Development and functional characterization of a tissue-engineered blood-air barrier model for in vitro applications.

IF 3.9

Nanomedicine (London, England) Pub Date : 2025-10-01 Epub Date: 2025-08-29 DOI:10.1080/17435889.2025.2552101

Neval Sevinc Ozdemir, Simal Yaren Sahin, Halime Kenar, Vasif Hasirci

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Abstract

Background: The blood-air barrier (BAB) of the lung is a critical interface responsible for gas exchange and protection against external attempts, and acts as a selective barrier. Developing in vitro models that replicate its structural and functional properties is essential in studying pulmonary diseases and their therapy.

Methods: In this study, a model consisting of alveolar epithelial (A549) and primary endothelial (pHUVEC) cells seeded on opposite sides of a thin (11 ± 4 μm), electrospun poly(ε-caprolactone) mesh of nanofibers (140-800 nm) to represent the basal membrane, and the interstitial matrix of the native BAB when coated with collagen type I, fibronectin, and laminin 511 proteins. The dense, nanofibrous architecture of the mesh enabled the formation of cellular monolayers on opposite sides, allowing gas and nutrient exchange for 14 days at air-liquid interface.

Results: The mesh had a Young's modulus of 8.0 ± 0.8 MPa, and upon coating with proteins, the water contact angles were decreased from 127.5°±2.6 to 94.4°±3.6. Epithelial and endothelial monolayers demonstrated tight junction formation as shown by ZO-1 and CD31 expression. TEER was measured as 44 ± 5.0 Ω·cm² with a permeability coefficient (P_app) of 2-5 × 10^-6 cm/s against fluorescein.

Conclusion: This study presents a physiologically relevant in vitro BAB model for respiratory research and therapies.

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用于体外应用的组织工程血气屏障模型的开发和功能表征。

背景：肺的血气屏障（BAB）是一个重要的界面，负责气体交换和保护免受外界的干扰，并作为一个选择性屏障。开发复制其结构和功能特性的体外模型对于研究肺部疾病及其治疗至关重要。方法：在本研究中，建立了一个由肺泡上皮细胞（A549）和原代内皮细胞（pHUVEC）细胞组成的模型，这些细胞分别分布在薄（11±4 μm）、静电纺聚（ε-己内酯）纳米纤维网（140-800 nm）的两侧，代表基膜，并在天然BAB的间质基质上涂有I型胶原蛋白、纤维连接蛋白和层粘连蛋白511。网状结构的致密、纳米纤维结构使细胞单层在相对的两侧形成，允许气体和营养物质在气液界面交换14天。结果：网状物的杨氏模量为8.0±0.8 MPa，涂覆蛋白质后，水接触角由127.5°±2.6减小到94.4°±3.6。ZO-1和CD31表达显示上皮和内皮单层紧密连接形成。测定TEER为44±5.0 Ω·cm2，对荧光素的渗透系数（Papp）为2-5 × 10-6 cm/s。结论：本研究为呼吸研究和治疗提供了一种与生理相关的体外BAB模型。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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Nanomedicine (London, England)

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