Concave pit-containing scaffold surfaces improve stem cell-derived osteoblast performance and lead to significant bone tissue formation.

PLoS ONE. 2007;2(6):e496

Authors: Graziano A, d'Aquino R, Cusella-De Angelis MG, Laino G, Piattelli A, Pacifici M, De Rosa A, Papaccio G

BACKGROUND: Scaffold surface features are thought to be important regulators of stem cell performance and endurance in tissue engineering applications, but details about these fundamental aspects of stem cell biology remain largely unclear. METHODOLOGY AND FINDINGS: In the present study, smooth clinical-grade lactide-coglyolic acid 85:15 (PLGA) scaffolds were carved as membranes and treated with NMP (N-metil-pyrrolidone) to create controlled subtractive pits or microcavities. Scanning electron and confocal microscopy revealed that the NMP-treated membranes contained: (i) large microcavities of 80-120 microm in diameter and 40-100 microm in depth, which we termed primary; and (ii) smaller microcavities of 10-20 microm in diameter and 3-10 microm in depth located within the primary cavities, which we termed secondary. We asked whether a microcavity-rich scaffold had distinct bone-forming capabilities compared to a smooth one. To do so, mesenchymal stem cells derived from human dental pulp were seeded onto the two types of scaffold and monitored over time for cytoarchitectural characteristics, differentiation status and production of important factors, including bone morphogenetic protein-2 (BMP-2) and vascular endothelial growth factor (VEGF). We found that the microcavity-rich scaffold enhanced cell adhesion: the cells created intimate contact with secondary microcavities and were polarized. These cytological responses were not seen with the smooth-surface scaffold. Moreover, cells on the microcavity-rich scaffold released larger amounts of BMP-2 and VEGF into the culture medium and expressed higher alkaline phosphatase activity. When this type of scaffold was transplanted into rats, superior bone formation was elicited compared to cells seeded on the smooth scaffold. CONCLUSION: In conclusion, surface microcavities appear to support a more vigorous osteogenic response of stem cells and should be used in the design of therapeutic substrates to improve bone repair and bioengineering applications in the future.

PMID: 17551577 [PubMed]

Molecular and biomechanical characterization of mineralized tissue by dental pulp cells on titanium.

J Dent Res. 2005 Jun;84(6):515-20

Authors: Nakamura H, Saruwatari L, Aita H, Takeuchi K, Ogawa T

The application of implant therapy is still limited, because of various risk factors and the long healing time required for bone-titanium integration. This study explores the potential for osseointegration engineering with dental pulp cells (DPCs) by testing a hypothesis that DPCs generate mineralized tissue on titanium. DPCs extracted from rat incisors positive for CD44, alkaline phosphatase activity, and mineralizing capability were cultured on polystyrene and on machined and dual-acid-etched (DAE) titanium. Tissue cultured on titanium with a Ca/P ratio of 1.4 exhibited plate-like morphology, while that on the polystyrene exhibited fibrous and punctate structures. Tissues cultured on titanium were harder than those on polystyrene, 1.5 times on the machined and 3 times on the DAE. Collagen I, osteopontin, and osteocalcin genes were up-regulated on titanium, especially the DAE surface. In conclusion, DPCs showing some characteristics of the previously identified dental pulp stem cells can generate mineralized tissue on titanium via the osteoblastic phenotype, which can be enhanced by titanium surface roughness.

PMID: 15914587 [PubMed - indexed for MEDLINE]

Genotyping, Orientalis-like Yersinia pestis, and plague pandemics.

Emerg Infect Dis. 2004 Sep;10(9):1585-92

Authors: Drancourt M, Roux V, Dang LV, Tran-Hung L, Castex D, Chenal-Francisque V, Ogata H, Fournier PE, Crubézy E, Raoult D

Three pandemics have been attributed to plague in the last 1,500 years. Yersinia pestis caused the third, and its DNA was found in human remains from the second. The Antiqua biovar of Y. pestis may have caused the first pandemic; the other two biovars, Medievalis and Orientalis, may have caused the second and third pandemics, respectively. To test this hypothesis, we designed an original genotyping system based on intergenic spacer sequencing called multiple spacer typing (MST). We found that MST differentiated every biovar in a collection of 36 Y. pestis isolates representative of the three biovars. When MST was applied to dental pulp collected from remains of eight persons who likely died in the first and second pandemics, this system identified original sequences that matched those of Y. pestis Orientalis. These data indicate that Y. pestis caused cases of Justinian plague. The two historical plague pandemics were likely caused by Orientalis-like strains.

PMID: 15498160 [PubMed - indexed for MEDLINE]

Molecular regulation of odontoblast activity under dentin injury.

Adv Dent Res. 2001 Aug;15:46-50

Authors: Magloire H, Romeas A, Melin M, Couble ML, Bleicher F, Farges JC

Pulp tissue responds to dentin damage by laying down a tertiary dentin matrix (reactionary or reparative) beneath the site of injury. Reactionary dentin is secreted by surviving odontoblasts in response to environmental stimuli, leading to an increase in metabolic activities of the cells. The inductive molecules that determine the success of the pulp healing may be released from the damaged dentin as well as from the pulp tissue subjacent to the injury. This paper will schematically consider two major growth factors probably implicated in the control of odontoblast activity: TGF beta-1 released from demineralized dentin and NGF from pulp. To analyze their role with an in vitro system that mimics the in vivo situation, we have used thick-sliced teeth cultured as described previously. The supply of factors was accomplished by means of a small tube glued onto the dentin. The tube was filled with TGF beta-1 (20 ng/mL) or NGF (50 ng/mL), and slices were cultured for 4 or 7 days. Results showed that TGF beta-1 binding sites are strongly detected on odontoblasts in the factor-rich zone. A strong expression of alpha 1(I) collagen transcripts was also detected. In the NGF-rich environment, p75NTR was re-expressed on odontoblasts and the transcription factor NF-kappa B activated. Modifications in the odontoblast morphology were observed with an atypical extension of the cell processes filled with actin filaments. These results suggest that odontoblasts respond to influences from both dentin and pulp tissue during pulp repair.

PMID: 12640739 [PubMed - indexed for MEDLINE]

New genes involved in odontoblast differentiation.

Adv Dent Res. 2001 Aug;15:30-3

Authors: Bleicher F, Couble ML, Buchaille R, Farges JC, Magloire H

The odontoblast phenotype has been mainly approached by the biochemical characterization of dentin matrix proteins and by extrapolation of the knowledge of bone cell biology, since dentin and bone share many similarities. In fact, direct investigations of the odontoblast phenotype have been hindered by the limited number of cells within the dental pulp and the difficulty in microdissection and isolation of a pure population of these cells. To overcome these obstacles, we previously developed a cell-culture system that promotes differentiation of human dental pulp cells into odontoblasts. This material now permits the study of odontoblasts through molecular biology techniques. Therefore, we constructed a cDNA library enriched for odontoblast-specific genes using the suppression subtractive hybridization technique (SSH). This library led us to identify new genes expressed by odontoblasts. In this paper, we will focus on some genes implied in various functions associated with odontoblast differentiation, such as cell polarization (MAP1B), dentin mineralization (PHEX, osteoadherin), and relationships between odontoblasts and nerve cells (reelin). Another important fact is that about 40% of the cDNA were unknown genes. Therefore, one can speculate that some of them will be odontoblast-specific, since, until now, only one gene (DSPP) presents this characteristic.

PMID: 12640735 [PubMed - indexed for MEDLINE]

The road to pulp biology research: a personal odyssey.

J Dent Res. 1998 Sep;77(9):1670-3

Authors: Stanley HR

PMID: 9759663 [PubMed - indexed for MEDLINE]

A new biological approach to vital pulp therapy.

Crit Rev Oral Biol Med. 1995;6(3):218-29

Authors: Rutherford B, Fitzgerald M

Molecular biology is providing opportunities to develop new strategies or agents for the treatment of a wide variety of diseases. The availability of large amounts of highly purified proteins produced by recombinant DNA techniques is an obvious example. Recent evidence has implicated proteins belonging to the bone morphogenetic protein (BMP) subgroup of the transforming growth factor beta supergene family in tooth formation and dentinogenesis. It has long been known that bone and dentin contain bone morphogenetic protein activity. Recently, recombinant human BMP-2, -4, and -7 (also known as OP-1), have been shown to induce reparative dentin formation in experimental models of large direct pulp exposures in permanent teeth. The manner in which these agents act appears unique. New reparative dentin replaces the stimulating agents applied directly to the partially amputated pulp. Hence, the new tissue forms contiguous with, largely superficial to, and not at the expense of the remaining vital pulp tissue. This suggests a therapeutic approach permitting the induction of a predetermined and controlled amount of reparative dentin. Additionally, OP-1 has been associated with the formation of reparative dentin after application to a freshly cut but intact layer of dentin. These findings may provide future clinicians with additional options for the treatment of substantially damaged or diseased vital teeth.

PMID: 8785262 [PubMed - indexed for MEDLINE]

Pulp biology research--is the frog still deaf?

J Dent Res. 1992 Oct;71(10):1752-3

Authors: Bakland LK

PMID: 1401435 [PubMed - indexed for MEDLINE]

How far can a frog jump? A current assessment of pulp biology research.

J Dent Res. 1988 Sep;67(9):1251

Authors: Cotton WR

PMID: 3166010 [PubMed - indexed for MEDLINE]

The neurobiology of facial and dental pain: present knowledge, future directions.

J Dent Res. 1987 May;66(5):962-81

Authors: Sessle BJ

This review outlines recent research which has identified critical neural elements and mechanisms concerned with the transmission of sensory information related to oral-facial pain, and which has also revealed some of the pathways and processes by which pain transmission can be modulated. The review highlights recent advances in neurobiological research that have contributed to our understanding of pain, how acute and chronic pain conditions can develop, and how pain can be controlled therapeutically. Each section of the review also identifies gaps in knowledge that still exist as well as research approaches that might be taken to clarify even further the mechanisms underlying acute and chronic oral-facial pain. The properties of the sense organs responding to a noxious oral-facial stimulus are first considered. This section is followed by a review of the sensory pathways and mechanisms by which the sensory information is relayed in nociceptive neurones in the brainstem and then transmitted to local reflex centers and to higher brain centers involved in the various aspects of the pain experience--namely, the sensory-discriminative, affective (emotional), cognitive, and motivational dimensions of pain. Reflex and behavioral responses to noxious oral-facial stimuli are also considered. The next section provides an extensive review of how these responses and the activity of the nociceptive neurones are modulated by higher brain center influences and by stimulation of, or alterations (e.g., by trauma) to, other sensory inputs to the brain. The neurochemical processes, involved in these modulatory mechanisms are also considered, with special emphasis on the role of neuropeptides and other neurochemicals recently shown to be involved in pain transmission and its control. The final section deals with recent findings of peripheral and central neural mechanisms underlying pain from the dental pulp.

PMID: 3301935 [PubMed - indexed for MEDLINE]