This Article Abstract Full Text Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Add to Saved Citations Download to citation manager Request Permissions Request Reprints Add to My Marked Citations Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Citing Articles via Scopus Google Scholar Articles by Batouli, S. Articles by Shi, S. Search for Related Content PubMed PubMed Citation Articles by Batouli, S. Articles by Shi, S. Pubmed/NCBI databases Substance via MeSH Medline Plus Health Information Stem Cells Social Bookmarking                         What's this?

Comparison of Stem-cell-mediated Osteogenesis and Dentinogenesis

S. Batouli^1,, M. Miura^1,, J. Brahim², T.W. Tsutsui¹, L.W. Fisher¹, S. Gronthos³, P. Gehron Robey¹ and S. Shi^1,*

¹ Craniofacial and Skeletal Diseases Branch,
² Clinical Research Core, National Institute of Dental and Craniofacial Research, National Institutes of Health, Department of Health and Human Services, Building 30, Room 222, NIDCR/NIH, 30 Convent Drive MSC-4320, Bethesda, MD 20892;
³ Mesenchymal Stem Cell Group, Division of Haematology, Institute of Medical and Veterinary Science, Frome Road Adelaide 5000, South Australia, Australia;

View larger version (125K): [in this window] [in a new window]   Figure 1. H&E staining of BMSSC and DPSC transplants. (A and a) BMSSC transplant at 2 wks post-transplantation showed connective tissue (CT) surrounding the HA/TCP carriers (HA). Bone regeneration (B) on the HA/TCP (HA) was found only on a very limited number of transplants. (B) At 4 wks post-transplantation, BMSSCs were able to generate bone (B) on the surface of the HA/TCP (HA) and had differentiated into osteoblasts (open arrows) and osteocytes (black arrows). (C) At 8 wks post-transplantation of the BMSSC transplant, a bone/marrow organ-like structure was generated, where osteoblasts (open arrows) lined the surface of bone tissue containing osteocytes (black arrows), and the connective tissue (CT) was replaced by hematopoietic marrow (HM). The interface between the hematopoietic marrow and connective tissue is indicated (yellow dashed line). (D) At 16 wks post-transplantation, the BMSSC transplant had developed into a mature bone/marrow organ, in which a significant amount of bone (B) was generated along with hematopoietic marrow elements (HM). (E and e) The DPSC transplant at 2 wks post-transplantation showed connective tissue (CT) surrounding the HA/TCP (HA). Similar to the BMSSC transplants, only very few transplants showed dentin (D) formation on the surface of the HA/TCP (HA). (F) After 4 wks of transplantation, DPSCs differentiated into odontoblasts (open arrows) responsible for new dentin (D) formation on the surface of the HA/TCP (HA). (G) At 8 wks post-transplantation of the DPSC transplant, a dentin/pulp complex was generated, at which odontoblasts (open arrows) lined the surface of the newly formed dentin (D), and the pulp-like tissue (Pulp) contained blood vessels and connective tissue. (H) At 16 wks post-transplantation, the DPSC transplant had continued to develop into a dentin/pulp complex, in which a significant amount of dentin (D) was generated adjacent to the pulp-like tissue (Pulp). Bar, 40 µm.

View larger version (143K): [in this window] [in a new window]   Figure 2. Human alu in situ hybridization of BMSSC and DPSC transplants. (A) Human BMSSCs (dark nuclear staining) were either attached to the surface of the HA/TCP (HA) or in the connective tissue compartment (CT) at 2 wks post-transplantation. (B) After 8 wks’ transplantation, BMSSCs differentiated into osteoblasts (open arrows) and osteocytes (black arrows) to generate bone (B) on the surface of the HA/TCP (HA). The number of BMSSCs in the connective tissue compartment (CT) was significantly diminished. (C) Human DPSCs (dark nuclei staining) either attached to the surface of the HA/TCP (HA) or resided in the connective tissue compartment (CT) at 2 wks post-transplantation. (D) At 8 wks post-transplantation, DPSCs differentiated into odontoblasts (open arrows) to generate dentin (D) on the surface of the HA/TCP (HA). The number of DPSCs present in the connective tissue compartment (CT) is comparable with the number present at 2 wks post-transplantation. Bar, 20 µm.

View larger version (104K): [in this window] [in a new window]   Figure 3. Immunohistochemical staining of bFGF and MMP-9 in BMSSC transplants. (A) BMSSC transplants showed that some cells were immunopositive for bFGF antibody staining (black arrows) in the connective tissue compartment (CT) surrounding the HA/TCP (HA) at 2 wks post-transplantation. (B) After 4 wks’ BMSSC transplantation, along with new bone (B) formation, the number of bFGF-positive cells (black arrows) was increased in the connective tissue compartment (CT). (C) After 8 wks’ BMSSC transplantation, the connective tissue (CT) showed a high-level expression of bFGF (black arrows). Newly formed hematopoietic marrow showed a negative staining for bFGF antibody staining. The interface between the hematopoietic marrow and connective tissue is indicated (yellow dashed line). (D) When hematopoietic marrow (HM) completely replaced the connective tissue at 16 wks post-transplantation, a very limited number of cells expressed bFGF in BMSSC transplants. (E) BMSSC transplant showed a negative staining for MMP9 antibody at 2 wks post-transplantation. (F) After 4 wks’ transplantation, MMP9 was expressed in the connective tissue compartment of BMSSC transplants (open arrows). (G and H) DPSC transplants showed a negative staining of bFGF prior to dentin regeneration at 2 wks post-transplantation (G) and after dentin formation at 8 wks post-transplantation (H). (I and J) VEGF was equally expressed on blood vessels of BMSSC (I) and DPSC (J) transplants at 4 wks post-transplantation (black arrows). The black dashed lines represent interfaces between HA/TCP and newly formed bone (I) or dentin (J). Bar, 20 µm.

View larger version (93K): [in this window] [in a new window]   Figure 4. Characterization of DPSC-mediated dentinogenesis in vivo. (A) Odontoblasts of human dental pulp show a positive immunostaining for DSP antibody (black arrows). Other cells and blood vessels (open arrows) within the dental pulp are negative to DSP antibody staining. (B) There is no DSP antibody immunostaining on human bone (B), osteoblasts (black arrows), and hematopoietic marrow elements (HM). (C) DPSC transplant at 8 wks post-transplantation. There is DSP-positive immunostaining on dentinogenic cells (open arrows) and tubular dentin (black arrows). Pulp-like tissue (P) contains blood vessel (BV) and connective tissue. Dashed line shows the interface between the hydroxyapatite carrier (HA) and the newly generated dentin (D). (D) At 8 wks post-transplantation, BMSSCs differentiated into osteoblasts (black arrows) to generate bone (B) and induced hematopoietic marrow formation (HM). There was no DSPP antibody immunostaining on newly formed bone and osteogenic cells. (E) BMSSCs were loaded onto the acid-treated dentin and cultured for 12 hrs. Then BMSSC/dentin complexes were transplanted into immunocompromised mice. There was no mineralized tissue regeneration after 8 wks’ transplantation. (F) Newly formed reparative dentin-like structure (ND) attached to the surfaces of human dentin in DPSC/dentin transplants. There were blood vessels (BV) and connective tissue (CT) associated with dentinogenic cells (black arrows). DPSCs were unable to generate organized tubular dentin structure; instead, they formed reparative dentin-like structure containing entrapped cells (triangle open arrows). (G) In DPSC/dentin transplants, dentinogenic cells (open arrows) and trapped cells (open triangles) within the newly formed reparative dentin-like structure (ND) were immunoreactive to human DSP antibody. Human dentin scaffold (Dentin) showed a positive DSP immunostaining on peritubular structures (black arrows). Connective tissue (CT) contained blood vessels (BV). (H) DSP immunohistochemical staining of BMSSC/dentin transplant showed a positive staining on the peritubular dentin (black arrows). Connective tissue (CT) showed a negative immunostaining for DSP antibody. (I) Immunohistochemical staining of human-specific anti-mitochondria antibody showed that human DPSCs differentiated into dentinogenic cells lining the surfaces (open arrows) or trapped within (open triangles) the newly generated reparative dentin-like tissue (ND). DPSCs were also found within the connective tissue compartment (CT) at 8 wks post-transplantation. (J) Negative control of immunohistochemical staining on DPSC/dentin transplant with pre-immuno serum. Bar, 40 µm in panels A and B, 20 µm in C-J. E and F were stained with hematoxylin & eosin.

Journal of Dental Research, Vol. 82, No. 12, 976-981 (2003)
DOI: 10.1177/154405910308201208


CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    Twitter    What's this?