Silvana B. Gonçalves, DDS, MS, Zhihong Dong, MD, PhD, Clovis M

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Tooth Slice–Based Models for the Study of Human Dental Pulp Angiogenesis Silvana B. Gonçalves, DDS, MS, Zhihong Dong, MD, PhD, Clovis M. Bramante, DDS, MS, Graham R. Holland, BSc, BDS, PhD, Anthony J. Smith, BSc, BDS, PhD, Jacques E. Nör, DDS, MS, PhD Journal of Endodontics Volume 33, Issue 7, Pages 811-814 (July 2007) DOI: 10.1016/j.joen.2007.03.012 Copyright © 2007 American Association of Endodontists Terms and Conditions

Figure 1 Tooth slices and pulp angiogenesis. Diagram depicting the overall scheme for the model systems used here; 1.5-mm thick tooth slices were prepared from the cervical region of human third molars. These tooth slices were either cultured in vitro or transplanted into subcutaneous pockets in immunodeficient mice. Journal of Endodontics 2007 33, 811-814DOI: (10.1016/j.joen.2007.03.012) Copyright © 2007 American Association of Endodontists Terms and Conditions

Figure 2 Effect of VEGF on in vitro angiogenesis in human dental pulps. (A, C, E) Tooth slices were left untreated or (B, D, F) cultured in the presence of 50 ng/mL rhVEGF165. (A, B) Hematoxylin/eosin staining of representative dental pulps after 7 days in culture (200×). (C-F) Representative microscopic fields of Factor VIII immunohistochemistry for identification of blood vessels (red staining) at (C, D) 200× or (E, F) 400×. Scale bar for panels A-F, 50 μm. (G) Graph depicting the mean (± SD) number of Factor VIII–positive blood vessels from 25 microscopic fields per condition. Asterisk indicates statistical significance (p < 0.001). Journal of Endodontics 2007 33, 811-814DOI: (10.1016/j.joen.2007.03.012) Copyright © 2007 American Association of Endodontists Terms and Conditions

Figure 3 Effect of VEGF on in vitro apoptosis in human dental pulps. Tooth slices were left (A, C) untreated or (B, D) cultured in the presence of 50 ng/mL rhVEGF165. (A, B) Hematoxylin/eosin staining of representative dental pulps after 7 days in culture (400×). (C-E) Representative microscopic fields of TUNEL staining for identification of apoptotic cells (dark blue staining) at 400× (C, D). Scale bar for panels A-D, 50 μm. (E) Graph depicting the mean (±SD) number of apoptotic cells from 25 microscopic fields per condition. Journal of Endodontics 2007 33, 811-814DOI: (10.1016/j.joen.2007.03.012) Copyright © 2007 American Association of Endodontists Terms and Conditions

Figure 4 Tooth-slice based model of human dental pulp angiogenesis. (A) Hematoxylin/eosin staining of a representative dental pulp after 7 days in a murine host (400×). (B) Representative microscopic field of Factor VIII immunohistochemistry for identification of blood vessels (red staining) at 200×. (C) Representative microscopic field of TUNEL staining for identification of apoptotic cells (dark blue staining) at 200×. Arrows point to TUNEL-positive (apoptotic) cells. Scale bar for panels A-C, 50 μm. Journal of Endodontics 2007 33, 811-814DOI: (10.1016/j.joen.2007.03.012) Copyright © 2007 American Association of Endodontists Terms and Conditions