Today, the search for solutions to reduce wound infection and restore wound receptivity also reduces its side effects which are a difficult problem in medical science research. The greatest options for this purpose are hydrogel dressings since they are compatible with tissue and have an antibacterial effect on wound healing. Chronic wounds represent a significant burden on people and healthcare systems worldwide. Bacteria often enter such skin wounds, causing irritation and complicating the healing process. In addition, bacteria cause infection, which inhibits rejuvenation and the production of collagen. This study is aimed at developing novel chitosan (CS)-hydrolyzed starch nanocomposite (HS/Ch-NC) loaded with ciprofloxacin to enhance its skin retention and wound healing efficacy and anti-biofilm efficacy. Drug-loading on the (HS/Ch-NC) and encapsulation efficiency was 55.2% and 97.2%, respectively. The activity of HS-NC loaded with ciprofloxacin as anti-biofilm activity by 72% and 63% against Enterobacter aerogenes and Pseudomonas aeruginosa, respectively. The obtained (HS/Ch-NC) loaded with ciprofloxacin is a promising candidate for the development of improved bandage materials, as cell viability and proliferation was assessed using an SRB assay with half-maximal inhibitory concentrations (IC) at 119.1 µg/ml. In vitro scratch wound healing assay revealed significant (p ≤ 0.05) acceleration in wound closure at 24 h enhanced by 56.04% 24-h and 100% 72-h post-exposure to (HS/Ch-NC) loaded ciprofloxacin, compared to the negative control. In vivo skin retention study revealed that (HS/Ch-NC)-loaded ciprofloxacin showed 3.65-fold higher retention, respectively, than ciprofloxacin. Thus, our study assumes that ciprofloxacin-loaded HS-NC is a potential delivery system for enhancing ciprofloxacin skin retention and wound healing activity.