining with AE5 confirmed the conjunctival phenotype of the ocular surface repair but also documented the incomplete model. The allogeneic stern cell transplants had varying results. One rabbit had a suppurative infection and lost the graft. Reparative surgery failed in 2 of the rabbits, failed partially in 3 of the rabbits, was partially successful in 3 others, and was successful in 1 rabbit at 28 days. Histologic and immunopathologic study documented successful growth of corneal epithelium onto the recipient surface. CONCLUSIONS: 1. Presumed corneal epithelial stem cells can be harvested safely from the limbus and expanded successfully in vitro. 2. Expanded corneal epithelial cell cultures can be grown onto various carriers, but currently denuded amniotic membrane seems to be the best carrier for ocular surface repair. 3. Expanded corneal epithelial cell transplants appear to resurface damaged ocular surfaces successfully, but cellular tracking and further confirmation are required. 4. Expanded allogeneic corneal epithelial cell transplants are technically possible and may represent alternative treatment modalities for selected ocular surface problems. 5. These techniques potentially offer a new method of restoring a normal ocular surface while minimizing the threat of damage or depletion to the contralateral or sibling limbal corneal epithelial stem cells. 6. The rabbit model was probably incomplete and should be interpreted with caution. The complete eradication of all corneal epithelial stem cells from any eye is difficult, making confirmation of such work challenging. 7. The results of the rabbit model suggest that allogeneic grafts may restore a nearly normal ocular epithelial surface to certain ocular surface injuries.

Adult↗
PubMed2004

[Molecular mechanism of pathological keratinization in severe ocular surface diseases].

PURPOSE: To investigate the molecular mechanism of pathological keratinization in severe ocular surface diseases based on the process of epidermal physiological keratinization and the gene expression profile of ocular surface epithelium. MATERIALS AND METHODS: We examined conjunctiva covering corneas which had ocular surface diseases in which pathological keratinization was seen. We then investigated the expression of epidermal keratinization-related proteins and clusterin, which is the most abundant gene transcript in human corneal epithelium. RESULTS: Semiquantitative reverse transcription-polymerase chain reaction(RT-PCR) and immunohistochemistry showed that transglutaminase 1, involucrin, filaggrin, and cytokeratin 1/10 expression was upregulated in keratinized conjunctiva compared to normal conjunctiva. On the other hand, the level of clusterin expression in the diseased ocular surfaces was significantly lower than normal. We also concluded that clusterin expression may depend on the severity of the pathological keratinization. CONCLUSION: Various keratinization-related proteins and clusterin are most likely to be involved in the pathogenesis of cicatrizing ocular surface diseases.

Clusterin↗
PubMed2005

CCR 4 and CCR 5 expression in conjunctival specimens as differential markers of T(H)1/ T(H)2 in ocular surface disorders.

BACKGROUND: Accurate inflammatory mechanisms in chronic ocular surface diseases (OSDs) cannot routinely be assessed. New techniques for investigating ocular surface inflammatory pathways are of major importance. OBJECTIVE: To investigate the expressions of CCR 4 and CCR 5, known to be related to the T(H)2 and T(H)1 systems, respectively, and HLA-DR in conjunctival impression cytology specimens from patients with chronic OSDs. METHODS: In this case-controlled study, impression cytology specimens were taken in a series of patients with vernal keratoconjunctivitis (n=21), giant papillary conjunctivitis (n=6), or keratoconjunctivitis sicca (KCS; n=17), or receiving topical antiglaucoma treatments (n=31), and from 20 normal subjects. Conjunctival cells were incubated with mAbs to CCR 4, CCR 5, CD45, and HLA-DR to quantify conjunctival inflammation in a masked manner using flow cytometry. RESULTS: HLA-DR was higher in the glaucoma and KCS groups than in allergic and normal eyes. CCR 4 was overexpressed in allergy and glaucoma, whereas CCR 5 was higher in the KCS and glaucomatous groups. CD45 was expressed by only few cells in all groups, with almost no significant differences. CCR 4 expression was negatively correlated with CCR 5 and HLA-DR, whereas CCR 5 was positively correlated with HLA-DR. CONCLUSION: This study confirms the overexpression of chemokine receptors by the conjunctival epithelium in OSDs. CCR 4 and CCR 5 expression may vary according to the immune pathway involved. Accurate mechanisms in ocular surface inflammatory reactions-that is, those related to the T(H)1 or T(H)2 systems-could be differentiated by CCR 4/CCR 5 profiles. Our results also suggest that long-term use of topical treatments may stimulate both systems.

Adolescent↗
PubMed2006

[Advances in the diagnosis and therapy for diseases of the ocular surface: dry eye and ocular allergies].

The 77th meeting of the Association for Research in Vision and Ophthalmology (ARVO) presented the latest developments in clinical research and practice across a range of ophthalmic disciplines. Of particular interest was the current status of research in dry eye and ocular allergy. With these conditions being very common to ophthalmologists, and the fact that they are difficult to differentiate due to the number of overlapping symptoms, there remains a need for more rapid and accurate methods for their diagnosis. Therefore, a number of new tools were presented that have the potential for use in clinical trials and practice to improve the diagnosis and assessment of dry eye and ocular allergy. Some of these tools have been developed based on a greater understanding of the pathophysiology of dry eye and ocular allergy, and the various molecular pathways involved. Indeed, a greater understanding of the molecular mechanisms involved in the pathogenesis of dry eye and ocular allergy has identified potential targets for the treatment of these conditions. Continuing research into new agents can provide ophthalmologists with a number of options that will ultimately benefit the patient.

Clinical Trials as Topic↗
PubMed2005

Spontaneous ocular surface inflammation and goblet cell disappearance in I kappa B zeta gene-disrupted mice.

PURPOSE: The ocular surface epithelium is part of the mucosal defense system. Because transcription factor NF-kappa B in mucosal epithelial cells plays a central role in regulating the genes that govern the onset of mucosal inflammatory responses, we examined the role of a regulator of NF-kappa B, I kappa B zeta, in murine ocular surface inflammation. METHODS: The eyes of I kappa B zeta(-/-) mice were analyzed biomicroscopically and histologically. I kappa B zeta expression in normal mouse cornea and conjunctiva was examined by RT-PCR. The results were compared with those obtained in other tissues by real-time PCR. I kappa B zeta mRNA on the ocular surface and in other mucosal tissues was localized by in situ hybridization. RESULTS: I kappa B zeta(-/-) mice manifested chronic inflammation, specifically in the ocular surface, but not in other tissues. In normal mice, I kappa B zeta was expressed in a variety of mucosal tissues. The I kappa B zeta transcript was predominantly distributed in the epithelia of these tissues. As inflammatory symptoms progressed on the ocular surface of I kappa B zeta(-/-) mice, inflammatory cells, mainly CD45R/B220(+) and CD4(+) cells, intensely infiltrated the submucosa of the conjunctival epithelia. This infiltration was accompanied by an almost complete loss of goblet cells in the conjunctival epithelia. CONCLUSIONS: The authors postulate that I kappa B zeta in the ocular surface epithelia negatively regulates the pathologic progression of ocular surface inflammation.

Adaptor Proteins, Signal Transducing↗
PubMed2000

Improvement of the ocular surface using hypotonic 0.4% hyaluronic acid drops in keratoconjunctivitis sicca.

BACKGROUND: The ocular surface changes of keratoconjunctivitis sicca (KCS) could be the result of the effect of an altered tear film on the epithelial environment. PURPOSE: To evaluate the possibility of improving the environmental conditions of the ocular surface by lowering tear osmolarity, increasing tear film volume and stabilising the tear film. Also, to study the effect of such an improvement on the epithelial cells of the ocular surface. METHODS: One hundred and thirty-five patients with a diagnosis of KCS were treated on a randomised basis with either unpreserved hypotonic 0.4% hyaluronic acid (HHA) eye drops or 0.3% hydroxypropylmethylcellulose plus 0.1% Dextran 70 (HPMC) eye drops 6 times a day for 60 or 90 days. In all patients a Schirmer I test, break-up time (BUT), ocular surface staining with 1% Bengal Rose, or 2% fluorescein, as well as subjective symptoms, were recorded before and 15, 30 and 60 days after the beginning of the study. Patients were divided into three subgroups and the effect of the treatment was studied using three different techniques: the tear ferning test, conjunctival impression cytology and tear osmolarity measurement. RESULTS: Improvements in BUT, vital staining, Schirmer I and symptoms were recorded in both groups of treatment, with significant differences for patients treated with 0.4% HHA. On day 60, 30 min after installation: tear ferning patterns changed from 100% pathological (types III-IV) to 93% physiological (types I-II) in the 0.4% HHA group and from 100% pathological to 78% physiological in the 0.3% HPMC group (p < 0.01 between groups). Tear osmolarity shifted from 353 +/- 23 to 305 +/- 6 mosmol/l in the 0.4% HHA group and from 346 +/- 15 to 336 +/- 8 mosmol/l in the 0.3% HPMC group (p < 0.001 between groups). On day 90, the impression cytology score improved from 1.2 to 1.9 in the 0.4% HHA group while it did not change in the 0.3% HPMC group (p < 0.05 between groups). CONCLUSION: In KCS appropriate treatment with a hypotonic 0.4% HHA tear substitute can change the tear environment and results in improvement of the epithelial conditions of the ocular surface.

Adjuvants, Immunologic↗
PubMed1995

Effects of ocular surface area and blink rate on tear dynamics.

OBJECTIVE: To study the effects of exposed ocular surface area and blink rate on tear dynamics by the measurement of tear evaporation under a range of conditions. METHODS: Tear evaporation was measured in three gaze positions in 15 normal volunteers, and the ocular surface area was determined as a function of the width of the palpebral fissure. In 17 normal volunteers, the effect of blink rate on tear evaporation was assessed. RESULTS: The ocular surface area was 1.2 +/- 0.27, 2.2 +/- 0.39, and 3.0 +/- 0.33 cm2 with patients looking down, ahead, and up, respectively. The corresponding tear evaporation rates per eye were 7.0 +/- 3.5, 17.6 +/- 6.6, and 23.7 +/- 6.3 x 10(-7) g/s, respectively. The tear evaporation per square meter also increased proportionally with the ocular surface area. When the blink rate was changed from 10 to 60 per minute, the tear evaporation did not change in those individuals with evaporation rates more than 7.8 x 10(-7) g/s per square centimeter, whereas it did increase with the blink rate in those whose evaporation rates were lower. CONCLUSIONS: Ocular surface area and blink rate affect tear dynamics. Moderate palpebral fissure width and blink rate are necessary for the prevention of desiccation of the ocular surface.

Adult↗
PubMed1999

Limbal autografting: comparison of results in the acute and chronic phases of ocular surface burns.

PURPOSE: To compare outcomes of limbal autograft transplantation (LAT) in the acute and chronic phases of ocular surface burns. METHODS: Retrospective analysis of case records of 16 consecutive patients who underwent LAT for ocular surface burns, at our institute, between April 1994 and March 1997. RESULTS: Limbal autograft transplantation was successful in reconstructing the corneal surface and restoring ocular comfort in 15 (93.8%) eyes. Limbal autografting failed to reconstruct the ocular surface in one patient undergoing surgery 2 weeks after grade IV alkali burns. In 13 eyes with counting fingers or worse vision, functional success (visual acuity >20/400) was attained after LAT in nine (69.2%) eyes. Visual acuity > or = 20/80 was achieved in two (25%) of eight eyes undergoing surgery for a persistent epithelial defect (PED) and five of six (83.3%) eyes undergoing surgery after the epithelial defect had healed (p = 0.03). Nine patients underwent simultaneous superior and inferior limbal autografting. Mean epithelial healing time in six of these patients undergoing surgery in the acute phase of injury (<4 months) was 15+/-6.1 days. In three patients undergoing a similar procedure in the chronic phase of injury, the healing time was 8.3+/-6.7 days. CONCLUSIONS: Limbal autograft transplantation is successful in reconstructing the corneal surface and restoring ocular comfort after ocular surface burns. Surgery in the acute phase of injury (<4 months), in the presence of a PED, could result in delayed corneal reepithelialization and poorer visual prognosis. If performed in the acute phase of injury, LAT should be performed after adequate limbal vascularization and resolution of surface inflammation in the recipient eye, avoiding graft placement over ischemic limbus.

Acids↗
PubMed1984

Conjunctival goblet cell densities in ocular surface disease.

Goblet cell densities were determined in the eyes of normal subjects and in the eyes of patients with various ocular surface diseases using an impression cytological technique. For normal eyes goblet cell densities on the interpalpebral bulbar and inferior palpebral ocular surfaces were 443 (+/- 266 [+/- SD]) and 1,972 (+/- 862) cells millimeter, per square, respectively. All patients in the ocular surface disease groups had decreased goblet cell densities compared with subjects with normal eyes. Compared with normal eyes, eyes with keratoconjunctivitis sicca (KCS) demonstrated a 17% greater goblet cell loss on the interpalpebral bulbar compared with the inferior palpebral ocular surface, while eyes with blepharitis and secondary KCS demonstrated an 8% greater loss on the inferior palpebral surface compared with the interpalpebral bulbar ocular surface. Eyes with cicatricial ocular pemphigoid and Stevens-Johnson syndrome demonstrated greater than 95% goblet cell loss on both the interpalpebral bulbar and inferior palpebral ocular surfaces compared with normal eyes.

Adolescent↗
PubMed2006

Cell markers and the side population phenotype in ocular surface epithelial stem cell characterization and isolation.

The ocular surface is covered by tworapidly renewing and embryologically-related linings, the corneal and conjunctival epithelia. The long-term survival of thesetissues is ultimately dependent on their respective resident stem cells. In the corneal epithelium, the stem cells and their early precursors are exclusively circumscribed to the narrow vasscularize limbal rim that provides epithelial precursor cells to the critically transparent central cornea. Limbal damage causes an interruption of this essential cell supply and allows the invasion of the corneal surface by the conjunctival epithelium, an event that ultimately leads to corneal scarring. The limited supply of immunocompatible tissue is a major hindrance to efforts to develop effective procedures for ocular surface reconstruction. This review describes some of the current work and strategies being developed to achieve the isolation of the limbal stem cell and define its genetic, biochemical, and functional make-up. The study of isolated ocular surface stem cells will foster basic understanding of the environmentalrequisites for their survival and proliferation in a self-replicative mode, leading eventually to advances in therapeutic approaches.

ATP-Binding Cassette Transporters↗
PubMed1998

Adenovirus-mediated gene transfer to the ocular surface epithelium.

Gene transfer to the ocular surface epithelium is of potential therapeutic value. It was determined whether a reporter gene can be introduced into the ocular surface epithelium in vitro (human cell lines), ex vivo (human tissues), and in vivo (rats) by treating with a recombinant, replication-deficient, adenovirus type 5. Human and conjunctival cell lines were cultured with various multiplicities of infection (MOI; 3.2x10(-5)-5x10(-1)) of adenovirus vector (Ad5:Adex1CAlacZ) containing the reporter gene lacZ (1.3-2.0x10(4) PFU ml-1). The ex vivo study used human corneal and conjunctival tissues obtained from an eye bank and during surgery. Non-specific upregulation of inflammatory cytokines of conjunctival epithelium infected by Ad5 was assayed and its suppression by steroids. For the in vivo study, Ad5 (5x10(5) PFU, 5-10 microliter) was applied to the eyes of 8-12-week-old cotton rats, which were enucleated 24 and 48 hr later. The maximum lacZ expression in vitro was demonstrated in the corneal epithelial cell line at 7 days (1x10(-1) MOI) and conjunctival epithelial cell line at 2 days (4x10(-4) MOI). Furthermore, lacZ was also expressed in the superficial corneal and conjunctival epithelium in the ex vivo study. IL-6, IL-8, and ICAM-1 expression from conjunctival epithelium by Ad5 was significantly inhibited by treatment with betamethasone (BM). For the in vivo study, only the conjunctival epithelium demonstrated beta-Gal activity at 24 and 48 hr after application. These data indicate that adenovirus vector is capable of directly delivering gene to the corneal and conjunctival epithelium, suggesting a variety of possible gene therapy uses. The concomitant application of steroid eye drops may avoid inflammation.

Adenoviridae↗
PubMed2001

Reconstruction of ocular surface with heterologous limbal epithelium and amniotic membrane in a rabbit model.

PURPOSE: To report in vivo reconstruction of the ocular surface using amniotic membrane and heterologous transplants of epithelial limbal cells in rabbits with chemical burns. METHODS: After severe damage to the ocular surface with n-heptanol and keratectomy, 15 rabbits developed total limbal deficiency with conjunctival epithelialization, vascularization, and chronic inflammation. One month later, a complete keratectomy was performed in all eyes: 12 received additional transplantation of human amniotic membrane and heterologous limbal epithelial cells in a double amniotic membrane layer, 2 received amniotic membrane only, and 1 control eye received no procedure. RESULTS: After 1 month of follow-up, corneas in eight of the operated eyes presented minimal vascularization, without signs of rejection. Corneal surface reconstruction was demonstrated with the growth of new corneal-like epithelial phenotype and integration of amniotic membrane to the basal corneal surface. A superficial amniotic membrane (with the amnion side up as a dressing) peeled off after 7 to 10 days. The epithelialization with heterologous limbal epithelial cells was evident underneath. The other four operated eyes were followed for 6 months; the ocular surface was also stable with a corneal-like epithelial phenotype. CONCLUSION: Simultaneous transplantation of amniotic membrane and heterologous limbal epithelial cells in severe ocular surface disorders could restore ocular surface and may be useful in patients with severe bilateral limbal epithelial loss, giving new perspectives for the treatment of severe ocular surface disorders.

Amnion↗
PubMed2004

Concept and clinical application of cultivated epithelial transplantation for ocular surface disorders.

Corneal epithelial replacement using a tissue engineering technique holds much promise for ocular surface reconstruction in cases of corneal epithelial stem cell deficiency. However, even though an autologous cultivated corneal epithelial stem cell sheet is the safest and most reliable form of sheet, bilaterally affected ocular surface disorders cannot be treated by this method. To treat bilateral cases, we must choose either an allogeneic cultivated corneal epithelial sheet or an autologous cultivated oral mucosal epithelial sheet. In the case of the former, immunological reaction is a threat. Thus, understanding of the immunological background of ocular surface reconstruction using allogeneic tissues is essential. In the case of the latter, the transplanted sheet is not exactly the same as corneal epithelium, and understanding ocular surface epithelial biology is important. In this review, we summarize and explain the concept and clinical application of cultivated mucosal epithelial transplantation for ocular surface disorders.

Journal Article↗
PubMed2004

Character of ocular surface mucins and their alteration in dry eye disease.

At the ocular surface, three types of mucins are present. The large gel-forming mucin MUC5AC is expressed by conjunctival goblet cells. Some cells of the lacrimal gland acini express the small soluble mucin MUC7. The corneal and conjunctival epithelia express the membrane-associated mucins MUCs 1, 4, and 16. With the characterization of the mucin gene repertoire of the ocular surface epithelia, studies of the function of specific mucins, their gene regulation, and their alteration in ocular surface disease have begun. Current information suggests that all the mucins are hydrophilic and play a role in maintenance of water on the surface of the eye. The large secreted mucins represent the "janitorial service" that moves over the surface of the eye to wrap up and remove debris. The membrane-associated mucins form the glycocalyx, which provides a continuous barrier across the surface of the eye that prevents pathogen penetrance and has signaling capabilities that influence epithelial activity. Factors regulating mucin gene expression include retinoic acid, serum, and dexamethasone. Alteration in both secreted and membrane-associated mucins occur in drying ocular surface diseases. In Sjogren syndrome, MUC5AC expression is reduced, and in non-Sjogren dry eye, glycosylation of MUC16 appears to be altered. The pattern of expression of enzymes that glycosylate mucins is altered in ocular cicatricial pemphigoid. Therapies being evaluated for dry eye, including cyclosporine A, P2Y2 agonists, gefarnate, 15-(S)-HETE, and corticosteroids, may be efficacious due to their effect on mucin gene expression and secretion.

Journal Article↗
PubMed1984

Dynamics of ocular surface pH.

We studied ocular surface pH in 161 subjects. The mean pH for 133 normal volunteers was 7 . 11, SD 1 . 5. We found that older women had a more alkaline pH than other subjects, that the pH shifted from acid to alkaline during the day, that one hour of eyelid closure caused an acid shift in pH, and that pH recovered to baseline values within 40 minutes after acid drop instillation. We explored the mechanism of pH regulation, and we believe that pH changes could affect contact lens toleration, drug effectiveness, and clinical signs in disease processes.

Adult↗
PubMed2003

Ocular surface impression cytology.

Impression cytology, either using cellulose acetate strips or the Biopore membrane device, is a simple, noninvasive technique that aids in the diagnosis of several disorders of the ocular surface. These disorders include ocular surface squamous neoplasia, dry eye syndrome, limbal stem-cell deficiency, specific viral infections, vitamin A deficiency, allergic disorders, conjunctival melanosis, and malignant melanoma. Another advantage is the preservation of limbal stem cells, which occur in the basal layer of the limbal epithelium and are responsible for renewal of the corneal epithelium. The Biopore membrane device is particularly user friendly, with little expertise required and adequate specimens obtained in a very high percentage of cases. The most common applications in diagnostic ocular pathology are:(i) primary diagnosis and follow-up of ocular surface squamous neoplasia, including after therapy with topical mitomycin C. The sensitivity is high (78-87%); and (ii) dry eye syndrome where squamous metaplasia and/ or hyperkeratosis are noted. Certain limitations of the technique for diagnosis of squamous neoplasia include the fact that dysplasias are often keratinizing and may yield very few or even no dysplastic cells with impression cytology. Secondly, no definite cytologic criteria reliably distinguish invasive SCC of ocular surface from in situ disease. Other applications include the rapid specific diagnosis of ocular surface infections with herpes simplex, adeno-, and varicella zoster viruses. Impression cytology samples may also be used to obtain mRNA, cells for phenotyping by flow cytometry, and proteins for Western blotting for research studies.

Cytodiagnosis↗