Electrospun nanofibrous SF/P(LLA-CL) membrane: a potential substratum for endothelial keratoplasty

Chen, Junzhao; Yan, Chenxi; Zhu, Mengyu; Yao, Qinke; Shao, Chen; Lu, Wenjuan; Wang, Jing; Mo, Xiumei; Gu, Ping; Fu, Yao; Fan, Xianqun

doi:10.2147/ijn.s77706

Cited by 28 publications

(12 citation statements)

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“…Known as the immortalized cell population of human corneal endothelial cells, B4G12 cells are polygonal, strongly adherent forming a strict monolayer, and are characterized by junctional proteins, collagens, and Na + /K + ATPase 24 , 25 . B4G12 cells have been widely used in different research aspects, such as evaluating sub-stratums supporting growth of CECs 26 , 27 or studying the mechanism of factors promoting CECs proliferation 28 . But there are difficulties in application in clinics due to consideration of the long-term safety of B4G12 cells’ immortalized nature.…”

Section: Discussionmentioning

confidence: 99%

Therapy of corneal endothelial dysfunction with corneal endothelial cell-like cells derived from skin-derived precursors

Shen

Sun

Zhang

et al. 2017

Sci Rep

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Corneal endothelial dysfunction occurs when corneal endothelial cells (CECs) are dramatically lost and eventually results in vision loss. Corneal transplantation is the only solution at present. However, corneal transplantation requires a fresh human cornea and there is a worldwide shortage of donors. Therefore, finding new functional CECs to replace human CECs is urgent. Skin-derived precursors (SKPs) can be easily acquired and have multiple differential potential. We co-cultured human SKPs with B4G12 cells in serum-free medium and obtained abundant CEC-like cells which had similar morphology and characteristic to human CECs. CEC-like cells exerted excellent therapeutic effect when they were transplanted into rabbit and monkey corneal endothelial dysfunction models by injection method. This protocol enables efficient production of CEC-like cells from SKPs. The renewable cell source, novel derivation method and simple treatment strategy may lead to potential applications in cell replacement therapy for corneal endothelial dysfunction.Corneal endothelial cells (CECs) are a monolayer of hexagonal cells covering the posterior surface of the cornea, and serve as a barrier between the corneal stroma and the aqueous humor. The tight junction as well as the ionic "pump" functions of CECs are important factors that maintain corneal transparency 1 . Many pathological factors such as trauma, surgery, and inflammation may cause dramatic loss of endothelial cell density, resulting in corneal endothelial dysfunction which is characterized by corneal edema, bullous keratopathy, and loss of visual acuity 2 . Human CECs have limited proliferative capacity in vivo and cannot be subcultured for more than a few passages in vitro 3,4 . Corneal transplantation is the only solution at present. However, corneal transplantation requires a fresh human cornea and there is a worldwide shortage of donors 5 . Therefore, finding new functional CECs to replace human CECs is a new and potential direction for future clinical application.Skin-derived precursors (SKPs) were discovered by Tomas 6,7 , and they have been a research hotspot in recent years. SKPs can be isolated in large quantities from many parts of the skin in rodents and humans. As they are multipotent, they can be differentiated into several functional cell types 8 , and are able to suppress the allogeneic activation of T-lymphocytes resulting in an improved health status of animals suffering from a graft-versus-host reaction 9 . Moreover, SKPs were demonstrated to be embryonic neural-crest-related precursors and share many properties with neural crest stem cells 10

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Section: Discussionmentioning

confidence: 99%

Therapy of corneal endothelial dysfunction with corneal endothelial cell-like cells derived from skin-derived precursors

Shen

Sun

Zhang

et al. 2017

Sci Rep

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“…To evaluate whether collagen/PLCL scaffolds altered the cell cycle, the B4G12 human corneal endothelial cell line (Creative Bioarray, Shirley, NY, USA) was seeded onto the scaffolds in a 24-well plate and cultured in a humidified environment at 37°C with 5% CO 2 for 3 days ( 18 ). The cells were then harvested using 0.25% trypsin (Thermo Fisher Scientific, Inc., Waltham, MA, USA), followed by washing with pre-cooled (4°C) PBS, and then fixed in pre-cooled (4°C) 70% ethanol for 1 h. Fixed cells were treated using propidium iodide (PI, Abcam, Cambridge, MA, USA).…”

Section: Methodsmentioning

confidence: 99%

Electrospun collagen/poly(L-lactic acid‑co‑ε‑caprolactone) scaffolds for conjunctival tissue engineering

Yao

Zhang

et al. 2017

Exp Ther Med

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Conjunctival injuries are general but intractable ocular surface diseases, the sequelae of which are particularly challenging to treat. A promising therapy for conjunctival injuries is to employ biodegradable scaffolds to deliver conjunctival epithelial cells for repairing damaged or diseased conjunctiva. In the present study, an ultrathin porous nanofibrous scaffold was fabricated by using collagen and poly(L-lactic acid-co-ε-caprolactone) (PLCL) and displayed a thickness of 20 µm, with a high porosity and an average fiber diameter of 248.83±26.44 nm. Conjunctival epithelial cells seeded on the scaffolds proliferated well and had a high cell viability. Reverse-transcription quantitative PCR showed the expression of conjunctival epithelial cell-specific genes; in addition, there was no significant difference in the inflammatory gene expression between cells grown on collagen/PLCL scaffolds and tricalcium phosphate scaffolds. After co-culture for 2 weeks in vitro, epithelial cell stratification was observed using hematoxylin and eosin staining, exhibiting three to four epithelial-cell layers. In conclusion, these results suggested that collagen/PLCL scaffolds have potential application for repairing conjunctival epithelial coloboma.

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“…In another study, monolayers of the human corneal endothelial cell line B4G12 could be successfully formed using blends of nanofibrous structures made of silk fibroin and poly(l-lactic acid-co-ε-caprolactone) (P(LLA-CL)) [75]. The authors stated that the combination of natural and synthetic materials offered by silk:(P(LLA-CL)) constructs could provide both biocompatibility and mechanical properties necessary for the culture of endothelial cells.…”

Section: 11 X For Peer Review 7 Of 13mentioning

confidence: 99%

Nanoscale Topographies for Corneal Endothelial Regeneration

et al. 2021

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The corneal endothelium is the innermost layer of the cornea that selectively pumps ions and metabolites and regulates the hydration level of the cornea, ensuring its transparency. Trauma or disease affecting human corneal endothelial cells (hCECs) can result in major imbalances of such transport activity with consequent deterioration or loss of vision. Since tissue transplantation from deceased donors is only available to a fraction of patients worldwide, alternative solutions are urgently needed. Cell therapy approaches, in particular by attempting to expand primary culture of hCECs in vitro, aim to tackle this issue. However, existing cell culture protocols result in limited expansion of this cell type. Recent studies in this field have shown that topographical features with specific dimensions and shapes could improve the efficacy of hCEC expansion. Therefore, potential solutions to overcome the limitation of the conventional culture of hCECs may include recreating nanometer scale topographies (nanotopographies) that mimic essential biophysical cues present in their native environment. In this review, we summarize the current knowledge and understanding of the effect of substrate topographies on the response of hCECs. Moreover, we also review the latest developments for the nanofabrication of such bio-instructive cell substrates.

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Electrospun nanofibrous SF/P(LLA-CL) membrane: a potential substratum for endothelial keratoplasty

Cited by 28 publications

References 50 publications

Therapy of corneal endothelial dysfunction with corneal endothelial cell-like cells derived from skin-derived precursors

Therapy of corneal endothelial dysfunction with corneal endothelial cell-like cells derived from skin-derived precursors

Electrospun collagen/poly(L-lactic acid‑co‑ε‑caprolactone) scaffolds for conjunctival tissue engineering

Nanoscale Topographies for Corneal Endothelial Regeneration

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