Effects Of Enamel Matrix Proteins Research Articles

Combining enamel matrix proteins with mechanical stimuli potentiates human periodontal ligament fibroblasts proliferation and periodontium remodeling.

Collagen I (Col-I) and matrix metalloproteinase-1 (MMP-1) have been implicated in the regeneration and remodeling of the periodontium. Studies have shown that enamel matrix proteins (EMPs) and mechanical stimuli can promote the synthesis and degradation, respectively, of Col-I and MMP-1. However, the effects of the combination of EMPs and mechanical stimuli on human periodontal ligament are not known. Our aim was to test the combined effects of EMPs and mechanical stimuli on the proliferation of human periodontal ligament fibroblasts (HPDLFs) and Col-I and MMP-1 mRNA expression. Primary HPDLFs were isolated using an enzyme digestion method. To select the optimum EMP concentration and the optimum magnitude and loading time of mechanical stimuli, HPDLFs were stimulated with gradient concentration of EMPs (0 µg/mL, 25 µg/mL, 50 µg/mL, 100 µg/mL and 200 µg/mL) and mechanical stimuli (0 kPa, 25 kPa, 50 kPa, 100 kPa, and 200 kPa for 0 h, 3 h, 6 h, 12 h, and 24 h), respectively. The cell proliferative response was tested by the MTT assay. The impact of EMPs combined with mechanical stimuli on Col-I and MMP-1 mRNA expression were measured by reverse transcription polymerase chain reaction. 100 µg/mL of EMPs and a 50 kPa mechanical stimulus were chosen as the optimum parameters due to the higher proliferation rates than other doses. The combination of 100 µg/mL of EMPs and a 50 kPa mechanical stimulus significantly stimulated HPDLFs proliferation and increased Col-I and MMP-1 expression levels compared with incubation with two factors alone. We concluded that the combination of EMPs and mechanical stimulus have synergistic effects on cell growth, cell number, collagen turnover, and periodontium remodeling.

Enamel matrix proteins regulate hypoxia-induced cellular biobehavior and osteogenic differentiation in human periodontal ligament cells

Hypoxia is a crucial microenvironment for inflamed periodontal tissue and periodontal wound healing. Enamel matrix proteins (EMPs) potentially can promote the formation of new periodontium. The effects of EMPs on periodontal ligament cells under hypoxia, however, remain unclear. We investigated the effects of EMPs on cellular biobehavior and osteogenic differentiation of human periodontal ligament cells (hPDLCs) under hypoxia. Under cobalt chloride (CoCl2)-induced hypoxia, cellular biobehavior of hPDLCs, including proliferation, attachment, spreading, and migration with or without EMPs, was evaluated by 3-(4, 5-dimethylthiazol- 2-yl)-2, 5-diphenyl tetrazolium bromide (MTT), cell counting, spreading area measurement and wound scratch assay. The osteogenic activity of hPDLCs was assessed using alkaline phosphatase (ALP) and alizarin red S staining (ARS). The expressions of osteogenic genes including runt related transcription factor 2 (Runx2), ALP, osteocalcin (OCN) and collagen type I (Col-I) were detected using real time quantitative PCR, western blot and immunocytochemistry assays. The biobehavior and osteogenic differentiation of hPDLCs were inhibited significantly under hypoxia. EMPs have no effect on cell proliferation under mimicked hypoxia. EMPs partly reversed the inhibitory effects of hypoxia, however, for other cellular biobehavior including attachment, spreading and migration, and markedly up-regulated osteogenic differentiation activities including ALP, mineralization ability and the expressions of osteogenic genes such as Runx2, ALP, osteocalcin, and collagen type I in hPDLCs under hypoxia. EMPs attenuate the hypoxic injury to cellular biobehavior and osteogenic differentiation in hPDLCs under hypoxia.

Effects of enamel matrix proteins on adherence, proliferation and migration of epithelial cells: A real-time in vitro study.

Enamel matrix derivative (EMD) can mimic odontogenic effects by inducing the proliferation and differentiation of connective tissue progenitor cells, stimulating bone growth and arresting epithelial cells migration. To the best of our knowledge, there is no data indicating that any active component of EMD reduces epithelial cell viability. The present study examines the impact of commercial lyophilized EMD, porcine recombinant amelogenin (prAMEL; 21.3 kDa) and tyrosine-rich amelogenin peptide (TRAP) on the adherence, proliferation and migration of human epithelial cells in real-time. The tongue carcinoma cell line SCC-25 was stimulated with EMD, porcine recombinant AMEL and TRAP, at concentrations of 12.5, 25 and 50 µg/ml. Cell adherence, migration and proliferation were monitored in real-time using the xCELLigence system. No significant effects of EMD on the morphology, adhesion, proliferation and migration of SCC-25 cells were observed. However, porcine recombinant AMEL had a dose-dependent inhibitory effect on SCC-25 cell proliferation and migration. Predominantly, no notable differences were found between control and TRAP-treated cells in terms of cell adhesion and migration, a decrease in proliferation was observed, but this was not statistically significant. EMD and its active components do not increase the tongue cancer cell viability.

The Effects of Enamel Matrix Proteins and Dentin Collagen on the Attachment of Periodontal Ligament Cells

The Effects of Enamel Matrix Proteins and Dentin Collagen on the Attachment of Periodontal Ligament Cells

Clinical concepts for regenerative therapy in furcations.

Furcation involvements present one of the greatest challenges in periodontal therapy because furcation-involved molar teeth respond less favorably to conventional periodontal therapy compared with noninvolved molar or nonmolar teeth. Various regenerative procedures have been proposed and applied with the aim of eliminating the furcation defect or reducing the furcation depth. An abundance of studies and several systematic reviews have established the effectiveness of membrane therapy (guided tissue regeneration) for buccal Class II furcation involvement of mandibular and maxillary molars compared with open flap surgery. Bone grafts/substitutes may enhance the results of guided tissue regeneration. However, complete furcation closure is not a predictable outcome. Limited data and no meta-analyses are available on the effects of enamel matrix proteins for furcation regeneration. Enamel matrix protein therapy has demonstrated clinical improvements in the treatment of buccal Class II furcation defects in mandibular molars; however, complete closure of the furcation lesion is achieved only in a minority of cases. Neither guided tissue regeneration nor enamel matrix protein therapy have demonstrated predictable results for approximal Class II and for Class III furcations. Promising preclinical data from furcation regeneration studies in experimental animals is available for growth factor- and differentiation factor-based technologies, but very limited data are available from human clinical studies. Although cell-based therapies have received considerable attention in regenerative medicine, their experimental evaluation in the treatment of periodontal furcation lesions is at a very early stage of development. In summary, the indications and the limitations for currently available treatment modalities for furcation defects are well established. New regenerative treatments are clearly needed to improve the predictability of a complete resolution of furcation defects.

Effects of enamel matrix proteins in combination with a bovine-derived natural bone mineral for the repair of bone defects

Previously, the use of enamel matrix derivative (EMD) in combination with a natural bone mineral (NBM) was able to stimulate periodontal ligament cell and osteoblast proliferation and differentiation. Despite widespread use of EMD for periodontal applications, the effects of EMD on bone regeneration are not well understood. The aim of the present study was to test the ability of EMD on bone regeneration in a rat femur defect model in combination with NBM. Twenty-seven rats were treated with either NBM or NBM + EMD and assigned to histological analysis at 2, 4, and 8 weeks. Defect morphology and mineralized bone were assessed by μCT. For descriptive histology, hematoxylin and eosin staining and Safranin O staining were performed. Significantly more newly formed trabecular bone was observed at 4 weeks around the NBM particles precoated with EMD when compared with NBM particles alone. The drilled control group, in contrast, achieved minimal bone regeneration at all three time points (P < 0.05). The present results may suggest that EMD has the ability to enhance the speed of new bone formation when combined with NBM particles in rat osseous defects. These findings may provide additional clinical support for the combination of EMD with bone graft for the repair of osseous and periodontal intrabony defects.

Effects of enamel matrix proteins on multi-lineage differentiation of periodontal ligament cells in vitro

The adult periodontal ligament (PDL) is considered to contain progenitor cells that are involved in the healing of periodontal wounds. Treatment with enamel matrix derivative (EMD), a heat-treated preparation derived from enamel matrix proteins (EMPs), has been shown to be of some clinical benefit in eliciting periodontal regeneration in vivo. Although there is extensive information available about the effects of EMD on periodontal regeneration, the precise influence of this material on alveolar bone and the formation of blood vessels and proprioceptive sensory nerves, prominent features of functionally active periodontal tissue, remain unclear. The aim of the present study was therefore to examine the effects of EMD on the ability of human periodontal ligament cells (HPCs) to undergo multi-lineage differentiation in vitro. Our results showed that HPCs treated with EMD under non-selective growth conditions did not show any evidence of osteogenic, adipogenic, chondrogenic, neovasculogenic, neurogenic and gliogenic “terminal” differentiation. In contrast, under selective lineage-specific culture conditions, EMD up-regulated osteogenic, chondrogenic and neovasculogenic genes and “terminal” differentiation, but suppressed adipogenesis, neurogenesis and gliogenesis. These findings thus demonstrate for the first time that EMD can differentially modulate the multi-lineage differentiation of HPCs in vitro.

Differential Effect of Amelogenin Peptides on Osteogenic DifferentiationIn Vitro: Identification of Possible New Drugs for Bone Repair and Regeneration

Enamel matrix proteins (EMP) have been shown to promote regeneration of periodontal ligament and root cementum, and sometimes to enhance the differentiation of bone-forming cells in vitro and new bone growth in vivo. However, the inconsistent and unpredictable effects of EMP that have been reported for bone regeneration may be due to the highly variable composition of this heterogeneous material, which is comprised mainly of amelogenin and amelogenin-derived peptides. The present study has therefore examined the effects of naturally occurring low-molecular-weight (LMW) and high-molecular-weight (HMW) fractions of Emdogain(®) (EMD; Institut Straumann, Basel, Switzerland), a commercially available form of EMP, on osteogenic differentiation of bone precursor cells in vitro. In addition, the effects of chemically synthesized specific components of LMW and HMW-namely, the tyrosine-rich amelogenin peptide (TRAP), a specific amelogenin isoform derived by proteolytic clipping, and a leucine-rich amelogenin peptide (LRAP), an isoform derived by alternative splicing-on bone-forming cell activity were also investigated. Our findings demonstrate that while TRAP suppressed the formation of bone-like mineralized nodules, LRAP upregulated osteogenic differentiation. Furthermore, synthetically produced TRAP and its unique C-terminal 12 amino acid sequence (TCT) also suppressed bone-forming cells, whereas LRAP and its unique C-terminal 23 amino acid sequence (LCT) markedly enhanced terminal differentiation of bone-forming cells. These findings suggest that the differential effects of amelogenin-derived peptide sequences present in EMP could be of potential clinical value, with the novel bioactive TCT peptide as a useful tool for limiting pathological bone cell growth and the unique LCT sequence having therapeutic benefits in the treatment of periodontal and orthopedic diseases.

Effects of enamel matrix proteins on proliferation, differentiation and attachment of human alveolar osteoblasts

Enamel matrix proteins (EMPs) have been demonstrated to promote periodontal regeneration. However, effects of EMPs on human alveolar osteoblasts (hAOBs), up to now, have still been unclear. The purpose of this study was to investigate influence of EMPs on proliferation, differentiation and attachment of hAOBs in vitro. EMPs were extracted using the acetic acid method, hAOBs were obtained and cultured in vitro. Cell proliferation, alkaline phosphatase (ALP) activity, mRNA expression of osteogenic markers and cell attachment were measured in the absence and in the presence of EMPs (50, 100 and 200 μg/ml). EMPs increased proliferation of hAOBs; however, they inhibited ALP activity and mRNA expression of osteogenic markers (collagen I, ALP, runt-related protein 2, osteocalcin, bone sialoprotein and osteopontin). Meanwhile, EMPs hindered hAOBs' attachment. These effects occurred in EMPs concentration-dependent manner. These results indicate that EMPs may inhibit osteoblastic differentiation and attachment to prevent ankylosis and allow other cell types to regenerate periodontal tissues.

Ectopic study of tissue-engineered bone complex with enamel matrix proteins, bone marrow stromal cells in porous calcium phosphate cement scaffolds, in nude mice

This study aimed to investigate the potential of enamel matrix proteins (EMPs) on promoting osteogenic differentiation of porcine bone marrow stromal cells (pBMSCs), as well as new bone formation capabilities, in a tissue-engineered bone complex scaffold of EMPs, pBMSCs and porous calcium phosphate cement (CPC). Effects of EMPs on pBMSCs in vitro was first determined by alkaline phosphatase (ALP) activity, von Kossa staining assay and mRNA expression of ALP, bone sialoprotein (BSP) and osteocalcin (OCN) genes. Next, an ectopic new bone formation test was performed in a nude mouse model with four groups: CPC scaffold alone; CPC scaffold + EMPs; CPC scaffold + pBMSCs; and CPC scaffold + EMPs + pBMSCs, for 2 or 4 weeks. ALP activity, von Kossa assay and mRNA expressions of ALP, BSP and OCN genes were all significantly higher with 150 μg/ml EMP treatment in vitro. In nude mice, new bone formation was detected only in the CPC scaffold + EMPs + pBMSCs group at 2 weeks. At 4 weeks, in the tissue-engineered construct there was significantly higher bone formation ability than other groups. EMPs promoted osteogenic differentiation of pBMSCs, and the tissue-engineered complex of EMPs, pBMSCs and CPC scaffold may be a valuable alternative to be used in periodontal bone tissue engineering and regeneration.

The effect of enamel matrix proteins on the spreading, proliferation and differentiation of osteoblasts cultured on titanium surfaces

Modifications of implant surface topography and chemistry have proven a means to enhance osseointegration, a process that ensures the stability of bone-contacting devices, including titanium dental implants. The commercial product Emdogain ® is an enamel matrix derivative (EMD) extracted from porcine teeth commonly used in periodontal surgery, where it has been shown to potentiate regeneration of bone. The aim of the present study was to evaluate the effect of EMD on the attachment, proliferation and differentiation of osteoblasts on titanium surfaces in vitro. Pickled (smooth) and SLA (roughened) titanium discs were coated with EMD or left uncoated. Primary rat calvarial osteoblasts were cultured on each surface from 1 h to 4 weeks. EMD significantly increased cell spreading and proliferation at time points ranging from 3 to 7 days on both topographies. Alkaline phosphatase activity was significantly increased on EMD-coated titanium compared with titanium alone. Moreover, there was a 6 fold increase in levels of mRNA encoding bone sialoprotein and osteocalcin in osteoblasts cultured on EMD-coated titanium surfaces compared with uncoated surfaces. We conclude that coating of titanium with EMD enhances the proliferation and differentiation of osteoblasts irrespective of the titanium substratum topography.

Effect of combined therapeutical methods on healing of intrabony defects in regenerative periodontal surgery

Several methods are available to enhance the healing and regeneration of periodontal tissues after surgical therapy of intrabony defects. The main indications for the use of combined regenerative procedures are the extent and morphology of the osseous lesions. The six studies of the present dissertation focused on the clinical effect of different barrier techniques, bone substitutes, enamel matrix derivatives and a growth factors containing adjuvant used in various combinations on the healing of severe periodontal intrabony impairments. Synthetic, xenogenetic and autologous materials were used in these randomized clinical studies. Mechanical barriers (polytetrafluoroethylene and collagen membranes) for GTR, biological barriers/enamel matrix proteins (EMD), synthetic (β-TCP) and xenogeneic (NBM) bone grafts and autologous platelet-rich plasma (PRP) were combined in the test and control groups of the trials. The main clinical variable was the clinical attachment level (CAL) and the subsidiary one was the probing pocket depth (PPD), estimated at baseline and after one year. The summation of the results after the statistical analysis takes cognizance of the followings: a) Each of the eleven regenerative methods evaluated (ten combined procedures) leads to significant CAL gain and PPD decrease. b) Using β-TCP or NBM with EMD or with PRP + GTR and GTR’s, the difference between the parameters of the test and control groups were not statistically significant. c) It was confirmed in four studies that the addition of PRP to graft materials has not increased significantly the positive outcomes independent of the type of barrier or graft. d) Adding platelet-rich plasma to natural bone mineral, no benefit was observed from the point of view of the clinical variables. e) The polypeptide proteins of the platelet-rich plasma do not enhance the clinical regenerative effect of enamel matrix proteins. In conclusion, the option of the periodontal surgeons between these methods depends mainly on the defect morphology, the patient’s approach to the different types of materials, the medical concept of the physician, the technical possibilities and the clinical experience of the periodontist.

Biological mediators and periodontal regeneration: a review of enamel matrix proteins at the cellular and molecular levels

Despite a large body of clinical and histological data demonstrating beneficial effects of enamel matrix proteins (EMPs) for regenerative periodontal therapy, it is less clear how the available biological data can explain the mechanisms underlying the supportive effects of EMPs. To analyse all available biological data of EMPs at the cellular and molecular levels that are relevant in the context of periodontal wound healing and tissue formation. A stringent systematic approach was applied using the key words "enamel matrix proteins" OR "enamel matrix derivative" OR "emdogain" OR "amelogenin". The literature search was performed separately for epithelial cells, gingival fibroblasts, periodontal ligament cells, cementoblasts, osteogenic/chondrogenic/bone marrow cells, wound healing, and bacteria. A total of 103 papers met the inclusion criteria. EMPs affect many different cell types. Overall, the available data show that EMPs have effects on: (1) cell attachment, spreading, and chemotaxis; (2) cell proliferation and survival; (3) expression of transcription factors; (4) expression of growth factors, cytokines, extracellular matrix constituents, and other macromolecules; and (5) expression of molecules involved in the regulation of bone remodelling. All together, the data analysis provides strong evidence for EMPs to support wound healing and new periodontal tissue formation.

Effect of enamel matrix proteins on the treatment of intrabony defects: a split-mouth randomized controlled trial study

The objective of this split-mouth, double-blind, randomized controlled trial was to compare the clinical effect of treatment of 2- or 3-wall intrabony defects with open flap debridement (OFD) combined or not with enamel matrix proteins (EMP). Thirteen volunteers were selected with one pair of or more intrabony defects and probing pocket depth (PPD) > or = 5 mm. All individuals received instructions regarding oral hygiene and were submitted to scaling and root planing. Each participant received the two treatment modalities: test sites were treated with OFD and EMP, and control sites received only OFD. After 6 months, a significant reduction was observed in PPD for the EMP group (from 6.42 +/- 1.08 mm to 2.67 +/- 1.15 mm) and for the OFD group (from 6.08 +/- 1.00 mm to 2.00 +/- 0.95 mm) (p < 0.0001), but with no significant difference between groups (p = 0.13). A significant gain in relative attachment level (RAL) was observed in both groups (EMP: from 13.42 +/- 1.88 mm to 10.75 +/- 2.26 mm, p < 0.001; OFD: from 12.42 +/- 1.98 mm to 10.58 +/- 2.23 mm, p = 0.013), but with no significant difference between groups (p = 0.85). Gingival recession (GR) was higher in the EMP group (from 1.08 +/- 1.50 mm to 2.33 +/- 1.43 mm; p = 0.0009) than in the OFD group (from 0.66 +/- 1.15 mm to 1.16 +/- 1.33 mm; p = 0.16), but this difference was not significant (p = 0.06). In conclusion, the results showed that OFD combined with EMP was not able to improve treatment of intrabony defects compared to OFD alone.

The effect of enamel matrix proteins and deproteinized bovine bone mineral on heterotopic bone formation

To evaluate the osteoinductive potential of deproteinized bovine bone mineral (DBBM) and an enamel matrix derivative (EMD) in the muscle of rats. Sixteen rats were used in this study. The animals were divided in three groups. Group A: a pouch was created in one of the pectoralis profundis muscles of the thorax of the rats and DBBM particles (Bio-Oss) were placed into the pouch. Healing: 60 days. Group B: a small pouch was created on both pectoralis profundis muscles at each side of the thorax midline. In one side, a mixture of EMD (Emdogain) mixed with DBBM was placed into one of the pouches, whereas in the contralateral side of the thorax the pouch was implanted with DBBM mixed with the propylene glycol alginate (PGA--carrier for enamel matrix proteins of EMD). Healing: 60 days. Group C: the same procedure as group B, but with a healing period of 120 days. Qualitative histological analysis of the results was performed. At 60 days, the histological appearance of the DBBM particles implanted alone was similar to that of the particles implanted together with EMD or PGA at both 60 and 120 days. The DBBM particles were encapsulated into a connective tissue stroma and an inflammatory infiltrate. At 120 days, the DBBM particles implanted together with EMD or PGA exhibited the presence of resorption lacunae in some cases. Intramuscular bone formation was not encountered in any group. The implantation of DBBM particles alone, combined with EMD or its carrier (PGA) failed to exhibit extraskeletal, bone-inductive properties.

Application of Periodontal Tissue Engineering Using Enamel Matrix Derivative and a Human Fibroblast‐Derived Dermal Substitute to Stimulate Periodontal Wound Healing in Class III Furcation Defects

Enamel matrix derivative (EMD) has been shown to promote several aspects of periodontal regeneration in vitro and in vivo. Recently, a bioengineered tissue (DG) was developed to promote wound healing of chronic skin ulcers. This pilot study sought to assess the effects of EMD and DG, alone or in combination, on periodontal wound healing in surgically created Class III furcation defects. Six female baboons received bilateral ostectomy of approximately 10 mm around the first and second mandibular molars to achieve Class III, subclass C furcation defects. Wire ligatures and cotton pellets were left in place for 2 months to maintain the depth of the defects and promote plaque accumulation. Each furcally involved molar was then assigned to one of four treatments: open flap debridement (OFD), OFD plus EMD, OFD plus DG, or OFD plus DG and EMD. This resulted in six total sites per treatment group. Seven months after defect creation and 5 months after treatment, and after no oral hygiene, tissue blocks of the mandible were taken for blinded histometric analysis to assess parameters of periodontal regeneration adjacent to furcal root surfaces and from the mid-furcal aspect (i.e., new bone, new connective tissue attachment, new epithelial attachment, and new cementum formation). Histometric analysis demonstrated differential regenerative responses with respect to treatment within each animal. However, statistically significant differences between treatments from all six animals were not observed (P >0.20, mixed-model analysis of variance). EMD-treated sites presented mildly positive regenerative results and no negative responses. Both DG only and combination therapy demonstrated similar or less than positive responses relative to OFD controls. The descriptive analysis may suggest a positive effect of enamel matrix proteins and a negative effect of DG used alone or in combination with enamel matrix proteins on the regeneration of Class III furcation defects in baboons.

Effects of Emdogain on osteoblast gene expression

Emdogain (EMD) is a protein extract purified from porcine enamel and has been introduced in clinical practice to obtain periodontal regeneration. EMD is composed mainly of amelogenins (90%), while the remaining 10% is composed of non-amelogenin enamel matrix proteins such as enamelins, tuftelin, amelin and ameloblastin. Enamel matrix proteins seem to be involved in root formation. EMD has been reported to promote proliferation, migration, adhesion and differentiation of cells associated with healing periodontal tissues in vivo. How this protein acts on osteoblasts is poorly understood. We therefore attempted to address this question by using a microarray technique to identify genes that are differently regulated in osteoblasts exposed to enamel matrix proteins. By using DNA microarrays containing 20,000 genes, we identified several upregulated and downregulated genes in the osteoblast-like cell line (MG-63) cultured with enamel matrix proteins (Emd). The differentially expressed genes cover a broad range of functional activities: (i) signaling transduction, (ii) transcription, (iii) translation, (iv) cell cycle regulation, proliferation and apoptosis, (v) immune system, (vi) vesicular transport and lysosome activity, and (vii) cytoskeleton, cell adhesion and extracellular matrix production. The data reported are the first genome-wide scan of the effect of enamel matrix proteins on osteoblast-like cells. These results can contribute to our understanding of the molecular mechanisms of bone regeneration and as a model for comparing other materials with similar clinical effects.

The effect of the amelogenin fraction of enamel matrix proteins on fibroblast-mediated collagen matrix reorganization

Enamel matrix proteins (EMP), extracted from developing porcine teeth, promote not only periodontal regeneration but also cutaneous wound healing presumably via the amelogenin fraction. Because it is unclear whether the effect of EMP can be ascribed to amelogenins, we compared EMP with recombinant amelogenin in the relaxed dermal equivalent (DE) in vitro model for early wound contraction. EMP and recombinant porcine amelogenin (rP172) at 1 mg/ml were incorporated into DEs composed of human dermal fibroblasts and a type I collagen matrix. The area reduction, as a measure of contraction, as well as fibroblast numbers and TGF- β1 levels, were quantified over 7 days in culture in the presence of 10% foetal bovine serum. Both EMP and recombinant amelogenin increased contraction ( p < 0.0 0 5 ) and fibroblast numbers ( p < 0.0 0 5 ) compared with controls (acetic acid vehicle and 1 mg/ml porcine serum albumin) and the positive control TGF- β1 added at 10 ng/ml. Increased contraction with EMP and recombinant amelogenin was most pronounced after the first day of incubation and was associated with elevated ( p < 0.0 0 5 ) TGF- β1 levels in conditioned medium. In conclusion, the amelogenin component of EMP augmented fibroblast-driven collagen matrix remodelling, at least partially, by increasing the endogenous production of TGF- β1. These effects of EMP/amelogenin may be beneficial for cutaneous wound healing.

Effects of enamel matrix proteins on tissue formation along the roots of human teeth

Enamel matrix-derived proteins (EMD) are thought to trigger the formation of acellular extrinsic fibre cementum (AEFC), while other reports indicate that EMD may have osteogenic potential. The aim of the present study was to characterize the tissues developing on the root surface following application of EMD. Twelve human periodontitis-affected teeth, scheduled for extraction, were treated with EMD. Two to 6 weeks later, the teeth were extracted, demineralized and processed for embedding in acrylic and epoxy resins. New tissue formation was analysed by light and transmission electron microscopy. New tissue formation on the root was observed in the notch and on both scaled and unscaled root surfaces distant of the notch area in six defects. The newly formed tissues on the root were thick, collagenous, devoid of extrinsic fibres, and had an irregular surface contour. The presence of electron-dense, organic material in the collagenous matrix indicated at least partial mineralization. Embedded cells were numerous and the cells on the matrix surface were very large in size. Abundant rough endoplasmic reticulum and a prominent Golgi complex were evident. The presence of a split between the treated root surfaces and the newly formed tissue was a common observation, as was the presence of bacteria and host cells in the interfacial gap. Following treatment with EMD, a bone-like tissue resembling cellular intrinsic fibre cementum may develop on the root surfaces, instead of AEFC. Furthermore, EMD may both induce de novo formation of a mineralized connective tissue on scaled root surfaces and stimulate matrix deposition on old native cementum. Interfacial bonding appeared to be weak after 6 weeks of healing.

Periodontal regeneration in intrabony defects after application of enamel matrix proteins with guided tissue regeneration: An experimental study in dogs

The aim of this study was to evaluate the effects of enamel matrix proteins (EMP) at the early stage of wound healing in the periodontal tissues by a combination treatment with guided tissue regeneration (GTR). Intrabony defects were produced surgically at the distal aspects of both mandibles in six beagle dogs. At 12 weeks following the surgery, the defects were exposed using a full thickness mucoperiosteal flap procedure. Subsequently, the defects were treated by the following treatments: a control group treated with GTR alone, and an experimental group treated with a combination of GTR and EMP. After one, two, four and eight weeks of the treatment, the animals were sacrificed, and sections of the tissue were stained and evaluated microscopically. After one and two weeks, the proliferating cell nuclear antigen (PCNA)-positive cell ratios of the experimental group were significantly greater than that of the control group. After 2 and 4 weeks, new bone and new cementum formation in the experimental group were significantly greater than those in the control group (P < 0.05). However, after 8 weeks, no statistical difference was found between the two groups in new bone or cementum formation. The study results suggest that a maturation of periodontal ligament cells might contribute, during the early stage of periodontal healing, to stimulate a proliferation of periodontal ligament cells.