Department of Surgery, National Taiwan University Hospital and College of Medicine, Taipei 100, Taiwan.
Institute of Biomedical Engineering, College of Medicine and College of Engineering, National Taiwan University, Taipei 100, Taiwan.
Int J Mol Sci. 2021 Jun 16;22(12):6451. doi: 10.3390/ijms22126451.
The clinical application of human platelet lysate (HPL) holds promise for tissue regeneration, and the development of an efficient vehicle for its delivery is desired. Chitosan-based hydrogels are potential candidates, but they often exhibit weak mechanical properties. In this study, a chitosan/gelatin (CS-GE) hydrogel crosslinked by glyoxal was fabricated for sustained release of HPL. The influence of HPL on Hs68 fibroblast and human umbilical vein endothelial cell (HUVEC) culture was evaluated, and we found that supplementing 5% HPL in the medium could significantly improve cell proliferation relative to supplementing 10% fetal bovine serum (FBS). Moreover, HPL accelerated the in vitro wound closure of Hs68 cells and facilitated the tube formation of HUVECs. Subsequently, we fabricated CS-GE hydrogels crosslinked with different concentrations of glyoxal, and the release pattern of FITC-dextrans (4, 40 and 500 kDa) from the hydrogels was assessed. After an ideal glyoxal concentration was determined, we further characterized the crosslinked CS-GE hydrogels encapsulated with different amounts of HPL. The HPL-incorporated hydrogel was shown to significantly promote the proliferation of Hs68 cells and the migration of HUVECs. Moreover, the release pattern of transforming growth factor-β1 (TGF-β1) and platelet-derived growth factor-BB (PDGF-BB) from hydrogel was examined in vitro, demonstrating a sustained release profile of the growth factors. Finally, the chick chorioallantoic membrane assay revealed that HPL encapsulation in the hydrogel significantly stimulated angiogenesis in ovo. These results demonstrate the great potential of the crosslinked CS-GE hydrogel to serve as an effective delivery system for HPL to promote tissue regeneration.
人血小板裂解液（HPL）的临床应用有望促进组织再生，因此需要开发一种有效的递送载体。壳聚糖基水凝胶是潜在的候选材料，但它们通常表现出较弱的机械性能。在本研究中，制备了一种通过乙二醛交联的壳聚糖/明胶（CS-GE）水凝胶，用于 HPL 的持续释放。评估了 HPL 对 Hs68 成纤维细胞和人脐静脉内皮细胞（HUVEC）培养的影响，发现与补充 10%胎牛血清（FBS）相比，在培养基中补充 5%的 HPL 可显著促进细胞增殖。此外，HPL 加速了 Hs68 细胞的体外伤口闭合，并促进了 HUVEC 的管状形成。随后，我们制备了不同浓度乙二醛交联的 CS-GE 水凝胶，并评估了 FITC-葡聚糖（4、40 和 500 kDa）从水凝胶中的释放模式。确定理想的乙二醛浓度后，我们进一步表征了包封不同量 HPL 的交联 CS-GE 水凝胶。结果表明，HPL 包封水凝胶显著促进了 Hs68 细胞的增殖和 HUVEC 的迁移。此外，还在体外研究了转化生长因子-β1（TGF-β1）和血小板衍生生长因子-BB（PDGF-BB）从水凝胶中的释放模式，结果表明生长因子呈持续释放的特征。最后，鸡胚绒毛尿囊膜试验显示，HPL 包封在水凝胶中显著刺激了鸡胚中的血管生成。这些结果表明交联 CS-GE 水凝胶作为 HPL 递送系统在促进组织再生方面具有巨大潜力。
