Cementum Regeneration Research Articles

CXXC5 Mediates P. gingivalis-suppressed Cementoblast Functions Partially via MAPK Signaling Network

Porphyromonas (P.) gingivalis associates tightly with periodontal diseases and it is also a dominant pathogen of periapical periodontitis. However, the influence of P. gingivalis on cementoblasts, root surface cells pivotal in the apical areas, and the possible involvement of other molecules remain largely elusive. CXXC5 is a nuclear protein that regulates gene expression as well as cell growth, differentiation, and apoptosis. In this study, P. gingivalis repressed the mineralization capacity of cementoblasts by inducing inflammatory reactions and inhibiting cell differentiation. Intriguingly, the expression of CXXC5 decreased in P. gingivalis-treated OCCM-30 cells and apical periodontitis models but gradually increased during mineralization. Furthermore, RNA interference of CXXC5 significantly inhibited cementoblast differentiation, represented by decline of bone-associated markers Osterix, osteocalcin (OCN), and alkaline phosphatase (ALP). CXXC5 overexpression facilitated differentiation, and therefore attenuated the P. gingivalis-repressed effects on OCCM-30 cells. In addition, Erk1/2, p38, and PI3K-Akt were inactivated by silencing CXXC5 and activated upon its overexpression, whereas Wnt/β-catenin exhibited an opposite trend. The employment of specific inhibitors revealed that the CXXC5-dependent promotions of cementoblast differentiation were partially abrogated by p38 and PI3K-Akt inhibitors but were exacerbated by inhibiting Erk1/2. Overall, our experiment demonstrated a novel function of CXXC5 in the regeneration of impaired cementum caused by P. gingivalis invasion and suggested that MAPK signaling network balances the facilitation effects of CXXC5 in cementoblast differentiation.

Use of Autologous Stem Cells for the Regeneration of Periodontal Defects in Animal Studies: a Systematic Review and Meta-Analysis.

ABSTRACTObjectivesTo overview preclinical animal trials and quantify the effect size that stem cell therapy has on the regeneration of periodontal tissue complex.Material and MethodsA systematic MEDLINE (PubMed) online library search was conducted for preclinical animal studies in vivo, using autologous periodontal ligament, dental pulp, cementum, alveolar periosteal, gingival margin or adipose stem cell types for periodontal tissue complex regeneration purposes. Studies had to be published between 2007.09.01 and 2017.09.01 in the English language.ResultsOnline library search yielded 2099 results. After the title, abstract and full-text screening ten studies fit inclusion criteria and were pooled into meta-analysis. Overall the stem cell regenerative therapy had a statistically significant positive influence on the periodontal tissue regeneration when compared to the control groups. The biggest influence was made to the regeneration of cementum (standardised mean difference [SMD] 2.25 [95% confidence interval (CI) = 1.31 to 3.2]) while the smallest influence was made to the alveolar bone (SMD 1.47 [95% CI = 0.7 to 2.25]) the effect size for periodontal ligament regeneration was (SMD 1.8 [95% CI = 1 to 2.59]). Subgroup analysis showed statistically significant (P < 0.05) differences between different cell types in the alveolar bone and cementum regeneration groups and in alveolar bone group in relation to scaffold materials.ConclusionsStem cell therapy has a positive impact on periodontal tissue complex regeneration. Such therapy has the biggest influence on cementum regeneration meanwhile alveolar bone regeneration is influenced by the least amount. However more and less diverse preclinical studies are needed to have a greater statistical power in future meta-analyses.

Response of periodontium to mineral trioxide aggregate and Biodentine: a pilot histological study on humans.

The aim of this study was to investigate for the first time the histological response of human periodontium to mineral trioxide aggregate (MTA) and Biodentine. Six patients scheduled for implant full-arch rehabilitation were randomly assigned to one of the two test groups: MTA or Biodentine treatment. For each patient, two teeth scheduled for strategic extraction were randomly assigned either to the test or to the control treatment. A lateral perforation was drilled on the root and either repaired with MTA/Biodentine or filled with gutta-percha(control). Three months later, the teeth were extracted along with the coronal third of the alveolar bone and a portion of gingival tissue, while performing implant placement, and processed for histological analysis. Biodentine resulted in less extrusion into the periodontal environment. All the materials showed good biocompatibility. A new mineralized cementum-like tissue incorporating periodontal fibres was visible in all cases treated with MTA. A small amount of new mineralized tissue was found in two Biodentine cases but not in control cases. Biodentine resulted in less damage to the periodontal ligament. Bioactivity and biocompatibility of MTA were confirmed in human models. Biodentine proved to be biocompatible, but it seems not to induce cementum regeneration.

Aligned Fibers Fabricated by Near-Field Electrospinning Influence the Orientation and Differentiation of hPDLSCs for Periodontal Regeneration.

The periodontal ligament (PDL) is a specific connective tissue composed of organized and aligned collagenous fibers that fix the tooth root in the alveolar bone. The alignment of PDL fibers and their function have been neglected in the past, as many studies investigated the regeneration of the periodontium, including alveolar bone and cementum regeneration. In this study, we fabricated biodegradable aligned fibers (ε-caprolactone/collagen) by near-field electrospinning (NFE) to control the arrangement of human periodontal ligament stem cells (hPDLSCs), aiming to guide the oriented regeneration of the periodontal ligament. Compared with random electrospun fibers, the in vitro study investigated the effects of nanotopography on stem cell differentiation of hPDLSCs. The hPDLSCs were identified by flow cytometry, and the multipotency of hPDLSCs was confirmed by successful osteogenic induction. The hPDLSCs were co-cultured with aligned and random fibers. The cell morphology was observed by confocal micrograph and scanning electron micrograph, which showed that aligned fibers could guide the orientation and elongation of hPDLSCs. The expression of periodontal ligament-related genes was higher when cultured with aligned fibers than when cultured with random fibers. In conclusion, via near-field electrospinning, aligned biodegradable fibers were prepared and guided the orientation arrangement of hPDLSCs, providing a better microenvironment for periodontal ligament regeneration. This technology might be further used in the regeneration of tissues in a given direction.

Effects of the incorporation of ε-aminocaproic acid/chitosan particles to fibrin on cementoblast differentiation and cementum regeneration.

Cementum, the mineralized layer on root dentin surfaces, functions to anchor fibrous connective tissues on tooth-root surfaces with the collagenous Sharpey's fibers integration, of which are essential for periodontal functioning restoration in the complex. Through the cementum-responsible fiber insertions on tooth-root surfaces, PDLs transmit various mechanical responses to periodontal complexes against masticatory/occlusal stimulations to support teeth. In this study, periodontal tissue regeneration was enhanced by use of modified fibrin biomaterial which significantly promoted cementogenesis within the periodontal complex with structural integration by collagenous Sharpey's fiber insertions in vivo by controlling fibrin degradation and consequent cementoblast apoptosis. Furthermore, the modified fibrin could improve repair and regeneration of tooth roof-of-furcation defects, which has spatial curvatures and geometrical difficulties and hardly regenerates periodontal tissues.

Efficacy of stem cells on periodontal regeneration: Systematic review of pre-clinical studies.

This systematic review aims to evaluate mesenchymal stem cells (MSC) periodontal regenerative potential in animal models. MEDLINE, EMBASE and LILACS databases were searched for quantitative pre-clinical controlled animal model studies that evaluated the effect of local administration of MSC on periodontal regeneration. The systematic review was conducted according to the Preferred Reporting Items for Systematic Reviews and Meta-Analyses statement guidelines. Twenty-two studies met the inclusion criteria. Periodontal defects were surgically created in all studies. In seven studies, periodontal inflammation was experimentally induced following surgical defect creation. Differences in defect morphology were identified among the studies. Autogenous, alogenous and xenogenous MSC were used to promote periodontal regeneration. These included bone marrow-derived MSC, periodontal ligament (PDL)-derived MSC, dental pulp-derived MSC, gingival margin-derived MSC, foreskin-derived induced pluripotent stem cells, adipose tissue-derived MSC, cementum-derived MSC, periapical follicular MSC and alveolar periosteal cells. Meta-analysis was not possible due to heterogeneities in study designs. In most of the studies, local MSC implantation was not associated with adverse effects. The use of bone marrow-derived MSC for periodontal regeneration yielded conflicting results. In contrast, PDL-MSC consistently promoted increased PDL and cementum regeneration. Finally, the adjunct use of MSC improved the regenerative outcomes of periodontal defects treated with membranes or bone substitutes. Despite the quality level of the existing evidence, the current data indicate that the use of MSC may provide beneficial effects on periodontal regeneration. The various degrees of success of MSC in periodontal regeneration are likely to be related to the use of heterogeneous cells. Thus, future studies need to identify phenotypic profiles of highly regenerative MSC populations.

Tri-Layered Nanocomposite Hydrogel Scaffold for the Concurrent Regeneration of Cementum, Periodontal Ligament, and Alveolar Bone.

A tri-layered scaffolding approach is adopted for the complete and concurrent regeneration of hard tissues-cementum and alveolar bone-and soft tissue-the periodontal ligament (PDL)-at a periodontal defect site. The porous tri-layered nanocomposite hydrogel scaffold is composed of chitin-poly(lactic-co-glycolic acid) (PLGA)/nanobioactive glass ceramic (nBGC)/cementum protein 1 as the cementum layer, chitin-PLGA/fibroblast growth factor 2 as the PDL layer, and chitin-PLGA/nBGC/platelet-rich plasma derived growth factors as the alveolar bone layer. The tri-layered nanocomposite hydrogel scaffold is cytocompatible and favored cementogenic, fibrogenic, and osteogenic differentiation of human dental follicle stem cells. In vivo, tri-layered nanocomposite hydrogel scaffold with/without growth factors is implanted into rabbit maxillary periodontal defects and compared with the controls at 1 and 3 months postoperatively. The tri-layered nanocomposite hydrogel scaffold with growth factors demonstrates complete defect closure and healing with new cancellous-like tissue formation on microcomputed tomography analysis. Histological and immunohistochemical analyses further confirm the formation of new cementum, fibrous PDL, and alveolar bone with well-defined bony trabeculae in comparison to the other three groups. In conclusion, the tri-layered nanocomposite hydrogel scaffold with growth factors can serve as an alternative regenerative approach to achieve simultaneous and complete periodontal regeneration.

Photoactivated disinfection using indocyanine green photosensitizer as an adjunct to regenerative periodontal therapy

Regenerative periodontal surgical procedures attempt to restore lost periodontal structure and functional attachment through the regeneration of cementum, periodontal ligament, and alveolar bone. Although meticulous debridement using surgical instrumentation reduces the bacterial burden, it fails to kill the periopathogens. To overcome this shortcoming, the development of alternative adjunctive or exclusive antibacterial therapeutic strategies, therefore, becomes important in the evolution of methods to control microbial growth in the oral cavity. The following case report explores the use of antimicrobial photodynamic therapy as an adjunct to regenerative periodontal therapy for the management of intrabony defects which was evaluated over a period of 6 months.

Periodontal Tissue Regeneration Using Syngeneic Adipose-Derived Stromal Cells in a Mouse Model.

Current treatment of periodontitis is still associated with a high degree of variability in clinical outcomes. Recent advances in regenerative medicine by mesenchymal cells, including adipose stromal cells (ASC) have paved the way to improved periodontal regeneration (PD) but little is known about the biological processes involved. Here, we aimed to use syngeneic ASCs for periodontal regeneration in a new, relevant, bacteria‐induced periodontitis model in mice. Periodontal defects were induced in female C57BL6/J mice by oral gavage with periodontal pathogens. We grafted 2 × 105 syngeneic mouse ASCs expressing green fluorescent protein (GFP) (GFP+/ASC) within a collagen vehicle in the lingual part of the first lower molar periodontium (experimental) while carrier alone was implanted in the contralateral side (control). Animals were sacrificed 0, 1, 6, and 12 weeks after treatment by GFP+/ASC or vehicle graft, and microscopic examination, immunofluorescence, and innovative bio‐informatics histomorphometry methods were used to reveal deep periodontium changes. From 1 to 6 weeks after surgery, GFP+ cells were identified in the periodontal ligament (PDL), in experimental sites only. After 12 weeks, cementum regeneration, the organization of PDL fibers, the number of PD vessels, and bone morphogenetic protein‐2 and osteopontin expression were greater in experimental sites than in controls. Specific stromal cell subsets were recruited in the newly formed tissue in ASC‐implanted periodontium only. These data suggest that ASC grafting in diseased deep periodontium, relevant to human pathology, induces a significant improvement of the PDL microenvironment, leading to a recovery of tooth‐supporting tissue homeostasis. Stem Cells Translational Medicine 2017;6:656–665

A Novel Regenerative Technique Combining Bone Morphogenetic Protein-2 With Fibroblast Growth Factor-2 for Circumferential Defects in Dog Incisors.

Periodontal regeneration of incisors is necessary for esthetic recovery. A novel regenerative method combining bone morphogenetic protein (BMP)-2 and fibroblast growth factor (FGF)-2 was developed. The purpose of this study is to evaluate periodontal healing, including root coverage, in circumferential defects of incisors. Fifty incisors in five beagles were used. After circumferential defects were surgically created, each group, consisting of ten recipient sites, received: 1) a double layer with FGF-2 (2 μg)/collagen as inner layer and BMP-2 (4 μg)/collagen as outer layer (FB-DL group); 2) collagen impregnated with both FGF-2 (2 μg) and BMP-2 (4 μg) (FB-M group); 3) BMP-2 (4 μg)/collagen (B group); 4) FGF-2 (4 μg)/collagen (F group); or 5) collagen (C group). Dogs were sacrificed 8 weeks post-surgery, and healing was evaluated histologically. The three groups treated with BMP-2 showed enhanced new bone formation compared with control and F groups (P < 0.05). Furthermore, connective tissue attachment with cementum regeneration in the FB-DL group was significantly greater than in FB-M and B groups (P <0.05). Ankylosis in the FB-DL group was significantly less than in FB-M and B groups (P <0.05). Gingival recession was inhibited significantly better in FB-DL and FB-M groups compared with control and B groups. These data support development of a double-layer method combining BMP-2 and FGF-2 as a therapeutic approach to periodontal regeneration at incisors with horizontal circumferential defects.

Controlled release of recombinant human cementum protein 1 from electrospun multiphasic scaffold for cementum regeneration

Periodontitis is a major cause for tooth loss, which affects about 15% of the adult population. Cementum regeneration has been the crux of constructing the periodontal complex. Cementum protein 1 (CEMP1) is a cementum-specific protein that can induce cementogenic differentiation. In this study, poly(ethylene glycol) (PEG)-stabilized amorphous calcium phosphate (ACP) nanoparticles were prepared by wet-chemical method and then loaded with recombinant human CEMP1 (rhCEMP1) for controlled release. An electrospun multiphasic scaffold constituted of poly(ε-caprolactone) (PCL), type I collagen (COL), and rhCEMP1/ACP was fabricated. The effects of rhCEMP1/ACP/PCL/COL scaffold on the attachment proliferation, osteogenic, and cementogenic differentiations of human periodontal ligament cells, (PDLCs) were systematically investigated. A critical size defect rat model was introduced to evaluate the effect of tissue regeneration of the scaffolds in vivo. The results showed that PEG-stabilized ACP nanoparticles formed a core-shell structure with sustained release of rhCEMP1 for up to 4 weeks. rhCEMP1/ACP/PCL/COL scaffold could suppress PDLCs proliferation behavior and upregulate the expression of cementoblastic markers including CEMP1 and cementum attachment protein while downregulating osteoblastic markers including osteocalcin and osteopontin when it was cocultured with PDLCs in vitro for 7 days. Histology analysis of cementum after being implanted with the scaffold in rats for 8 weeks showed that there was cementum-like tissue formation but little bone formation. These results indicated the potential of using electrospun multiphasic scaffolds for controlled release of rhCEMP1 for promoting cementum regeneration in reconstruction of the periodontal complex.

Metabotropic glutamate receptor 1 promotes cementoblast proliferation via MAP kinase signaling pathways

ABSTRACTPurpose/aim: Glutamate is one of the signaling molecules responsible for transmission in the central nervous system. Periodontal ligament (PDL) cells were recently reported to express metabotropic glutamate receptors (mGluRs). However, the functions of mGluR signaling in PDL cells or PDL-related cells remain largely unknown. The aim of this study was to investigate the expression and function of mGluRs in PDL-related cells. Materials and methods: OCCM-30 cells, immortalized murine cementoblasts, were stimulated with l-glutamate or mGluRs antagonists. The cells’ proliferative response was evaluated using a colorimetric assay and gene expression was assessed using real-time polymerase chain reaction. The nuclear translocation of cyclin D1 was evaluated by immunohistochemistry. Results: l-Glutamate promoted the proliferation of OCCM-30 cells, which expressed mGluR1, but not mGluR5. Dihydroxyphenylglycine (DHPG), an agonist of group I mGluRs (mGluR1 and mGluR5), also promoted cell proliferation, and this was inhibited by LY456236, an mGluR1 antagonist. DHPG increased the expression of cyclin D1, a key regulator of cell proliferation, and its nuclear translocation. DHPG also increased the expression of Bcl2A1, an antiapoptotic oncogene and simultaneously reduced the expression of Bax, a pro-apoptotic marker. Furthermore, the DHPG-induced proliferation of OCCM-30 cells was reduced by pretreatment with SB203580, SP600125, and PD98059, inhibitors of p38, JNK, and ERK1/2, respectively. Conclusions: These findings indicate that activation of mGluR1 expressed by OCCM-30 cells induces cell proliferation in a manner that is dependent on mitogen-activated protein kinase pathways and that cyclin D1 and Bcl2A1/Bax may be involved. Our results provide useful information for elucidating the mechanisms underlying cementum homeostasis and regeneration.

Periodontal ligament stem/progenitor cells with protein-releasing scaffolds for cementum formation and integration on dentin surface

ABSTRACTPurpose/Aim: Cementogenesis is a critical step in periodontal tissue regeneration given the essential role of cementum in anchoring teeth to the alveolar bone. This study is designed to achieve integrated cementum formation on the root surfaces of human teeth using growth factor–releasing scaffolds with periodontal ligament stem/progenitor cells (PDLSCs). Materials and methods: Human PDLSCs were sorted by CD146 expression, and characterized using CFU-F assay and induced multi-lineage differentiation. Polycaprolactone scaffolds were fabricated using 3D printing, embedded with poly(lactic-co-glycolic acids) (PLGA) microspheres encapsulating connective tissue growth factor (CTGF), bone morphogenetic protein-2 (BMP-2), or bone morphogenetic protein-7 (BMP-7). After removing cementum on human tooth roots, PDLSC-seeded scaffolds were placed on the exposed dentin surface. After 6-week culture with cementogenic/osteogenic medium, cementum formation and integration were evaluated by histomorphometric analysis, immunofluorescence, and qRT-PCR. Results: Periodontal ligament (PDL) cells sorted by CD146 and single-cell clones show a superior clonogenecity and multipotency as compared with heterogeneous populations. After 6 weeks, all the growth factor–delivered groups showed resurfacing of dentin with a newly formed cementum-like layer as compared with control. BMP-2 and BMP-7 showed de novo formation of tissue layers significantly thicker than all the other groups, whereas CTGF and BMP-7 resulted in significantly improved integration on the dentin surface. The de novo mineralized tissue layer seen in BMP-7-treated samples expressed cementum matrix protein 1 (CEMP1). Consistently, BMP-7 showed a significant increase in CEMP1 mRNA expression. Conclusion: Our findings represent important progress in stem cell–based cementum regeneration as an essential part of periodontium regeneration.

Effects of short-term inflammatory and/or hypoxic pretreatments on periodontal ligament stem cells: in vitro and in vivo studies

In this study, we extensively screened the in vitro and in vivo effects of PDLSCs following short-term inflammatory and/or hypoxic pretreatments. We found that the 24-h hypoxic pretreatment of PDLSCs significantly enhanced cell migration and improved cell surface CXCR4 expression. In addition, hypoxia-pretreated PDLSCs exhibited improved cell colony formation and proliferation. Cells that were dually stimulated also formed more colonies compared to untreated cells but their proliferation did not increase. Importantly, the hypoxic pretreatment of PDLSCs enhanced cell differentiation as determined by elevated RUNX-2 and ALP protein expression. In this context, the inflammatory stimulus impaired cell OCN protein expression, while dual stimuli led to decreased RUNX-2 and OCN mRNA levels. Although preconditioning PDLSCs with inflammatory and/or hypoxic pretreatments resulted in no differences in the production of matrix proteins, hypoxic pretreatment led to the generation of thicker cell sheets; the inflammatory stimulus weakened the ability of cells to form sheets. All the resultant cell sheets exhibited clear bone regeneration following ectopic transplantation as well as in periodontal defect models; the amount of new bone formed by hypoxia-preconditioned cells was significantly greater than that formed by inflammatory stimulus- or dual-stimuli-treated cells or by nonpreconditioned cells. The regeneration of new cementum and periodontal ligaments was only identified in the hypoxia-stimulus and no-stimulus cell groups. Our findings suggest that PDLSCs that undergo short-term hypoxic pretreatment show improved cellular behavior in vitro and enhanced regenerative potential in vivo. The preconditioning of PDLSCs via combined treatments or an inflammatory stimulus requires further investigation.

Sealing Ability of MTA Used in Perforation Repair of Permanent Teeth; Literature Review.

There were several materials used to seal different types of perforation defects. MTA is one of these restorative materials that is considered the most effective, biocompatible, non-toxic, and non-irritant; promote bone healing and cementum regeneration. The objective of this article was to review and summarize the sealing ability of MTA compared with the other materials used for sealing different types of root perforations of permanent teeth. A literature search was conducted using Medline, accessed via the National Library of Medicine Pub Med from 2005 to 2015 searching for articles related to sealing ability of MTA. This study found that factors affecting prognosis are the size, site of the perforation and time elapsed as well as the repair material. MTA is an important filling material to be used for sealing different types of perforations when perforated sites sealed immediately with MTA.

Global proteome profiling of dental cementum under experimentally-induced apposition

Dental cementum (DC) covers the tooth root and has important functions in tooth attachment and position. DC can be lost to disease, and regeneration is currently unpredictable due to limited understanding of DC formation. This study used a model of experimentally-induced apposition (EIA) in mice to identify proteins associated with new DC formation. Mandibular first molars were induced to super-erupt for 6 and 21days after extracting opposing maxillary molars. Decalcified and formalin-fixed paraffin-embedded mandible sections were prepared for laser capture microdissection. Microdissected protein extracts were analyzed by liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS), and the data submitted to repeated measure ANOVA test (RM-ANOVA, alpha=5%). A total of 519 proteins were identified, with 97 (18.6%) proteins found exclusively in EIA sites and 50 (9.6%) proteins exclusively expressed in control sites. Fifty six (10.7%) proteins were differentially regulated by RM-ANOVA (p<0.05), with 24 regulated by the exclusive effect of EIA (12 proteins) or the interaction between EIA and time (12 proteins), including serpin 1a, procollagen C-endopeptidase enhancer, tenascin X (TNX), and asporin (ASPN). In conclusion, proteomic analysis demonstrated significantly altered protein profile in DC under EIA, providing new insights on DC biology and potential candidates for tissue engineering applications. SignificanceDental cementum (DC) is a mineralized tissue that covers the tooth root surface and has important functions in tooth attachment and position. DC and other periodontal tissues can be lost to disease, and regeneration is currently unpredictable due to lack of understanding of DC formation. This study used a model of experimentally-induced apposition (EIA) in mice to promote new cementum formation, followed by laser capture microdissection (LCM) and liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS) proteomic analysis. This approach identified proteins associated with new cementum formation that may be targets for promoting cementum regeneration.

Influence of enamel matrix derivative on healing of root surfaces after bonding treatment and intentional replantation of vertically fractured roots.

The objectives of this study were to histopathologically evaluate cementum regeneration on root surfaces when an enamel matrix derivative (EMD) was used to bond a vertically fractured root and to evaluate the effectiveness of EMD in inhibiting root resorption. A total of 40 roots from 24 maxillary premolars in beagles were used. The root was vertically fractured using a chisel and mallet. Super-Bond was then used to bond the fractured root. In the experimental group, the root surface was treated with ethylenediaminetetraacetic acid and an EMD. The control group received no treatment. The root was then replanted in its original location. Histopathological observation and measurement using image-analyzing software were carried out after 8 weeks. In the experimental group, shallow resorption cavities developed on the root planed surfaces with new acellular cementum appearing over them. In the control group, no new cementum was seen on the planed surfaces, and there was connective tissue joining the roots. In some of the samples, resorption and multinucleated giant cells were seen. The experimental group showed a significantly larger volume of cementum formation (P < 0.001), and the volume of root resorption was significantly smaller (P = 0.004). When bonding and replanting tooth roots after a vertical fracture, the application of EMD was effective in regenerating cementum on root surfaces from which periodontal ligament had been lost in the area around the fracture line and in reducing the incidence of root resorption.

Sequential process in brain-derived neurotrophic factor-induced functional periodontal tissue regeneration.

We recently demonstrated that brain-derived neurotrophic factor (BDNF) promotes periodontal tissue regeneration. The purpose of this study was to establish an essential component of a rational approach for the clinical application of BDNF in periodontal regenerative therapy. Here, we assessed the sequence of early events in BDNF-induced periodontal tissue regeneration, especially from the aspect of cementum regeneration. Brain-derived neurotrophic factor was applied into experimental periodontal defects in Beagle dogs. The localization of cells positive for neurotrophic tyrosine kinase, receptor, type 2, proliferating cell nuclear antigen, osteopontin, integrin αVβ3, and integrin α2β1 was evaluated by immunohistochemistry. The effects of BDNF on adhesion of cultured human periodontal ligament cells was examined by an in vitro study. The results suggest that BDNF could induce rapid cementum regeneration by stimulating adhesion, proliferation, and differentiation of periodontal ligament cells in the early regenerative phase, resulting in enhancement of periodontal tissue regeneration.

Effects of the incorporation of epsilon-aminocaproic acid/chitosan particles to fibrin on cementoblast differentiation and cementum regeneration

Effects of the incorporation of epsilon-aminocaproic acid/chitosan particles to fibrin on cementoblast differentiation and cementum regeneration

Periodontal healing with a preameloblast-conditioned medium in dogs.

The predictability of conventional periodontal treatments for damaged periodontal tissue is limited, particularly on the regeneration of new cementum. As signaling molecules, a range of growth factors has been used to promote periodontal regeneration on periodontal ligament (PDL) and cementum defects. A preameloblast-conditioned medium (PA-CM) was prepared from cultured murine apical bud cells, which can differentiate into ameloblasts. We examined the effect of PA-CM on PDL cells and cementoblasts invitro and evaluated histologically the effects of PA-CM on the regeneration of experimentally induced periodontal defects invivo. In vitro, the effects of PA-CM on the migration of human PDL cells were examined using a scratch wound healing assay and a transwell assay. The differentiation and mineralization potential of PA-CM-treated human PDL cells and murine cementoblastic OCCM-30 cells was examined by real-time polymerase chain reaction and Alizarin red-S staining. In vivo, six mongrel dogs (12-16kg; 6-8mo old) were used. Twenty-four roots were replanted with either, (i) only periodontal defects (n=12; control group), or (ii) periodontal defects and PA-CM treatment (n=12; experimental group). In the experimental group, the PDL and cementum between notches was removed using a Gracey curette and soaked in 0.08mL water containing 80μg of a PA-CM for 2min. The dogs were killed at 4 and 8wk post-surgery. The invitro results showed that PA-CM stimulated the migration of PDL cells and promoted the differentiation and mineralization of PDL cells and cementoblasts. Real-time polymerase chain reaction analysis revealed stronger expression of Runx2, Osx, OC, Bsp and Cap mRNAs in the PA-CM-treated PDL cells and cementoblasts than those in the control cells. In vivo, newly formed PDL-like tissue and cementum-like tissue were observed partially between the root surfaces and newly formed bone in the experimental group. The regenerated PDL-like tissue in the experimental group was significantly higher than that in the control group at 8wk (p<0.05). The replacement resorption on the experimental group was significantly lower than that in the control group at 8wk (p<0.05). In addition, the amount of newly formed cementum-like tissue in the experimental group was significantly higher than that in the control group at 4 and 8wk (p<0.05). These results suggest that PA-CM has the potential to regenerate periodontal tissues in PDL and cementum defects.