Caballero-Aguilar Lilith M, Duchi Serena, Quigley Anita, Onofrillo Carmine, Di Bella Claudia, Moulton Simon E
ARC Centre of Excellence for Electromaterials Science, Faculty of Science, Engineering and Technology, Swinburne University of Technology, Victoria 3122, Australia.
BioFab3D, Aikenhead Centre for Medical Discovery, St Vincent's Hospital, Melbourne, Australia.
MethodsX. 2021 Mar 28;8:101324. doi: 10.1016/j.mex.2021.101324. eCollection 2021.
The encapsulation of growth factors is an important component of tissue engineer- ing. Using microspheres is a convenient approach in which the dose of factors can be regulated by increasing or decreasing the number of encapsulated microspheres. Moreover, microspheres offer the possibility of delivering the growth factors directly to the target site. However, the fabrication of microspheres by traditional emulsion methods is largely variable due to the experimental procedure. We have developed a protocol using a commercially available microfluidic system that allows formation of tunable particle-size droplets loaded with growth factors. The methodology includes a guide for preparing an alginate-growth factors solution followed by the specific set-up needed for using the microfluidic system to form the microspheres. The pro- cedure also includes a unique post-crosslinking process without pH modification. These methods allow the preservation of integrity and bioactivity of the growth factors tested (BMP-6 and TGF -3) and their subsequent sustained delivery.•The protocol can be tuned to form particles of various sizes.•The gentle post-crosslinking process allows conformational integrity of various bioactive molecules.
生长因子的封装是组织工程的一个重要组成部分。使用微球是一种便捷的方法,通过增加或减少封装微球的数量可以调节因子的剂量。此外,微球提供了将生长因子直接递送至靶位点的可能性。然而,由于实验过程,传统乳液法制备微球的过程在很大程度上是可变的。我们开发了一种使用市售微流控系统的方案,该系统允许形成负载生长因子的可调粒径液滴。该方法包括制备藻酸盐 - 生长因子溶液的指南,以及使用微流控系统形成微球所需的特定设置。该过程还包括一个无需调节pH的独特后交联过程。这些方法能够保留所测试生长因子(骨形态发生蛋白-6和转化生长因子-β3)的完整性和生物活性,并实现其后续的持续释放。•该方案可进行调整以形成各种尺寸的颗粒。•温和的后交联过程可保持各种生物活性分子的构象完整性。