In-silico engineering of an antimicrobial growth factor fusion protein to prevent post-laser infection in hemorrhoidal wounds

Aim: Post laser wound infection is a recurrent complication after hemorrhoidal surgery mainly because polymicrobial flora is colonized in the anorectal environment and the mucosal repair is impaired. This study aims to overcome the drawbacks of current therapeutic options and to provide a novel generation of prophylactic biomolecule with antimicrobial activities, monitored to meet the specific conditions of the microenvironment of anorectal wound repair.Material and Methods: To overcome this two-fold problem of infection and slow wound healing, a current study developed an in-silico bioengineering intervention to design and characterize a new antimicrobial- growth factor fusion protein fusing selected antimicrobial peptide (AMPs) with the fibroblast growth factor 10 (FGF10) domain.Results: The Antimicrobial Peptide Database was searched with four cationic AMPs, which included AP00608, AP02570, AP03795, and AP00952, and screened using AllerTOP v2.0 and ToxiPred, which confirmed their non-allergenicity and non-toxicity. The obtained engineered fusion construct possessed desirable physicochemical properties (32.84 kDa; pI 10.19; instability index 38.52; GRAVY -0.33; solubility 0.48) which implies high stability and hydrophilicity. Secondary structure prediction showed 27.34% alpha-helices and 16.26% beta-strands, whereas AlphaFold probability and ERRAT (94.28% proved structural accuracy). ClusPro 2.0 showed good binding affinities to FGFR2b (1NUN) and FGFR2 (1EV2) with energies of -1161.9 kcal mol -1 and -1144.4 kcal mol -1 respectively. Following 500 Desmond molecular dynamics simulations determined the thermodynamic stability, compact folding, and the maintenance of hydrogen bonds within the complexes.Conclusion: All these findings suggest that the proposed fusion construct has a potential to serve as a dual-acting therapeutic - preventing microbial colonization and stimulating epithelial repair in post-laser hemorrhoidal wounds. This computational research therefore provides the basis of experimental validation and translational development of multifunctional wound-healing biologics in the future.