Cameron M. L. Clokie

Closure of critical sized defects with allogenic and alloplastic bone substitutes.

PurposeThis study evaluates bone regeneration of critical sized cranial vault defects in New Zealand white rabbits using four commercially available bone substitutes: OsteoSet (calcium sulphate pellets), DynaGraft Putty (demineralized bone matrix delivered in a poloxmer excipient), Norian CRS, and Bone Source (two commercially available calcium phosphate cements). Materials and MethodsCritical sized defects 15 mm in diameter were created bilaterally in the parietal bones of 30 adult male New Zealand White rabbits. They were divided into three groups with ten animals in each. Bone healing was assessed clinically, radiographically, and histomorphometrically. Group 1 had calcium sulfate bioimplant on one side of the calvarium and an unfilled defect on the contralateral side. Group 2 had DBM putty on one side and Poloxamer gel on the contralateral side. Group 3, the Calcium phosphate cements (CPC), had Norian CRS on one side and Bone Source on the contralateral side. Five animals in each group were killed at 6 weeks and 12 weeks post operatively. ResultsAll unfilled defects healed with fibrous scar, as did the Plaster of Paris and the poloxamer gel defects. Defects reconstructed with the demineralized bone matrix putty healed with bone throughout the entire defect. This was obvious clinically and radiographically where the defects appeared completely filled with a dense radiopaque tissue. The six-week group displayed new bone formation (87.1%) surrounding the remaining allogeneic particles. Resorption was evidenced by the presence of osteoclastic activity and by the significant decrease in the size of the demineralized bone particles. By 12 weeks, the demineralized bone putty bioimplant was almost completely replaced by new bone (95.5%). Both calcium phosphate cement groups (Norian CRS and Bone Source) had identical patterns of healing. They clinically were visible and firm and uniformly radiopaque with little evidence of new bone formation. Histologically the cement remained unresorbed with little new bone with in the defect at 12 weeks. ConclusionsThe utilization of a demineralized bone matrix putty appeared to allow for complete closure of critical sized calvarial defects in New Zealand white rabbits with viable new bone at 12 weeks.

Successful mandibular reconstruction using a BMP bioimplant.

A bone morphogenetic protein bioimplant was used for primary reconstruction of a 6-cm mandibular discontinuity defect, after a segmental resection of an ameloblastoma. Radiographic evidence of new bone induction was seen at 3 and 9 months, postoperatively. A biopsy was taken at 9 months demonstrated viable new bone formation at the bioimplant site. This is the first reported case using a bone morphogenetic protein bioimplant in a human, followed by histological confirmation of new bone.

Differences between the sexes in post-surgical pain.

Abstract It has been shown that women have a lower pain threshold and lower tolerance to some forms of experimental pain then men. However, the evidence that clinical pain is perceived differently by the two sexes is not yet as strong. The placement of intraoral implants is a highly controlled surgical procedure that we have used to investigate this possibility. Forty‐eight edentulous (without teeth) subjects (27 females), aged from 35 to 63 years, received two titanium implants in the anterior mandible under local anesthesia. After the surgery, subjects completed a pain diary three times each day, rating pain intensity and unpleasantness on 100 mm visual analog scales (VAS). Once a day, they chose verbal descriptors from the McGill Pain Questionnaire (MPQ). Age of subjects, duration of surgery, the amount of local anesthetic used and the amount of pain medication taken were not statistically different for the two groups (P≥0.32). Results showed that the senior surgeon produced significantly less pain than a 4th year resident (P=0.04). Although there were no significant differences between sexes for mean daily ratings of intensity or unpleasantness over time (P≥0.10), most women experienced the highest intensity of pain during the day, while most men had higher pain in the evening (P=0.025). Also, the relative unpleasantness (unpleasantness/intensity ratio) increased significantly with time for males, but not for females (P=0.016). Males and females did not differ in the total number of words chosen from the MPQ (P=0.61), or in the averaged Pain Rating Index (PRI) (P=0.53). However, women used significantly more evaluative words than men (P=0.04), suggesting that woman found the overall intensity greater. These results indicate that women find post‐surgical pain more intense than males, but that men are more disturbed than women by low levels of pain that last several days.

Closure of rabbit calvarial critical-sized defects using protective composite allogeneic and alloplastic bone substitutes.

This study evaluated the repair of critical-sized cranial vault defects in thirty New Zealand white rabbits using various allogeneic and alloplastic bone substitutes designed to provide mechanical protection to the brain as well as osteoinductivity. The strategies employed included demineralized bone matrix (DBM), a putty used in combination with a rigid resorbable plating system as a protective covering and calcium phosphate cement (CPC) combined with native partially purified bone morphogenetic protein (BMP). Bilateral critical-sized defects measuring 15 mm in diameter were created in the parietal bones of 30 adult male New Zealand white rabbits. They were divided into three groups with ten animals in each. Group 1 had one defect left unfilled as a control while autogenous bone was placed in the defect on the other side. In Group 2 a rigid resorbable copolymer membrane, Lactosorb (Lorenz Surgical, Jacksonville, Florida), was placed over both defects to cover them and protect the underlying tissues. The pericranial aspect of one defect was left unfilled while the other defect was filled with DBM putty. Group 3 had a CPC, Mimix (Lorenz Surgical, Jacksonville, Florida), placed into one of the defects while the defect on the other side was filled with the same CPC in combination with BMP in a concentration of 25 mg/mL. Bone healing was assessed clinically, radiographically, and histomorphometrically. All unfilled controlled defects, the defects covered with the resorbable Lactosorb membrane and those filled with calcium phosphate cement alone, healed with a fibrous scar. Defects reconstructed with DBM putty in combination with the resorbable Lactosorb membrane and calcium phosphate in combination with BMP healed with bone bridging the entire defect. This was obvious radiographically where the defects appeared completely filled with a dense radiopaque tissue. Histological analysis demonstrated that specimens where DBM putty was used in combination with the resorbable Lactosorb membrane had 67.7% new bone fill at 6 weeks and 84.0% at 12 weeks. Resorption of DBM particles was evidenced by the presence of osteoclastic activity and by the significant decrease in the size of the demineralized bone particles. In the calcium phosphate groups where BMP was added to the bioimplant there was 45.8% new bone formation at 12 weeks. The utilization of a composite consisting of DBM with resorbable Lactosorb membrane or a composite of calcium phosphate cement composite with BMP promoted complete closure of critical-sized calvarial defects in New Zealand white rabbits with viable new bone at 12 weeks. The complete bone bridging observed with these composites suggests that they could be used to enhance the protection of intracranial contents following craniofacial surgical procedures.

In search of the ideal bone morphogenetic protein delivery system: in vitro studies on demineralized bone matrix, purified, and recombinant bone morphogenetic protein.

The clinical use of recombinant bone morphogenetic protein (rBMP) is limited by the lack of a suitable delivery system. The bone morphogenetic protein (BMP) delivery system provided by nature is highly effective, and by studying purified BMP (BMP/NCP) and demineralized bone matrix (DBM), it may be possible to learn how to emulate natures success. The current study used an in vitro muscle cell model to study the activity of BMP/NCP and DBM and the effects of extracellular matrix on BMP activity. C2C12 cells transiently exposed to recombinant human BMP-4 (rhBMP-4) rapidly increased their alkaline phosphatase (AP) activity to day 5, after which it steadily declined. Cells exposed to BMP/NCP or DBM continued to increase their AP activity over the 14-day culture. If BMP/NCP was treated to remove a 22-kd protein, it became water-soluble and exhibited a similar activity pattern to rhBMP-4. Cells cultured on collagen type I, fibronectin, and hyaluronic-coated surfaces demonstrated increased AP activity when exposed to rhBMP-4 or BMP/NCP compared with cells cultured on bovine serum albumin or poly-l-lysine. These results suggest that the natural BMP delivery system operates both by binding to the BMP molecule and slowly releasing it into the extracellular milieu and by interacting with the responding cells through cell-matrix receptors to enhance the cellular response to BMP.

A Within-subject Comparison of Mandibular Long-bar and Hybrid Implant-supported Prostheses: Psychometric Evaluation and Patient Preference

Although it has been shown that patients are more satisfied with prostheses supported by implants than with conventional dentures, there have been few direct comparisons of the various designs of implant-supported prostheses. This within-subject crossover clinical trial was designed to compare two forms of removable prostheses which are frequently prescribed for the edentulous mandible: a long-bar overdenture supported by 4 implants and a two-implant hybrid overdenture. Sixteen completely edentulous subjects were given a new maxillary conventional denture: Ten of them received the mandibular long-bar prosthesis first and six the hybrid. After a two-month adaptation period, psychometric measures of various aspects of the prostheses and physiological tests of masticatory efficiency were carried out over three weeks. The mandibular prostheses were then changed and the procedures repeated. At the end of the study, subjects were asked to choose the mandibular prosthesis that they wished to keep, and final psychometric measures were taken. In this paper, the results of the psychometric assessment and patient preference are presented. Subjects assessed factors such as general satisfaction, quality of life, stability, retention, comfort, esthetics, ease of cleaning, speaking, and chewing, and how well-chewed foods were before being swallowed. Most of the factors except ease of cleaning and speaking were rated significantly better with long-bar overdentures than with hybrid ones. These results are consistent with the fact that all subjects chose long-bar overdentures, reporting stability, ease of chewing, and comfort as the most important factors influencing their choice. These results suggest that, although subjects assign high ratings for most factors to hybrid overdentures, they find long-bar overdentures to be significantly more stable, comfortable, and easier for chewing.

A Within-subject Comparison of Mandibular Long-bar and Hybrid Implant-supported Prostheses: Evaluation of Masticatory Function

Sixteen edentulous subjects participated in a within-subject crossover clinical trial to test the hypotheses that a long-bar overdenture attached to 4 implants gives greater patient satisfaction and masticatory efficiency than a two-implant hybrid overdenture. All subjects were given a new maxillary conventional denture. Ten received mandibular long-bar overdentures first and six the hybrid overdentures. Two months later, psychometric assessments and functional tests were repeated 3 times at one-week intervals. The mandibular prosthesis was then changed, and recordings were repeated after another 2 months. Mandibular movements and electromyographic activity of jaw muscles were recorded while subjects chewed standard-sized pieces of 5 foods: bread, cheese, apple, sausage, and carrot. Measurements included masticatory time, cleaning time (the time between the end of mastication and the last swallow), and duration and amplitude of masticatory cycles and phases. Multilevel analyses were performed. No significant differences in masticatory time were found between prostheses for any test food. However, cleaning time for carrot [estimated mean of difference (A) ± SE: 1.6 sec ± 0.7] and bread (A = 1.0 sec ± 0.4) was slightly but significantly longer for subjects wearing long-bar overdentures. Cycle duration was longer with the long-bar overdenture only for subjects chewing carrot. The opening phase was shorter and the closing phase longer with the long-bar overdenture for almost all test foods. Vertical amplitude was significantly less with the long-bar overdenture for cheese (A = -2.6 mm ± 1.1), apple (A = -2.6 mm ± 1.0), and sausage (A = -2.9 mm ± 1.3). These results suggest that mastication with the 2 prostheses is equally efficient, although clearance of some foods from the mouth is longer with the long-bar overdentures. They also indicate that patients adapt their masticatory movements to the characteristics of different prostheses.

Role of bovine bone morphogenetic proteins in bone matrix protein and osteoblast-related gene expression during rat bone marrow stromal cell differentiation.

Bone morphogenetic proteins (BMPs) are known to promote osteogenesis, and clinical trials are currently underway evaluating the ability of certain BMPs to promote bone graft and fracture healing. To observe the mechanism of osteoinductive and bone formation, 100 ug of bovine BMP was tested during osteogenic differentiation of rat bone marrow stromal cells (MSCs) and C2C12 line culture for 14 and 28 days. We examined alkaline phosphatase (ALP) by assay, immunohistochemical studies for bone matrix proteins, and mRNA expression of bone matrix proteins and osteoblast-related analysis by reverse-transcription polymerase chain reaction. ALP activity in MSC cultures was elevated by bovine BMP by two to fivefold (P < 0.05-0.001). DNA and protein content increased over 14 days. BMP significantly increased the mRNA expression of type I collagen, ALP, osterix, osteocalcin, osteopontin, vascular endothelial growth factor (VEGF), platelet-derived growth factor (PDGF)-A, and parathyroid hormone receptor time dependently during the osteoblastic differentiation. There was no markedly enhanced mRNA expression of bone sialoprotein (BSP) and glyceraldehyde-3-phosphate dehydrogenase compared with that of control. Immunohistochemical results also showed BMP increased immunoreactive positivity of type I collagen, osteocalcin, osteonectin, osteopontin, and BSP during the C2C12 differentiation. These data indicated that BMP enhances our ability to stimulate the differentiation of osteoblast-like cells and increases osteoinductivity, bone matrix protein formation and mineralization, angiogenesis, and chondrogenesis during osteoblast progenitor cell differentiation in vitro and that the role of chondrogenic is weak.

Bisphosphonate-associated Osteonecrosis of the Jaw in Ontario: A Survey of Oral and Maxillofacial Surgeons

Objective. Osteonecrosis of the jaw (ONJ) in association with use of bisphosphonate (BP) has been described primarily in cancer patients receiving high-dose intravenous BP. The frequency of the condition in patients with osteoporosis appears to be low. We evaluated the frequency of BP-associated ONJ in Ontario in the cancer population and in those receiving BP for osteoporosis and metabolic bone disease. Methods. A survey developed by representatives of the Ontario Society of Oral and Maxillofacial Surgeons was mailed to Ontario oral and maxillofacial surgeons (OMFS) in December 2006, asking oral surgeons to provide information on cases of ONJ seen in the previous 3 calendar years (2004 to 2006). OMFS were subsequently contacted by telephone if they had not responded or if they had reported cases of ONJ. The frequency of ONJ in association with BP use was estimated from the number of patients with filled prescriptions for BP in Ontario between 2004 and 2006. The cumulative incidence of ONJ was calculated separately for patients using intravenous (IV) BP for cancer treatment and for patients using oral or IV BP for osteoporosis or other metabolic bone disease. Results. Between 2004 and 2006, 32 ONJ cases were identified. Nineteen patients received IV BP for cancer treatment and 13 patients received oral or IV BP for osteoporosis or metabolic bone disease. Over a 3-year period the cumulative incidence of BP-associated ONJ was 0.442% of cancer patient observations (442 per 100,000) and 0.001% of osteoporosis or other metabolic bone disease observations (1.04 per 100,000). The relative risk of low dose IV/oral BP-associated ONJ was 0.002 (95% CI 0.001, 0.005) compared to high-dose IV BP. Other risk factors for ONJ were present in all cases in whom detailed assessment was available. The median duration of exposure to BP was 42 months (range 36 to 120 mo) and 42 months (range 11 to 79 mo) in osteoporosis patients and cancer patients, respectively. Conclusion. Over a 3-year period, the cumulative incidence for BP-associated ONJ was 0.442% of cancer patient observations (442 per 100,000) and 0.001% of osteoporosis or metabolic bone disease observations (1.04 per 100,000). This study provides an approximate frequency of BP-associated ONJ in Canada. These data need to be quantified prospectively with accurate assessment of coexisting risk factors.

Bone morphogenetic protein excipients : Comparative observations on Poloxamer

Clinicians await the availability of synthetic bioimplants that will replace the need for autogeneic bone grafts in bone reconstructive surgery. For more than a decade, researchers have evaluated delivery vehicles for the tissue morphogen bone morphogenetic protein. The object of this investigation was to measure induced bone development when bone morphogenetic protein was delivered by human tendon collagen, human demineralized bone matrix, hydroxyapatite, a composite of human tendon collagen and human demineralized bone matrix (tendon collagen + demineralized bone matrix), Poloxamer 407, and a composite of human demineralized bone matrix and Poloxamer 407. Sixty-three adult male Swiss Webster mice (Harlan Sprague-Dawley, Indianapolis, Ind.) received 126 implants. The animals were divided into seven groups of nine animals, depending on carrier (six carriers plus the positive control group) used. Each animal received a bone morphogenetic protein-enhanced carrier in one hindquarter muscle mass, with the contralateral leg being implanted with the carrier alone. Implants were evaluated by quantitative radiomorphometry validated by histologic methods. Radiographically, no significant differences were identified among any of the implants evaluated (p > 0.05). Histomorphometric analysis demonstrated that Poloxamer 407 was significantly (p < 0.05) better at delivering bone morphogenetic protein than the other carriers involved in this investigation. The new bone developed in a tubular or spherical shape. Interaction of endogenous and exogenous delivery systems seems to be essential for optimal transmission of bone morphogenetic protein. The importance of the excipient to deliver bone morphogenetic protein and develop a bone morphogenetic protein concentration gradient has been emphasized by other investigators and confirmed by our research on poloxamer. With further research on the physicochemical mechanisms of localization and transmission of bone morphogenetic protein, it may be possible to avoid hazardous operations with autogeneic bone.