Yonsei Med J.  2005 Feb;46(1):51-60. 10.3349/ymj.2005.46.1.51.

Therapeutic Effects of Holmium-166 Chitosan Complex in Rat Brain Tumor Model

• Huh R
• Park YS
• Lee JD
• Chung YS
• Park YG
• Chung SS
• Chang JW

• KMID: 2158112
• DOI: http://doi.org/10.3349/ymj.2005.46.1.51

Abstract

This study examined the effectiveness of Holmium-166 (Ho-166) chitosan complex therapy for a malignant glioma. Cultured C6 glioma cells (100, 000 in 5microliter) were injected into the caudate/putamen of 200 - 250 gram Wistar rats. Five days later, a Ho-166 chitosan complex was injected into the same site of the glioma injection. Four injection doses were administered: the control group received PBS 10microliter, group 1 received an injection of 100micro Ci (10microliter), group 2 received an injection of 50microCi (5microliter), and group 3 received an injection of 10micro Ci (1microliter). The average tumor volume for each group was 1.385 mm3 for the control group, 0.036 mm3 for group 1, 0.104 mm3 for group 2, and 0.111 mm3 for group 3. Compared with the control group, the size of the tumors in groups 1, 2 and 3 was reduced by an average of 97.4%, 92.5% and 91.9%, respectively. The Kaplan-Meier survival curve of group 2 was the longest, followed by groups 3, group 1 and the control. The mean survival was 22.8, 59, 60, and 44.6 days for the control group and groups 3, 2 and 1, respectively. H-E staining revealed that group 2 yielded the best results in the destruction of the malignant glioma. TUNEL staining and immunohistochemical studies indicated apoptotic features. The Ho-166 chitosan complex proved to be effective in destroying the malignant glioma.

Keyword

Holmium-166 chitosan complex; malignant glioma; brachytherapy; therapeutic effects

MeSH Terms

Animals
Brachytherapy
Brain Neoplasms/mortality/pathology/*radionuclide imaging
Cell Line, Tumor
Chitin/analogs & derivatives/*pharmacology
Disease Models, Animal
Glioma/mortality/pathology/*radionuclide imaging
Holmium/*pharmacology
Radioisotopes/*pharmacology
Rats
Rats, Wistar
Research Support, Non-U.S. Gov't

Figure

• 

  Fig. 1 Gross images for each group. In the control group, a successful tumor implant in the left putamen can be observed. In group 1, cavitation (arrows) can be observed in spots where the Ho-166 chitosan complex was over-administered. In groups 2 and 3, the implanted tumors are substantially reduced in size due to destruction by Ho-166 chitosan complex.

• 

  Fig. 2 Comparison of the tumor volume for each group. The average tumor volume for the control group was 1.385 mm3. For group 1, it was 0.036 mm3, for group 2, it was 0.104 mm3, and for group 3 it was 0.111 mm3. The ratio of the tumor volume against the control volume was 97.4% for group 1, 92.5% for group 2, and 91.1% for group 3, showing a gradual reduction.

• 

  Fig. 3 Survival curve for each group. Curves for groups 2 and 3 extended further than the control and group 1, and this extension was statistically significant (p < 0.001).

• 

  Fig. 4 H&E stain images. Under 100 × and 400 × magnification, malignant cerebral tumors with abundant nucleus and cytoplasm could be observed in the control group. In group 3, some tumor cells destroyed by Ho-166 chitosan complex could be observed, but the areas with active tumor cells were larger than in the other groups. In group 2, necrosis of the tumor cells were observed more extensively. In group 1, cavities (almost all cells completely destroyed with no structure) could be observed around the centers of administration (marked with arrow). Destruction spread even into normal tissues, and tumor cells showed the appearance of coagulation necrosis.

• 

  Fig. 5 TUNEL stain images. In all experimental groups with the Ho-166 chitosan complex administration, there were positive results with the nucleus being stained (arrowed). In group 3, positive reactions were observed even in the normal cells around a tumor cell where it appeared to be going through radionecrosis.

• 

  Fig. 6 Comparison of TUNEL-stain-positive rats. The average positive rates were 19.6% for group 3, 30.4% for group 2, 36.7% for group 1.

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  Fig. 7 Bax stain results. In group 1, extensive staining of the cytoplasm could be observed around the tumor destruction as the center. In group 2, similar observations were made. However, in group 3, there appeared to be barely any staining.

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