Detection of Formation of Recombinant Human Keratinocyte Growth Factor Loaded Chitosan Nanoparticles Based on its Optical Properties

Abstract

Background: Recombinant human keratinocyte growth factor (rHuKGF) is a protein used to treat oral mucositis caused by radio and chemotherapy in patients with hematologic malignancy. The rHuKGF is available in the form of intravenous bolus injection. In this study, new formulation of rHuKGF-loaded chitosan nanoparticles was developed to improve patient compliance. Methods: Chitosan nanoparticles (CNPs) loaded with rHuKGF were prepared by ionic gelation method. The tripolyphosphate (TPP) cross-linked with chitosan molecules at pH >5.0 and form the nanoparticles. An infrared spectroscopic technique was conducted to confirm the formation of nanoparticles as a result of ionotropic interaction between TPP and chitosan. Zeta Sizer was used to determine the size, polydispersity index (PdI) and zeta potential of the prepared nanoparticles. The morphological characteristics of CNPs were measured by field emission scanning electron microscope. During the formation of CNPs, the rHuKGF was entrapped in the nanoparticles. The loading capacity of rHuKGF in CNPs was observed to be dependent on how much amount of rHuKGF/TPP solution was added to convert all the chitosan molecules to form nanoparticles. A double beam UV/Vis spectroscopic method was used to detect the formation of these rHuKGF loaded CNPs based on their optical properties. Results: The produced rHuKGF-loaded CNPs were colorless, cloudy, and positively charged monodisperse with a spherical shape. The prepared CNPs have particles size of 119 ± 74.62 nm, surface charge of +20.3 ± 6.46 mV and 0.217 polydispersity index. The shape of prepared CNPs was found to be spherical using field emission scanning electron microscope (FESEM). The interfacial polyelectrolyte complexation between TPP and chitosan was confirmed by comparing the FTIR spectra of TPP, chitosan, physical mixture of chitosan and TPP and CNPs. The loading capacity of the rHuKGF in CNPs was found to be 93.3 ± 2.02%. The formation of rHuKGF loaded CNPs was detected by double beam UV/Vis Spectroscopy at 232.2 nm. Conclusion: The results of the current work were utilized for designing a continuous monitoring and detection system for the formation of CNPs. The outcomes of this technique are useful to avoid the loss of rHuKGF during nanoparticle formation and improving the loading capacity of CNPs.