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Octacalcium phosphate combined with collagen orthotopically enhances bone regeneration.

Octacalcium phosphate (OCP) is resorbable bone regenerative material, but its brittleness makes it difficult to maintain its shape without restraint. We have engineered a scaffold constructed of synthetic OCP and porcine collagen sponge (OCP/Collagen) and investigated whether OCP/Collagen composite could improve bone regeneration. To examine this hypothesis, bone regeneration by the implantation of OCP/Collagen was compared with those by OCP and collagen. Radiographic and histological examination was performed and the percentage of newly formed bone (n-Bone%) in the defect was determined by a histomorphometrical analysis. OCP/Collagen, OCP, or collagen was implanted into the critical-sized defects in rat crania and fixed at 2, 4, or 8 weeks after implantation. OCP/Collagen improved the handling performance than the granules of OCP, and synergistically enhanced the bone regeneration beyond expectation, which were composed of bone nucleation by OCP and cell infiltration by collagen. Histomorphometrical analysis showed that n-Bone% +/- standard error treated with OCP/Collagen (48.4 +/- 5.14) was significantly higher than those with OCP (27.6 +/- 4.04) or collagen (27.4 +/- 5.69) in week 8. The present study suggests that the combination OCP with collagen elicited the synergistic effect for bone regeneration.

Animals↗

VEGF scaffolds enhance angiogenesis and bone regeneration in irradiated osseous defects.

UNLABELLED: Bone regeneration is challenging in sites where the blood supply has been compromised by radiation. We examined the potential of a growth factor (VEGF) delivery system to enhance angiogenesis and bone formation in irradiated calvarial defects. VEGF-releasing polymers significantly increased blood vessel density and vascular perfusion in irradiated defects and increased bone formation relative to control conditions. INTRODUCTION: Radiation therapy causes damage to tissues and inhibits its regenerative capacity. Tissue injury from radiation is in large part caused by a compromised vascular supply and reduced perfusion of tissues. The aim of this study was to determine if delivery of vascular endothelial growth factor (VEGF) from a biodegradable PLGA (copolymer of D,L-lactide and glycolide) scaffold could enhance neovascularization and bone regeneration in irradiated osseous defects. MATERIALS AND METHODS: An isolated area of the calvarium of Fisher rats was irradiated (12 Gy) 2 weeks preoperatively, and two 3.5-mm osseous defects were created in this area, followed by the placement of PLGA scaffolds or VEGF scaffolds (PLGA scaffolds with incorporated VEGF) into the defects. Laser Doppler perfusion imaging was performed to measure perfusion of these areas at 1, 2, and 6 weeks. Implants were retrieved at 2, 6, and 12 weeks, and histologic and muCT analyses were performed to determine neovascularization and bone regeneration. RESULTS: Histological analyses revealed statistically significant increases in blood vessel formation (>2-fold) and function (30%) within the VEGF scaffolds compared with PLGA scaffolds. Additionally, evaluation of bone regeneration through bone histomorphometric and muCT analyses revealed significantly greater bone coverage (26.36 +/- 6.91% versus 7.05 +/- 2.09% [SD]) and increased BMD (130.80 +/- 58.05 versus 71.28 +/- 42.94 mg/cm(3)) in VEGF scaffolds compared with PLGA scaffolds. CONCLUSIONS: Our findings show that VEGF scaffolds have the ability to enhance neovascularization and bone regeneration in irradiated osseous defects, outlining a novel approach for engineering tissues in hypovascular environments.

Animals↗

Evaluation of a bioabsorbable physical barrier for guided bone regeneration. Part II. Material and a bone replacement graft.

Part I of the evaluation of a prototype bioabsorbable physical barrier composed of a copolymer of lactide and glycolide for treatment of bone defects in the guided bone regeneration procedure indicated that the prototype bioabsorbable physical barrier did not possess sufficient spacemaking characteristics to prevent collapse of the barrier into the defect or against the threads of the titanium implants. The purpose of Part II was to evaluate this bioabsorbable physical barrier in combination with a supporting material to prevent barrier collapse. Posterior mandibular teeth in three dogs were extracted and allowed to heal for 3 months. This produced localized alveolar ridge defects with a narrow buccolingual width. Six titanium threaded implants were placed in the right and left mandibles of each dog so that nonspacemaking dehiscencetype defects were produced. Two defects in each animal were randomly treated with the prototype bioabsorbable physical barrier and decalcified freeze-dried bone allograft; two defects were treated with a nonbioabsorbable expanded polytetrafluoroethylene barrier with decalcified freeze-dried bone allograft; and one defect each was treated with prototype bioabsorbable physical barrier alone or by flap access with no barrier or bone replacement graft. The results demonstrated that both the bioabsorbable and the nonbioabsorbable barrier combined with decalcified freeze-dried bone allograft produce comparable amounts of new bone with percent bone-to-implant contact, height, width, and area. Defects treated with the prototype bioabsorbable physical barrier alone or no barrier demonstrated unfavorable results. It is suggested that a bone replacement graft is indicated when treating defects with a nonspacemaking morphology.

Alveolar Bone Loss↗

Combinatorial gene therapy for bone regeneration: cooperative interactions between adenovirus vectors expressing bone morphogenetic proteins 2, 4, and 7.

Bone morphogenetic proteins (BMPs) have demonstrated effectiveness as bone regeneration agents whether delivered as recombinant proteins or via gene therapy. Current gene therapy approaches use vectors expressing single BMPs. In contrast, multiple BMPs are coordinately expressed during bone development and fracture healing. Furthermore, BMPs likely exist in vivo as heterodimeric molecules having enhanced biological activity. In the present study, we test the hypothesis that gene therapy-based bone regeneration can be enhanced by expressing combinations of BMPs. For in vitro studies, mesenchymal cell lines were transduced with individual adenoviruses containing BMP2, 4, or 7 cDNA under control of a CMV promoter (AdBMP2, 4, 7) or virus combinations. Significantly, combined transduction with AdBMP2 plus AdBMP7 or AdBMP4 plus AdBMP7 resulted in a synergistic stimulation of osteoblast differentiation. This synergy is best explained by formation of BMP2/7 and 4/7 heterodimers. To test in vivo biological activity, fibroblasts were transduced with specific virus combinations and implanted into C57BL6 mice. Consistent with in vitro results, strong synergy was observed using combined AdBMP2/BMP7 treatment, which induced twofold to threefold more bone than would be predicted based on the activity of individual AdBMPs. These studies show that dramatic enhancement of osteogenesis can be achieved using gene therapy to express specific combinations of interacting regenerative molecules.

Adenoviridae↗

Morphology of peri-implant regenerated bone, in sheep's tibia, by means of guided tissue regeneration.

BACKGROUND: No data on the resistance to functional loads of bone regenerated by means of guided tissue regeneration, can be found. In this paper the new-formed bone is analyzed, in cases that see the surgical creation of defects in testing animals. The morphological aspects of perimplant bone, regenerated by means of bone substitutes or not, were evaluated. METHODS: A coronal defect was created by means of a counter-sink bur. The following step provided for osseointegrated fixtures, with a machined or blasted (TiOblast) surface, to be installed. Some bone defects were filled with Bio-Oss, and covered with resorbable membrane (Vycril), others were left unfilled. The animals were sacrificed after different periods of time (24 and 45 days). Some thick sections (200 microns) underwent then microradiography and were examined with a microscope, under transmitted and polarized light. Other sections, thin (5 microns), were coloured with toluidine blue, Gomori's Stain and Solochrome Cyanine/Congo red. RESULTS: Data confirm the excellent behaviour of bovine bone and of TiO blasted fixtures. The experimental results bring into evidence that, in cases of Guided Bone Regeneration (GBR) by means of membrane, the use of fixtures with a blasted surface (TiOblast), and of Bio-Oss as bone substitute, made osseointegration easier. CONCLUSIONS: Results, moreover, suggest that in case of implant surgery with GBR, in order to allow the maturation of regenerated bone, it's healing time must be prolonged.

Animals↗

Acellular dermal matrix graft as a membrane for guided bone regeneration: a case report.

Guided bone regeneration has been successfully used to treat several types of bone defects. Problems associated with the procedure, such as premature exposure of the membranes to the oral cavity and, consequently, contamination, may occur. The acellular dermal matrix graft material is presently used to treat several soft tissue problems. It could be used for guided bone regeneration with the advantage of forming soft tissue while acting as a barrier membrane. A patient in need of guided bone regeneration for the staged placement of an implant was treated by using the acellular dermal matrix graft material as a barrier membrane. Healing progressed uneventfully with the formation of adequate new bone and an increase in the width of keratinized tissue.

Alveolar Bone Loss↗

Periodontal repair in dogs: a bioabsorbable calcium carbonate coral implant enhances space provision for alveolar bone regeneration in conjunction with guided tissue regeneration.

BACKGROUND: Collapse or compression of a barrier device into a periodontal defect or onto the root surface compromises outcomes following guided tissue regeneration (GTR). Bone biomaterials have been suggested to support regeneration of alveolar bone and to improve space provision with GTR devices. The objective of this study was to evaluate space provision, alveolar bone, and cementum regeneration following use of a bioabsorbable, calcium carbonate biomaterial in conjunction with GTR. METHODS: Routine, critical size, 5 to 6 mm, supraalveolar, periodontal defects were created in 5 young adult beagle dogs. Alternate jaw quadrants in consecutive animals received GTR and the coral biomaterial (cGTR) or GTR alone. The animals were euthanized 4 weeks postsurgery and tissue blocks processed for histometric analysis. RESULTS: The coral implant particles were surrounded by newly-formed bone or immersed in connective tissue and appeared to resorb and be replaced by bone. There was limited, if any, appreciable cementum regeneration. Space provision was enhanced in cGTR compared to GTR sites (6.1 +/- 1.6 versus 2.4 +/- 0.8 mm2; P<0.05). Bone regeneration (height) was significantly increased in cGTR compared to GTR sites averaging 1.9 +/- 0.6 and 1.2 +/- 0.6 mm, respectively (P<0.05). Bone regeneration (area) was 2-fold greater in cGTR sites compared to the GTR control (3.3 +/- 1.8 versus 1.4 +/- 0.5 mm2), however the difference was not statistically significant (P>0.05). CONCLUSIONS: The coral implant significantly enhanced space provision for GTR while alveolar bone formation appeared to be enhanced by its use. Increased healing intervals are needed to fully understand the biologic value of the coral implant as an adjunct to GTR.

Absorbable Implants↗

Success and failure rates of osseointegrated implants in function in regenerated bone for 6 to 51 months: a preliminary report.

Six hundred twenty-six titanium plasma-sprayed cylindrical implants that had been functioning in regenerated bone for a period of up to 51 months were evaluated according to Albrektsson's criteria. These implants had been placed either with concomitant guided bone regeneration procedures or in bony ridges that had previously been augmented. According to Albrektsson's criteria, absolute success rates of 97.6% overall, 98.8% in the maxilla, and 98.2% in the mandible were recorded, as were cumulative success rates of 94.9% in the maxilla and 91.9% in the mandible. The overall cumulative success rate was 93.8%. One patient, who underwent protracted chemotherapy 14 months after implant function began, accounted for 33% (3 of 9) of the total implant loss in the study. If this patient is excluded from the study, the cumulative success rates are 94.9% (maxilla), 96.7% (mandible), and 95.8% (overall). These findings support the theory that regenerated bone will achieve osseointegration with titanium plasma-sprayed cylindrical implants, and that this osseointegration can be maintained under function over time.

Adolescent↗

Tissue engineering for bone regeneration using differentiated alveolar bone cells in collagen scaffolds.

Regeneration of osseous defects by a tissue-engineering approach provides a novel means of treatment utilizing cell biology, materials science, and molecular biology. In this study the concept of tissue engineering was tested with collagen type I matrices seeded with cells with osteogenic potential and implanted into sites where osseous damage had occurred. Explant cultures of cells from human alveolar bone and gingiva were established. When seeded into a three-dimensional type I collagen-based scaffold, the bone-derived cells maintained their osteoblastic phenotype as monitored by mRNA and protein levels of the bone-related proteins including bone sialoprotein, osteocalcin, osteopontin, bone morphogenetic proteins 2 and 4, and alkaline phosphatase. These in vitro-developed matrices were implanted into critical-size bone defects in skulls of immunodeficient (SCID) mice. Wound healing was monitored for up to 4 weeks. When measured by microdensitometry the bone density within defects filled with osteoblast-derived matrix was significantly higher compared with defects filled with either collagen scaffold alone or collagen scaffold impregnated with gingival fibroblasts. New bone formation was found at all the sites treated with the osteoblast-derived matrix at 28 days, whereas no obvious new bone formation was identified at the same time point in the control groups. In situ hybridization for the human-specific Alu gene sequence indicated that the newly formed bone tissue resulted from both transplanted human osteoblasts and endogenous mesenchymal stem cells. The results indicate that cells derived from human alveolar bone can be incorporated into bioengineered scaffolds and synthesize a matrix, which on implantation can induce new bone formation.

Alveolar Process↗

[Filling skull defects with regenerating bone].

A destruction method was elaborated in dog experiments which allowed bone defects in the skull to be repaired with regenerating bone. The bone is ground to fillings, moistened with the recipient's blood, and implanted in the bone defect in the skull onto the dura mater which is necessary for bone regeneration. The bone fillings dissolve within 7 days, their cells perish and the substances released from them induce osteogenesis in the cells of the immature connective tissue. Fresh bone filings may be used for auto-, allo- and heteroplasty. They preserve their osteo-inducing property after freezing at--78 degrees C and are inactivated after being autoclaved and treated with 0.5% formaldehyde solution or diocide solution. Lyophilization reduces the osteo-inducing property of bone fillings.

Adolescent↗

Effects of guided bone regeneration around commercially pure titanium and hydroxyapatite-coated dental implants. II. Histologic analysis.

The purpose of this study was to determine which treatment of a large osseous defect adjacent to an endosseous dental implant would produce the greatest regeneration of bone and degree of osseointegration: barrier membrane therapy plus demineralized freeze-dried bone allograft (DFDBA), membrane therapy alone, or no treatment. The current study histologically assessed changes in bone within the healed peri-implant osseous defect. In a split-mouth design, 6 implants were placed in edentulous mandibular ridges of 10 mongrel dogs after preparation of 6 cylindrical mid-crestal defects, 5 mm in depth, and 9.525 mm in diameter. An implant site was then prepared in the center of each defect to a depth of 5 mm beyond the apical extent of the defect. One mandibular quadrant received three commercially pure titanium (Ti) screw implants (3.75 x 10 mm), while the contralateral side received three hydroxyapatite (HA) coated root-form implants (3.3 x 10 mm). Consequently, the coronal 5 mm of each implant was surrounded by a circumferential defect approximately 3 mm wide and 5 mm deep. The three dental implants in each quadrant received either DFDBA (canine source) and an expanded polytetrafluoroethylene membrane (ePTFE), ePTFE membrane alone, or no treatment which served as the control. Clinically, the greatest increase in ridge height and width was seen with DFDBA/ePTFE. Histologically, statistically significant differences in defect osseointegration were seen between treatment groups (P < 0.0001: DFDBA/ePTFE > ePTFE alone > control). HA-coated implants had significantly greater osseointegration within the defect than Ti implants (P < 0.0001). Average trabeculation of newly formed bone in the defect after healing was significantly greater for HA-coated implants than for titanium (P < 0.0001), while the effect on trabeculation between treatments was not significantly different (P = 0.14). Finally, there were significantly less residual allograft particles in defect areas adjacent to HA-coated implants than Ti implants (P = 0.0355). The use of HA-coated implants in large size defects with DFDBA and ePTFE membranes produced significantly more osseointegration histologically than other treatment options and more than Ti implants with the same treatment combinations. The results of this study indicate that, although the implants appeared osseointegrated clinically after 4 months of healing, histologic data suggest that selection of both the implant type and the treatment modality is important in obtaining optimum osseointegration in large size defects.

Alveolar Bone Loss↗

Effects of guided bone regeneration around commercially pure titanium and hydroxyapatite-coated dental implants. I. Radiographic analysis.

The purpose of this study was to determine which treatment of a large osseous defect adjacent to an endosseous dental implant would produce the greatest regeneration of bone and degree of osseointegration: barrier membrane therapy plus demineralized freeze-dried bone allograft (DFDBA), membrane therapy alone, or no treatment. The current study assessed radiographic density changes in bone within the healed peri-implant osseous defect. In a split-mouth design, 6 implants were placed in edentulous mandibular ridges of 10 mongrel dogs after preparation of 6 cylindrical mid-crestal defects, 5 mm in depth and 9.525 mm in diameter. An implant site was then prepared in the center of each defect to a depth of 5 mm beyond the apical extent of the defect. One mandibular quadrant received three commercially pure titanium (Ti) screw implants (3.75 X 10 mm), while the contralateral side received three hydroxyapatite (HA) coated root-form implants (3.3 X 10 mm). Consequently, the coronal 5 mm of each implant was surrounded by a circumferential defect approximately 3 mm wide and 5 mm deep. The three dental implants in each quadrant received either DFDBA (canine source) and an expanded polytetrafluoroethylene membrane (ePTFE), ePTFE membrane alone, or no treatment (control). Standardized radiographs were taken at 1 week and 4 months post-implant placement. Computer-assisted densitometric image analysis (CADIA) was performed at 6 areas of interest (coronal, middle, and apical defect areas mesial and distal to each implant) for each of the implant sites. Significantly greater increase in bone density was obtained using DFDBA/ePTFE compared to ePTFE alone or the controls; likewise, ePTFE alone resulted in greater bone density change than the controls. There were no significant differences in radiographic bone density adjacent to Ti versus HA-coated implants. When 3 dogs having postoperative membrane complications were eliminated from the analysis, the results were similar with the exception that defects adjacent to Ti implants had significantly less density gain when compared to HA-coated implants. The results of this study indicate the use of DFDBA/ePTFE in large surgically-created defects promotes a denser healing of bone adjacent to implants when measured radiographically than either ePTFE alone or no treatment.

Absorptiometry, Photon↗

Controlled bone regeneration: the ultimate process in bone repair.

The main goal of bone healing, besides the healing of the bone, is ensuring that the dynamic system of repair is under biologic control. To do so, the soft tissue has to be kept isolated to avoid any herniation into the bony defect, that would produce and interfere with the bone interface or collapse the original defect resulting in a relapse.

Adolescent↗

Bone regeneration with recombinant human bone morphogenetic protein-2 (rhBMP-2) using absorbable collagen sponges (ACS): influence of processing on ACS characteristics and formulation.

The effects of variability in three parameters (mass, cross-linking with CH2O, and EtO sterilization) of three surgically implantable absorbable collagen sponges (ACS) were studied. Sponges soaked with recombinant human bone morphogenetic protein-2 (rhBMP-2) solution were analyzed for pH, conductivity, and rhBMP-2 precipitation. A method using trinitrobenzenesulfonic acid was developed to quantify the free amino groups of the collagen sponge. With up to 240 min exposure to CH2O, the amount of free amino groups was reduced to 80%. In comparison, the denaturation temperature as determined by differential scanning calorimetry (DSC) after the sponges were soaked with phosphate-buffered saline, increased from 48 to 55 degrees C, indicating stronger interactions due to cross-linking. Subsequent sterilization with EtO caused a marked decrease in the amount of free amino groups (approximately 33% of nonsterilized controls) independent of previous CH2O treatment. However, the denaturation temperature was on average 5 degrees C lower in sterilized sponges than in nonsterilized material. In contrast to CH2O exposure, the strong reaction with EtO appeared to weaken the collagen structure. Resistance of the sponge to collagenase correlated with the degree of collagen cross-linking but was slightly reduced by sterilization. In addition, the pH of ACS soaked with water was substantially increased by sterilization. Protein precipitation was a function of pH and salt concentration but there was no effect due to collagen alone. Results indicated that ACS weight has to be limited to avoid rhBMP-2 precipitation.

Absorbable Implants↗

Spontaneous bone regeneration in surgically induced bone defects in young rabbits.

Defects of the femoral shaft (15%, 20%, 25%, and 30% of the femoral length) with intact periosteum were made in young rabbits to investigate differences in union time. The mean union time was 7.3, 7.1, 7.4, and 7.0 weeks, respectively and there was no significant difference ( >0.05) between the groups. The mean healing indices were 6.7, 4.6, 3.9, and 3.1 weeks/cm, respectively and these were significantly different ( <0.05) between the 15% bone defect group and the other bone defect groups. These results suggested that union time was not affected by the amount of bone defect. Femoral lengthening of 20% was performed to compare the quality of callus with those of the femoral defect of 20%. The mean healing indices of defect and lengthening were 4.6 and 5.7, respectively and the difference was significant ( = 0.004). The callus stiffness of femoral defect was higher ( = 0.02) than that after femoral lengthening whereas the bone mineral density of the callus showed no significant difference ( = 0.37) between two groups. The method of filling bone defect with callus generated from the intact periosteum may be a new therapeutic option for the reconstruction of large bone defects in children when other treatments are not available.

Age Factors↗

Bone regeneration at rabbit skull defects treated with transforming growth factor-beta1 incorporated into hydrogels with different levels of biodegradability.

OBJECT: Skull bone regeneration induced by transforming growth factor-beta1 (TGFbeta1)-containing gelatin hydrogels (TGFbeta1-hydrogels) was investigated using a rabbit skull defect model. Different strengths of TGFbeta1 were examined and compared: different TGFbeta1 doses in gelatin hydrogels with a fixed water content, different water contents in gelatin hydrogels with a fixed TGFbeta1 dose, and TGFbeta1 in solution form. In addition, regenerated skull bone was observed over long time periods after treatment. METHODS: Soft x-ray, dual energy x-ray absorptometry, and histological studies were performed to assess the time course of bone regeneration at a 6-mm-diameter skull defect in rabbits after treatment with TGFbeta1-hydrogels or other agents. The influence of TGFbeta1 dose and hydrogel water content on skull bone regeneration by TGFbeta1-hydrogels was evaluated. Gelatin hydrogels with a water content of 95 wt% that incorporated at least 0.1 microg of TGFbeta1 induced significant bone regeneration at the rabbit skull defect site 6 weeks after treatment, whereas TGFbeta1 in solution form was ineffective, regardless of dose. The in vivo degradability of the hydrogels, which varied according to water content, played an important role in skull bone regeneration induced by TGFbeta1 -hydrogels. In our hydrogel system, TGFbeta1 is released from hydrogels as a result of hydrogel degradation. When the hydrogel degrades too quickly, it does not retain TGFbeta1 or prevent ingrowth of soft tissues at the skull defect site and does not induce bone regeneration at the skull defect. It is likely that hydrogel that degrades too slowly physically impedes formation of new bone at the skull defect. Following treatment with 0.1-microg TGFbeta1-hydrogel (95 wt%), newly formed bone remained at the defect site without being resorbed 6 and 12 months later. The histological structure of the newly formed bone was similar to that of normal skull bone. Overgrowth of regenerated bone and tissue reaction were not observed after treatment with TGFbeta1 -hydrogels. CONCLUSIONS: A TGFbeta1-hydrogel with appropriate biodegradability will function not only as a release matrix for the TGFbeta1, but also as a space provider for bone regeneration. The TGFbeta1-hydrogel is a promising surgical tool for skull defect repair and skull base reconstruction.

Absorptiometry, Photon↗