Di Meo Celeste, Stellavato Antonietta, d'Agostino Maria, D'Agostino Antonella, Schiraldi Chiara, La Gatta Annalisa
Department of Experimental Medicine, Section of Biotechnology, Medical Histology and Molecular Biology, University of Campania "Luigi Vanvitelli", via L. De Crecchio 7, 80138 Naples, Italy.
Int J Biol Macromol. 2024 Dec;282(Pt 5):137125. doi: 10.1016/j.ijbiomac.2024.137125. Epub 2024 Oct 30.
The effect of hyaluronan (HA) molecular weight (MW) and concentration (c) on key features of HA-formulations was systematically studied, in vitro, exploring the widest range/number of MW/c to date. Nine pharmaceutical grade HA samples (60-2500 kDa) were hydrodynamically characterized using Size Exclusion Chromatography-Triple Detector Array (SEC-TDA) also providing conformational data. HAs aqueous solutions (thirteen concentrations in the range 0.1-32 g/L) were tested for dynamic viscosity (η). η dependence on MW/c was analyzed providing mathematical correlations not only for the specific zero-shear viscosity, but also for the critical shear rate and the shear-thinning-extent. Besides confirming the dilute and semi-dilute c-regimes for the HAs, a third concentrated regime was evidenced for the 220-2500 kDa samples. Data on how MW affects the dependence of viscosity parameters on c and vice-versa were provided. The 60-90 kDa HAs proved stable to thermal sterilization and enzymatically catalyzed hydrolysis, while the 220-2500 kDa samples depolymerized to an extent depending, beyond concentration, on MW. HA size did not significantly affect fibroblasts behavior: under the conditions here tested, the HAs similarly sustained human dermal fibroblasts growth and wound healing also showing comparable effect on collagen-I, elastin and hyaluronan synthase-1 expression. Overall, results valuably contribute to the understanding of the HA MW/c impact on the rheological, stability and biochemical features of the final formulations, also providing mathematical correlations allowing for their optimization towards specific performance.
系统地研究了透明质酸（HA）分子量（MW）和浓度（c）对HA制剂关键特性的影响，这是在体外进行的，探索了迄今为止最广泛的MW/c范围/数量。使用尺寸排阻色谱-三检测器阵列（SEC-TDA）对九个医药级HA样品（60-2500 kDa）进行了流体动力学表征，该方法还提供了构象数据。测试了HA水溶液（0.1-32 g/L范围内的十三种浓度）的动态粘度（η）。分析了η对MW/c的依赖性，不仅提供了特定零剪切粘度的数学相关性，还提供了临界剪切速率和剪切变稀程度的数学相关性。除了确认HA的稀溶液和半稀溶液c区域外，还证明了220-2500 kDa样品存在第三种浓溶液区域。提供了关于MW如何影响粘度参数对c的依赖性以及反之亦然的数据。60-90 kDa的HA对热灭菌和酶催化水解具有稳定性，而220-2500 kDa的样品会发生一定程度的解聚，其程度除了取决于浓度外，还取决于MW。HA的大小对成纤维细胞行为没有显著影响：在本文测试的条件下，HA同样维持人真皮成纤维细胞的生长和伤口愈合，对I型胶原蛋白、弹性蛋白和透明质酸合酶-1的表达也显示出类似的效果。总体而言，研究结果对理解HA的MW/c对最终制剂的流变学、稳定性和生化特性的影响有重要贡献，还提供了数学相关性，有助于针对特定性能对其进行优化。
