Preliminary study on dental pulp stem cell-mediated pulp regeneration in canine immature permanent teeth.

The health of human teeth depends on the integrity of the hard tissue and the activity of the pulp and periodontal tissues, which are responsible for nutritional supply. Without the nourishing of the pulp tissue, the possibility of tooth fracture can increase. In immature permanent teeth, root development may be influenced as well. This study explored the potential of using autologous dental pulp stem cells (DPSCs) to achieve pulp regeneration in a canine pulpless model. The establishment of the pulpless animal model involved pulp extirpation and root canal preparation of young permanent incisor teeth in beagles. Autologous DPSCs were obtained from extracted first molars and expanded ex vivo to obtain a larger number of cells. The biological characteristics of canine DPSCs (cDPSCs) were analyzed both in vitro and in vivo by using the same method as used in human DPSCs. cDPSCs were transplanted into the pulpless root canal with Gelfoam as the scaffold, and root development was evaluated by radiographic and histologic analyses. cDPSCs with rapid proliferation, multiple differentiation capacity, and development potential were successfully isolated and identified both in vitro and in vivo. After they were transplanted into the pulpless root canal with Gelfoam as the scaffold, DPSCs were capable of generating pulp-like tissues containing blood vessels and dentin-like tissue. Thickening of the root canal wall was also observed. This study demonstrates the feasibility of using stem cell-mediated tissue engineering to realize pulp regeneration in immature teeth.

Wang Y ，Zhao Y ，Jia W ，Yang J ，Ge L ... -  《-》

Investigation of dental pulp stem cells isolated from discarded human teeth extracted due to aggressive periodontitis.

Recently, human dental pulp stem cells (DPSCs) isolated from inflamed dental pulp tissue have been demonstrated to retain some of their pluripotency and regenerative potential. However, the effects of periodontal inflammation due to periodontitis and its progression on the properties of DPSCs within periodontally compromised teeth remain unknown. In this study, DPSCs were isolated from discarded human teeth that were extracted due to aggressive periodontitis (AgP) and divided into three experimental groups (Groups A, B and C) based on the degree of inflammation-induced bone resorption approaching the apex of the tooth root before tooth extraction. DPSCs derived from impacted or non-functional third molars of matched patients were used as a control. Mesenchymal stem cell (MSC)-like characteristics, including colony-forming ability, proliferation, cell cycle, cell surface antigens, multi-lineage differentiation capability and in vivo tissue regeneration potential, were all evaluated in a patient-matched comparison. It was found that STRO-1- and CD146-positive DPSCs can be isolated from human teeth, even in very severe cases of AgP. Periodontal inflammation and its progression had an obvious impact on the characteristics of DPSCs isolated from periodontally affected teeth. Although all the isolated DPSCs in Groups A, B and C showed decreased colony-forming ability and proliferation rate (P < 0.05), the decreases were not consistent with the degree of periodontitis. Furthermore, the cells did not necessarily show significantly diminished in vitro multi-differentiation potential. Only DPSCs from Group A and the Control group formed dentin-like matrix in vivo when cell-seeded biomaterials were transplanted directly into an ectopic transplantation model. However, when cell-seeded scaffolds were placed in the root fragments of human teeth, all the cells formed significant dentin- and pulp-like tissues. The ability of DPSCs to generate dental tissues decreased when the cells were isolated from periodontally compromised teeth (P < 0.05). Again, increased periodontal destruction was not necessarily followed by a decrease in the amount of dentin- and pulp-like tissue formed. These findings provide preliminary evidence that periodontally compromised teeth might contain putative stem cells with certain MSC properties, as long as the vitality of the pulp has not been totally damaged. Whether these cells can serve as a source of autologous multipotent MSCs for clinical regenerative therapies warrants further investigation with larger sample sizes and various types of periodontitis.

Sun HH ，Chen B ，Zhu QL ，Kong H ，Li QH ，Gao LN ，Xiao M ，Chen FM ，Yu Q ... -  《-》

Experimental formation of dentin-like structure in the root canal implant model using cryopreserved swine dental pulp progenitor cells.

The purpose of the present study was to present histological and immunohistochemical evidence showing the regenerative capacity of swine dental pulp stem cells (S-DPSCs) seeded on organic or synthetic scaffolds and implanted as hybrid root implants in the jaw bone of minipigs. Immature permanent incisor teeth and unerupted premolars at the early root-forming stage were extracted from three 7-month-old minipigs, and mesenchymal stem/progenitor cells were isolated from dental pulp. Cells were cryopreserved in liquid nitrogen. A year later, new permanent incisor and premolar teeth were extracted; pulp tissue was removed; and pieces of root canals of the extracted teeth, containing collagen or Poly(lactic-co-glycolic acid) scaffolds seeded with the autologous cryopreserved DPSCs, were implanted into the fresh post-extraction socket of the mini pig jaw. The resulting constructs were harvested after 6 and 10 weeks and evaluated by histological and immunohistochemical analyses. Six weeks postoperatively, the central canal space of the root implants showed degrading scaffold material. New extracellular matrix had been deposited in a polar predentin-like pattern on the canal dentinal walls by cuboidal nonpolarized cells. Ten weeks postoperatively, newly formed organic matrix had been consistently deposited on the canal walls. The presence of a continuous layer of polarized cells showing typical columnar morphology adjacent to the newly deposited organic matrix was evident. The interactions of S-DPSCs with the dentin matrix of roots implanted in the jawbone of minipigs constitute a model to study in vivo organization and differentiation potential of DPSCs.

Kodonas K ，Gogos C ，Papadimitriou S ，Kouzi-Koliakou K ，Tziafas D ... -  《-》

Responses of immature permanent teeth with infected necrotic pulp tissue and apical periodontitis/abscess to revascularization procedures.

To report several types of response of immature permanent teeth with infected necrotic pulp tissue and either apical periodontitis or abscess to revascularization procedures. Twenty immature permanent teeth with infected necrotic pulp tissue and either apical periodontitis or abscesses from 20 patients were included. The teeth were isolated with rubber dam, and pulp chambers was accessed through the crowns. The canals were gently irrigated with 5.25% sodium hypochlorite with minimal mechanical debridement. Calcium hydroxide was used as an inter-appointment intracanal medicament and placed into the coronal half of the canal space. After resolution of clinical signs and symptoms, bleeding was induced into the canal space from the periapical tissues using K-files. The coronal canal space was sealed with a mixture of mineral trioxide aggregate (MTA) and saline solution. The access cavity was filled with composite resin. These immature permanent teeth with infected necrotic pulp tissue and apical periodontitis/abscesses were followed up from 6 to 26 months. Five types of responses of these immature permanent teeth with infected necrotic pulp tissue and apical periodontitis/abscess to revascularization procedures were observed: type 1, increased thickening of the canal walls and continued root maturation; type 2, no significant continuation of root development with the root apex becoming blunt and closed; type 3, continued root development with the apical foramen remaining open; type 4, severe calcification (obliteration) of the canal space; type 5, a hard tissue barrier formed in the canal between the coronal MTA plug and the root apex. Based on this case series, the outcome of continued root development was not as predictable as increased thickening of the canal walls in human immature permanent teeth with infected necrotic pulp tissue and apical periodontitis/abscess after revascularization procedures. Continued root development of revascularized immature permanent necrotic teeth depends on whether the Hertwig's epithelial root sheath survives in case of apical periodontitis/abscess. Severe pulp canal calcification (obliteration) by hard tissue formation might be a complication of internal replacement resorption or union between the intracanal hard tissue and the apical bone (ankylosis) in revascularized immature permanent necrotic teeth.

Chen MY ，Chen KL ，Chen CA ，Tayebaty F ，Rosenberg PA ，Lin LM ... -  《-》

Effects of morphogen and scaffold porogen on the differentiation of dental pulp stem cells.

Dental pulp tissue engineering is an emerging field that can potentially have a major impact on oral health. However, the source of morphogens required for stem cell differentiation into odontoblasts and the scaffold characteristics that are more conducive to odontoblastic differentiation are still unclear. This study investigated the effect of dentin and scaffold porogen on the differentiation of human dental pulp stem cells (DPSCs) into odontoblasts. Poly-L-lactic acid (PLLA) scaffolds were prepared in pulp chambers of extracted human third molars using salt crystals or gelatin spheres as porogen. DPSCs seeded in tooth slice/scaffolds or control scaffolds (without tooth slice) were either cultured in vitro or implanted subcutaneously in immunodefficient mice. DPSCs seeded in tooth slice/scaffolds but not in control scaffolds expressed putative odontoblastic markers (DMP-1, DSPP, and MEPE) in vitro and in vivo. DPSCs seeded in tooth/slice scaffolds presented lower proliferation rates than in control scaffolds between 7 and 21 days (p < 0.05). DPSCs seeded in tooth slice/scaffolds and transplanted into mice generated a tissue with morphological characteristics similar to those of human dental pulps. Scaffolds generated with gelatin or salt porogen resulted in similar DPSC proliferation. The porogen type had a relatively modest impact on the expression of the markers of odontoblastic differentiation. Collectively, this work shows that dentin-related morphogens are important for the differentiation of DPSC into odontoblasts and for the engineering of dental pulp-like tissues and suggest that environmental cues influence DPSC behavior and differentiation potential.

Demarco FF ，Casagrande L ，Zhang Z ，Dong Z ，Tarquinio SB ，Zeitlin BD ，Shi S ，Smith AJ ，Nör JE ... -  《-》