Enzyme-Mediated Crosslinking of Silk Fibroin/ N-Acetylcysteine-Modified Chitosan/ Hydroxyapatite Hydrogel Composites for Subchondral Bone Regeneration - additional experiments

This dataset contains the results of comprehensive physicochemical, morphological, structural, and biological analyses of hydrogel composites based on silk fibroin, N-acetylcysteine-modified chitosan, and hydroxyapatite. The investigated materials were prepared using an enzymatic cross-linking approach and evaluated as composite hydrogel systems for potential application in subchondral bone regeneration. The data include the results of additional experiments conducted during the peer-review process and comprise: (1) direct biocompatibility assessment, including absorbance measurements and fluorescence microscopy observations; (2) alkaline phosphatase (ALP) activity measurements, together with total protein quantification at the corresponding time points; (3) Alizarin Red S mineralization assays, including absorbance measurements and microscopic observations; (4) ELISA measurements of osteocalcin (OCN) and RUNX2, normalized to total protein content determined using the BCA assay; (5) rheological measurements; (6) scanning electron microscopy (SEM) observations; and (7) assessment of superoxide anion (O₂•⁻) generation in THP-1 cells using the nitroblue tetrazolium (NBT) assay. This research was funded by the National Science Centre, Poland, under the project “Hierarchical approaches to osteochondral tissue engineering” (grant no. UMO-2022/45/B/ST8/02557).本数据集包含基于丝素蛋白、N-乙酰半胱氨酸修饰壳聚糖与羟基磷灰石的水凝胶复合材料的全面理化、形貌、结构及生物学分析结果。所研究的材料采用酶促交联法制备，作为复合水凝胶体系被评估用于软骨下骨再生的潜在应用。本数据集涵盖同行评审阶段补充开展的实验结果，具体包括：(1) 直接生物相容性评估，包含吸光度测定与荧光显微镜观测；(2) 碱性磷酸酶（alkaline phosphatase, ALP）活性检测，以及对应时间点的总蛋白定量分析；(3) 茜素红S矿化试验，包含吸光度测定与显微镜观测；(4) 骨钙蛋白（osteocalcin, OCN）与RUNX2的ELISA检测，结果以BCA法测定的总蛋白含量进行归一化处理；(5) 流变学性能测试；(6) 扫描电子显微镜（scanning electron microscopy, SEM）观测；(7) 采用硝基蓝四唑（nitroblue tetrazolium, NBT）法检测THP-1细胞中超氧阴离子（O₂•⁻）的生成情况。 本研究由波兰国家科学中心资助，项目名称为“骨软骨组织工程的分级构建策略”（项目编号：UMO-2022/45/B/ST8/02557）。