(T1130-02-08) Intestinal Organoid as a Predictive Model for the Assessment of Nano-Drug Delivery Systems

Purpose: In the current study, we studied the role of intestinal organoids as a predictive model to assess the efficiency of the nano-drug delivery system. Therefore, we generated organoid models from mice's small intestines and studied the effect of surface functionalization of nanoformulations, such as protein nanoparticles (Zein) with PEG molecules and/or folic acid (FA), on the stability and enhancement of the accumulation and residence time which will consequently improve drug transport of hydrophobic drugs such as paclitaxel.

Methods: Crypts were isolated from mice using the previously reported method [98, 99]. All animal procedures were conducted with the approval of Philipps University of Marburg, Institutional Animal Care, and Use Committee. For the uptake studies, the intestinal organoids were seeded into ibidi 8 well μ Slides and covered with culture medium. imaging was captured by confocal laser scanning microscopy (CLSM). For the in vivo biodistribution analysis, Lyophilized formulations loaded with DIR (IR-thermosensitive dye) were dispersed in PBS and given via oral gavage at a single dose. Fluorescence of Dir was Ddetected, using PhotonIMAGER™ OPTIMA . In vivo pharmacokinetics was performed on albino rabbits and concentrations of paclitaxel in blood plasma concentration was analyzed using HPLC.

Results: PEGylated and folate-conjugated nanoparticles loaded with coumarin, in contact with organoid models, resulted in a higher accumulation of the folate-tethered nanoparticles over 24 h. Additionally, these findings in ex-vivo organoids provided initial evidence that the FA-tagged nanoparticle could be used to achieve better bioavailability enhancement of paclitaxel, a prerequisite for the success of the in vivo assessment studies. Furthermore, to validate the above-predicted results, in vivo biodistribution and pharmacokinetic studies were performed. The optical imaging after the oral administration of the Dir-loaded formulations in biodistribution analysis revealed several folds increase in residence time for the FA-tailored nanoparticles over 24 h compared to the pegylated particles. More interestingly, the pharmacokinetics outcomes empathized that the plasma concentration of paclitaxel in the case of FA-targeted PEGylated particles has considerably higher absorption post-gavage which stands for the elevated AUC of 7.6 folds relative to the free paclitaxel and was detectable in rabbits' plasma over the course of 72 h.

Conclusion: In conclusion, the in vivo validation of the predicted results from organoids suggested that these models are promising for the optimization of formulations- based drug delivery systems which offer an early-stage predicting approach for bioavailability studies.

References: 1. S. Ayehunie, T. Landry, Z. Stevens, A. Armento, P. Hayden, M. Klausner, Human Primary Cell-Based Organotypic Microtissues for Modeling Small Intestinal Drug Absorption, Pharm. Res. 35 (2018) 72.
2. R.I. Mahato, A.S. Narang, L. Thoma, D.D. Miller, Emerging trends in oral delivery of peptide and protein drugs, Crit. Rev. Ther. Drug Carrier Syst. 20 (2003) 153–214.
3. A.M. Abdelsalam, A. Somaida, G. Ambreen, A.M. Ayoub, I. Tariq, K. Engelhardt, P. Garidel, I. Fawaz, M.U. Amin, M. Wojcik, U. Bakowsky, Surface tailored zein as a novel delivery system for hypericin: Application in photodynamic therapy, Mater. Sci. Eng. C. 129 (2021) 112420.

Fig. 1. Demonstrative images for organoid models generated from mice's small intestines as a predictive tool for oral delivery enhancement. After 24 h of incubation with nanoformulations encapsulating coumarin 6 (green), the output results showed the efficient accumulation of Z-PEG-FA nanoparticles compared to the non-targeted ones.


Fig. 2. In vivo optical biodistribution. Fluorescence images of rats (n = 3 ± SD) after 2, 4, and 24 h of oral administration of DIR-loaded Z-PEG and Z-PEG-GA formulations (A) Fluorescence images of isolated GIT after the same time intervals. (B) The quantitative average NIR signals were obtained from rats’ GIT at the different time points (***: p < 0.001).


Fig. 3. Oral Pharmacokinetic study. (A) The plasma concentration (Cp) versus time curves of orally administered free paclitaxel and the paclitaxel loaded in Z-PEG and Z-PEG-FA formulations (B) The average folds enhancement in the Area under the concentration/time curve in rabbits' plasma for Z-PEG and Z-PEG-FA formulations relative to free paclitaxel (n = 3 ± SD).