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Article ; Online: Cornea-Specific Human Adipose Stem Cell-Derived Extracellular Matrix for Corneal Stroma Tissue Engineering.

Puistola, Paula / Kethiri, Abhinav / Nurminen, Antti / Turkki, Johannes / Hopia, Karoliina / Miettinen, Susanna / Mörö, Anni / Skottman, Heli

ACS applied materials & interfaces

2024 Volume 16, Issue 13, Page(s) 15761–15772

Abstract: Utilizing tissue-specific extracellular matrices (ECMs) is vital for replicating the composition of native tissues and developing biologically relevant biomaterials. Human- or animal-derived donor tissues and organs are the current gold standard for the ... ...

Abstract	Utilizing tissue-specific extracellular matrices (ECMs) is vital for replicating the composition of native tissues and developing biologically relevant biomaterials. Human- or animal-derived donor tissues and organs are the current gold standard for the source of these ECMs. To overcome the several limitations related to these ECM sources, including the highly limited availability of donor tissues, cell-derived ECM offers an alternative approach for engineering tissue-specific biomaterials, such as bioinks for three-dimensional (3D) bioprinting. 3D bioprinting is a state-of-the-art biofabrication technology that addresses the global need for donor tissues and organs. In fact, there is a vast global demand for human donor corneas that are used for treating corneal blindness, often resulting from damage in the corneal stromal microstructure. Human adipose tissue is one of the most abundant tissues and easy to access, and adipose tissue-derived stem cells (hASCs) are a highly advantageous cell type for tissue engineering. Furthermore, hASCs have already been studied in clinical trials for treating corneal stromal pathologies. In this study, a corneal stroma-specific ECM was engineered without the need for donor corneas by differentiating hASCs toward corneal stromal keratocytes (hASC-CSKs). Furthermore, this ECM was utilized as a component for corneal stroma-specific bioink where hASC-CSKs were printed to produce corneal stroma structures. This cost-effective approach combined with a clinically relevant cell type provides valuable information on developing more sustainable tissue-specific solutions and advances the field of corneal tissue engineering.
MeSH term(s)	Animals ; Humans ; Tissue Engineering/methods ; Corneal Stroma/metabolism ; Cornea ; Extracellular Matrix/chemistry ; Biocompatible Materials/metabolism ; Adipose Tissue ; Stem Cells ; Tissue Scaffolds ; Bioprinting/methods
Chemical Substances	Biocompatible Materials
Language	English
Publishing date	2024-03-21
Publishing country	United States
Document type	Journal Article
ISSN	1944-8252
ISSN (online)	1944-8252
DOI	10.1021/acsami.3c17803
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Article ; Online: Current perspectives of limbal-derived stem cells and its application in ocular surface regeneration and limbal stem cell transplantation.

Singh, Vivek / Tiwari, Anil / Kethiri, Abhinav Reddy / Sangwan, Virender Singh

Stem cells translational medicine

2021 Volume 10, Issue 8, Page(s) 1121–1128

Abstract: Limbal stem cells are involved in replenishing and maintaining the epithelium of the cornea. Damage to the limbus due to chemical/physical injury, infections, or genetic disorders leads to limbal stem cell deficiency (LSCD) with partial or total vision ... ...

Abstract	Limbal stem cells are involved in replenishing and maintaining the epithelium of the cornea. Damage to the limbus due to chemical/physical injury, infections, or genetic disorders leads to limbal stem cell deficiency (LSCD) with partial or total vision loss. Presently, LSCD is treated by transplanting limbal stem cells from the healthy eye of the recipient, living-related, or cadaveric donors. This review discusses limbal-derived stem cells, the importance of extracellular matrix in stem cell niche maintenance, the historical perspective of treating LSCD, including related advantages and limitations, and our experience of limbal stem cell transplantation over the decades.
MeSH term(s)	Cell Transplantation ; Cells, Cultured ; Corneal Diseases/therapy ; Epithelium, Corneal ; Humans ; Limbus Corneae ; Stem Cell Transplantation
Language	English
Publishing date	2021-05-05
Publishing country	England
Document type	Journal Article ; Research Support, Non-U.S. Gov't ; Review
ZDB-ID	2642270-0
ISSN	2157-6580 ; 2157-6580
ISSN (online)	2157-6580
ISSN	2157-6580
DOI	10.1002/sctm.20-0408
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Article ; Online: Development and validation of a reliable rabbit model of limbal stem cell deficiency by mechanical debridement using an ophthalmic burr.

Singh, Vijay Kumar / Kethiri, Abhinav Reddy / Pingali, Tejaswini / Sahoo, Abhishek / Salman, Mohd / Koduri, Madhuri Amulya / Prasad, Deeksha / Bokara, Kiran Kumar / Basu, Sayan / Singh, Vivek

Experimental eye research

2023 Volume 236, Page(s) 109667

Abstract: A simple and reproducible method is necessary to generate reliable animal models of limbal stem cell deficiency (LSCD) for assessing the safety and efficacy of new therapeutic modalities. This study aimed to develop and validate a rabbit model of LSCD ... ...

Abstract	A simple and reproducible method is necessary to generate reliable animal models of limbal stem cell deficiency (LSCD) for assessing the safety and efficacy of new therapeutic modalities. This study aimed to develop and validate a rabbit model of LSCD through mechanical injury. The corneal and limbal epithelium of New Zealand White rabbits (n = 18) were mechanically debrided using an ophthalmic burr (Algerbrush II) with a 1.0-mm rotating head after 360° conjunctival peritomy. The debrided eyes were serially evaluated for changes in corneal opacity, neo-vascularization, epithelial defect and corneal thickness using clinical photography, slit lamp imaging, fluorescein staining, and anterior segment optical coherence tomography scanning (AS-OCT). Following this, an assessment of histopathology and phenotypic marker expression of the excised corneas was conducted. The experimental eyes were grouped as mild (n = 4), moderate (n = 10), and severe (n = 4) based on the grade of LSCD. The moderate group exhibited abnormal epithelium, cellular infiltration in the stroma, and vascularization in the central, peripheral, and limbal regions of the cornea. The severe group demonstrated central epithelial edema, peripheral epithelial thinning with sparse goblet cell population, extensive cellular infiltration in the stroma, and dense vascularization in the limbal region of the cornea. A significant decrease in the expression of K12 and p63 (p < 0.0001) was observed, indicating the loss of corneal epithelium and limbal epithelial stem cells in the LSCD cornea. This study demonstrates that the Alger brush-induced mechanical debridement model provides a reliable model of LSCD with comprehensive clinic-pathological features and that is well suited for evaluating novel therapeutic and regenerative approaches.
MeSH term(s)	Rabbits ; Animals ; Limbus Corneae/metabolism ; Debridement ; Limbal Stem Cells ; Cornea/metabolism ; Epithelium, Corneal/metabolism ; Corneal Diseases/pathology
Language	English
Publishing date	2023-09-26
Publishing country	England
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	80122-7
ISSN	1096-0007 ; 0014-4835
ISSN (online)	1096-0007
ISSN	0014-4835
DOI	10.1016/j.exer.2023.109667
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Article ; Online: Histopathological Characteristics of Limbal Stem Cell Deficiency Secondary to Chronic Vernal Keratoconjunctivitis.

Donthineni, Pragnya R / Varma, Shobhit / Kethiri, Abhinav / Shanbhag, Swapna / Mishra, Dilip K / Singh, Vivek / Basu, Sayan

Cornea

2021 Volume 41, Issue 6, Page(s) 722–728

Abstract: Purpose: To describe the histopathological characteristics of limbal stem cell deficiency (LSCD) due to chronic vernal keratoconjunctivitis (VKC).: Methods: This retrospective study included 14 eyes of 13 patients who underwent simple limbal ... ...

Abstract	Purpose: To describe the histopathological characteristics of limbal stem cell deficiency (LSCD) due to chronic vernal keratoconjunctivitis (VKC). Methods: This retrospective study included 14 eyes of 13 patients who underwent simple limbal epithelial transplantation for total LSCD from 2017 to 2018. The histological characteristics of the excised fibrovascular pannus were compared between 2 groups of 7 eyes, each with LSCD due to VKC and chemical burns (CB). Histological characteristics and type of inflammation were studied using special stains and immunohistochemistry. Fisher exact test was used to detect the statistical significance of the histological differences between both groups. Results: Epithelial hypertrophy, epithelial downgrowth, and eosinophilic infiltration were noted in all eyes in the VKC group (7/7, 100%). Epithelial hypertrophy was noted in 3 of the 7 (42.8%) eyes in the CB group, whereas epithelial downgrowth and eosinophilic infiltrates were absent. The average chronic inflammatory score of the pannus (5.28) was higher in VKC than in CB (3.85; P = 0.1080). The presence of goblet cells was higher in the CB group (5/7, 1.4%) than in the VKC group (3/4, 2.8%), although not statistically significant. Other histological differences between the groups were not statistically significant. Conclusions: The histopathological features of LSCD in VKC reveal some distinctive characteristics. These include the presence of epithelial downgrowth, eosinophilic infiltration, and epithelial solid and cystic implants. Although this information may be used to establish the diagnostic criteria for VKC as the cause of LSCD, further studies are needed to elucidate the reasons behind these unique findings.
MeSH term(s)	Burns, Chemical/pathology ; Conjunctivitis, Allergic/complications ; Conjunctivitis, Allergic/diagnosis ; Corneal Diseases/diagnosis ; Corneal Neovascularization/pathology ; Graft vs Host Disease/pathology ; Humans ; Inflammation/pathology ; Limbus Corneae/pathology ; Retrospective Studies ; Scleral Diseases/pathology ; Stem Cells/pathology
Language	English
Publishing date	2021-06-08
Publishing country	United States
Document type	Journal Article
ZDB-ID	604826-2
ISSN	1536-4798 ; 0277-3740
ISSN (online)	1536-4798
ISSN	0277-3740
DOI	10.1097/ICO.0000000000002775
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Article ; Online: Long term observation of ocular surface alkali burn in rabbit models: Quantitative analysis of corneal haze, vascularity and self-recovery.

Kethiri, Abhinav Reddy / Singh, Vijay Kumar / Damala, Mukesh / Basu, Sayan / Rao, Ch Mohan / Bokara, Kiran Kumar / Singh, Vivek

Experimental eye research

2021 Volume 205, Page(s) 108526

Abstract: Limbal Stem Cell Deficiency (LSCD), caused due to corneal injury, primarily by chemical/alkali burns, leads to compromised vision. Recently, several animal models of corneal alkali burn injury have become available. The majority of the studies with these ...

Abstract	Limbal Stem Cell Deficiency (LSCD), caused due to corneal injury, primarily by chemical/alkali burns, leads to compromised vision. Recently, several animal models of corneal alkali burn injury have become available. The majority of the studies with these animal models start interventions soon after the injury. However, in the clinical setting, there is a considerable delay before the intervention is initiated. Detailed knowledge of the molecular, histopathological, and clinical parameters associated with the progression of the injury leading to LSCD is highly desirable. In this context, we set out to investigate clinical, histopathological parameters of ocular surface alkali burn over a long period of time, post-injury. Limbal stem cell-deficient animal models of rabbits were created by alkali burn using sodium hydroxide, which was then assessed for their progression towards LSCD by grading the alkali burn, corneal haze, and vascularization. Additionally, cells present on the corneal surface after the burn was investigated by histology and immunophenotyping. Grading of rabbit eyes post-alkali burn had shown complete conjunctivalization in 80% (n = 12/15) of the rabbits with the alkali burn grade score of 3.88 ± 0.29 in three months and remained stable at four months (4.12 ± 0.24). However, ocular surface showed self-healing in 20% (n = 3/15) of the rabbits with a score of 1.67 ± 0.34 in four months irrespective of similar alkali injury. These self-healing corneas exhibited decreased opacity score from 2.51 ± 0.39 to 0.66 ± 0.22 (p = 0.002) and regressed vascularity from 1.66 ± 0.41 to 0.66 ± 0.33 in one to nine months, respectively. Restoration of the corneal phenotype (CK3+) was observed in central and mid-peripheral regions of the self-healing corneas, and histology revealed the localization of inflammatory cells to the peripheral cornea when compared to conjunctivalized and scarred LSCD eyes. Our study shows the essentiality to consider the time required for surgical intervention after the corneal alkali injury in rabbit models as evident from their tendency to self-heal and restore corneal phenotype without therapy. Such information on the possibility of self-healing should be useful in further studies as well as determining interventional timings and strategy during clinical presentation of corneal alkali burns.
MeSH term(s)	Animals ; Burns, Chemical/physiopathology ; Caustics/toxicity ; Conjunctiva/physiopathology ; Cornea/physiopathology ; Corneal Injuries/physiopathology ; Corneal Neovascularization/physiopathology ; Corneal Opacity/physiopathology ; Disease Models, Animal ; Eye Burns/chemically induced ; Eye Burns/physiopathology ; Follow-Up Studies ; Limbus Corneae/cytology ; Rabbits ; Recovery of Function/physiology ; Sodium Hydroxide/toxicity ; Stem Cell Transplantation ; Wound Healing/physiology
Chemical Substances	Caustics ; Sodium Hydroxide (55X04QC32I)
Language	English
Publishing date	2021-03-02
Publishing country	England
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	80122-7
ISSN	1096-0007 ; 0014-4835
ISSN (online)	1096-0007
ISSN	0014-4835
DOI	10.1016/j.exer.2021.108526
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Article ; Online: Optimizing the role of limbal explant size and source in determining the outcomes of limbal transplantation: An in vitro study.

Kethiri, Abhinav Reddy / Basu, Sayan / Shukla, Sachin / Sangwan, Virender Singh / Singh, Vivek

PloS one

2017 Volume 12, Issue 9, Page(s) e0185623

Abstract: Purpose: Simple limbal epithelial transplantation (SLET) and cultivated limbal epithelial transplantation (CLET) are proven clinical techniques for treating limbal stem cell deficiency (LSCD). However, the ideal size and number of the limbal explants ... ...

Abstract	Purpose: Simple limbal epithelial transplantation (SLET) and cultivated limbal epithelial transplantation (CLET) are proven clinical techniques for treating limbal stem cell deficiency (LSCD). However, the ideal size and number of the limbal explants required for transplantation has not been clearly elucidated. This in vitro study aimed to determine the optimal limbal explant size required for complete corneal epithelialization by characterizing the cell expansion. Methods: Limbal explants obtained from both live and cadaveric biopsies were cultured on the denuded amniotic membrane. Explant size and the explant cell outgrowth (expansion) were measured using ImageJ software with respect to days. Cultures were characterized by assessing the rate of proliferation of cells with 5-bromo-2'-deoxyuridine (BrdU) assay along with the expression of different stem cell markers (ABCG2, p63α), corneal epithelial (CK3+12) and adherens junction molecules (E-Cadherin) by immunofluorescence. Results: Explants from live biopsies had 80% growth potential in vitro whereas 40% of the cadaveric tissue failed to grow. Minimum explant sizes of 0.3 mm2 for live and ≥0.5 mm2 for cadaveric tissue had a mean expansion areas of 182.39±17.06 mm2 and 217.59±16.91 mm2 respectively suggesting adequate growth potential of the explants. Mean total percentage of proliferative cells was 31.80±3.81 in live and 33.49±4.25 in cadaveric tissue expansion. The expression was noted to be similar in cells cultured from cadaveric compared to cells cultured from live limbal tissue with respect to ABCG2, p63α, CK(3+12) and E-cadherin. Conclusion: Our findings show that a minimal amount of 0.3 mm2 live tissue would be sufficient for ample limbal cell expansion in vitro. Cadaveric explants <0.5 mm2 had poor growth potential. However, larger explants (≥ 0.5 mm2) had growth rate and proliferative potential similar to the live tissue. These findings could prove to be critical for clinical success especially while attempting cadaveric limbal transplantation. This study provides a novel clinical strategy for enhancing efficacy of the limbal transplantation surgery and opens the probability of even using the cadaveric tissue by considering the size of explant.
MeSH term(s)	Bromodeoxyuridine/metabolism ; Cadaver ; Cell Proliferation ; Corneal Transplantation ; Epithelium, Corneal/cytology ; Fluorescent Antibody Technique ; Humans ; In Vitro Techniques ; Limbus Corneae/cytology
Chemical Substances	Bromodeoxyuridine (G34N38R2N1)
Language	English
Publishing date	2017
Publishing country	United States
Document type	Journal Article
ISSN	1932-6203
ISSN (online)	1932-6203
DOI	10.1371/journal.pone.0185623
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Article ; Online: Tuning Electrospun Substrate Stiffness for the Fabrication of a Biomimetic Amniotic Membrane Substitute for Corneal Healing.

Paterson, Thomas E / Dhowre, Hala S / Villanueva, Danilo / Holland, Joseph W / Reddy Kethiri, Abhinav / Singh, Vivek / Claeyssens, Frederik / MacNeil, Sheila / Ortega Asencio, Ilida

ACS applied bio materials

2021 Volume 4, Issue 7, Page(s) 5638–5649

Abstract: Corneal blindness is the fourth most common cause of vision impairment worldwide with a high incidence in global south countries. A recently developed surgical technique for treating corneal blindness is simple limbal epithelial transplantation (SLET), ... ...

Abstract	Corneal blindness is the fourth most common cause of vision impairment worldwide with a high incidence in global south countries. A recently developed surgical technique for treating corneal blindness is simple limbal epithelial transplantation (SLET), which uses small pieces of healthy limbal tissue (limbal explants) delivered to the damaged eye using the human amniotic membrane (AM) as a carrier. SLET relies on the use of tissue banks for the AM that reduces the availability of the technique. Replacing the AM with a synthetic membrane is key to making SLET more accessible to those who need it. Previous research has demonstrated the suitability of electrospun poly(lactide-
MeSH term(s)	Amnion/transplantation ; Animals ; Biomimetics ; Blindness ; Cornea ; Rabbits ; Swine ; Wound Healing
Language	English
Publishing date	2021-06-29
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ISSN	2576-6422
ISSN (online)	2576-6422
DOI	10.1021/acsabm.1c00436
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Article ; Online: Inflammation, vascularization and goblet cell differences in LSCD: Validating animal models of corneal alkali burns.

Kethiri, Abhinav Reddy / Raju, Enoch / Bokara, Kiran Kumar / Mishra, Dilip Kumar / Basu, Sayan / Rao, Ch Mohan / Sangwan, Virender Singh / Singh, Vivek

Experimental eye research

2019 Volume 185, Page(s) 107665

Abstract: Limbal stem cell deficiency (LSCD) is one of the serious cause of visual impairment and blindness with loss of corneal clarity and vascularization. Factors such as ocular burns (acids, lime, thermal), genetic disorders or infections results in the loss ... ...

Abstract	Limbal stem cell deficiency (LSCD) is one of the serious cause of visual impairment and blindness with loss of corneal clarity and vascularization. Factors such as ocular burns (acids, lime, thermal), genetic disorders or infections results in the loss of limbal stem cells leading to LSCD. Reliable animal models of LSCD are useful for understanding the pathophysiology and developing novel therapeutic approaches. The purpose of the present study was to validate small and large animal models of LSCD by immunohistochemcal, clinical and histopathological comparison with human. The animal models of LSCD were created by topical administration of sodium hydroxide on the ocular surface of C57BL/6 mice (m, n = 12) and New Zealand white rabbits (r, n = 12) as per the standard existing protocol. Human corneal specimens (h, n = 12) were obtained from tissue bank who had chemical burn-induced LSCD. All samples were either paraffin embedded or frozen in cryogenic medium and the sections were processed for Hematoxylin-Eosin and Periodic Acid-Schiff staining to analyse the morphology and histopathological features of the corneal surface such as vascularization, inflammation, presence of goblet cells, epithelial hyperplasia and keratinization. Immunofluorescence was performed to distinguish between corneal (CK3+), conjunctival (CK19+) and epidermal (CK10+) epithelial phenotype. Histological analysis of corneal specimens from the three groups showed the presence of goblet cells (h:83%, m:50%, r:50%, p = 0.014), epithelial hypertrophy (h:92%, m:50%, r:66.6%, p = 0.04), epithelial hyperplasia (h:50%, m:17%, r:17%, p = 0.18), intra epithelial edema (h:42%, m:33%, r:100%, p = 0.02), stromal inflammation (h:100%, m:67%, r:67%, p = 0.01) and stromal vascularization (h:100%, m:50%, r:67%), in varying proportions. Immunostaining showed presence of total LSCD (CK19 + and/or CK10+, CK3-) in 92% of human and 50% of animal specimens. While partial LSCD (CK19 + and/or CK10+, CK3+) was seen in 8% of human and 50% of animal specimens. Our study shows the significant differences in the extent of vascularization, inflammation, epithelial thickness and goblet cell formation in mice and rabbit models of LSCD when compared to post-chemical burn LSCD in human corneas. In both mice and rabbit models complete LSCD developed in only 50% of cases and this important fact needs to be considered when working with animal models of LSCD.
MeSH term(s)	Animals ; Burns, Chemical/metabolism ; Burns, Chemical/pathology ; Corneal Diseases/metabolism ; Corneal Diseases/pathology ; Corneal Neovascularization/metabolism ; Corneal Neovascularization/pathology ; Disease Models, Animal ; Epithelial Cells/metabolism ; Epithelium, Corneal ; Eye Burns/chemically induced ; Eye Burns/metabolism ; Female ; Fluorescent Antibody Technique, Indirect ; Goblet Cells/metabolism ; Goblet Cells/pathology ; Humans ; Immunophenotyping ; Inflammation/metabolism ; Inflammation/pathology ; Keratin-19/metabolism ; Keratin-3/metabolism ; Keratitis/metabolism ; Keratitis/pathology ; Limbus Corneae/metabolism ; Limbus Corneae/pathology ; Male ; Mice ; Mice, Inbred C57BL ; Mucins/metabolism ; Rabbits ; Sodium Hydroxide/toxicity
Chemical Substances	Keratin-19 ; Keratin-3 ; Mucins ; Sodium Hydroxide (55X04QC32I)
Language	English
Publishing date	2019-05-13
Publishing country	England
Document type	Comparative Study ; Journal Article ; Research Support, Non-U.S. Gov't ; Validation Study
ZDB-ID	80122-7
ISSN	1096-0007 ; 0014-4835
ISSN (online)	1096-0007
ISSN	0014-4835
DOI	10.1016/j.exer.2019.05.005
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Article ; Online: Simple Limbal Epithelial Transplantation: Long-Term Clinical Outcomes in 125 Cases of Unilateral Chronic Ocular Surface Burns.

Basu, Sayan / Sureka, Shraddha P / Shanbhag, Swapna S / Kethiri, Abhinav R / Singh, Vivek / Sangwan, Virender S

Ophthalmology

2016 Volume 123, Issue 5, Page(s) 1000–1010

Abstract: Purpose: This study describes the long-term clinical outcomes of autologous simple limbal epithelial transplantation (SLET), a relatively new technique of limbal stem cell transplantation.: Design: This was a single-center prospective interventional ... ...

Abstract	Purpose: This study describes the long-term clinical outcomes of autologous simple limbal epithelial transplantation (SLET), a relatively new technique of limbal stem cell transplantation. Design: This was a single-center prospective interventional cases series. Participants: This study included 125 patients, 65 adults and 60 children who developed unilateral limbal stem cell deficiency (LSCD) after suffering with ocular surface burns and underwent SLET between 2010 and 2014. Methods: A 1-clock hour limbal biopsy sample was obtained from the unaffected eye. At the same sitting, the recipient eye was surgically prepared and the donor tissue was divided into small pieces and transplanted using an amniotic membrane scaffold with fibrin glue. Main outcome measures: The diagnosis and outcome in every case was validated by 5 independent masked assessors. The primary outcome measure was restoration of a completely epithelized, stable, and avascular corneal surface. The secondary outcome measure was improvement in visual acuity. Complications, risk factors for failure, and immunohistochemistry analysis of corneas that underwent SLET also were described. Results: At a median postoperative follow-up of 1.5 years (range, 1-4 years), 95 of 125 eyes (76%; 95% confidence interval, 68.5%-83.5%) maintained a successful outcome. Kaplan-Meier analysis revealed a comparable survival probability at 1 year of 80% in adults and 72% in children (P = 0.304). Two-line improvement in visual acuity was seen in 75.2%, and 67% of successful cases attained 20/60 or better vision (P < 0.0001). Progressive conjunctivalization occurred in 18.4% of eyes. The clinical factors associated with failure were identified as acid injury, severe symblepharon, SLET combined with keratoplasty, and postoperative loss of transplants (P ≤ 0.0075). Success rates were comparable among faculty and trainees (P = 0.71). Immunohistochemistry revealed successful regeneration of normal corneal epithelium (CK3(+)/12(+)) without admixture of conjunctiva cells (Muc5AC(-)/CK19(-)) and replenishment of limbal stem cell (ΔNp63α(+)/ABCG2(+)) reserve. Conclusions: Autologous SLET is an effective, reliable and replicable technique for long-lasting corneal regeneration and vision restoration in unilateral chronic ocular surface burns. Simple limbal epithelial transplantation is probably preferable to other techniques of limbal stem cell transplantation, particularly where cell cultivation facilities are unavailable.
MeSH term(s)	Adolescent ; Adult ; Burns, Chemical/physiopathology ; Burns, Chemical/surgery ; Child ; Chronic Disease ; Epithelial Cells/transplantation ; Epithelium, Corneal/physiology ; Eye Burns/chemically induced ; Eye Burns/surgery ; Female ; Follow-Up Studies ; Graft Rejection/diagnosis ; Humans ; Limbus Corneae/cytology ; Male ; Prospective Studies ; Regeneration/physiology ; Risk Factors ; Stem Cell Transplantation ; Transplantation, Autologous ; Visual Acuity/physiology ; Wound Healing/physiology ; Young Adult
Language	English
Publishing date	2016
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	392083-5
ISSN	1549-4713 ; 0161-6420
ISSN (online)	1549-4713
ISSN	0161-6420
DOI	10.1016/j.ophtha.2015.12.042
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Article ; Online: Differential Expression of Stem Cell Markers in Ocular Surface Squamous Neoplasia.

Mishra, Dilip Kumar / Veena, Uppala / Kaliki, Swathi / Kethiri, Abhinav Reddy / Sangwan, Virender S / Ali, Mohammed Hasnat / Naik, Milind N / Singh, Vivek

PloS one

2016 Volume 11, Issue 9, Page(s) e0161800

Abstract: Ocular Surface Squamous Neoplasm (OSSN) is the neoplasia arising from the conjunctiva, cornea and limbus. OSSN ranges from mild, moderate, severe dysplasia, carcinoma in situ (CIS) to squamous cell carcinoma (SCC). Recent findings on cancer stem cells ... ...

Abstract	Ocular Surface Squamous Neoplasm (OSSN) is the neoplasia arising from the conjunctiva, cornea and limbus. OSSN ranges from mild, moderate, severe dysplasia, carcinoma in situ (CIS) to squamous cell carcinoma (SCC). Recent findings on cancer stem cells theory indicate that population of stem-like cell as in neoplasia determines its heterogeneity and complexity leading to varying tumor development of metastatic behavior and recurrence. Cancer stem cell markers are not much explored in the cases of OSSN. In the present study, we aim to evaluate the expression of stem cells using stem cell markers mainly p63, ABCG2, c-KIT (CD117) and CD44 in OSSN tissue, which could have prognostic significance. The present study tries for the first time to explore expression of these stem markers in the cases of OSSN. These cases are subdivided into two groups. One group comprises of carcinoma in situ (n = 6) and the second group comprises of invasive carcinoma (n = 6). The mean age at presentation was 52 years; with 53 years for CIS group and 52 years for SCC group. From each group section from the paraffin block were taken for the IHC staining of p63, c-Kit, ABCG2 and CD44. Our experiments show high expression of P63 and CD44 in the cases of CIN and SCC. Both CIS and SCC displayed positive staining with p63, with more than 80% cells staining positive. However minimal expression of c-kit in both CIN and SCC. But surprisingly we got high expression of ABCG2 in cases of carcinoma in situ as compared to that of invasive squamous cell carcinoma. More than 50% of cells showed CD44 positivity in both CIS and SCC groups. Our results show for the first time that these four stem cells especially the limbal epithelium stem cells play a vital role in the genesis of OSSN but we need to explore more cases before establishing its clinical and biological significance.
MeSH term(s)	Adult ; Aged ; Aged, 80 and over ; Biomarkers, Tumor/metabolism ; Carcinoma, Squamous Cell/diagnosis ; Carcinoma, Squamous Cell/metabolism ; Carcinoma, Squamous Cell/pathology ; Eye Neoplasms/diagnosis ; Eye Neoplasms/metabolism ; Eye Neoplasms/pathology ; Female ; Gene Expression Regulation, Neoplastic ; Humans ; Male ; Middle Aged ; Neoplastic Stem Cells/metabolism ; Prognosis
Chemical Substances	Biomarkers, Tumor
Language	English
Publishing date	2016-09-01
Publishing country	United States
Document type	Journal Article
ZDB-ID	2267670-3
ISSN	1932-6203 ; 1932-6203
ISSN (online)	1932-6203
ISSN	1932-6203
DOI	10.1371/journal.pone.0161800
Database	MEDical Literature Analysis and Retrieval System OnLINE

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