Transplantation of human corneal endothelial cells using functional biomaterials: Poly(N-isopropylacrylamide) and gelatin

Wen Ming Hsu, Ko Hua Chen, Jui Yang Lai, Ging Ho Hsiue

研究成果: 雜誌貢獻文章

7 引文 (Scopus)

摘要

Purpose: To evaluate the feasibility of human corneal endothelial cell (HCEC) transplantation by harvesting the HCEC sheet on a thermoresponsive surface, delivering it with a gelatin disc, and testing it in a rabbit model. Methods: Cultivated human adult HCECs labeled with a red fluorescent dye (PKH26) were seeded on a poly(N-isopropylacrylamide) (PNIPAAM)-grafted surface culture dish at 37°C. After reaching confluence, the HCEC monolayer was detached by reducing the incubation temperature to 20°C and immediately delivered by means of a 7-mm gelatin disc to the rabbit's cornea denuded with endothelial cells (HCEC group, n = 8). The morphology, viability, pump, and barrier functions of the harvested HCEC were evaluated. Traumatized rabbit corneas with only the gelatin disc graft (gelatin disc group, n = 4) and without any transplantation (wound group, n = 4) were the sham controls. Surgical corneas of each group underwent histological and clinical evaluations including corneal thickness, intraocular pressure (IOP), and corneal clarity at different time points during a follow-up period of 12 weeks. Results: Cell morphology, viability, densities, Na+/K+ ATPase, and zonula occluden-1 (ZO-1) of the cultivated HCEC monolayer were similar with those of native HCECs. After endothelial removal, corneas of three groups turned severely edematous and opaque. In the HCEC group, the clarity of cornea recovered within 2 weeks with a corneal thickness of 552 ± 18 μm, which was significantly less than those (>1,000 μm) of the control groups (p <0.05). Histological examinations showed that the PKH26-labeled HCECs were spread over the Descemet's membrane with tight junction formation in the HCEC group, but none in the control group. The postoperative IOP of all three groups was within normal limits. Conclusion: This study provides a novel strategy for reconstruction of corneal endothelial cells using cultured adult HCECs and functional biomaterials including PNIPAAM and gelatin.
原文英語
頁(從 - 到)56-64
頁數9
期刊Journal of Experimental and Clinical Medicine(Taiwan)
5
發行號2
DOIs
出版狀態已發佈 - 四月 2013

指紋

Corneal Transplantation
Biocompatible Materials
Gelatin
Endothelial Cells
Cornea
Tight Junctions
Rabbits
Intraocular Pressure
poly-N-isopropylacrylamide
Descemet Membrane
Control Groups
Cell Transplantation
Fluorescent Dyes
Cell Survival
Transplantation
Transplants
Temperature

ASJC Scopus subject areas

  • Medicine(all)

引用此文

Transplantation of human corneal endothelial cells using functional biomaterials : Poly(N-isopropylacrylamide) and gelatin. / Hsu, Wen Ming; Chen, Ko Hua; Lai, Jui Yang; Hsiue, Ging Ho.

於: Journal of Experimental and Clinical Medicine(Taiwan), 卷 5, 編號 2, 04.2013, p. 56-64.

研究成果: 雜誌貢獻文章

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title = "Transplantation of human corneal endothelial cells using functional biomaterials: Poly(N-isopropylacrylamide) and gelatin",
abstract = "Purpose: To evaluate the feasibility of human corneal endothelial cell (HCEC) transplantation by harvesting the HCEC sheet on a thermoresponsive surface, delivering it with a gelatin disc, and testing it in a rabbit model. Methods: Cultivated human adult HCECs labeled with a red fluorescent dye (PKH26) were seeded on a poly(N-isopropylacrylamide) (PNIPAAM)-grafted surface culture dish at 37°C. After reaching confluence, the HCEC monolayer was detached by reducing the incubation temperature to 20°C and immediately delivered by means of a 7-mm gelatin disc to the rabbit's cornea denuded with endothelial cells (HCEC group, n = 8). The morphology, viability, pump, and barrier functions of the harvested HCEC were evaluated. Traumatized rabbit corneas with only the gelatin disc graft (gelatin disc group, n = 4) and without any transplantation (wound group, n = 4) were the sham controls. Surgical corneas of each group underwent histological and clinical evaluations including corneal thickness, intraocular pressure (IOP), and corneal clarity at different time points during a follow-up period of 12 weeks. Results: Cell morphology, viability, densities, Na+/K+ ATPase, and zonula occluden-1 (ZO-1) of the cultivated HCEC monolayer were similar with those of native HCECs. After endothelial removal, corneas of three groups turned severely edematous and opaque. In the HCEC group, the clarity of cornea recovered within 2 weeks with a corneal thickness of 552 ± 18 μm, which was significantly less than those (>1,000 μm) of the control groups (p <0.05). Histological examinations showed that the PKH26-labeled HCECs were spread over the Descemet's membrane with tight junction formation in the HCEC group, but none in the control group. The postoperative IOP of all three groups was within normal limits. Conclusion: This study provides a novel strategy for reconstruction of corneal endothelial cells using cultured adult HCECs and functional biomaterials including PNIPAAM and gelatin.",
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T1 - Transplantation of human corneal endothelial cells using functional biomaterials

T2 - Poly(N-isopropylacrylamide) and gelatin

AU - Hsu, Wen Ming

AU - Chen, Ko Hua

AU - Lai, Jui Yang

AU - Hsiue, Ging Ho

PY - 2013/4

Y1 - 2013/4

N2 - Purpose: To evaluate the feasibility of human corneal endothelial cell (HCEC) transplantation by harvesting the HCEC sheet on a thermoresponsive surface, delivering it with a gelatin disc, and testing it in a rabbit model. Methods: Cultivated human adult HCECs labeled with a red fluorescent dye (PKH26) were seeded on a poly(N-isopropylacrylamide) (PNIPAAM)-grafted surface culture dish at 37°C. After reaching confluence, the HCEC monolayer was detached by reducing the incubation temperature to 20°C and immediately delivered by means of a 7-mm gelatin disc to the rabbit's cornea denuded with endothelial cells (HCEC group, n = 8). The morphology, viability, pump, and barrier functions of the harvested HCEC were evaluated. Traumatized rabbit corneas with only the gelatin disc graft (gelatin disc group, n = 4) and without any transplantation (wound group, n = 4) were the sham controls. Surgical corneas of each group underwent histological and clinical evaluations including corneal thickness, intraocular pressure (IOP), and corneal clarity at different time points during a follow-up period of 12 weeks. Results: Cell morphology, viability, densities, Na+/K+ ATPase, and zonula occluden-1 (ZO-1) of the cultivated HCEC monolayer were similar with those of native HCECs. After endothelial removal, corneas of three groups turned severely edematous and opaque. In the HCEC group, the clarity of cornea recovered within 2 weeks with a corneal thickness of 552 ± 18 μm, which was significantly less than those (>1,000 μm) of the control groups (p <0.05). Histological examinations showed that the PKH26-labeled HCECs were spread over the Descemet's membrane with tight junction formation in the HCEC group, but none in the control group. The postoperative IOP of all three groups was within normal limits. Conclusion: This study provides a novel strategy for reconstruction of corneal endothelial cells using cultured adult HCECs and functional biomaterials including PNIPAAM and gelatin.

AB - Purpose: To evaluate the feasibility of human corneal endothelial cell (HCEC) transplantation by harvesting the HCEC sheet on a thermoresponsive surface, delivering it with a gelatin disc, and testing it in a rabbit model. Methods: Cultivated human adult HCECs labeled with a red fluorescent dye (PKH26) were seeded on a poly(N-isopropylacrylamide) (PNIPAAM)-grafted surface culture dish at 37°C. After reaching confluence, the HCEC monolayer was detached by reducing the incubation temperature to 20°C and immediately delivered by means of a 7-mm gelatin disc to the rabbit's cornea denuded with endothelial cells (HCEC group, n = 8). The morphology, viability, pump, and barrier functions of the harvested HCEC were evaluated. Traumatized rabbit corneas with only the gelatin disc graft (gelatin disc group, n = 4) and without any transplantation (wound group, n = 4) were the sham controls. Surgical corneas of each group underwent histological and clinical evaluations including corneal thickness, intraocular pressure (IOP), and corneal clarity at different time points during a follow-up period of 12 weeks. Results: Cell morphology, viability, densities, Na+/K+ ATPase, and zonula occluden-1 (ZO-1) of the cultivated HCEC monolayer were similar with those of native HCECs. After endothelial removal, corneas of three groups turned severely edematous and opaque. In the HCEC group, the clarity of cornea recovered within 2 weeks with a corneal thickness of 552 ± 18 μm, which was significantly less than those (>1,000 μm) of the control groups (p <0.05). Histological examinations showed that the PKH26-labeled HCECs were spread over the Descemet's membrane with tight junction formation in the HCEC group, but none in the control group. The postoperative IOP of all three groups was within normal limits. Conclusion: This study provides a novel strategy for reconstruction of corneal endothelial cells using cultured adult HCECs and functional biomaterials including PNIPAAM and gelatin.

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KW - Poly (N-isopropylacrylamide)

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