Antibacterial Vitamin K3 Carnosine Peptide-Laden Silk Fibroin Electrospun Fibers for Improvement of Skin Wound Healing in Diabetic Rats

Abstract

The utilization of a multifunctional bioactive molecule functionalized electrospun dressing in tissue repair and regenerative function is a prominent therapeutic strategy for preparing efficient biomaterials to promote chronic wound healing. Designing robust and highly efficient antibacterial agents in resistance against microbes and bacterial infections is a key challenge for accelerating diabetic wound healing until today. In this study, we developed a vitamin K3 carnosine peptide (VKC)laden silk fibroin electrospun scaffold (SF-VKC) for diabetic wound healing. The structural confirmation of synthesized VKC was characterized by 1 H NMR, 13 C NMR, electrospray ionization mass spectrometry (ESI-MS), and attenuated total reflection Fourier transform infrared (ATR-FTIR) spectroscopy analysis, and the cell viability of VKC was evaluated by the CCK-8 assay in HFF1 and NIH 3T3 cells. VKC shows excellent cell viability on both cell lines, and the VKC and SF-VKC electrospun mats exhibited excellent antibacterial activity against both Gram-positive (Staphylococcus aureus) and Gram-negative (Escherichia coli and Pseudomonas aeruginosa) bacteria. Prepared SF and SF-VKC fibrous mats were well characterized, and the SF-VKC nanofiber mat presented good biodegradability, adhesiveness, unique mechanical property, expedient water uptake property, sustained drug release, and excellent biocompatibility for chronic wound healing. The in vitro tissue engineering study depicted excellent cell migration and cell−cell interaction in the NIH 3T3 cells over the VKCimpregnated silk fibroin (SF-VKC) mat. A higher population of cell migration was observed in cells' denuded area (scratched region) compared to the native SF fibrous mat. Interestingly, our results demonstrated that the prepared VKC-impregnated SF mat had potentially promoted the STZ-induced diabetic wound healing in a shorter period than the pure SF mat. Thus, obtained in vitro and in vivo outcomes suggest that the VKC-laden SF electrospun fibrous mat could be a better and inexpensive fibrous antibacterial biomaterial to elicit earlier re-epithelialization and efficient matrix remodeling for accelerating chronic infected wound reconstruction in skin diabetic wound healing applications.

Scaffolds in the microbial resistant era: Fabrication, materials, properties and tissue engineering applications

“…Kandhasamy et al. developed silk fibroin electrospun fiber mats containing antibacterial Vitamin K3 carnosine peptides for diabetic wound healing applications [ 299 ]. The fiber mats presented good biodegradability, adhesiveness and sustained drug release.…”

“…Some studies, however, did not include biocompatibility assays [ 279 , 284 , 285 , 288 ]. Fibroblasts [ 293 , 294 , 299 , 300 , 302 , 304 ], fibroblast-like cells [ 301 ], epidermal [ 296 ], endothermal [ 297 ] and keratinocytes [ 298 , 303 ] were used as cell models to evaluate the cytotoxicity of antibacterial scaffolds prepared with antibacterial polymers, peptides and scaffolds incorporating carbon nanomaterials. The approaches reported by the in vivo assays also showed good wound healing efficacy [ 293 , 299 , 302 , 303 ].…”

“…Fibroblasts [ 293 , 294 , 299 , 300 , 302 , 304 ], fibroblast-like cells [ 301 ], epidermal [ 296 ], endothermal [ 297 ] and keratinocytes [ 298 , 303 ] were used as cell models to evaluate the cytotoxicity of antibacterial scaffolds prepared with antibacterial polymers, peptides and scaffolds incorporating carbon nanomaterials. The approaches reported by the in vivo assays also showed good wound healing efficacy [ 293 , 299 , 302 , 303 ]. Finally, biocompatibility of antibacterial scaffolds prepared by combining strategies and alternative methods has been evaluated using fibroblasts (both human and murine) [ [307] , [308] , [309] , 312 , 314 , 320 ], fibroblast-like cells [ 316 , 321 , 322 ], epithelial cells [ 306 ], osteoblasts [ 312 ], and also Schwann cells [ 318 ] and red blood cells [ 322 ].…”

Effect of vitamin K on wound healing: A systematic review and meta-analysis based on preclinical studies

“…In addition, the different forms and adjuvants of vitamin K will cause high heterogeneity. In the two studies conducted by Kandhasamy et al (2021), vitamin K was brought into silk fibroin electrospun fibers and compared with silk fibroin electrospun fibers and PBS. In these studies, the strain and body weight of rats may also cause heterogeneity.…”

“…applied different concentrations of vitamin K for intervention. In this meta-analysis, studies with vitamin K as the intervention method were included, ignoring the form and adjuvant of vitamin K. For example, in the study ofKandhasamy et al (2021), silk fibroin electrospun fibers in vitamin K were considered in the intervention method. The included studies used Eucerin, 1% phenytoin, phosphatebuffered saline (PBS), etc.…”