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.