Covalent immobilization of biomolecules on stent materials through mussel adhesive protein coating to form biofunctional films

Abstract

It is widely accepted that surface biofunctional modification may be an effective approach to improve biocompatibility and confer new bioactive properties on biomaterials. In this work, mussel adhesive protein (MAP) was applied as a coating on 316L stainless steel substrates (316L SS) and stents, and then either immobilized VEGF or CD34 antibody were added to create biofunctional films. The properties of the MAP coating were characterized by scanning electron microscope (SEM), atomic force microscope (AFM) and water contact angle test. Universal tensile testing showed that the MAP coating has adequate adhesion strength on a 316L stainless steel material surface. Subsequent cytotoxicity and hemolysis rate tests showed that the MAP coatings have good biocompatibility. Moreover, using N-(3-Dimethylaminopropyl)-N`-ethylcarbodiimide hydrochloride and Nhydroxysulfosussinimide (EDC/NHS) chemistry, VEGF and CD34 antibody were immobilized on the MAP coatings. The amount and immobilized yield of VEGF on the MAP coatings were analyzed by enzyme-linked immuno-assays (ELISA). Finally, an endothelial cells culture showed that the VEGF biofunctional film can promote the viability and proliferation of endothelial cells. An in vitro CD34 + cells capturing test also verified the bioactive properties of the CD34 antibody coated stents. These results showed that the MAP coatings allowed effective biomolecule immobilization, providing a promising platform for vascular device modification.

The emerging translational potential of GDF11 in chronic wound healing

“…Wang et al. recently reported the increased retention of VEGF on mussel-adhesion protein (MAP)-coated stents via carbodiimide chemistry [ 74 ]. However, covalent linking may cause protein denaturation and impairment of overall bioactivity.…”

Applying Principles of Regenerative Medicine to Vascular Stent Development

“…For all regenerative purposes and translational intents, current and future endeavors utilizing cell-capture mechanism may continue in three directions: 1) combining cell-capture molecules with other prohealing/proregenerative molecules; ( Chen et al, 2015 ; Zhang et al, 2015 ; Wang et al, 2020a ; Yang et al, 2020a ) 2) using more selective or specific molecules to recognize a subset of EPCs; ( Hirase, 2016 ; Tang et al, 2016 ) 3) exploiting EPC-derived secretomes such as those in the form of exosomes ( Zeng et al, 2021 ).…”

Biomaterials for protein delivery for complex tissue healing responses

“…47 Building upon this approach, Wang et al recently demonstrated enhanced retention of VEGF on mussel-adhesion protein (MAP)-coated stents through carbodiimide chemistry. 48 MAP, which is rich in 3,4-dihydroxy-L-phenylalanine (Dopa), has immense celladhesive capacity. 49 MAP substrates coated onto a silicon wafer were dip-coated in VEGF, which was immobilized using EDC/ NHS chemistry.…”

“…The tethered VEGF maintained its bioactivity, resulting in increased spreading of endothelial cells, while crosslinking of the MAP increased its retention on the stent. 48 Although covalent conjugation of proteins to materials prolongs local protein presentation within an implantation site, this strategy may reduce overall protein bioactivity by causing protein denaturation during processing or interfering with the ability of proteins to interact with cell integrins and receptors. [50][51][52][53][54][55] For example, modification of collagen I with EDC reduced cell adhesion by inhibiting cationic cell interactions with collagen's integrin binding domains.…”

“…Antibodies specific to proteins of interest can be covalently conjugated 87 or physically adsorbed to biomaterials. 48,88 Alternatively, high affinity molecules can be developed to achieve extremely specific protein-material interactions and can be produced inexpensively compared to antibodies. [89][90][91] Some notable strategies include peptides that directly bind to the protein of interest, 88,90 the synthesis of fusion proteins, consisting of a peptide-or ECM-binding domain and functional protein domain, [91][92][93][94][95][96] and DNA aptamers that contain a protein sequestering component and a cell adhesive component that provides localized cargo release.…”