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PubMed1994

Guided bone regeneration for implant placement using absorbable collagen membranes: case presentations.

The current research on the application of guided tissue regeneration principles is focused on the use of absorbable matrix barriers in bone augmentation procedures in extraction sites and around implants. This paper presents two cases in which a new absorbable collagen membrane is used for guided bone regeneration. In the first case, freeze-dried demineralized bone was used with the membrane to place an implant in a fresh extraction site with a grave osseous defect. In the second case, the collagen membrane provided the augmentation of a narrow ridge where three implants were placed, without the use of allograft. In both cases, the barrier membrane demonstrated its applicability for bone regeneration with only one surgical procedure, without complications. The learning objective of this article is to update the knowledge of absorbable barrier membranes from the clinical experience of the authors.

Adult↗
PubMed2007

Effects of ipriflavone on augmented bone using a guided bone regeneration procedure.

: This study investigated the effects of ipriflavone (IP) on augmented bone using a guided bone regeneration (GBR) procedure. In 15 rabbits, two titanium caps were placed into calvarial bone for GBR. The animals were divided into three groups: the No-IP (no intake of IP), Post-IP (IP orally, 10 mg/kg/day after GBR), and Pre-IP (IP intake beginning before GBR) groups. One cap was removed from each rabbit after 3 months, and the remaining site was a control. One month after one cap removal, all the animals were euthanized, and histologic and histomorphometric analyses were performed. In all of the groups, the newly generated tissue was of varying size, and it consisted of thin pieces of mineralized bone and large marrow spaces with fat cells and some hematopoietic cells. In all of the control sites, the newly generated tissue was noted and almost filled the space under the cap. There was a significant difference between groups No-IP and Pre-IP (93.8+/-4.6% vs. 98.5+/-0.8%, P<0.05). The tissue generated at the test sites in all of the groups was resorbed, and its original shape and volume were not maintained 1 month after one cap removal. In particular, the greatest percentage, approximately 20% of the newly generated tissue, was resorbed in the No-IP group (93.8+/-4.6% vs. 73.9+/-3.7%, P<0.05), and approximately 11% and 15% in groups Post-IP and Pre-IP, respectively. The relative amount of mineralized bone generated at the control and test sites was significantly larger in groups Post-IP and Pre-IP when compared with group No-IP, except for the test site between groups No-IP and Post-IP (P<0.05). Therefore, the amount of mineralized tissue generated appeared to increase with an increase in the total IP dose. Within the limitations of this rabbit experimental model, we conclude that the daily intake of IP before or after GBR inhibits the resorption of augmented tissue and would be useful for improving the quality of newly generated bone beyond the skeletal envelope.

Adipose Tissue↗
PubMed1995

Localized ridge augmentation using guided bone regeneration. II. Surgical procedure in the mandible.

The principle of guided bone regeneration can be applied for localized ridge augmentation in a staged approach. The surgical procedure for the mandible is presented through three case reports. Incision technique and flap design, utilization of autogenous bone grafts as a membrane-supporting device and osteoconductive scaffold, proper placement of barrier membranes and their stabilization with miniscrews, and wound closure are all emphasized. Furthermore, factors essential for achieving predictable results with barrier membranes for localized ridge augmentation and the benefits of combining barrier membranes with autogenous bone grafts are discussed.

Alveolar Ridge Augmentation↗
PubMed2006

Bioabsorbable scaffold for in situ bone regeneration.

A non-porous poly-DL-lactide tubular chamber filled by demineralised bone matrix (DBM) and bone marrow stromal cells (BMSC) in combination, was evaluated as a scaffold for guided bone regeneration (GBR) in an experimental model using the rabbit radius. The tubular chamber had an internal diameter of 4.7 mm, a wall thickness of 0.4 mm and a length of 18 mm. Autologous BMSC were obtained, under general anaesthesia from rabbit iliac crest and isolated by centrifugation technique. Allogenic DBM was obtained from cortico-cancellous bone of rabbits. In general anaesthesia, a 10-mm defect was bilaterally created in the radii of 10 rabbits. On the right side (experimental side) the defect was bridged with the chamber filled with both BMSC and DBM. On the left side (control side) the defect was treated by positioning DBM and BMSC between the two stumps. At an experimental time of 4 months histology and histomorphometry demonstrated that the presence of a tubular chamber significantly improved bone regrowth in the defect The mean thickness of newly-formed bone inside the chamber was about 56.7+/-3.74% of the normal radial cortex, in comparison with 46.7+/-10.7% when DBM and BMSC without the chamber were placed in the defect, P<0.05). These results confirmed the effectiveness of the chamber as a container for factors promoting bone regeneration.

Absorbable Implants↗
PubMed1998

The bone growing chamber: a new model to investigate spontaneous and guided bone regeneration of artificial defects in the human jawbone.

Spontaneous bone repair and regeneration of jawbone defects have been insufficiently studied in the dental literature. The present study analyzes a new human model designed to evaluate the basis for spontaneous bone regeneration in human jawbones. Hollow titanium cylinders, termed "bone growing chambers," were prepared with commercially pure titanium. Ten volunteers undergoing routine implant surgery were enlisted. A properly calibrated drill was used to prepare the bone-growing-chamber bed. The bone growing chamber was inserted inside the bone defect, and care was taken to submerge the cylinder at the level of the bone crest. After an adequate healing period, the bone growing chambers were retrieved with a small quantity of peripheral bone using a calibrated trephine bur. The retrieved specimens were processed to obtain thin undecalcified ground sections. The stable bone growing chambers showed bone tissue inside the growing space. The maturity of the regenerated bone was related to the time of removal. The bone growing chamber provides a well-defined space that is easy to prepare and to retrieve; its dimensions are always identical and it allows quantitative measurements of bone regeneration inside the chamber space.

Alveolar Process↗
PubMed2005

Biological efficacy of silk fibroin nanofiber membranes for guided bone regeneration.

The favorable biological properties of silk fibroin (SF) nanofiber membrane make it a good candidate for clinical applications as a device in bone and periodontal regenerative therapy. The purpose of this study is to evaluate the biocompatibility of the SF nanofiber membrane, and to examine its effect on bone regeneration in a rabbit calvarial model. To examine the biocompatibility of the electrospun SF membrane, we investigated cell proliferation, morphology, and differentiation. The bone regenerative efficacy of the membrane was evaluated in the calvarial defect of rabbits. The cell numbers and osteocalcin production labels were significantly increased in accordance with culture period. Cells had a stellate shape and broad cytoplasmic extensions on the membrane. The cells showed activity of ALPase that was comparable to culture dishes, and were calcified similarly to culture dishes. In in vivo tests, a complete bony union across the defects was observed after 8 weeks. At 12 weeks, the defect had completely healed with new bone. In conclusion, the SF nanofiber membrane was shown to possess good biocompatibility with enhanced bone regeneration and no evidence of any inflammatory reaction. These results strongly suggest that the SF membrane should be useful as a tool for guided bone regeneration.

3T3 Cells↗
PubMed1998

Growth factors and bone regeneration. Implications of barrier membranes.

Insufficient or absence of bone healing is a frequent problem within all surgical fields. This often necessitates treatment by autogenous bone grafting. Recently, two new techniques to promote bone healing were introduced, the osteopromotive membrane technique, and local delivery of growth-stimulatory factors, both with a high rate of success in preclinical experiments. The aims of the present series of investigations were to further develop the membrane barrier technique, both by itself as well as in combination with local delivery of growth factors, in animal experiments. During membrane-promoted bone formation, the membrane porosity was found to be of importance for the initial rate of bone formation as well as for the performance of the material in the tissue. In contrast, the final amount of bone was not affected. In a well-known bone healing model, the 5 mm in diameter 'critical size defect' at the rat mandibular ramus, the efficacy of rhBMP-2, rhTGF-beta 1 and rhFGF-2 to promote bone regeneration alone and in combination with barrier membranes was evaluated. Under both conditions, rhBMP-2 was found to be an efficient promoter of bone healing. rhFGF-2 had some stimulatory effect both with and without barrier membranes, whereas rhTGF-beta 1 was found to have a minor stimulatory effect by itself, but in combination with barrier membranes it was inhibitory. These observations were interpreted as being the result of an effect of the growth factors at different levels of the osteoblastic lineage; rhBMP-2 being an inducer of osteoblastic cells from stem cells, whereas rhTGF-beta 1 may primarily act on already committed cells. In contrast, rhFGF-2 may have stimulatory effect at different levels of the lineage. Based on the positive results obtained by the combination of rhBMP-2 and barrier membranes in the rat mandible, this combination was then applied to (i) rabbit radius defects; and (ii) a rat calvarial osteoneogenesis model. In the long bone model, membranes by themselves were insufficient to promote bone healing, but the combination resulted in complete regeneration. In the osteoneogenesis model, the combination of barrier membranes and rhBMP-2 resulted in a 100% increase in the final amount of achievable bone. In the last study, rhFGF-2 (no barrier membranes) was shown to enhance revitalization of autoclaved autogenous bone grafts, a procedure clinically used in craniofacial reconstruction mainly after tumor surgery. The combined use of rhBMP-2 and barrier membranes has great potential to be a useful treatment for improving bone healing and might be an alternative to bone grafting.

Administration, Topical↗
PubMed1985

Short- and long-term effects of irradiation on bone regeneration.

The aim of the present study is to quantify bone-regenerative capacity directly and 1 year after administration of 15 Gy 60Co irradiation. A titanium implant, the bone growth chamber, which in nonirradiated cases becomes filled with newly formed bone over a 4-week period, was inserted into each tibial metaphysis of 20 rabbits. In 10 animals the chambers were installed directly after irradiation, while in 10 other rabbits the implants were installed 1 year after the 60Co trauma. In both groups the bone-forming capacity on the irradiated side was compared to that of the contralateral, nonirradiated, control tibia. The amount of bone formed was determined by microradiography and microdensitometry. It was found that bone regeneration was depressed by 70.9 percent within a 4-week period after irradiation. At a follow-up of 1 year, the average depression of bone-forming capacity was only 28.9 percent. This means a recovery by a factor of almost 2.5. The clinical implications of these findings are discussed.

Animals↗
PubMed2006

Evaluation of an in situ formed synthetic hydrogel as a biodegradable membrane for guided bone regeneration.

The aim of the present study was to test whether or not the application of an in situ formed synthetic hydrogel made of polyethylene glycol (PEG) used as a biodegradable membrane for guided bone regeneration will result in the same amount of bone regeneration as with the use of an expanded polytetrafluoro-ethylene (ePTFE) membrane. In eight New Zealand White rabbits, four evenly distributed 6 mm diameter defects were drilled into the calvarial bone. Three treatment modalities were evenly distributed among the 32 defects: hydroxyapatite (HA)/tricalciumphosphate (TCP) granules covered at the outer and inner surface with a PEG membrane (test), HA/TCP granules covered at the outer and inner surface with an ePTFE membrane (positive control) and HA/TCP granules alone without membranes (negative control). After 4 weeks, the animals were sacrificed and the calvarial bones were removed. The area fraction of newly formed bone was determined by histomorphometrical analysis of the vertical sections from the middle of the defect and by micro-computed tomography of the entire defect. Multiple regression analysis (SAS GLM) was used to model the amount of new bone formation. The quantitative histomorphometric analysis clearly revealed higher values of newly formed bone for the two membrane groups compared with the negative control group. The average area fractions of newly formed bone measured within the former defect amounted to 20.3+/-9.5% for the PEG membrane, 18.9+/-9.9% for the ePTFE membrane, and 7.3+/-5.3% for the sites with no membrane. The micro-computed tomography also showed higher values of new bone formation for the PEG and for the ePTFE groups compared with the negative control group. The GLM revealed a highly significant effect of the treatment on the amount of bone formation (P=0.0048). The values for the negative control group were significantly lower than the ones found in the PEG membrane group (P=0.0017), whereas the ePTFE membrane group showed no significant difference from the PEG membrane group. It is concluded that the PEG membrane can be used successfully as a biodegradable barrier membrane in the treatment of non-critical-size defects in the rabbit skull, and leads to similar amounts of bone regeneration as an ePTFE membrane.

Absorbable Implants↗
PubMed1991

[Risk of fracture of regenerated bone and metal implant following bone lengthening osteotomy].

Many problems may arise, jeopardize and spoil the outcome of lengthening osteotomies. Fracture of the plate and the newly formed bone bridging the gap between the fragments is only one of the possible complications. Overload is its cause in all the cases. Following the theory of Pauwels the paper analyses the high stresses in hollow bone and the relationship to their principles of construction. Accurate knowledge of bone stressing and correct assessment of bone strength in the X-rays may help to reduce the incidence of fatigue fractures after limb lengthening.

Bone Lengthening↗
PubMed2004

Investigation of microstructural features in regenerating bone using micro computed tomography.

We illustrate some of the uses of micro-computed tomography (micro-CT) to study tissue-engineered bone using a micro-CT facility for imaging and visualizing biomaterials in three dimensions (3-D). The micro-CT is capable of acquiring 3D X-ray CT images made up of 2000(3) voxels on specimens up to 5 cm in extent with resolutions down to 2 microm. This allows the 3-D structure of tissue-engineered materials to be imaged across orders of magnitude in resolution. This capability is used to examine an explanted, tissue-engineered bone material based on a polycaprolactone scaffold and autologous bone marrow cells. Imaging of the tissue-engineered bone at a scale of 1 cm and resolutions of 10 microm allows one to visualize the complex ingrowth of bone into the polymer scaffold. From a theoretical viewpoint the voxel data may also be used to calculate expected mechanical properties of the tissue-engineered implant. These observations illustrate the benefits of tomography over traditional techniques for the characterization of bone morphology and interconnectivity. As the method is nondestructive it can perform a complimentary role to current histomorphometric techniques.

Animals↗
PubMed2003

Allogeneic mesenchymal stem cells regenerate bone in a critical-sized canine segmental defect.

BACKGROUND: Mesenchymal stem cells from adult bone marrow are multipotent cells capable of forming bone, cartilage, and other connective tissues. In a previous study, we demonstrated that autologous mesenchymal stem cells could repair a critical-sized bone defect in the dog. The objective of this study was to determine whether the use of allogeneic mesenchymal stem cells could heal a critical-sized bone defect in the femoral diaphysis in dogs without the use of immunosuppressive therapy. METHODS: A critical-sized segmental bone defect, 21 mm in length, was created in the mid-portion of the femoral diaphysis of twelve adult dogs that weighed between 22 and 25 kg. Each defect was treated with allogeneic mesenchymal stem cells loaded onto a hollow ceramic cylinder consisting of hydroxyapatite-tricalcium phosphate. A complete mismatch between donor stem cells and recipient dogs was identified by dog leukocyte antigen typing prior to implantation. The healing response was evaluated histologically and radiographically at four, eight, and sixteen weeks after implantation. The radiographic and histological results at sixteen weeks were compared with the historical data for the control defects, which included defects that had been treated with a cylinder loaded with autologous mesenchymal stem cells, defects treated with a cylinder without mesenchymal stem cells, and defects that had been left untreated (empty). The systemic immune response was evaluated by the analysis of recipient serum for production of antibodies against allogeneic cells. RESULTS: For defects treated with allogeneic mesenchymal stem cell implants, no adverse host response could be detected at any time-point. Histologically, no lymphocytic infiltration occurred and no antibodies against allogeneic cells were detected. Histologically, by eight weeks, a callus spanned the length of the defect, and lamellar bone filled the pores of the implant at the host bone-implant interface. Fluorescently labeled allogeneic cells were also detected. At sixteen weeks, new bone had formed throughout the implant. These results were consistent with those seen in implants loaded with autologous cells. Implants loaded with allogeneic or autologous stem cells had significantly greater amounts of bone within the available pore space than did cell-free implants at sixteen weeks (p < 0.05). CONCLUSIONS: The results of this study demonstrated that allogeneic mesenchymal stem cells loaded on hydroxyapatite-tricalcium phosphate implants enhanced the repair of a critical-sized segmental defect in the canine femur without the use of immunosuppressive therapy. No adverse immune response was detected in this model.

Animals↗
PubMed2001

[Ultrastructural and Ca/P changes of the regenerated bone after mandibular distraction in goats].

OBJECTIVE: The aim of this study was to examine the ultra structural and the changes of Ca/P elements of the new bone resulting from mandibular distraction osteogenesis. METHODS: Bilateral mandibular osteotomies were performed in six goats. The mandibles in all animals were lengthened 10 mm using a custom made distracter. Each 2 goats were sacrificed respectively at 2, 4 and 8 weeks after completion of distraction. The distracted callus were harvested and processed for SEM evaluation and Ca/P ratio analysis. RESULTS: The newly generated bone in distraction gap was confirmed by SEM examination, and the contents of Ca, P were found to be elevated with the maturity of distracted callus. CONCLUSION: This study suggested that newly formed bone by mandibular distraction osteogenesis in goats could be remodeled to normal bone.

Animals↗
PubMed2006

[Expression of transforming growth factor beta1 and insulin-like growth factor I in the regenerated bones after low frequency micro movement].

OBJECTIVE: To investigate the expression of transforming growth factor beta1 (TGF-beta1) and insulin-like growth factor I (IGF-I) in new bone after low frequency micro movement. METHODS: Fifteen female sheep from Shandong province were involved in the study and their bilateral tibias transversely osteotomized in the middle shafts with a defect of 2 mm. The hind limbs were fixed with unilateral external fixators connected to a controlled micro movement device. Ten days after osteotomy, one hind limb of each sheep randomly was selected to perform micro movement at an amplitude of 0. 25 mm and a frequency of 1 Hertz, 30 min a day for 4 weeks (micro movement group). The other hind limb served as the control group. Five sheep were sacrificed at 3, 4 and 6 weeks after osteotomy, respectively, and specimens were harvested for detecting the expression of TGF-beta1 and IGF-I by immunohistochemistry and RT-PCR. RESULTS: Immunohistochemistry: In the third postoperative week in the micro movement group, the expression of TGF-beta1 was detected in different areas of new chondrocytes at the margin of callus, mainly in proliferating area, and IGF-I expressed in osteoblasts at the margin of endochondral ossification area, calcified and mature chondrocytes and osteocytes. There was seldom expression of IGF-I and little expression of TGF-beta1 in the corresponding area in the control one. In the 4th postoperative week in the micro movement group, the expression of TGF-beta1 diminished gradually with the mature of new bone and be located in extracellular matrix and osteoblasts around ossified areas; The expression of IGF-I reached the peak and be located mainly in osteoblasts of new bone surface, maturing osteocytes and calcifying osteoid. But there was little expression of them in the control group. In the sixth postoperative week in the micro movement group, there was a little expression of IGF-I expression but little expression of TGF-beta1; there was nearly no expression of them in the control group. In the micro movement group, the absorbance values of TGF-beta1 at 3 and 4 weeks and of IGF-I at 3, 4 and 6 weeks were significantly higher than those in control group (P<0. 05). RT-PCR: In the third and fourth postoperative weeks in the micro movement group, there was higher expression of mRNA of TGF-beta1 and TGF-I than those in control group; in the sixth postoperative week, the expression diminished gradually, but was higher than that in control group. The absorbance values of TGF-beta1 at 3 and 4 weeks and IGF-I at 3, 4 and 6 weeks were significantly higher than those of control group (P<0. 05). CONCLUSION: Low frequency and controlled micro movement in the early stage of the fracture healing can promote the expression of TGF-beta1 and IGF-I. They worked together to regulate the process of the endochondral ossification, while in the late stage the differentiation of osteocytes and mineralization of osteoid were regulated mainly by IGF-I, which played an important role in regulating the cell biological behavior during micro movement.

Animals↗
PubMed1991

Neutral proteases in regenerating bone.

Proteolytic enzymes acting at physiologic pH (neutral proteases) are involved in both the formation and modeling of new bone and the remodeling of mature bone. In endochondral ossification systems such as growth-plate calcification, fracture healing, osteophyte formation, and demineralized bone matrix-induced osteogenesis, neutral proteases are predominantly involved in modifying proteins and proteoglycans in the extracellular matrix in preparation for calcification. These enzymes are of low molecular weight (below 30,000 Mr), are poorly charged, metal ion dependent, and appear to become active only after being released from chondrocytes. These neutral proteases may be distributed to the extracellular matrix in association with matrix vesicles that are derived from chondrocyte plasma membranes. A similar mechanism of calcification may also exist during malignant osteogenesis in an osteosarcoma; however, the cell producing the neutral protease in this lesion is the osteoblast and the matrix being synthesized is osteoid. In remodeling bone, osteoblasts secrete neutral collagenase (as an inactive enzyme) and produce not only additional proteases capable of activating the collagenase but also a collagenase inhibitor. Osteoblast collagenase or neutral protease may act to remove unmineralized osteoid from bone surfaces, thus facilitating its subsequent degradation by osteoclasts. The production of all these factors by osteoblasts appears to be regulated by calciotropic hormones (e.g., parathyroid hormone, 1,25-dihydroxyvitamin D, and calcitonin), possibly in a concerted fashion. Other possible functions of neutral proteases involve direct actions on cells or on specific molecules (growth factors) residing in the extracellular matrix.

Animals↗