Biological and structural properties of curcumin-loaded graphene oxide incorporated collagen as composite scaffold for bone regeneration

Abstract

Introduction: To address the challenges related to bone defects, including osteoinductivity deficiency and post-implantation infection risk, this study developed the collagen composite scaffolds (CUR-GO-COL) with multifunctionality by integrating the curcumin-loaded graphene oxide with collagen through a freeze-drying-cross-linking process.

Methods: The morphological and structural characteristics of the composite scaffolds were analyzed, along with their physicochemical properties, including water absorption capacity, water retention rate, porosity, in vitro degradation, and curcumin release. To evaluate the biocompatibility, cell viability, proliferation, and adhesion capabilities of the composite scaffolds, as well as their osteogenic and antimicrobial properties, in vitro cell and bacterial assays were conducted. These assays were designed to assess the impact of the composite scaffolds on cell behavior and bacterial growth, thereby providing insights into their potential for promoting osteogenesis and inhibiting infection.

Results: The CUR-GO-COL composite scaffold with a CUR-GO concentration of 0.05% (w/v) exhibits optimal biological compatibility and stable and slow curcumin release rate. Furthermore, in vitro cell and bacterial tests demonstrated that the prepared CUR-GO-COL composite scaffolds enhance cell viability, proliferation and adhesion, and offer superior osteogenic and antimicrobial properties compared with the CUR-GO composite scaffold, confirming the osteogenesis promotion and antimicrobial effects.

Discussion: The introduction of CUR-GO into collagen scaffold creates a bone-friendly microenvironment, and offers a theoretical foundation for the design, investigation and utilization of multifunctional bone tissue biomaterials.

Keywords: antibacterial; bone regeneration; collagen; curcumin; graphene oxide.

FIGURE 1

Schematic illustration of the fabrication of composite scaffold and their application in vitro.

FIGURE 2

Morphology and structure of the CUR-GO-COL composite scaffold. Macroscopic view and SEM images of the COL scaffold (A, B) and CUR-GO-COL scaffold (C, D). (E) XRD patterns, (F) FTIR spectra, and (G) Raman spectra obtained from the COL, GO-COL and CUR-GO-COL scaffolds.

FIGURE 3

Characterization of the CUR-GO-COL composite scaffold. (A) Water absorption, (B) water retention and (C) porosity obtained from COL, CUR-GO-COL1, CUR-GO-COL2, CUR-GO-COL3 and CUR-GO-COL4 scaffolds. (n = 3; *, P < 0.05; **, P < 0.01; ***P < 0.001 compared with the COL group). (D) CCK-8 assay of cell viability on the COL and CUR-GO-COL scaffolds. (n = 4; *, P < 0.05; **, P < 0.01; ***P < 0.001 compared with the COL group). (E) The degradation performance of the COL, GO-COL and CUR-GO-COL scaffolds. (F) Curcumin release from the CUR-GO-COL scaffolds.

FIGURE 4

Growth ability of BMSCs cells cultured in vitro. (A) Live/dead staining images of BMSCs cultured with the COL, GO-COL and CUR-GO-COL scaffolds. (B) Immunofluorescence images of BMSCs morphology on the COL, GO-COL and CUR-GO-COL scaffolds BMSCs nuclei stained with DAPI (blue) and the cytoskeleton stained with FITC (red). (C) CCK-8 assay of cell viability on the COL, GO-COL and CUR-GO-COL scaffolds at 1 and 4 days (n = 4).

FIGURE 5

Osteogenic differentiation ability of BMSCs co-cultured with three groups of scaffolds. (A) ALP staining. (B) ALP activity analysis. (C) ARS staining. (D) Semi-quantitative analysis. (n = 3; *, P < 0.05; **, P < 0.01; ***, P < 0.001).

FIGURE 6

Effect of three groups of scaffolds on bacterial growth. Growth curves (A) of (E) coli and (B) S.aureus. (n = 3) Plate diffusion results for (C) E. coli and (D) S.aureus.