Locally delivered rhBMP-2 enhances bone ingrowth and gap healing in a canine model.

The purpose of the present study was to determine if recombinant human bone morphogenetic protein-2 (rhBMP-2) enhances bone ingrowth into porous-coated implants and gap healing around the implants. In the presence of a 3-mm gap between the implant and host bone, porous-coated implants were placed bilaterally for four weeks in the proximal humeri of skeletally mature, adult male dogs. In three treatment groups, the test implant was treated with HA/TCP and rhBMP-2 in buffer at a dose of 100 microg/implant (n=5), 400 microg/implant (n=6), or 800 microg/implant (n=5) and placed in the left humerus. In these same animals, an internal control implant was treated only with HA/TCP and buffer and placed in the right humerus. These groups were compared with a previously reported external control group of seven animals in which no growth factor was delivered [J. Orthop. Res. 19 (2001) 85]. The BMP treated implants in the two lower dose groups had significantly more bone ingrowth than the external controls with the greatest effect in the 100 g/implant group (a 3.5-fold increase over the external control, p=0.008). All three dose groups had significantly more bone formation in the 3-mm gap surrounding the BMP treated implants than the external controls with the greatest effect in the 800 microg group (2.9-fold increase, p<0.001). Thus, application of rhBMP-2 to a porous-coated implant stimulated local bone ingrowth and gap healing. The enhancement of bone formation within the implant (bone ingrowth) was inversely related to dose.

Recombinant human bone morphogenetic protein-2 accelerates healing in a rabbit ulnar osteotomy model.

BACKGROUND: Approximately 5% to 20% of fractures have delayed or impaired healing. Therefore, it is desirable to develop new therapies to enhance fracture-healing that can be used in conjunction with traditional treatment methods. The purpose of this study was to evaluate the ability of a single application of recombinant human bone morphogenetic protein-2 to accelerate fracture-healing in a rabbit ulnar osteotomy that heals spontaneously. METHODS: Bilateral mid-ulnar osteotomies (approximately 0.5 to 1.0 mm wide) were created in seventy-two skeletally mature male rabbits. The limbs were assigned to one of three groups: those treated with an absorbable collagen sponge containing recombinant human bone morphogenetic protein-2, those treated with an absorbable collagen sponge containing buffer, and those left untreated. In the first two groups, an 8 20-mm strip of absorbable collagen sponge containing either 40 g of recombinant human bone morphogenetic protein-2 or buffer only was wrapped around the osteotomy site. The rabbits were killed at two, three, four, or six weeks after surgery. In addition, twenty-four age-matched rabbits were used to provide data on the properties of intact limbs. The retention of recombinant human bone morphogenetic protein-2 at the osteotomy site was determined with scintigraphic imaging of (125)I-labeled recombinant human bone morphogenetic protein-2. After the rabbits were killed, the limbs were scanned with peripheral quantitative computed tomography to assess the area and mineral content of the mineralized callus. The limbs were then tested to failure in torsion, and undecalcified specimens were evaluated histologically. RESULTS: Gamma scintigraphy of (125)I-recombinant human bone morphogenetic protein-2 showed that 73% +/- 6% (mean and standard deviation) of the administered dose was initially retained at the fracture site. Approximately 37% +/- 10% of the initial dose remained at the site one week after surgery, and 8% +/- 7% remained after two weeks. The mineralized callus area was similar in all groups at two weeks, but it was 20% to 60% greater in the ulnae treated with recombinant human bone morphogenetic protein-2 than in either the ulnae treated with buffer or the untreated ulnae at three, four, and six weeks (p < 0.05). Biomechanical properties were similar in all groups at two weeks, but they were at least 80% greater in the ulnae treated with recombinant human bone morphogenetic protein-2 at three and four weeks than in either the ulnae treated with buffer (p < 0.005) or the untreated ulnae (p < 0.01). By four weeks, the biomechanical properties of the ulnae treated with recombinant human bone morphogenetic protein-2 were equivalent to those of the intact ulnae, whereas the biomechanical properties of both the ulnae treated with buffer and the untreated ulnae had reached only approximately 45% of those of the intact ulnae. At six weeks, the biomechanical properties were similar in all groups and were equivalent to those of the intact ulnae. The callus geometry and biomechanical properties of the ulnae treated with buffer were equivalent to those of the untreated ulnae at all time-points. CONCLUSIONS AND CLINICAL RELEVANCE: These findings indicate that treatment with an absorbable collagen sponge containing recombinant human bone morphogenetic protein-2 enhances healing of a long-bone osteotomy that heals spontaneously. Specifically, osteotomies treated with recombinant human bone morphogenetic protein-2 healed 33% faster than osteotomies left untreated. The results of this study provide a rationale for testing the ability of recombinant human bone morphogenetic protein-2 to accelerate healing in patients with fractures requiring open surgical management.

Mandibular reconstruction using bone morphogenetic protein 2: long-term follow-up in a canine model.

OBJECTIVE: To determine the degree of bone resorption and stability of 3-cm, full-thickness canine mandibular defects reconstructed with recombinant human bone morphogenetic protein 2 (rhBMP-2) and a bioerodible particle carrier followed for 30 months after reconstruction. STUDY DESIGN: Nine dogs, divided into three groups, underwent reconstruction of surgically created 3-cm, full-thickness defects of the body of the mandible. METHODS: Mandibular reconstruction was performed via a combined intraoral and extraoral approach. Using standard plating techniques, a unilateral full-thickness, 3-cm defect was created in the body of the mandible. After stabilizing the defects with titanium reconstruction plates, test implants composed of rhBMP-2 and poly(lactide-co-glycolide) bioerodible particles were placed in the mandibular defects of six animals. Reconstruction plates were removed from test animals at 10 weeks. Three short-term test animals were sacrificed 3 months after reconstruction. Three long-term test animals were sacrificed 30 months after reconstruction to determine the degree of resorption and long-term stability of the rhBMP-2-induced bone. Control implants (carrier without rhBMP-2) were used in three animals and were sacrificed at 3 months. At 9 months, long-term animals were advanced to a solid diet. Masticatory function and body weight were monitored periodically to assess diet tolerance. Roentgenographic photodensitometry was performed on serial dental roentgenograms of the reconstructed segments to determine bone density and the degree of bone resorption over 30 months. After sacrifice, reconstructed segments were harvested and embedded in plastic for histological analysis and histomorphometry to determine the percentage of the defect replaced by mineralized bone (area density) and degree of resorption from 3 to 30 months after reconstruction. The main outcome measures were bone density and bone height determined from serial roentgenograms and percentage of the reconstructed segment replaced by mineralized bone (area density) determined from histomorphometry. RESULTS: Control animals without rhBMP-2 showed no evidence of bone formation across the defect. Histological examination revealed good bone formation in two of three of the short-term test animals with a mean area density of 41.0%. The long-term test animals treated with rhBMP-2 demonstrated good bone formation that was comparable to that of normal host bone by 3 months. The roentgenographic photodensity measurements stabilized at 5 months without evidence of persistent bone resorption. The height of the reconstructed segment (rhBMP-2-induced bone) initially decreased, then stabilized by 11 months after reconstruction with no indication of resorption or failure. Histological examination of the long-term test animals revealed good bone formation across the mandibular defects. However, there were localized areas of thinning of the cortical bone as compared with the short-term test animals sacrificed at 3 months. Histological examination verified the loss of height of the bone in the reconstructed segments. The area density (mean) of the long-term test animals was 56.5%. Despite the decrease in height of the induced bone, there was an increase in area density of the bone over time. CONCLUSIONS: This study demonstrated that rhBMP-2 in a bioerodible particle carrier induced new host bone formation across critical-size mandibular defects. The newly formed bone successfully integrated with existing host bone creating a stable union capable of withstanding the forces of masticatory function in a canine. There was some evidence of early bone resorption (thinning of the cortical bone and decrease in height) in the rhBMP-2-induced bone. The rhBMP-2-induced bone stabilized by 11 months after reconstruction and no further resorption was noted. The percentage of area of the defect replaced by rhBMP-2-induced bone (area density) increased over 30 months. (ABSTRACT TRUNCATED)

Periodontal repair in dogs: recombinant human bone morphogenetic protein-2 significantly enhances periodontal regeneration.

This study evaluated bone and cementum regeneration following periodontal reconstructive surgery using recombinant human bone morphogenetic protein-2 (rhBMP-2) in six beagle dogs. Surgically created mandibular supraalveolar premolar tooth defects in contralateral jaw quadrants were randomly assigned to receive rhBMP-2 or control vehicle. Clinical defect height was prepared to 5 mm. rhBMP-2 was applied with synthetic bioerodable particles and autologous blood using 20 micrograms rhBMP-2 per 100 microliters implant volume. Flaps were advanced to submerge the teeth and sutured. The dogs were sacrificed 8 weeks postsurgery. Histometric recordings included defect height, height and area of alveolar bone regeneration, height of cementum regeneration, root resorption, and ankylosis. Group means, standard deviations, and P values are shown (Student t test; n = 6). Histometric defect height for rhBMP-2 and control defects was 3.7 +/- 0.3 and 3.9 +/- 0.4 mm, respectively (P = 0.446). Height of alveolar bone regeneration amounted to 3.5 +/- 0.6 and 0.8 +/- 0.6 mm for rhBMP-2 and control defects, respectively (P = 0.000). Corresponding values for bone area were 8.4 +/- 4.5 and 0.4 +/- 0.5 mm2, respectively (P = 0.006). Cementum regeneration was observed in all experimental defects (17/17) and in 15 out of 17 controls, averaging 1.6 +/- 0.6 and 0.4 +/- 0.3 mm for rhBMP-2 and control defects, respectively (P = 0.005). Small amounts of root resorption were seen in rhBMP-2 defects, whereas controls exhibited substantial resorption (0.2 +/- 0.1 and 1.1 +/- 0.3 mm, respectively; P = 0.001).(ABSTRACT TRUNCATED AT 250 WORDS)

Radiographic analysis of regenerated bone around endosseous implants in the canine using recombinant human bone morphogenetic protein-2.

Ideal endosseous implant placement involves a congruent bony housing in close apposition to the implant surface. Clinical situations are encountered, however, in which the entire implant surface cannot be in close apposition to bone. In these instances, bone grafting materials are generally used to regenerate bone around the implant. In this study, a biologically active bone differentiation factor, recombinant human bone morphogenetic protein-2 (rhBMP-2), was used with two different carriers to regenerate bone around implants in standardized critical-sized defects in the canine mandible. Half of the sites had a nonresorbable membrane placed over the defect. Longitudinal standardized radiographs were obtained to assess the amount of bone regeneration on the mesial and distal of the implants after 4 and 12 weeks of healing. Ninety-six implants were placed in 12 fox-hounds. Bone fill was determined by linear measurement of bone on the radiographs, and changes in bone density were evaluated by computer-assisted densitometric image analysis of discrete areas adjacent to the implant. After 4 weeks of healing, nonmembrane sites had significantly greater bone height than membrane-protected sites. Following 12 weeks of healing, sites treated with rhBMP-2 had significantly greater bone formation than untreated sites. Sites treated with rhBMP-2 and a membrane had the greatest bone fill, followed by sites treated with rhBMP-2 but no membrane. Sites without rhBMP-2, whether with or without a membrane, had less bone fill than sites with rhBMP-2. At 12 weeks, sites with a membrane resulted in significantly more gain in bone density than sites without a membrane. Furthermore, sites treated with a collagen carrier resulted in greater gains in bone density than sites treated with a polylactide/glycolide carrier. The results from this study demonstrate by radiographic evidence that new bone formation in critical-sized defects around implants is dependent on time after defect treatment, the type of carrier used, the use of a barrier membrane, and the presence of rhBMP-2. In addition, these findings suggest that rhBMP-2, a bone differentiation factor, can significantly stimulate bone formation around endosseous dental implants.

Periodontal repair in dogs: evaluation of rhBMP-2 carriers.

This study evaluated candidate carriers for recombinant human bone morphogenetic protein-2 (rhBMP-2)-driven periodontal regeneration. Previous experiments indicated the ability of rhBMP-2 in a particulate delivery system to result in substantial regeneration of bone and periodontal regeneration. In the present study, canine demineralized bone matrix (DBM), bovine deorganified crystalline bone matrix (Bio-Oss), an absorbable collagen sponge (ACS) of type I bovine collagen, poly(D,L-lactide-co-glycolide) microparticles (PLGA), and polylactic acid granules (Drilac) were tested for their ability to support rhBMP-2 (0.2 mg/mL implant volume)-driven periodontal regeneration. The implants were tested in routine critical size canine supra-alveolar periodontal defects with transgingival tooth positioning. Contralateral defects in six beagle dogs were semirandomly assigned to receive: DBM/rhBMP-2, DBM (no rhBMP-2), Bio-Oss/rhBMP-2, ACS/rhBMP-2, PLGA/rhBMP-2, or Drilac/rhBMP-2. Animals were sacrificed 8 weeks postsurgery, and block sections of the defects were processed for light microscopy. Substantial bone regeneration was observed in all defects implanted with rhBMP-2. Other measures of periodontal healing, including cementum regeneration and presence of ankylosis, were more variable between the implants. DBM and Bio-Oss performed well as carriers for rhBMP-2-driven periodontal regeneration, although other impediments to their clinical use exist. This study indicates that qualities of the carrier system, including its space-maintaining capacity can affect the ability of rhBMP-2 to regenerate both alveolar bone and periodontal attachment.

Our own experience with CT angiography in early diagnosis of cerebral vascular malformations.

Computed tomography (CT) and intraarterial cerebral angiography are essential methods in early diagnosis of cerebral vascular malformations. In recent years however non-invasive or minimally invasive methods like MR angiography and CT angiography (CTA), which could potentially replace angiography, have been developed. The aim of presented study is to demonstrate our own experience in application of CTA in early diagnosis of cerebral vascular malformations. The material consists of 86 CTA examinations performed shortly after non-traumatic intracranial haemorrhage. Angiographic correlation has been available in 23 patients and surgical one--in 31 cases. CTA studies began with serio-CT to select the optimal time between contrast injection and CT scanning. After that 100-120 ml of non-ionic contrast medium was injected intravenously (5 ml per sec.) and spiral CT acqusition was performed with the delay calculated on the basis of the serio-CT. The obtained images were postprocessed on the workstation using always MIP and in many cases also SSD and VRT reconstructions. Vascular malformations have been diagnosed in 44 of 86 CTA studies including 38 patients with aneurysms (total number 51) and 6 patients with AVMs. In 17 cases the diameter of the aneurysm did not exceed 5 mm. In all surgical cases the CTA diagnosis of the aneurysm has been confirmed. There was however one false-negative case. On the other hand in 7 patients CTA revealed the small aneurysm, despite unclear angiographic appearance. In 2 of 6 patients with CTA suspicion of AVM this diagnosis has been excluded either by angiography or surgery. Comparison of CTA and angiography in 22 aneurysms showed in 17 cases superiority of CTA in evaluation of aneurysmal neck and the relationship between aneurysm and adjacent vessels (especially with VRT and SSD reconstructions). In patients with AVMs however the evaluation of supplying and draining vessels was better with angiography. On the base of our material we can conclude that CTA is very efficient in detecting and evaluating the aneurysms. We believe that CTA can replace angiography if it reveals aneurysm in a site corresponding with location of haemorrhage on CT. In patients with suspicion of AVM value of CTA is doubtful and angiography remains the method of choice.

[The effect of small doses of 7.5% NaCl on brain bulk during elective craniotomies]. / Wplyw malych dawek 7.5% NaCl na objetosc mózgu podczas planowych kraniotomii.

In 16 patients (ASA I i II) aged 16-76 years (48 +/- 15; mean +/- SD) operated on because of intracranial expanding mass, the effect of hypertonic saline (7.5%--1 ml/kg b.w.) on brain bulk (BB) was evaluated. Patients were anaesthetised with a slight hypocarbia (PaCO2 = 33.3 +/- 3.5 mmHg). BB was scored after opening the dura (T0) and 15 min. (T15) after hypertonic saline (HS) infusion. Five points BB scale was used. Brain bulk reduction (D BB) was calculated as a difference: BB15-BB0. Tomographic signs of intracranial expansion (TSIE) in preoperative CT were scored using the scale from 5 to 15 points assessing (1 to 3 points) the size of mass lesion, the size of perifocal oedema, midline shift, displacement of ventricles and basal cisterns compression. Systolic (SBP), diastolic (DBP) blood pressures and heart rate (HR) were monitored. Serum natrium (SNa), kalium (SK) and osmolarity (Sosm) were measured at T0, T15 and Tp0 (one hour after operation). Student's t-test, Wilcoxon test and Spearman correlation were used for statistical analysis. P < 0.05 was considered as statistically significant. HS caused significant decrease in BB (p = 0.002). In 12 patients with a solid brain tumor a negative correlation between BB and TSIE was found (r = -0.68). A slight but significant decreases in SBP and DBP at Tp0 and T15 as well as decrease in HR due to HS were stated. SNa and Sosm increased at T15 and reminded elevated at Tpo. We can conclude that 7.5% saline in a dose of 1 ml/kg b.w. reduces brain bulk during craniotomy in patients with supratentorial mass lesions. In patients with a solid brain tumor this effect correlates negatively with a size of expanding mass. A slight changes in blood pressure and heart rate due to HS as well as moderate decrease in SK are within limits of clinical acceptance.

Long-term survival of concurrent meniscus allograft transplantation and repair of the articular cartilage: a prospective two- to 12-year follow-up report.

We describe 119 meniscal allograft transplantations performed concurrently with articular cartilage repair in 115 patients with severe articular cartilage damage. In all, 53 (46.1%) of the patients were over the age of 50 at the time of surgery. The mean follow-up was for 5.8 years (2 months to 12.3 years), with 25 procedures (20.1%) failing at a mean of 4.6 years (2 months to 10.4 years). Of these, 18 progressed to knee replacement at a mean of 5.1 years (1.3 to 10.4). The Kaplan-Meier estimated mean survival time for the whole series was 9.9 years (sd 0.4). Cox's proportional hazards model was used to assess the effect of covariates on survival, with age at the time of surgery (p = 0.026) and number of previous operations (p = 0.006) found to be significant. The survival of the transplant was not affected by gender, the severity of cartilage damage, axial alignment, the degree of narrowing of the joint space or medial versus lateral allograft transplantation. Patients experienced significant improvements at all periods of follow-up in subjective outcome measures of pain, activity and function (all p-values < 0.05), with the exception of the seven-year Tegner index score (p = 0.076).

[Use of multiplanar reconstruction of CT images (MPR) in diagnosis of low back pain syndromes]. / Zastosowanie weiloplaszczyznowych przeksztalcen obrazu KT (MPR) w diagnostyce zaspolów bólowych ledzwiowo-krzyzowych.

The authors describe program of multiplanar reconstruction of CT images (MPR) and discuss its usefulness in diagnosing low back pain syndromes. The program was especially useful in differentiation between herniated disc and osteophytes, in spondylolisthesis and in the cases of asymmetric spine. MPR enables also imaging of pathological spinal lesions in the operative view which is helpful for planning of the surgery. In the authors' opinion CT with MPR is not inferior and in some authors aspects even superior to MR imaging.

BMP-2 for intramuscular bone induction: effect in squirrel monkeys is dependent on implantation site.

Demineralized bone matrix (DBM) reproducibly induces extraskeletal bone formation in rodents, but its effects in dogs and primates are negative or uncertain. In previous studies on the squirrel monkey, DBM did not induce bone, although the same implants were effective in rats. DBM implants augmented with recombinant human bone morphogenetic protein-2 (rhBMP-2) induced intramuscular bone formation in squirrel monkeys. However, the amount of induced bone was often minimal and sometimes absent. One explanation of this weak and unpredictable effect could be reactions to cellular components of the allogeneic DBM that was used as carrier. Therefore, we now repeated the experiment, using bovine type 1 collagen as carrier. 48 collagen discs (10 mm diameter), containing 0, 10, 40 or 200 microgram rhBMP-2, were implanted in 6 monkeys and 6 rats. The BMP-2 implants induced dose-dependent amounts of intramuscular bone in rats whereas, in squirrel monkeys, macroscopic bone induction occurred in less than half of the BMP-2 implants. There was no dose-dependency. Whether bone was formed or not was significantly influenced by individual variation among the monkeys, and by implant location within the muscle. Implants close to the muscle aponeurosis more often induced bone than did purely intramuscular ones. In this small series, we could not demonstrate a significant effect of BMP-2, as compared to control implants. Presumably there were too few cells expressing BMP-2 receptors in the only minimally traumatized muscle of these monkeys.

[Statistical relations between blood and cerebrospinal fluid serology in patients with syphilis]. / Statistische Beziehungen zwischen Blut- und Liquorserologie bei Syphilispatienten.

In any case of latent syphilis without conclusive history, an examination of the cerebrospinal fluid (CSF) is generally recommended. The present paper deals with the question whether particular findings obtained by serologic tests on the blood can supply sufficient proof to rule out any specific involvement of the central nervous system in an individual patient. Serologic investigation on 148 blood-CSF sample pairs revealed striking correlations between several blood and CSF parameters on a high level of statistical significance. Based on blood serology data alone, a multi-dimensional, non-parametric statistical test (the k-nearest-neighbor method) was able to rule out specific involvement of the CSF in more than one third of the patients. We consider our approach useful in certain cases where CSF examination should rather be avoided, especially in patients with impaired general condition due to other diseases. Yet, in any case of neurological irregularity, CSF examination should generally be carried out regardless of the results of blood serology.

[The value of CT in diagnosis of low back pain syndromes]. / Wartosc KT w diagnostyce zespolów bólowych ledzwiowo- krzyzowych.

The authors discuss results of CT studies in 300 patients with low back pain syndromes. Apart from herniated discs (56.3% of cases) pain could be often explained by degenerative changes of facet joints or bulging disc leading to spinal canal stenosis. In the group of patients operated on due to disc herniation correlation between CT and surgical results has 85.4%. In the authors' opinion CT is an effective, non-invasive diagnostic method in patients with low back pain syndromes.

Correlation of hexokinase content and basal energy metabolism in discrete regions of rat brain.

Brain hexokinase (ATP:D-hexose 6-phosphotransferase, EC 2.7.1.1) levels in seven regions of rat brain were estimated by photometric measurement of immunofluorescence in cryostat-cut sections. When compared with basal rates of glucose metabolism in these regions, estimated by the 6-[14C]glucose method, a significant correlation was observed. Thus, hexokinase content reflects metabolic energy demands.