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采用压缩胶原和激光穿孔的静电纺丝垫构建组织工程角膜。

Tissue-engineered cornea constructed with compressed collagen and laser-perforated electrospun mat.

机构信息

Macromolecular Platforms for Translational Medicine and Bio-Manufacturing Laboratory, Tsinghua-Berkeley Shenzhen Insititute, Shenzhen, 518055, P.R. China.

Biomanufacturing Engineering Laboratory, Graduate School at Shenzhen, Tsinghua University, Shenzhen, 518055, P.R. China.

出版信息

Sci Rep. 2017 Apr 20;7(1):970. doi: 10.1038/s41598-017-01072-0.

Abstract

While Plastic Compressed (PC) collagen technique is often used to fabricate bioengineered constructs, PC collagen gels are too weak to be sutured or conveniently handled for clinical applications. To overcome this limitation, electrospun poly (lactic-co-glycolide) (PLGA) mats, which have excellent biocompatibility and mechanical properties, were combined with PC collagen to fabricate sandwich-like hybrid constructs. By laser-perforating holes with different sizes and spacings in the electrospun mats to regulate the mechanical properties and light transmittance of the hybrid constructs, we produced hybrid constructs with properties very suitable to apply in corneal tissue engineering. The maximum tensile stress of the optimal hybrid construct was 3.42 ± 0.22 MPa. The light transmittance of the hybrid construct after perforation was approximately 15-fold higher than before, and light transmittance increased gradually with increasing time. After immersing into PBS for 7 days, the transmittance of the optimal construct changed from 63 ± 2.17% to 72 ± 1.8% under 500 nm wavelength. The live/dead staining, cell proliferation assay and immunohistochemistry study of human corneal epithelial cells (HCECs) and human keratocytes (HKs) cultured on the optimal hybrid construct both demonstrated that the cells adhered, proliferated, and maintained their phenotype well on the material. In addition, after culturing for 2 weeks, the HCECs could form stratified layers. Thus, our designed construct is suitable for the construction of engineered corneal tissue.

摘要

虽然塑料压缩(PC)胶原技术常用于制造生物工程构建物,但 PC 胶原凝胶太弱,无法缝合或方便地处理用于临床应用。为了克服这一限制,将具有优异的生物相容性和机械性能的电纺聚(乳酸-共-乙醇酸)(PLGA)垫与 PC 胶原结合,制造三明治状混合构建物。通过在电纺垫上用不同大小和间距的激光穿孔来调节混合构建物的机械性能和透光率,我们制造了非常适合应用于角膜组织工程的混合构建物。最佳混合构建物的最大拉伸应力为 3.42±0.22 MPa。混合构建物穿孔后的透光率比穿孔前提高了约 15 倍,并且随着时间的增加透光率逐渐增加。将最佳构建物浸入 PBS 中 7 天后,在 500nm 波长下,最佳构建物的透光率从 63±2.17%变为 72±1.8%。对在最佳混合构建物上培养的人角膜上皮细胞(HCEC)和人角膜基质细胞(HK)进行的活/死染色、细胞增殖测定和免疫组织化学研究均表明,细胞在材料上良好地粘附、增殖并保持其表型。此外,培养 2 周后,HCEC 可以形成分层层。因此,我们设计的构建物适合构建工程化的角膜组织。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c170/5430529/71f26cee9eef/41598_2017_1072_Fig1_HTML.jpg

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