Microfluidic 3D Cell Culture: Potential Application of Collagen Hydrogels with an Optimal Dose of Bioactive Glasses

Faezeh Ghobadi, Maryam Saadatmand, Sara Simorgh, Peiman Brouki Milan
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Abstract

We engineered a microfluidic platform to study the effects of bioactive glass nanoparticles (BGNs) on cell viability under static culture. We incorporated different concentrations of BGNs (1%, 2%, and 3% w/v) in collagen hydrogel (with a concentration of 3.0 mg/mL). The microfluidic chip's dimensions were optimized through fluid flow and mass transfer simulations. Collagen type I extracted from rat tail tendons was used as the main material, and BGNs synthesized by the sol-gel method were used to enhance the mechanical properties of the hydrogel. The extracted collagen was characterized using FTIR and SDS-PAGE, and BGNs were analyzed using XRD, FTIR, DLS, and FE-SEM/EDX. The structure of the collagen-BGNs hydrogels was examined using SEM, and their mechanical properties were determined using rheological analysis. The cytotoxicity of BGNs was assessed using the MTT assay, and the viability of fibroblast (L929) cells encapsulated in the collagen-BGNs hydrogel inside the microfluidic device was assessed using a live/dead assay. Based on all these test results, the L929 cells showed high cell viability in vitro and promising microenvironment mimicry in a microfluidic device. Collagen3-BGNs3 (Collagen 3 mg/mL + BGNs 3% (w/v)) was chosen as the most suitable sample for further research on a microfluidic platform.
微流控三维细胞培养:具有最佳生物活性玻璃剂量的胶原水凝胶的潜在应用
我们设计了一个微流控平台来研究生物活性玻璃纳米粒子(BGNs)在静态培养条件下对细胞活力的影响。我们在胶原水凝胶(浓度为 3.0 mg/mL)中加入了不同浓度的 BGNs(1%、2% 和 3% w/v)。通过流体流动和传质模拟优化了微流控芯片的尺寸。主要材料是从大鼠尾部肌腱中提取的 I 型胶原蛋白,并使用溶胶-凝胶法合成的 BGN 增强水凝胶的机械性能。利用傅立叶变换红外光谱和 SDS-PAGE 对提取的胶原蛋白进行了表征,并利用 XRD、傅立叶变换红外光谱、DLS 和 FE-SEM/EDX 对 BGNs 进行了分析。利用扫描电镜观察了胶原蛋白-BGNs 水凝胶的结构,并利用流变分析测定了它们的力学性能。使用 MTT 试验评估了 BGNs 的毒性,并使用活/死试验评估了微流控装置中胶原蛋白-BGNs 水凝胶包裹的成纤维细胞(L929)的存活率。根据所有测试结果,L929 细胞在体外显示出很高的细胞活力,并有望在微流控装置中模拟微环境。Collagen3-BGNs3(Collagen 3mg/mL + BGNs 3%(w/v))被选为最适合在微流控平台上进一步研究的样品。
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