Effects of local nitric oxide release on human mesenchymal stem cell attachment and proliferation on gelatin hydrogel surface

Abstract

Nitric oxide plays important roles in cardiovascular homeostasis, immune responses and wound repair. Therefore, polymers that release nitric oxide locally at the surface exhibit improved biocompatibility for biomedical implants through reducing neointimal hyperplasia and thrombosis caused by blood vessel wall damage. The objective of this article was to fabricate a nitric oxide–releasing gelatin hydrogel that can continuously generate nitric oxide at a physiologically relevant level and inhibit cell attachment and proliferation. The nitric oxide donor, S-nitroso-N-acetylpenicillamine (SNAP), was successfully conjugated to the gelatin hydrogel, which showed a rapid nitric oxide release in the first 2 h and then a slower but sustained release in the next 70-h period. Human mesenchymal stem cells (hMSCs), as a model cell line with wide biomedical applications, were used to examine the cell attachment and proliferation of the nitric oxide–releasing gelatin hydrogel. Compared with the control gelatin, the nitric oxide–releasing gelatin hydrogel demonstrated a 0·35 times lower hMSCs attachment at 6 h and a 3·15 times lower hMSCs proliferation after 72-h incubation. Moreover, hMSCs on nitric oxide–releasing gelatin exhibited a rounder cell shape and covered less cellular area than their counterparts on the control gelatin hydrogel. This gelatin hydrogel with local nitric oxide release at physiological level provides a promising therapeutic approach in enhancing the performance of biomedical implants.

Increasing Mechanical Strength of Gelatin Hydrogels by Divalent Metal Ion Removal

“…The purified gelatin solution was obtained as previously described36. Briefly, Chelex 100® resin was added into the unpurified gelatin solution (5 g/100 ml) and continuously stirring for 24 hours at 60°C.…”

Nitric oxide regulates cell behavior on an interactive cell‐derived extracellular matrix scaffold

“…The influence of NO on cell proliferation, migration, and differentiation have been studied using different NO donors on various substrates such as tissue culture plastic, hydrogel, and polymer nanofibers. 25,27,28 However, none of these systems provide a naturally occurring ECM-rich microenvironment that can dynamically interact with cells and mediating molecules involved in tissue healing process. Highly aligned cell-derived ECM scaffolds recapitulate both the in vivo three-dimensional (3D) nanofibrous anisotropic structure and biochemical microenvironment, therefore can serve as an ideal platform to study the in vitro cell growth, adhesion, and interaction.…”

“…The influence of NO on cell proliferation, migration, and differentiation have been studied using different NO donors on various substrates such as tissue culture plastic, hydrogel, and polymer nanofibers . However, none of these systems provide a naturally occurring ECM‐rich microenvironment that can dynamically interact with cells and mediating molecules involved in tissue healing process.…”

“…The NO release under physiological conditions was measured as described in our previous publication . To do this, the samples were immersed in cell culture medium at 37°C in 5% CO 2 incubator.…”

“…The NO release under physiological conditions was measured as described in our previous publication. 25 To do this, the samples were immersed in cell culture medium at 378C in 5% CO 2 incubator. The supernatant was collected at specific time points for nitrite concentration quantification via the Griess assay kit (Life Technologies) with absorbance recorded on a Versamax tunable microplate reader (Molecular Devices, Sunnyvale, CA).…”