1. LOCALIZED SURFACE PLASMON RESONANCE
Nanomedicine Application of Alternative Therapy Cancer Treatment in Animal Models with High-Grade Gliomas CT-2A
Oscar Casanova Carvajal1,2, Ana Lorena Urbano Bojorge1,2, Patricia Galán Montano1, Cristina Sánchez López de Pablo1,2,
Francisco del Pozo Guerrero1,2, Ricardo Martínez Murillo3, José Javier Serrano Olmedo1,2
1Center for Biomedical Technology, Technical University of Madrid, Madrid, Spain
2Biomedical Research Networking Center Bioengineering, Biomaterials, Nanomedicine (CIBER-BBN), Zaragoza, Spain
3Instituto Cajal, Spanish National Research Council, Madrid, Spain
Introduction
Cancer is one of the leading causes of death worldwide. It is expected that annual cases of cancer will increase from 14 million in 2012 to 22 million in the next two decades according to the World Health Organization [1]. On the other hand, the estimates of the Spanish Society of Medical Oncology for 2015 point out new cases only in Spain [2]. Tumors of glial cells, anaplastic astrocytoma and glioblastoma account for approximately 38% of primary brain tumors [3]. At present, despite the development of existing treatments such as chemotherapy, radiation therapy and neurosurgery these tumors tend to breed again, leading to a high mortality rate. The Center for Biomedical Technology (CTB) of the Technical University of Madrid (UPM) in cooperation with the Cajal Institute of the Spanish Council for Scientific Research (CSIC), research has been done with an application of nanomedicine using therapy through optical hyperthermia, taking as the starting point the results of in vitro experimentation with prior glioblastoma cells, using gold nanorods with laser at a wavelength of 808 nanometers, a wavelength in which tissue penetration is more efficient [4,5].
Material and Methods
Variable in each test
First Experiment
Second Experiment
Times Therapy 3 4
Nº injected dose 1
Nº Mouse 5 6
Mouse Classification
Mouse 1
Control
Control NPs
Mouse 2
Laser control 40 min no NPs
Laser control 25 min no NPs
Mouse 3
Therapy 15 min
Mouse 4
Therapy 25 min
Mouse 5
Therapy 40 min
Mouse 6
---
Healthy mouse
For this, it was used a syngeneic mouse model of glioblastoma by injecting of CT-2A astrocytoma cells into the caudate-putamen of C57BL/6 mice, as previously reported [6]. Five mice were used and distributed as follows: Control nanoparticles (NPs), NPs + laser therapy 15 minutes, plus 25 minutes NPs laser therapy, laser therapy and tumor control without NPs.
IN VIVO EXPERIMENTS
LOCALIZED SURFACE PLASMON RESONANCE
Light energy
Thermal energy
HYPERTHERMIA
LOCALIZED CELL DEATH
Stereotactic intratumoral injection of GNRs (λspr = 808 nm)
Therapy evaluation by quantifying tumoral volumes (MRI)
External irradiation with NIR Laser source
(808 nm)
CW laser
(MDL H808, CNILaser)
Extraction of NPs in the mouse brain
Stereotactic surgical methodology implemented
Sample preparation process concentrated AuNRs
Results
First Experiment
Second Experiment
The results of these experiments, in which the mice population was not large enough from a statistical point of view, showed comparing the individual control NPs versus animal therapy NPs + 15 laser minutes groups, a reduction of tumor growth (volume) of 15.99% which implies a reduction in the volume of 82,97 mm3.
Volume
Mouse 1
Mouse 2
Mouse 3
Mouse 4
Mouse 5
Control
Laser control
Therapy
15 min
Therapy 25 min
Therapy 40 min
Brain volume mm3
389
380
372
394
388
Tumor volume mm3 13 8 5 11 6
MEASURED VOL. IN MRI
Mouse 1
Mouse 2
Mouse 3
Mouse 4
Mouse 5
Control NPs
Laser control 25´
Therapy
15 min
25 min
CONTROL
Brain Vol. (mm3 )
506,2
493,0
472,3
475,3
501,4
Tumor Vol. (mm3 )
3 days post Therapy
19,3
8,0
18,9
18,5
9,9
Tumor Vol. (mm3 )
15 days post Therapy
146,4
137,3
63,5
212,8
90,7
Size difference observed in tumors from control. Percent by volume
Mouse 2: Laser control
38,5 %
Mouse 3: Therapy 15´
61,5 %
Mouse 4: Therapy 25´
15,4 %
Mouse 5: Therapy 40´
53,8 %
Mouse 3 – Therapy 15 min
Mouse 4 - Therapy 25 min
Conclusions
The observations of tumor volumes were made using Nuclear Magnetic Resonance Image.
The NPs remain in the injection site. Small fractions may migrate to neighboring regions.
Mouse 4 (Therapy of 25 minutes) suffered stroke pollution, so is not considered.
Future work will seek to optimize the intracranial stereotactic surgical method implemented in these experiments with mice as well as to increase the number of individuals to produce statistics that effectively validate this application.
References
[1] World Cancer Report World Health Organization. In
[2] Sociedad Española de Oncología Médica In
[3] Mehta M, Vogelbaum MA, Chang S, et al. Neoplasms of the central nervous system. In DeVita VT Jr, Lawrence TS, Rosenberg SA. Cancer: Principles and Practice of Oncology. 9th ed. Philadelphia, Pa: Lippincott Williams & Wilkins, 2011, pp
[4] Cristina Sánchez de Pablo. Contribución al Estudio del Sistema de Hipertermia Óptica y su Aplicación en Muestras Biológicas Tesis Doctoral.
[5] Fernández Cabada, T., Sánchez López de Pablo, C., et al. Induction of cell death in a glioblastoma line by hyperthermic therapy based on gold nanorods. International Journal of Nanomedicine, 7, 1511–1523. (2012) doi: /IJN.S28470.
[6] R. Martínez Murillo, A. Martínez. Standardization of an orthotopic mouse brain tumor model following transplantation of CT-2A astrocytoma cells. Histology and Histopathology Cellular and Molecular Biology. 2007, pp
Contact
Parque Científico y Tecnológico de la UPM
Campus de Montegancedo Pozuelo de Alarcón, Madrid, España,
Oscar Casanova Carvajal
Phone:
Bioinstrumentation Laboratory
Hyperthermia Unit Centre for Biomedical Technology (CTB)