Department of Experimental Medicine, Section of Biotechnology, Medical Histology and Molecular Biology, University of Campania Luigi Vanvitelli, 80138 Naples, Italy.
Department of Environmental, Biological and Pharmaceutical Sciences and Technologies, University of Campania L. Vanvitelli, 81100 Caserta, Italy.
Int J Mol Sci. 2022 Feb 1;23(3):1686. doi: 10.3390/ijms23031686.
Chondroitin obtained through biotechnological processes (BC) shares similarities with both chondroitin sulfate (CS), due to the dimeric repetitive unit, and hyaluronic acid (HA), as it is unsulfated. In the framework of this experimental research, formulations containing BC with an average molecular size of about 35 KDa and high molecular weight HA (HHA) were characterized with respect to their rheological behavior, stability to enzymatic hydrolysis and they were evaluated in different skin damage models. The rheological characterization of the HHA/BC formulation revealed a G' of 92 ± 3 Pa and a G″ of 116 ± 5 Pa and supported an easy injectability even at a concentration of 40 mg/mL. HA/BC preserved the HHA fraction better than HHA alone. BTH was active on BC alone only at high concentration. Assays on scratched keratinocytes (HaCaT) monolayers showed that all the glycosaminoglycan formulations accelerated cell migration, with HA/BC fastening healing 2-fold compared to the control. In addition, in 2D HaCaT cultures, as well as in a 3D skin tissue model HHA/BC efficiently modulated mRNA and protein levels of different types of collagens and elastin remarking a functional tissue physiology. Finally, immortalized human fibroblasts were challenged with TNF-α to obtain an in vitro model of inflammation. Upon HHA/BC addition, secreted IL-6 level was lower and efficient ECM biosynthesis was re-established. Finally, co-cultures of HaCaT and melanocytes were established, showing the ability of HHA/BC to modulate melanin release, suggesting a possible effect of this specific formulation on the reduction of stretch marks. Overall, besides demonstrating the safety of BC, the present study highlights the potential beneficial effect of HHA/BC formulation in different damage dermal models.
通过生物技术过程获得的软骨素（BC）由于二聚重复单元与硫酸软骨素（CS）相似，并且由于其未硫酸化而与透明质酸（HA）相似。在这项实验研究中，对含有平均分子量约为 35 kDa 和高分子量 HA（HHA）的 BC 的配方进行了流变特性、对酶水解的稳定性以及在不同皮肤损伤模型中的评估。HHA/BC 配方的流变特性表明 G'为 92 ± 3 Pa，G″为 116 ± 5 Pa，即使在 40 mg/mL 的浓度下也支持易于注射。HA/BC 比 HHA 单独保留 HHA 部分更好。BTH 仅在高浓度下对 BC 单独有效。对划痕角质形成细胞（HaCaT）单层的测定表明，所有糖胺聚糖配方均加速了细胞迁移，与对照相比，HA/BC 使愈合速度加快了 2 倍。此外，在 2D HaCaT 培养物以及 3D 皮肤组织模型中，HHA/BC 有效地调节了不同类型胶原蛋白和弹性蛋白的 mRNA 和蛋白质水平，突出了功能性组织生理学。最后，永生化人成纤维细胞用 TNF-α 进行挑战，以获得体外炎症模型。添加 HHA/BC 后，分泌的 IL-6 水平降低，并且有效建立了 ECM 生物合成。最后，建立了 HaCaT 和黑素细胞的共培养物，表明 HHA/BC 能够调节黑色素释放，表明这种特定配方可能对减少妊娠纹有影响。总体而言，除了证明 BC 的安全性外，本研究还强调了 HHA/BC 配方在不同损伤皮肤模型中的潜在有益作用。
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