transforming-growth-factor-beta and Furcation-Defects

Tissue engineering is the emerging field of science developing techniques for fabrication of new tissues for replacement based on principles of cell and developmental biology and biomaterials. Morphogenesis is the cascade of pattern formation and the attainment of form of the various organs and the organism as a whole. The periodontium consist of the periodontal ligament, cementum, and alveolar bone. Bone has considerable potential for regeneration and therefore is a prototypic model for tissue engineering. The three main ingredients for tissue engineering are regulatory signals, responding stem cells, and extracellular matrix. Recent advances in molecular biology of the bone morphogenetic proteins (BMPs) have set the stage for tissue engineering of bone and related tissues, including the periodontium. Bone-derived BMPs, with a collagenous matrix as carrier, induced cementum and alveolar bone regeneration in surgically created furcation defects in the primate. It is noteworthy that there was morphogenesis of periodontal ligament and a faithful insertion of Sharpey's fibers into cementum. In the same furcation model, recombinant human osteogenic protein-1 (rhOP-1, also known as BMP-7), in conjunction with the collagenous carrier, induced extensive cementogenesis with insertion of Sharpey's fibers into the newly formed cementum. The observation that BMPs induce cementogenesis and periodontal ligament formation indicates that these proteins may have multiple functions in vivo not limited to cartilage and bone induction. The rapid advances in the molecular biology of BMPs and their receptors bode well for novel strategies to engineer the regeneration of the periodontal tissues.

Topics: Alveolar Process; Animals; Biocompatible Materials; Biomedical Engineering; Bone Morphogenetic Protein 7; Bone Morphogenetic Proteins; Collagen; Dental Cementum; Extracellular Matrix; Furcation Defects; Guided Tissue Regeneration, Periodontal; Humans; Molecular Biology; Morphogenesis; Periodontal Ligament; Periodontium; Recombinant Proteins; Regeneration; Signal Transduction; Stem Cells; Transforming Growth Factor beta

This study was an initial evaluation of the use of a bovine-derived bone protein (BP) extract that contains various growth factors combined with decalcified freeze-dried bone allograft (DFDBA) as regenerative treatment for class II mandibular furcations. Twenty-five patients were divided into 5 groups according to the dosage of BP present per mg of DFDBA to be grafted: (1) 0.00 microgram/mg, (2) 3.13 micrograms/mg, (3) 6.25 micrograms/mg, (4) 12.5 micrograms/mg, and (5) 25.0 micrograms/mg. Surgical exploration of the furcation defects was performed followed by grafting with BP/DFDBA. Results at 6 months showed that attachment gain in the treated furcation areas was greatest in Groups 4 and 5, suggesting that BP has the potential to increase the effects of DFDBA in gaining clinical attachment in mandibular class II furcations.

Topics: Adult; Aged; Animals; Bone Morphogenetic Proteins; Bone Regeneration; Bone Transplantation; Cattle; Dental Plaque Index; Double-Blind Method; Female; Follow-Up Studies; Furcation Defects; Gingival Recession; Humans; Male; Mandibular Diseases; Middle Aged; Observer Variation; Periodontal Attachment Loss; Periodontal Index; Periodontal Pocket; Prospective Studies; Reproducibility of Results; Statistics as Topic; Surgical Flaps; Transforming Growth Factor beta; Transplantation, Homologous

The quantity of regenerated bone induced by recombinant human bone morphogenetic protein-2 (rhBMP2) is encouraging, but sometimes the quality is inferior. Recombinant human beta-nerve growth factor (rh beta-NGF) plays a major role in bone remodeling. This study evaluates the quality and quantity of regenerated bone in periodontal regeneration following topical application of the two growth factors to Class III furcation defects.. Thirty-six inflamed Class III furcation defects were created in six beagle dogs at sites of mandibular premolars 2, 3, and 4, and then biodegradable hydrogel incorporating rhBMP2 and rh beta-NGF was topically applied to the defects. The groupings were as follows: G1, untreated (control group A); G2, carrier alone (control group B); G3, 0.4% rhBMP2 + carrier; G4, 2% rh beta-NGF + carrier; G5, 0.4% rhBMP2 + 2% rh beta-NGF + carrier; and G6, 0.2% rhBMP2 + 1% rh beta-NGF + carrier. Eight weeks after application, the quality and quantity of regenerated tissue were evaluated by scanning electron microscopy observation, calcium/phosphorus ratio analysis, and histologic evaluation.. The regenerated bone in G5 exhibited the highest calcium/phosphorus ratio among all groups and showed a denser structure with more calcified substances on the collagen fiber surface than that in the other groups. Histomorphometric analysis revealed that 0.4% rhBMP2 + 2% rh beta-NGF promoted the highest percentage of periodontal regeneration among all groups.. The results of this pilot study suggest that a topical application of rhBMP2 and rh beta-NGF may improve the quality and quantity of regenerated bone in artificially created Class III furcation defects of beagle dogs.

Topics: Administration, Topical; Animals; Bone Morphogenetic Protein 2; Bone Morphogenetic Proteins; Bone Regeneration; Calcium; Dogs; Drug Carriers; Drug Combinations; Female; Furcation Defects; Guided Tissue Regeneration, Periodontal; Humans; Hydrogel, Polyethylene Glycol Dimethacrylate; Inflammation; Nerve Growth Factor; Phosphates; Pilot Projects; Recombinant Proteins; Spectrometry, X-Ray Emission; Transforming Growth Factor beta

Osteogenic proteins of the transforming growth factor-beta superfamily induce periodontal tissue regeneration in animal models, including primates. To our knowledge, no studies have been performed in periodontal regeneration using the transforming growth factor-beta 3 isoform. In the present study, recombinant human transforming growth factor-beta 3 was examined for its ability to induce periodontal tissue regeneration in the nonhuman primate, Papio ursinus.. Class II furcation defects were surgically created bilaterally in the maxillary and mandibular molars of four adult baboons. Heterotopic ossicles, for transplantation to selected furcation defects, were induced within the rectus abdominis muscle by recombinant human transforming growth factor-beta 3. Forty days later, the periodontal defects were implanted with recombinant human transforming growth factor-beta 3 in Matrigel as the delivery system, with recombinant human transforming growth factor-beta 3 plus minced muscle tissue in Matrigel, or with the harvested recombinant human transforming growth factor-beta 3-induced ossicles. Sixty days after periodontal implantation, the animals were killed and the specimens harvested. Histological analysis on undecalcified sections measured the area and volume of new alveolar bone and the coronal extension of newly formed alveolar bone and cementum.. Morphometric analyses showed pronounced periodontal regeneration in experimental defects compared with controls. Substantial regeneration was observed in defects implanted with fragments of heterotopically induced ossicles and with recombinant human transforming growth factor-beta 3 plus minced muscle tissue.. Recombinant human transforming growth factor-beta 3 in Matrigel significantly enhanced periodontal tissue regeneration in the nonhuman primate, P. ursinus.

Topics: Alveolar Process; Animals; Bone Morphogenetic Protein 7; Bone Morphogenetic Proteins; Bone Regeneration; Furcation Defects; Guided Tissue Regeneration, Periodontal; Humans; Molar; Ossification, Heterotopic; Papio ursinus; Protein Isoforms; Rectus Abdominis; Transforming Growth Factor beta; Transforming Growth Factor beta3

Periodontal regeneration under application of bone morphogenetic protein (BMP) is compromised by ankylosis. Ankylosis disappearance following application of BMP has been observed in the case of a small defect, which might be beneficial change for periodontal regeneration. However, the histological observation of ankylosis disappearance has not been demonstrated in a large defect. The purpose of this present study was to confirm resolution of ankylosis during periodontal regeneration by recombinant human BMP-2 (rhBMP-2) applied to class III furcation defects.. Class III furcation defects were created in the premolars of six adult cats. The rhBMP-2 material, prepared by applying rhBMP-2 to a combination of polylactic acid-polygricolic copolymer and gelatin sponge (PGS; 0.33 microg rhBMP-2/mm(3) PGS) or control material containing only PGS, was implanted into each defect. The cats were killed at 3, 6 or 12 weeks after surgery and serial sections were prepared for histological and histometrical observation.. Ankylosis was observed in some of the rhBMP-2/PGS group at 3 and 6 weeks, but not at 12 weeks. At 6 weeks, osteoclast-like cells were visible in the rhBMP-2/PGS group with ankylosis. Residual PGS was evident between the bone and root surface in the rhBMP-2/PGS group without ankylosis at 3 weeks.. Resolution of ankylosis by osteoclast-like cells possibly occurred under application of rhBMP-2. Residual PGS might play an important role in preventing ankylosis formation.

Topics: Absorbable Implants; Animals; Bone Morphogenetic Protein 2; Bone Morphogenetic Proteins; Bone Regeneration; Cats; Dental Cementum; Drug Carriers; Furcation Defects; Gelatin; Humans; Implants, Experimental; Lactic Acid; Male; Osteoclasts; Periodontal Ligament; Polyglycolic Acid; Polylactic Acid-Polyglycolic Acid Copolymer; Polymers; Recombinant Proteins; Regeneration; Tooth Ankylosis; Transforming Growth Factor beta

Hydrogels are based on hydrophilic polymers which are cross-linked to prevent dissolution in water. Because hydrogels can contain large amounts of water, they are interesting devices for the delivery of protein drugs. In this contribution, biodegradable dextran-co-gelatin hydrogel microspheres (DG-MPs) are described which are based on physical interactions and are particularly suitable for the controlled delivery of pharmaceutically active proteins. The unique feature of this preparation system is that the hydrogel microsphere formation takes place in an all-aqueous solution, by which the use of organic solvents is avoided. We investigated the preparation and biological activities of recombinant human bone morphogenetic protein-2 (rhBMP2)-loaded dextran-co-gelatin hydrogel microspheres (rhBMP2-DG-MPs), which aimed to keep rhBMP2's biological activity and to achieve a long-term sustained release of rhBMP2. The microspheres' average diameter was about 20-40 microm and rhBMP2 release in vitro could be maintained for >10 days. Cytology studies showed that using rhBMP2-DG-MPs could promote the proliferation and osteoblastic differentiation of periodontal ligament cells better than using rhBMP2 aqueous solution. By a freeze-drying method, rhBMP2-DG-MPs could be adhered in chitosan membranes for guided tissue regeneration use, namely functionalized membranes. To evaluate bone regeneration induced by rhBMP2-DG-MPs, an animal experiment with canine class III furcation defects was adopted and the results indicated that using rhBMP2-DG-MPs incorporating scaffolds and functionalized membranes could gain more periodontal tissue regeneration than using scaffolds and general membranes soaked with concentrated rhBMP2 aqueous solution. Therefore, those studies demonstrate the potential of DG-MPs in the sustained delivery of low dosages of rhBMP2 to periodontal defects.

Topics: Animals; Bone Morphogenetic Protein 2; Bone Morphogenetic Proteins; Cell Proliferation; Cells, Cultured; Chitosan; Delayed-Action Preparations; Dextrans; Dogs; Female; Furcation Defects; Gelatin; Guided Tissue Regeneration, Periodontal; Humans; Hydrogels; Microspheres; Periodontal Ligament; Recombinant Proteins; Technology, Pharmaceutical; Transforming Growth Factor beta

Recombinant human bone morphogenetic protein-2 (rhBMP-2) has been shown to induce clinically relevant bone formation for orthopedic, craniofacial, and oral indications. It appears critical, in particular for onlay indications, that the associated carrier technology exhibits structural integrity to offset compressive forces in support of rhBMP-2-induced bone formation. The objective of this study was to evaluate a calcium phosphate (CP) cement, Ceredex, as a candidate carrier for rhBMP-2 in a defect model with limited osteogenic potential.. Bilateral, critical size, 6-mm, supra-alveolar, periodontal defects were created in six, adult, male, Hound Labrador mongrels. Three animals received rhBMP-2/Ceredex (rhBMP-2 at 0.20 and 0.40 mg/ml) in contralateral defect sites (implant volume/defect approximately 1 ml). One defect site in each of the three remaining animals received Ceredex without rhBMP-2 (control). The animals were euthanized at 12 weeks postsurgery for histologic and histometric analysis.. Mean induced bone height exceeded 80% of the defect height for supra-alveolar periodontal defects receiving rhBMP-2/Ceredex without major differences between rhBMP-2 concentrations compared with approximately 40% for the control. The newly formed bone, a mixture of lamellar and woven bone in fibrovascular tissue, circumscribed relatively large portions of the residual Ceredex biomaterial. Inflammatory lesions were associated with limited bone formation in some sites. From a periodontal perspective, sites receiving rhBMP-2/Ceredex exhibited increased cementum formation compared with control, but without a functionally oriented periodontal ligament, and increased ankylosis and root resorption. Control sites exhibited early wound failure and exposure, loss of the Ceredex biomaterial, and limited bone formation.. The Ceredex CP cement appears a potentially promising carrier technology for rhBMP-2 onlay indications. However, a slow resorption rate may prevent its wider use. This study does not support use of the rhBMP-2/Ceredex combination for periodontal indications.

Topics: Absorbable Implants; Animals; Bone Morphogenetic Protein 2; Bone Morphogenetic Proteins; Bone Regeneration; Calcium Phosphates; Cementogenesis; Dental Cements; Dogs; Drug Carriers; Furcation Defects; Humans; Male; Oral Surgical Procedures; Recombinant Proteins; Root Resorption; Tooth Ankylosis; Transforming Growth Factor beta

Native and recombinant human bone morphogenetic/osteogenic proteins (BMPs/ OPs) singly initiate bone induction in vivo. The finding of synchronous but spatially different BMPs/OPs expression during periodontal tissue morphogenesis suggests novel therapeutic approaches using morphogen combinations based on recapitulation of embryonic development. Twelve furcation defects prepared in the first and second mandibular molars of three adult baboons (Papio ursinus) were used to assess whether qualitative histological aspects of periodontal tissue regeneration could be enhanced and tissue morphogenesis modified by combined or single applications of recombinant hOP-1 and hBMP-2. Doses of BMPs/OPs were 100 microg of each protein per 1 g of insoluble collagenous bone matrix as carrier. Approximately 200 mg of carrier matrix was used per furcation defect. Undecalcified sections cut for histological analysis 60 d after healing of hOP-1-treated specimens showed substantial cementogenesis with scattered remnants of the collagenous carrier. hBMP-2 applied alone induced greater amounts of mineralized bone and osteoid when compared to hOP-1 alone or to combined morphogen applications. Combined applications of hOP-1 and hBMP-2 did not enhance alveolar bone regeneration or new attachment formation over and above the single applications of the morphogens. The results of this study, which is the first to attempt to address the structure-activity relationship amongst BMP/OP family members, indicate that tissue morphogenesis induced by hOP-1 and hBMP-2 is qualitatively different when the morphogens are applied singly, with hOP-1 inducing substantial cementogenesis. hBMP-2 treated defects, on the other hand, showed limited cementum formation but a temporal enhancement of alveolar bone regeneration and remodelling. The demonstration of therapeutic mosaicism in periodontal regeneration will require extensive testing of ratios and doses of recombinant morphogen combinations for optimal tissue engineering in clinical contexts.

Topics: Alveolar Process; Analysis of Variance; Animals; Bone Matrix; Bone Morphogenetic Protein 2; Bone Morphogenetic Protein 7; Bone Morphogenetic Proteins; Bone Regeneration; Bone Remodeling; Calcification, Physiologic; Dental Cementum; Disease Models, Animal; Drug Carriers; Furcation Defects; Humans; Linear Models; Male; Molar; Morphogenesis; Papio; Periodontium; Pilot Projects; Recombinant Proteins; Regeneration; Statistics as Topic; Structure-Activity Relationship; Transforming Growth Factor beta; Wound Healing

Osteogenic protein-1 (OP-1) is a member of the transforming growth factor beta superfamily and is a potent modulator of osteogenesis and bone cell differentiation. This preclinical study in dogs sought to assess the effects of OP-1 on periodontal wound healing in surgically created critical size Class III furcation defects. Eighteen male beagle dogs were subjected to the creation of bilateral mandibular 5 mm osseous defects. A split-mouth design was utilized which randomly assigned opposing quadrants to control therapy (surgery alone or collagen vehicle) or 1 of 3 ascending concentrations of OP-1 in a collagen vehicle (0.75 mg OP-1/g collagen, 2.5 mg/g, or 7.5 mg/g). Thus, 9 quadrants per test group received OP-1, 9 quadrants per control group received surgery alone, and 9 quadrants received collagen vehicle alone. Test articles were delivered by a surgeon masked to the treatment, and fluorogenic bone labels were injected at specified intervals post-treatment. Eight weeks after defect creation and OP-1 delivery, tissue blocks of the mandibulae were taken for masked histomorphometric analysis to assess parameters of periodontal regeneration (e.g., bone height, bone area, new attachment formation, and percent of defect filled with new bone). Histomorphometry revealed limited evidence of osteogenesis, cementogenesis, and new attachment formation in either vehicle or surgery-alone sites. In contrast, sites treated with all 3 concentrations of OP-1 showed pronounced stimulation of osteogenesis, regenerative cementum, and new attachment formation. Lesions treated with 7.5 mg/g of OP-1 in collagen regenerated 3.9+/-1.7 mm and 6.1+/-3.4 mm2 (mean +/-S.D.) of linear bone height and bone area, respectively. Furthermore, these differences were statistically different from both control therapies for all wound healing parameters (P < 0.0001). No significant increase in tooth root ankylosis was found among the treatment groups when compared to the surgery-alone group. We conclude that OP-1 offers promise as an attractive candidate for treating severe periodontal lesions.

Topics: Alveolar Process; Animals; Ankylosis; Bone Density; Bone Morphogenetic Protein 7; Bone Morphogenetic Proteins; Collagen; Dental Cementum; Disease Models, Animal; Dogs; Fluoresceins; Fluorescent Dyes; Furcation Defects; Humans; Male; Mandible; Osteogenesis; Periodontal Attachment Loss; Pharmaceutical Vehicles; Random Allocation; Recombinant Proteins; Regeneration; Single-Blind Method; Tooth Diseases; Tooth Root; Transforming Growth Factor beta; Wound Healing

This study evaluated alveolar bone and cementum regeneration following surgical implantation of recombinant human transforming growth factor-beta1 (rhTGF-beta1) in conjunction with guided tissue regeneration (GTR). Supraalveolar, critical size, periodontal defects were surgically created around the 3rd and 4th mandibular premolar teeth in right and left jaw quadrants in 5 beagle dogs. Alternate jaw quadrants in consecutive animals received rhTGF-beta1, in a CaCO3/hydroxyethyl starch carrier with GTR, or carrier with GTR alone (control). 20 microg of rhTGF-beta1 in buffer solution was incorporated into approximately 0.8 ml of carrier for each defect scheduled to receive rhTGF-beta1. Animals were sacrificed at week 4 postsurgery and tissue blocks were harvested and processed for histometric analysis. Clinical healing was generally uneventful. Minor membrane exposures were observed. Defects with membrane exposure displayed an inflammatory infiltrate underneath the membrane. Bone regeneration of trabecular nature, apparent in all animals, was generally limited to the very apical aspect of the defects. Cementum regeneration was limited without obvious differences between experimental conditions. Comparing rhTGF-beta1, to control defects, statistically significant differences were found for area (1.8+/-0.4 and 1.3+/-0.6 mm2, respectively; p<0.05) and density (0.3+/-0.1 and 0.2+/-0.03, respectively; p<0.05) of alveolar bone regeneration. Observed differences are small and represent a clinically insignificant potential for enhanced regeneration in this preclinical model. Within the limitations of study, it may be concluded that rhTGF-beta1 has a restricted potential to enhance alveolar bone regeneration in conjunction with GTR.

Topics: Alveolar Bone Loss; Alveolar Process; Animals; Bicuspid; Bone Regeneration; Calcium Carbonate; Dental Cementum; Dogs; Drug Carriers; Furcation Defects; Guided Tissue Regeneration, Periodontal; Humans; Hydroxyethyl Starch Derivatives; Membranes, Artificial; Periodontal Diseases; Polytetrafluoroethylene; Recombinant Proteins; Regeneration; Transforming Growth Factor beta

The purpose of this study was to analyse the effect of TFG-beta 1 on wound healing in standardized Class II furcation defects of 48 mandibular second premolar teeth in 24 sheep. The experimental design included a control group (carrier only, 25% pluronic F-127), and 2 experimental groups: group A (80 micrograms/ml TGF-beta 1 + carrier) and group B (80 micrograms/ml TGF-beta 1 + carrier covered with a barrier membrane). Sheep were killed either 2 wk or 6 wk after surgery. Mesiodistal sections of the decalcified specimens were quantified histologically using stereology. Percentage volumes of regenerated bone, fibrous connective tissue and cementum were calculated for each furcation defect. Mean values were analysed using multiple ANOVA; p values were calculated using paired and unpaired Student's t-tests. After 2 wk there was more bone in group B than either of the other 2 groups, but this was not statistically significant. By 6 wk more bone was present in group A than in the control group (p < 0.02) and also in group B when compared with both group A and the control group (p < 0.02 and p < 0.44), respectively. In the 4 wk between sampling significantly more bone had formed (group A < 0.05 and group B p < 0.003, respectively). A negative correlation existed between volumes of bone and fibrous connective tissue and no significant differences between the volumes of cementum were evident between any of the groups. This study demonstrated that TGF-beta 1 encouraged bone regeneration in Class II furcation defects in sheep, an effect enhanced by the presence of a barrier membrane. This is the first report on the use of TGF-beta 1 in conjunction with GTR in periodontal defects.

Topics: Alveolar Process; Analysis of Variance; Animals; Bicuspid; Bone Regeneration; Connective Tissue; Dental Cementum; Female; Furcation Defects; Guided Tissue Regeneration, Periodontal; Mandible; Membranes, Artificial; Periodontal Ligament; Random Allocation; Sheep; Transforming Growth Factor beta; Wound Healing

Recombinant human osteogenic protein-1 (hOP-1), a member of the bone morphogenetic protein family, was examined for its efficacy in periodontal regeneration. Twelve furcation defects, surgically prepared in the first and second mandibular molars, were treated with bovine insoluble collagenous matrix in conjunction with 0.0 (control), 100 and 500 mu g of recombinant hOP-1 per g of matrix. After 60 days of healing, histological and histometric analyses on serial, undemineralized sections cut at 7 mu m showed substantial cementogenesis on the exposed dentine of furcations treated with both doses of hOP-1 (p < 0.01 vs control). Foci of nascent mineralization were seen within the newly deposited cementoid along the coronal areas of hOP-1-treated defects. Within the furcations, there were substantial amounts of residual collagenous carrier, interspersed with a mineralized matrix having histological features of cementum. This mineralized cementum-like material was predominantly deposited around the carrier, and blended into newly formed cementum along the root surfaces. In the apical area, the cementum-like material and the remaining alveolar bony housing were not connected; indeed the two components were separated by a fibrovascular tissue that had numerous features of the periodontal ligament space. Formation and insertion of Sharpey's fibres into newly formed root cementum were also observed. It is likely that the expression of specific cell phenotypes by hOP-1 is regulated, in part, by the extracellular matrix microenvironment, including dentine. Thus, exposed dentine, in the presence of exogenous hOP-1 at the doses tested, may preferentially modulate the expression of the cementogenic phenotype. These findings in a non-human primate show that hOP-1, at the doses tested, induced cementogenesis on surgically denuded root surfaces, indicating a specific function during repair and regeneration of periodontal tissues.

Topics: Alveolar Process; Animals; Bone Morphogenetic Protein 7; Bone Morphogenetic Proteins; Cattle; Collagen; Dental Cementum; Dentin; Extracellular Matrix; Furcation Defects; Gene Expression Regulation; Humans; Papio; Periodontal Ligament; Periodontium; Phenotype; Recombinant Proteins; Regeneration; Tooth Calcification; Tooth Root; Transforming Growth Factor beta; Wound Healing