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Evaluation of the biodegradation and in vitro biocompatibility of Glutaraldehyde-crosslinked decellularized human umbilical artery scaffolds
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Abstract
Objectives: This study aimed to evaluate the biomechanical durability and cytocompatibility of decellularized umbilical artery (DUA) scaffolds treated with 0.1% glutaraldehyde (GA).
Methods: The 0.1% GA-treated decellularized umbilical artery scaffolds (G - DUA) were incubated in PBS (pH 7.4) at 37°C for 2, 4, and 8 weeks. Changes in tensile strength and remaining mass were compared with the untreated group (DUA). Biocompatibility was assessed using an indirect cytotoxicity test (MTT assay), a cell migration test (scratch wound healing assay), and by evaluating the direct adhesion of fibroblasts onto the G - DUA scaffold surface via H&E staining.
Results: Glutaraldehyde treatment demonstrated a pronounced effect on enhancing the durability of the scaffold. Regarding mass change, at week 4, the DUA group lost approximately 25% of its mass (75.2 ± 2.41% remaining), whereas the G - DUA group showed almost no change (101.94 ± 0.77%). This degradation continued to week 8, where the remaining mass of the DUA group was only 55.38 ± 2.18%, in contrast to 99.46 ± 1.361% for the G-DUA group (p < 0.0001). Similarly, in terms of mechanical strength, the tensile strength of the DUA group decreased by nearly half after 4 weeks (2.886 ± 0.67 N) and continued to decline sharply by week 8 (1 ± 0.31 N). Conversely, the G-DUA group maintained significantly better durability, with tensile strengths of 5.526 ± 0.67 N at week 4 and 4.4 ± 0.58 N at week 8, respectively (p < 0.0001). Regarding biocompatibility, the results indicated that the extract from G-DUA was not cytotoxic; cell viability rates exceeded 99% at all time points, comparable to the PTFE negative control group (p > 0.05). The G - DUA scaffold also did not adversely affect migration and adhesion of fibroblasts on its surface.
Conclusion: Glutaraldehyde treatment significantly enhances the mechanical strength and biodegradation resistance of decellularized human umbilical artery scaffolds without inducing in vitro cytotoxicity in fibroblasts.
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Keywords
Decellularized scaffold, Glutaraldehyde, Cross-linking, Biocompatibility
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