Nanofibrous Polycaprolactone-Nitazoxanide Membrane: a Novel Solution for Periodontitis

Objectives: Periodontitis affects half the global population and results in loss of tooth-supporting structures. Antimicrobial membranes are beneficial, but acquired resistance to antibiotics concerns demand safer alternatives. Nitazoxanide (NZ), a broad spectrum drug, is known to evade bacterial resistance by disrupting the pyruvate:ferredoxin oxidoreductase pathway. Utilizing the versatility of electrospinning, this study aimed to synthesize, characterize, and assess the cytocompatibility, osteogenic potential, and antimicrobial efficacy of NZ-loaded membranes for periodontal tissue regeneration.
Methods: Poly(ε-caprolactone) was dissolved in hexafluoroisopropanol and electrospun with NZ at 0 (N0, control), 2.5 (N2.5) and 5 (N5) % concentrations (w/w, with respect to polymer weight) (n=4). Morpho-chemical characterization was performed by scanning electron microscopy-energy dispersive spectroscopy and Fourier Transform Infrared spectroscopy. NZ release profile from membranes was determined by spectrophotometer (λ_max=444) for 14 days. Cytocompatibility of periodontal ligament stem cells was investigated through live and dead imaging and MTS assay. Mineralization potential was determined by ALP assay and Alizarin red S staining. Antimicrobial efficacy of the membranes was evaluated against Aggregatibacter actinomycetemcomitans and Fusobacterium nucleatum by counting viable bacteria and agar diffusion method. Statistical analysis was done by Kruskal Wallis test. α=0.05.
Results: Morpho-chemical characterization revealed average fiber diameter of 200 nm across all groups and confirmed incorporation of secondary amide groups and sulphur atoms of NZ molecule in both test groups. Drug release profile indicated an initial burst release within 24 hours. Qualitative and quantitative cell viability results exceeded 70% at days 3 and 5 in all groups. Both test groups presented notable ALP and Alizarin red S concentrations confirming that NZ had no adverse impact on mineralization potential. Agar diffusion assays depicted significant diameters of inhibition zones and no viable periodontopathogens were observed in any of the test groups.
Conclusions: Results indicate PCL-NZ membrane has promising potential in combating periodontopathogens and supporting mineralization.