Enhanced bone regeneration and cellular protection from oxidative stress using a vitamin C-based scaffold

To fabricate and investigate a novel VitC@HAP-HYA nanocomposite scaffold that combines the osteoconductive attributes of hydroxyapatite (HAP), the extracellular matrix-replicating properties of hyaluronic acid (HYA), and the osteoinductive/antioxidant effects of Vitamin C (VitC) for enhanced bone regeneration.

The composite was developed using an in-situ precipitation method. Physicochemical characterisation (FT-IR, XRD, and HR-TEM) was used to validate the composition, crystallinity, and shape. The examination of Simulated Body Fluid (SBF) evaluated bioactivity. The biological assessments used the MC3T3-E1 osteoblast-like cell line, including MTT and AO/PI tests for biocompatibility, DAPI labelling for cell colonization, Alkaline Phosphatase (ALP) activity to assess early osteogenic differentiation, and Alizarin Red S (ARS) staining for biomineralization analysis. Monitoring of intracellular Reactive Oxygen Species (ROS) evaluated its cytoprotective efficacy against oxidative stress.

FT-IR and XRD analyses confirmed the effective integration of amorphous VitC and HYA into HAP, yielding a stable 120 nm nanocomposite. The SBF study demonstrated fast production of a calcium phosphate layer, indicating elevated bioactivity. In cell culture, VitC@HAP-HYA demonstrated enhanced cell survival and colonization, as well as strong biocompatibility. It markedly improved early-stage osteogenic commitment and late-stage biomineralization compared with HAP and HAP-HYA controls. Moreover, the composite exhibited enhanced cytoprotective properties by inhibiting intracellular ROS in H2O2-treated cells.

The VitC@HAP-HYA composite effectively combines multifunctional qualities, demonstrating superior bioactivity, increased osteogenic potential, and vital cytoprotective features. These results robustly endorse its potential as a sophisticated framework for bone tissue engineering, including applications such as anti-cancer bone excision repair.