Facile design and development of nano-clustery graphene-based macromolecular protein hydrogel loaded with ciprofloxacin to antibacterial improvement for the treatment of burn wound injury

Abstract

Nowadays, awareness about the burn wound is often considered difficult due to bacterial and other organism infections. The facile and eco-friendly preparations of antibiotic-loaded hydrogel-based bio-composites have great attention in the field of wound dressing for burn wound therapy and nursing care. In the present investigation, we have developed ciprofloxacin (CF)-encapsulated graphene-silk fibroin macromolecular hydrogel dressings material with unique chemical and physical properties to achieve the desirable antibacterial efficacy and healing activity. The antibacterial activity of prepared hydrogel was evaluated against bacterial pathogens treated with different concentrations of CF, which have been provided improved antibacterial activity on burn wound infection. In vitro, cytocompatibility evaluations were performed to imply the suitability of hydrogel on fibroblast cells, which has been dramatically related to in vivo wound healing. Furthermore, an in vivo wound healing analysis was carried out using a rat to observe the capability of the CF-incorporated GH/SF hydrogel matrix. Thus, this investigation widely demonstrates the healing ability of prepared hydrogel matrix and could be a significant landmark in the research on burn wound healing applications.

Keywords: Antibacterial activity; Burn wound; Ciprofloxacin; Graphene nano; Hydrogel; Silk fibroin.

Fig. 1

The phase purity and crystallinity nature of the as-prepared samples were observed and exhibited XRD pattern analysis; a the results of graphene oxide (GO), bare graphene, silk fibroin (SF), and GO-loaded SF sample (GO/SF); b the XRD patterns of bare ciprofloxacin (CF) and ciprofloxacin-encapsulated GO/SF (CF@GH/SF) samples

Fig. 2

The surface morphological structure of the prepared silk fibroin hydrogel in the absence (a) and presence (b) of nano-graphene sheet was exhibited through scanning electron microscopic (SEM) analysis

Fig. 3

The chemical structure and intermolecular interactions between the distributed nano-formulations and bio-polymeric groups have been observed and demonstrated by the FTIR and Raman spectroscopic analysis; a FTIR spectroscopic investigations of prepared SF, GH, GO and CF-loaded GH/SF samples and b Raman spectroscopic analysis of prepared SF, GH, and SF/GH, respectively

Fig. 4

The thermal stability and nature of temperature on prepared materials were observed by thermal gravimetric analysis (TGA); TGA results exhibited the thermal degradation behaviors of prepared SF, GH, and GH/SF in the temperature from 0 to 700 °C

Fig. 5

The elemental presentations and nature of prepared hydrogel materials were observed by X-ray photoelectron spectrum (XPS) survey analysis; the presentation of carbon (C), nitrogen (N), and oxygen (O) elements into the prepared SF, GH, and GH/SF hydrogel samples was observed

Fig. 6

Antibacterial properties of prepared CF-loaded GH/SF hydrogel materials were investigated against clinically approved gram-positive (S. aureus) and gram-negative (E. coli) bacterial pathogens; a & b growth curve analysis of as-prepared CF@GH/SF hydrogel material in different concentrations and zone of inhibition; c & d results observation of prepared CF@GH/SF sample in different concentration

Fig. 7

In vitro cytocompatibility of prepared hydrogel materials was studied by the methods of quantitative observation of in vitro cell viability (MTT assay) and cell migration analysis in different time intervals; a microscopic visualization of in vitro cell migrations treated with control (PBS), GH/SF, and CF@GH/SF samples in different time durations (6, 12, and 18 h); b & c qualitative results of cell viability and cell proliferation in different time intervals

Fig. 8

In vivo burn wound healing was observed primarily by created burn wound injury and treated with prepared control (SF gel), GH/SF, and CF@GH/SF hydrogel samples in different postoperative days (3, 6, 12, and 18 days); The photovisualization exhibited that CF@GH/SF hydrogel sample has great healing ability when compared to GH/SF and control sample

Fig. 9

Histomorphological observation (hematoxylin–eosin staining microscopic study) of burn wound regeneration after treatment of prepared hydrogel dressings of GH/SF, CG@GH/SF, and control sample on the 3rd, 6th, 12th, and 18th day after postoperative; the images exhibited in different magnifications (10x and 40x)