A novel application of electrospun silk fibroin/poly(<scp>l</scp>-lactic acid-<i>co</i>-ε-caprolactone) scaffolds for conjunctiva reconstruction

Yao, Qinke; Hu, Yang; Yu, Fei; Zhang, Weijie; Fu, Yao

doi:10.1039/c7ra13551c

Cited by 22 publications

(12 citation statements)

References 39 publications

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“…Also, it is important to be careful with sterility to prevent bacterial or fungal contamination. The methods of cell isolation and cultivation have been described previously 40,41 . Briefly, conjunctiva was rinsed and soaked several times in phosphate buffered saline (PBS) containing penicillin, and incubated in Dispase II (neutral protease, grade II) (5 mg/mL, Roche Diagnostics GmbH, Manheim, Germany) at 4°C for 14 to 16 hours.…”

Section: Methodsmentioning

confidence: 99%

“…The methods of cell isolation and cultivation have been described previously. 40,41 Briefly, conjunctiva was rinsed and soaked sev- Collagenase A (0.001 mg/mL, Roche Diagnostics GmbH) and trypsin (Gibco, Fremont, California) for 15 to 20 minutes at 37 C. Cells were seeded on type I collagen-coated culture dishes and cultured with DMEM/F-12 (dulbecco's modified eagle medium, Gibco) supplemented with human keratinocyte growth factor (Peprotech, Rocky Hill, New Jersey), Y27632 (Sigma-Aldrich, St. Louis, Michigan), and B27 (Thermo Fisher Scientific, Waltham, Massachusetts).…”

Section: Human Conjunctival Epithelial Cell Isolation and Culturementioning

confidence: 99%

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Conjunctival reconstruction via enrichment of human conjunctival epithelial stem cells by p75 through the NGF-p75-SALL2 signaling axis

Yan

Chen

et al. 2020

Stem Cells Translational Medicine

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Severe conjunctival diseases can cause significant conjunctival scarring, which seriously limits eye movement and affects patients' vision. Conjunctival reconstruction remains challenging due to the lack of efficient methods for stem cells enrichment. This study indicated that p75 positive conjunctival epithelial cells (CjECs) were mainly located in the basal layer of human conjunctival epithelium and showed an immature differentiation state in vivo. The p75 strongly positive (p75++) CjECs enriched by immuno-magnetic beads exhibited high expression of stem cell markers and low expression of differentiated keratins. During continuous cell passage cultivation, p75 ++ CjECs showed the strongest proliferation potential and were able to reconstruct the conjunctiva in vivo with the most complete structure and function. Exogenous addition of NGF promoted the differentiation of CjECs by increasing nuclear localization of SALL2 in p75++ CjECs while proNGF played an opposite role. Altogether, p75 ++ CjECs present stem cell characteristics and exhibit the strongest proliferation potential so can be used as seed cells for conjunctival reconstruction, and NGF-p75-SALL2 signaling pathway was involved in regulating the differentiation of CjECs.

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Section: Methodsmentioning

confidence: 99%

Section: Human Conjunctival Epithelial Cell Isolation and Culturementioning

confidence: 99%

Conjunctival reconstruction via enrichment of human conjunctival epithelial stem cells by p75 through the NGF-p75-SALL2 signaling axis

Yan

Chen

et al. 2020

Stem Cells Translational Medicine

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“…Therefore, novel approaches have been developed with the advent of novel bioengineering technologies in order to try and prevent these drawbacks. Given the important role of goblet cells in the functionality of conjunctiva, it is fundamental that these novel engineering approaches ensure the restoration of this cell population, which is frequently compromised after injury [151].…”

Section: Bioengineering the Conjunctival Stem Cell Nichementioning

confidence: 99%

Biofabrication of Artificial Stem Cell Niches in the Anterior Ocular Segment

et al. 2021

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The anterior segment of the eye is a complex set of structures that collectively act to maintain the integrity of the globe and direct light towards the posteriorly located retina. The eye is exposed to numerous physical and environmental insults such as infection, UV radiation, physical or chemical injuries. Loss of transparency to the cornea or lens (cataract) and dysfunctional regulation of intra ocular pressure (glaucoma) are leading causes of worldwide blindness. Whilst traditional therapeutic approaches can improve vision, their effect often fails to control the multiple pathological events that lead to long-term vision loss. Regenerative medicine approaches in the eye have already had success with ocular stem cell therapy and ex vivo production of cornea and conjunctival tissue for transplant recovering patients’ vision. However, advancements are required to increase the efficacy of these as well as develop other ocular cell therapies. One of the most important challenges that determines the success of regenerative approaches is the preservation of the stem cell properties during expansion culture in vitro. To achieve this, the environment must provide the physical, chemical and biological factors that ensure the maintenance of their undifferentiated state, as well as their proliferative capacity. This is likely to be accomplished by replicating the natural stem cell niche in vitro. Due to the complex nature of the cell microenvironment, the creation of such artificial niches requires the use of bioengineering techniques which can replicate the physico-chemical properties and the dynamic cell–extracellular matrix interactions that maintain the stem cell phenotype. This review discusses the progress made in the replication of stem cell niches from the anterior ocular segment by using bioengineering approaches and their therapeutic implications.

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“…The most commonly8 used substrate is the amniotic membrane [ 127 , 136 , 137 ]. However, because the use of biological material may present some inconvenience, other substrates, such as collagen [ 138 , 139 , 140 , 141 , 142 ], fibrin [ 143 ], poly(L-lactic acid-co-ε-caprolactone) [ 142 , 144 ], gelatin-chitosan [ 145 ], silk fibroin [ 144 ], or the cadaveric, acellular dermis AlloDerm ® [ 146 ], have also been used. Although these conjunctival equivalents have not yet been used as cell culture models, some of them have the potential to provide this function.…”

Section: Three-dimensional In Vitro Models Of Anterior Eye Tissuesmentioning

confidence: 99%

Three-Dimensional Human Cell Culture Models to Study the Pathophysiology of the Anterior Eye

García-Posadas

Diebold

2020

Pharmaceutics

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In recent decades, the establishment of complex three-dimensional (3D) models of tissues has allowed researchers to perform high-quality studies and to not only advance knowledge of the physiology of these tissues but also mimic pathological conditions to test novel therapeutic strategies. The main advantage of 3D models is that they recapitulate the spatial architecture of tissues and thereby provide more physiologically relevant information. The eye is an extremely complex organ that comprises a large variety of highly heterogeneous tissues that are divided into two asymmetrical portions: the anterior and posterior segments. The anterior segment consists of the cornea, conjunctiva, iris, ciliary body, sclera, aqueous humor, and the lens. Different diseases in these tissues can have devastating effects. To study these pathologies and develop new treatments, the use of cell culture models is instrumental, and the better the model, the more relevant the results. Thus, the development of sophisticated 3D models of ocular tissues is a significant challenge with enormous potential. In this review, we present a comprehensive overview of the latest advances in the development of 3D in vitro models of the anterior segment of the eye, with a special focus on those that use human primary cells.

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A novel application of electrospun silk fibroin/poly(l-lactic acid-co-ε-caprolactone) scaffolds for conjunctiva reconstruction

Abstract: We present a promising scaffold with favorable mechanical and biological properties for conjunctival regeneration.

Cited by 22 publications

References 39 publications

Conjunctival reconstruction via enrichment of human conjunctival epithelial stem cells by p75 through the NGF-p75-SALL2 signaling axis

Conjunctival reconstruction via enrichment of human conjunctival epithelial stem cells by p75 through the NGF-p75-SALL2 signaling axis

Biofabrication of Artificial Stem Cell Niches in the Anterior Ocular Segment

Three-Dimensional Human Cell Culture Models to Study the Pathophysiology of the Anterior Eye

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