Weldon School of Biomedical Engineering, Purdue University, West Lafayette, IN, USA.
School of Mechanical Engineering, Purdue University, West Lafayette, IN, USA.
Colloids Surf B Biointerfaces. 2023 Feb;222:113123. doi: 10.1016/j.colsurfb.2023.113123. Epub 2023 Jan 3.
Therapeutic macromolecules possess properties such as size and electrostatic charge that will dictate their transport through subcutaneous (SC) tissue and ultimate bioavailability and efficacy. To improve therapeutic design, platforms that systematically measure the transport of macromolecules as a function of both drug and tissue properties are needed. We utilize a Transwell chamber with tunable collagen-hyaluronic acid (ColHA) hydrogels as an in vitro model to determine mass transport of macromolecules using non-invasive UV spectroscopy. Increasing hyaluronic acid (HA) concentration from 0 to 2 mg/mL within collagen gels decreases the mass transport of five macromolecules independent of size and charge and results in a maximum decrease in recovery of 23.3% in the case of bovine immunoglobulin G (IgG). However, in a pure 10 mg/mL HA solution, negatively-charged macromolecules bovine serum albumin (BSA), β-lactoglobulin (BLg), dextran (Dex), and IgG had drastically increased recovery by 20-40% compared to their performance in ColHA matrices. This result was different from the positively-charged macromolecule Lysozyme (Lys), which, despite its small size, showed reduced recovery by 3% in pure HA. These results demonstrate two distinct regimes of mass transport within our tissue model. In the presence of both collagen and HA, increasing HA concentrations decrease mass transport; however, in the absence of collagen, the high negative charge of HA sequesters and increases residence time of positively-charged macromolecules and decreases residence time of negatively-charged macromolecules. Through our approach, ColHA hydrogels serve as a platform for the systematic evaluation of therapeutic macromolecule transport as a function of molecular characteristics.
治疗性大分子具有大小和电荷等特性，这些特性将决定它们在皮下 (SC) 组织中的传输以及最终的生物利用度和疗效。为了改善治疗设计，需要有能够系统地测量大分子作为药物和组织特性函数的传输的平台。我们利用具有可调谐胶原蛋白-透明质酸 (ColHA) 水凝胶的 Transwell 室作为体外模型，使用非侵入性 UV 光谱法来确定大分子的质量传输。在胶原蛋白凝胶中，透明质酸 (HA) 浓度从 0 增加到 2 mg/mL，会降低五种大分子的质量传输，而与大小和电荷无关，牛免疫球蛋白 G (IgG) 的回收率最大降低 23.3%。然而，在纯 10 mg/mL HA 溶液中，带负电荷的大分子牛血清白蛋白 (BSA)、β-乳球蛋白 (BLg)、葡聚糖 (Dex) 和 IgG 的回收率与它们在 ColHA 基质中的性能相比，增加了 20-40%。与带负电荷的大分子溶菌酶 (Lys) 不同，尽管其尺寸较小，但在纯 HA 中的回收率降低了 3%。这些结果表明，在我们的组织模型中存在两种不同的质量传输状态。在同时存在胶原蛋白和 HA 的情况下，增加 HA 浓度会降低质量传输；然而，在没有胶原蛋白的情况下，HA 的高负电荷会隔离并增加带正电荷的大分子的停留时间，并降低带负电荷的大分子的停留时间。通过我们的方法，ColHA 水凝胶可作为一个平台，用于系统地评估治疗性大分子的传输，作为分子特性的函数。
