Final Report Summary - MUCOSINT (Multifunctional Composite Silica Nanotubes for Targeted Delivery)
The project objectives for the MUCOSINT project can be summarised in two sections:
1. mobility and integration objectives; and
2. experimental objectives.
Experimental objectives are:
a) fabrication of silica nano test tubes (SiNTs);
b) creating a responsive gel formulation and filling of SiNTs with this gel to form composite SiNTs (CoSiNTs);
c) biofunctionalisation and related characterisation of SiNTs with Folic acid (FA) moieties;
d) investigation of the in vitro performance of SiNTs with various compositions.
It can be easily concluded that the mobility and integration objectives of the MUCOSINT project are fulfilled. The researcher made the 3 proposed United States (US) university visits, participated in four international conferences and three national conferences, and made several presentations. He made various collaborations which yielded 9 journal publications within the project period. Among these, one article is related with the MUCOSINT project and the direct results of the project are in preparation for a full journal article. The project also greatly enhanced the reintegration of the researcher. He is currently a faculty member at the top biomedical engineering department in Turkey as an associate professor.
In terms of experimental progress, AAO membranes were synthesised by two-step anodisation1 and SiNTs of different dimensions were produced by the surface sol-gel method2 and characterised by electron microscopy. A responsive gel formulation was created using the following constituents: hydroxyethylmethacrylate (HEMA), poly(ethylene glycol) ethylether methacrylate (PEG-EEM), 2-aminoethylmethacrylate hydrochloride (AEM), trimethyloylpropane ethoxylate triacrylate as the crosslinker, doxorubicin (DOX) hydrochloride as the drug, 2,2-diethoxyacetophenone as the photoinitiator, and finally water and isopropanol as solvents for AEM, DOX and the initiator. After showing the pH responsive release of DOX from bulk gels, fabrication of gel-filled SiNTs, namely, CoSiNTs were conducted.
Obtaining drug release profiles from the CoSiNTs at different pH values was a problematic task either by the originally proposed UV-Vis or electrochemical detection of the released DOX although SiNTs with largest proposed dimension were utilised. Hence, the drug release performances of drug-filled SiNTs are solely based on cell viability studies. Prior to this in vitro analysis part, SiNTs were targeted by FA conjugation via carbodiimide activation3, and here, each step was characterised by X-Ray photoelectron spectroscopy (XPS), Fourier Transform infrared spectroscopy (FT-IR), as well as zeta potential measurements. After the optimisation of cell viability assay and incubation times, cell viability of tumour (SK-BR3 ) and healthy breast cell lines (MCF-12A) were investigated against SiNTs of various compositions and concentrations and further confirmed by apoptotic index calculations. The results demonstrate that MUCOSINTs (SiNT 5) are effective killers of tumor cells, larger viabilities are obtained for MUCOSINT treated healthy cells, and to induce cell death for both cell lines, the presence of DOX together with the targeting moiety are required. Formulations that do not contain DOX does not cause significant cell death (SiNT 1-3). Our results also suggest that when packaged within MUCOSINTs, much smaller DOX concentrations are required to cause comparable cell death that originates from the exposure to free drug.
1. mobility and integration objectives; and
2. experimental objectives.
Experimental objectives are:
a) fabrication of silica nano test tubes (SiNTs);
b) creating a responsive gel formulation and filling of SiNTs with this gel to form composite SiNTs (CoSiNTs);
c) biofunctionalisation and related characterisation of SiNTs with Folic acid (FA) moieties;
d) investigation of the in vitro performance of SiNTs with various compositions.
It can be easily concluded that the mobility and integration objectives of the MUCOSINT project are fulfilled. The researcher made the 3 proposed United States (US) university visits, participated in four international conferences and three national conferences, and made several presentations. He made various collaborations which yielded 9 journal publications within the project period. Among these, one article is related with the MUCOSINT project and the direct results of the project are in preparation for a full journal article. The project also greatly enhanced the reintegration of the researcher. He is currently a faculty member at the top biomedical engineering department in Turkey as an associate professor.
In terms of experimental progress, AAO membranes were synthesised by two-step anodisation1 and SiNTs of different dimensions were produced by the surface sol-gel method2 and characterised by electron microscopy. A responsive gel formulation was created using the following constituents: hydroxyethylmethacrylate (HEMA), poly(ethylene glycol) ethylether methacrylate (PEG-EEM), 2-aminoethylmethacrylate hydrochloride (AEM), trimethyloylpropane ethoxylate triacrylate as the crosslinker, doxorubicin (DOX) hydrochloride as the drug, 2,2-diethoxyacetophenone as the photoinitiator, and finally water and isopropanol as solvents for AEM, DOX and the initiator. After showing the pH responsive release of DOX from bulk gels, fabrication of gel-filled SiNTs, namely, CoSiNTs were conducted.
Obtaining drug release profiles from the CoSiNTs at different pH values was a problematic task either by the originally proposed UV-Vis or electrochemical detection of the released DOX although SiNTs with largest proposed dimension were utilised. Hence, the drug release performances of drug-filled SiNTs are solely based on cell viability studies. Prior to this in vitro analysis part, SiNTs were targeted by FA conjugation via carbodiimide activation3, and here, each step was characterised by X-Ray photoelectron spectroscopy (XPS), Fourier Transform infrared spectroscopy (FT-IR), as well as zeta potential measurements. After the optimisation of cell viability assay and incubation times, cell viability of tumour (SK-BR3 ) and healthy breast cell lines (MCF-12A) were investigated against SiNTs of various compositions and concentrations and further confirmed by apoptotic index calculations. The results demonstrate that MUCOSINTs (SiNT 5) are effective killers of tumor cells, larger viabilities are obtained for MUCOSINT treated healthy cells, and to induce cell death for both cell lines, the presence of DOX together with the targeting moiety are required. Formulations that do not contain DOX does not cause significant cell death (SiNT 1-3). Our results also suggest that when packaged within MUCOSINTs, much smaller DOX concentrations are required to cause comparable cell death that originates from the exposure to free drug.