Evaluation of the osteogenic potential of crocin-incorporated collagen scaffold on the bone marrow mesenchymal stem cells

Abstract

Objective: Present study aimed to evaluate the effect of crocin-loaded collagen scaffold on the osteogenic differentiation of rat bone marrow-derived mesenchymal stem cells (BM-MSCs). Significance: Different studies have been conducted to develop an efficient strategy to accelerate and improve the recovery process of bone defects. It was shown that crocin (CRO), extracted from saffron, could induce osteogenic differentiation of rat BM-MSCs. Scaffolds can also provide a 3-dimensional environment for migration, adhesion, growth, and proliferation of MSCs. Methods: Collagen (COL) scaffolds were fabricated through freeze-drying followed by cross-linking by dehydrothermal method. Then, crocin was incorporated into the scaffolds. Physicochemical characterization of the scaffolds was evaluated. Rat BM- MSCs were seeded on crocin-loaded collagen scaffolds and cultured for 14 days. Osteogenic differentiation was evaluated using alizarin red (ALZ) staining and alkaline phosphatase (ALP) activity assay and compared to the positive control group. Results: The average pore size of the collagen scaffolds was about 97 ± 6.7 µm. Formation of amide cross-links was confirmed by FTIR. The scaffolds were capable of uptaking water up to 50 times more than their initial dry weight and releasing above 90% of their uploaded crocin during 24 h. Collagen scaffolds containing crocin (25 and 50 μM) increased ALZ intensity (3.16 ± 0.3 and 7.32 ± 0.3 folds, respectively) and ALP activity (13.7 ± 1.1 and 12.2 ± 9.4 folds, respectively) in comparison with the positive control group. Conclusion: Crocin-loaded collagen scaffold could effectively enhance calcium deposition and ALP activity in BM-MSCs and therefore proposed as a good candidate to accelerate the healing process of vital bone defects.