Evidence Of New Bone Formation Research Articles

Embryonic Stem Cells Incorporate into Newly Formed Bone and do Not Form Tumors in an Immunocompetent Mouse Fracture Model

Embryonic stem (ES) cells are a uniquely self-renewing, pluripotent population of cells that must be differentiated before being useful for cell therapy. Since most studies utilize subcutaneous implantation to test the in vivo functionality of ES cell-derived cells, the objective of the current study was to develop an appropriate and clinically relevant in vivo implantation system in which the behavior and tumorigenicity of ES cell-derived cells could be effectively tested in a tissue-specific (orthotopic) site. Male ES cells were differentiated either into osteoblasts or chondrocytes using protocols that were previously developed and published by our laboratory. The differentiated cells were implanted into a burr-hole fracture created in the proximal tibiae of immunocompetent female mice, strain matched to the ES cell line. The ability of the differentiated ES cell-derived cells (bearing the Y chromosome) to incorporate into the newly formed bone was assessed by micro-computed tomography imaging and histochemistry. ES cells differentiated with either osteogenic or chondrogenic medium supplementation formed a soft tissue mass that disrupted the normal bone architecture by 4 weeks after implantation in some mice. In contrast, mice receiving osteoblastic cells that were differentiated in a three-dimensional type 1 collagen gel showed evidence of new bone formation at the defect site without evidence of tumor formation for up to 8 weeks after implantation. In this injury model, type 1 collagen is more effective than medium supplementation at driving more complete differentiation of ES cells, as evidenced by reducing their tumorigenicity. Overall, the current study emphasizes the importance of using an appropriate orthotopic implantation system to effectively test the behavior and tumorigenicity of the cells in vivo.

Systematic approach to treat chronic osteomyelitis through localized drug delivery system: Bench to bed side

Chronic osteomyelitis is a challenging setback to the orthopedic surgeons in deciding an optimal therapeutic strategy. Conversely, patients feel frustrated of the therapeutic outcomes and development of adverse drug effects, if any. Present investigation deals with extensive approach incorporating in vivo animal experimentation and human application to treat chronic osteomyelitis, using antibiotic loaded porous hydroxyapatite scaffolds. Micro- to macro-porous hydroxyapatite scaffolds impregnated with antibiotic ceftriaxone–sulbactam sodium (CFS) were fabricated and subsequently evaluated by in vivo animal model after developing osteomyelitis in rabbit tibia. Finally 10 nos. of human osteomyelitis patients involving long bone and mandible were studied for histopathology, radiology, pus culture, 3D CT etc. up to 8–18months post-operatively. It was established up to animal trial stage that 50N50H samples [with 50–55% porosity, average pore size 110μm, higher interconnectivity (10–100μm), and moderately high drug adsorption efficiency (50%)] showed efficient drug release up to 42days than parenteral group based on infection eradication and new bone formation. In vivo human bone showed gradual evidence of new bone formation and fracture union with organized callus without recurrence of infection even after 8months. This may be a new, alternative, cost effective and ideal therapeutic strategy for chronic osteomyelitis treatment in human patients.

Paper 284: Double Bundle Anterior Cruciate Ligament Reconstruction Using a Polylactide Carbonate (CALAXO) Osteoconductive Interference Screw

The purpose of this study was to describe our experience of the new Polylactide Carbonate (Calaxo) Osteoconductive interference screw (Smith & Nephew) when used for both femoral and tibial graft fixation in Double Bundle ACL reconstruction. Since May 2006, patients with an ACL deficient knee were reconstructed using the Double Bundle technique. All were followed prospectively and outcome data collected. Evidence of fixation failure was established subjectively by clinical examination (Pivot Shift) and objectively via KT-1000 arthrometer. Following ethical approval, post-operative CT scans (immediate and 12-18 months) were performed on five (a further 5 awaited) patients allowing assessment of tunnel dimensions/fill. Twenty nine patients (26 male, 3 female) with a mean age of 30 (range 18-47) were included. At last follow-up, no evidence of graft/fixation failure was found; KT-1000 mean side-side difference 1.4mm (range −3 to +6). All patients had a positive pivot shift preoperatively which was abolished postoperatively. Subjective IKDC score was significantly improved with a mean of 76.27% (SD 17.38) at last review compared with 54.02% (SD 17.86) preoperatively (t (22) = −5.37, p<0.05) One patient had a postoperative infection with no other complications reported. Radiologically the screws did not show complete resorption in 2 cases, with significant tunnel widening found on 3 cases. Some evidence of new bone formation was noted but not found to be significant. We have shown satisfactory clinical results with use of the Calaxo screw when used in Double Bundle ACL Reconstruction. However, radiologically the ostoeconductive properties of the screw shown in the pre-clinical animal model were not evident. This study helps illustrate that preclinical animal data cannot necessarily be translated into human practice. Surgeons should proceed with caution when using new biological implants tested purely in animal models. Although we have not had any cases of the reported adverse local soft tissue reaction associated with Calaxo, we support Smith & Nephew's decision to withdraw the screw from clinical use.

Determination of relative in vivo osteoconductivity of modified potassium fluorrichterite glass–ceramics compared with 45S5 bioglass

Potassium fluorrichterite (KNaCaMg(5)Si(8)O(22)F(2)) glass-ceramics were modified by either increasing the concentration of calcium (GC5) or by the addition of P(2)O(5) (GP2). Rods (2 × 4 mm) of stoichiometric fluorrichterite (GST), modified compositions (GC5 and GP2) and 45S5 bioglass, which was used as the reference material, were prepared using a conventional lost-wax technique. Osteoconductivity was investigated by implantation into healing defects in the midshaft of rabbit femora. Specimens were harvested at 4 and 12 weeks following implantation and tissue response was investigated using computed microtomography (μCT) and histological analyses. The results showed greatest bone to implant contact in the 45S5 bioglass reference material at 4 and 12 weeks following implantation, however, GST, GC5 and GP2 all showed direct bone tissue contact with evidence of new bone formation and cell proliferation along the implant surface into the medullary space. There was no evidence of bone necrosis or fibrous tissue encapsulation around the test specimens. Of the modified potassium fluorrichterite compositions, GP2 showed the greatest promise as a bone substitute material due to its osteoconductive potential and superior mechanical properties.

Interpretation of Primate Behavior, Ambulation and Biomechanics: Caveat Arthritis

Copyright: © 2012 Rothschild B. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Analysis of ambulation or biomechanics on the basis of functional morphology (anatomical shape of bones and location of muscle origins and insertions) seems predictive of potential behaviors [1-3]. It delineates possible range of limb motion and potential uses of the limb, and may seem all that is available for examination of the fossil primates. Actually, evidence of new bone formation at sites of muscle or tendon insertion (entheses) and osteoarthritis further identify how the limb is actually utilized [4-6]. The stress of limb activity promotes osteoblastic reaction [4], with visible evidence. The combination of these physical manifestations allows speculation as to organismal behavior. Establishing confidence (that a given speculation accurately portrays the life of a primate) requires an additional step: Observation of limb utilization in vivo in the natural setting. This requires observations in the wild. Performance of such observations in artificial environments allows more controlled study, but the character of the artificial environment must also be considered. Caya Santiago contains a phenomenal project, in which rhesus macaques have been given their own island. It is a natural habitat in a hurricane-susceptible region. In contrast to the two-three levels of canope in their natural environment [7], there is generally only one layer of canape on this Puerto Rican island. The macaques appear to spend more time on the ground than they do in their “home” environment. A behavior alteration, with osseous manifestations: They have a higher frequency of osteoarthritis than do their conspecifics in Africa [8-11]. Osteoarthritis is extremely rare in the natural habitat, with frequency less than 1% in primates [812]. There is another factor which may affect behavioral observations, both in generated habitats (e.g., Caya Santiago) and in the wild, what might be referred to as the behavioral analogy to the Heisenberg uncertainty issue in physics. Does the very fact of observation alter behavior? And, some investigators do more than just observe (e.g., Diane Fossey).

Poly-N-Acetyl Glucosamine Fibers Activate Bone Regeneration in a Rabbit Femur Injury Model

The purpose of this study was to evaluate the ability of a membrane material, consisting only of short poly-N-acetyl glucosamine (sNAG) nanofibers, to regenerate bone tissue after implantation into circular holes in the rabbit femur. Three circular holes were created in the femurs of five male New Zealand white rabbits. The holes were ∼ 2.0 mm in diameter. Three holes in the left femur were implanted with the comparative control substance (Bone Wax; Ethicon, Inc.); three holes in the right femur were implanted with the sNAG membrane test article. Animals were killed 4 weeks after surgery, and macroscopic evaluation of the implant sites was made. Hematoxylin and eosin histology was performed on both control and test sites. All control (bone wax) sites had visible holes (defects) at the 28-day end point of the study and showed no evidence of new bone formation. All the 15 sNAG test sites were found to have new bone tissue present in the bone hole defects. Hematoxylin and eosin histology of the sNAG-treated test sites showed the presence of osteoblasts, osteocytes, and trabecula of new bone formation at the 28-day end point of the study. The sNAG membrane test material activated the regeneration of new bone tissue in a rabbit femur bone model after 28 days of implantation, whereas the control bone wax material did not.

RhBMP-2 and -7 combined with absorbable collagen sponge carrier enhance ectopic bone formation: An in vivo bioassay

Abstract Background: Recombinant human bone morphogenetic proteins (rhBMPs) have been characterized especially chondrogenic and osteogenic activity both in vitro and in vivo studies. However, delivery of more than one growth factor by sustained release carrier to orthopedic site has yet been questionable in terms of efficacy and synergism. Objective: Evaluate osteoinductivity and synergistic effect of rhBMP-2 and -7 using absorbable collagen sponge (ACS) carrier system in vivo. Methods: cDNA of BMP-2 and -7 active domains were cloned and expressed in Escherichia coli BL21 StarTM (DE3) using pRSETc expression system. Then, the purified rhBMPs were loaded onto ACS and evaluated by in vivo rat subcutaneous bioassay. Two and eight weeks postoperatively, all treated groups were histologically verified for evidence of new bone formation and neovascularization by hematoxylin-eosin staining and light microscopy. Results: The Wistar rat treated with rhBMP-2 or -7/ACS exhibited new bone formation, compared to ACS control. The group treated with ACS supplemented with both rhBMP-2 and -7 significantly showed the osteoid matrix very well-organized into trabeculae-like structure with significant blood vessel invasion. Conclusion: The osteogenic induction of rhBMPs was combined with ACS carrier in the in vivo bioassay. In addition, the combination of both two potent recombinant osteoinductive cytokines, rhBMP-2 and -7, with ACS carrier demonstrated synergistic effect and might be a more promising and effective choice for therapeutic applications.

Histologic Analysis of Healing After Tooth Extraction With Ridge Preservation Using Mineralized Human Bone Allograft

Ridge preservation was developed as a therapy to prevent severe bone resorption after tooth extraction. The purpose of this study is to determine if there is any difference in the amount of new bone formation ≈3 months after extraction and ridge preservation compared to that after ≈6 months. Minimally traumatic extraction with ridge preservation using mineralized human bone allograft was performed at 38 single-rooted tooth sites in 33 subjects. Sixteen sites healed for an average of 14 weeks (early healing), whereas 22 sites were allowed to heal for an average of 27 weeks (delayed healing) before harvesting bone core samples. Histomorphometric analysis was performed to determine the percent of new bone formation, residual graft particles, and connective tissue/non-mineralized structures for each site. All specimens showed evidence of new bone formation, with most of the residual graft particles surrounded intimately by woven bone. No statistically significant differences in the amount of newly formed bone or residual graft particles were found between the two groups. Overall, the early healing group demonstrated a mean of 45.8% new bone, 14.6% residual graft material, and 39.6% connective tissue/non-mineralized tissue. The delayed healing group showed mean values of 45%, 13.5%, and 41.3%, respectively. The results of this study suggest that waiting ≈6 months after tooth extraction and ridge preservation using mineralized bone allograft does not provide a greater amount of new bone formation or less residual bone particles compared to that after only ≈3 months.

A new mode of clinical failure of porous tantalum rod

The area of osteonecrosis of the head of femur affected by the disease process varies from a small localized lesion to a global lesion. Without specific treatment 80% of the clinically diagnosed cases will progress, and most will eventually require arthroplasty. Therefore the goal is to diagnose and treat the condition in the earliest stage. A number of surgical procedures have been described to retard or prevent progression of the disease and to preserve the femoral head. An implant made of porous tantalum has been developed to function as a structural graft to provide mechanical support to the subchondral plate of the necrotic femoral head, and possibly allow bone growth into the avascular region. Porous tantalum implant failure with associated radiological progression of the disease is reported in the literature; however, there is no report of clinical failure of the implant without radiological progression of the disease. We report a case of clinical failure of porous tantalum implant, seven months after surgery without any radiological progression of the disease, and with histopathological evidence of new bone formation around the porous tantalum implant. The patient was succesfully treated by total hip arthroplasty.

Acinetobacter baumannii is not Associated with Osteomyelitis in a Rat Model: A Pilot Study

Multidrug resistant Acinetobacter baumannii (MDR AB) with and without Staphylococcus aureus (SA) is a commonly isolated organism in infected segmental bone defects in combat-related trauma in Iraq and Afghanistan. Although MDR AB in visceral infections is a therapeutic challenge, control of infection appears more common for combat-related osteomyelitis. Using a rat model, we explored the virulence of MDR AB in segmental bone defects alone and in combination with SA. Segmental defects in 60 rat femurs were created, stabilized, and inoculated with MDR AB alone and 60 with MDR AB and SA. We performed qualitative and quantitative bacteriology and radiographic assessments at 2, 4, and 8 weeks for MDR AB and at 1, 2, and 3 weeks for MDR AB and SA. Quantitative bacteriology revealed a 3- to 5-log decrease in MDR AB from the initial inoculum. After polymicrobial inoculation, only 10 of 60 animals had positive cultures for MDR AB, whereas 59 of 60 animals had positive cultures for SA. Recovered SA were 2 to 5 log greater than the initial inoculum, while there again was a 3- to 5-log decrease in MDR AB. MDR AB alone did not cause bony lysis, but there was radiographic evidence of new bone formation in 67% of the segmental defects. Osteolysis was noted with MDR AB and SA. MDR AB did not appear to cause or contribute to clinically apparent osteomyelitis in this pilot study. Resolution of infections in combat-related segmental bone defects inoculated with MDR AB may be attributable to low virulence. Additional studies are needed to confirm low virulence and bone formation with MDR AB.

Does Osgood-Schlatter Disease exist in the dog? A comment

tures occurring in terrier breeds (all but one fracture) and in particular the Staffordshire Bull Terrier (86% of affected dogs). In this breed TTAF was not a rare fracture, accounting for 3.3% of admissions in dogs less than one-year of age compared to 0.18% for other breeds. We agree with von Pfeil et al that TTAF is generally a result of a tensile trauma, commonly occurring following a fall or awkward jump. We suggested that the Staffordshire Bull Terrier was at particular risk due to its large quadriceps mass/ strength relative to the size/strength of its tibial tuberosity physis such that strong contraction of the quadriceps in combination with sudden stifle flexion (as may occur on landing from a fall) would result in TTAF. We also agree with von Pfeil et al that OSD in people should be considered a separate entity from TTAF in dogs. In none of the dogs in our series did we see evidence of newbone formation at the insertion of the patellar tendon on the tibial tuberosity as reported in OSD. Other conclusions from von Pfeil et al’s review however were not supported by our study. In the 65 fractures we reported on, three predominant fracture patterns were seen and these do not correlate with the classification system proposed by von Pfeil et al. The most common pattern was an isolated tibial tuberosity avulsion fracture with no involvement of the proximal tibial epiphysis (57% of fractures). The degree of displacement of the tibial tuberosity was variable. We also recognised TTAF in combination with variable degrees of elevation/ separation of the cranial aspect of the proximal tibial epiphysis (23%) and TTAF in combination with complete Salter-Harris type II fracture of the epiphysis, with the fracture line extending into the caudal metaphysis of the tibia (20%). In the dogs we reported on the majority of TTAF of all types occurred at fourto five-months of age, contrary to the suggestion of von Pfeil et al that certain types were seen most often in eightto 11-month-old dogs. We recognise of course that our results were dominated by a single medium-sized breed and that von Pfeil et al’s eight cases included large breed dogs. It should not be surprising that disparate study populations yielded different results. It is clear however that some of the conclusions drawn by von Pfeil et al, and also their proposed classification scheme, are not relevant to all dogs.

Influence of rhBMP-2 on the Healing Patterns Associated With Allograft Interbody Constructs in Comparison With Autograft

A prospective, nonblinded, multicenter study. To determine the patterns of allograft incorporation and new bone formation after ALIF using stand-alone threaded cortical allograft dowels with either autograft or rhBMP-2. Accelerated healing patterns of allograft dowels have been reported in an animal study and a small human study of ALIF with rhBMP-2 and bone dowels. We enrolled and randomly assigned 131 patients with lumbar spondylosis who were undergoing single-level ALIF with allograft dowels to either the investigational group who received rhBMP-2 (79 patients) or the control group who received autologous bone graft (52 patients). Independent radiologists used plain radiographs and computed tomography scans to evaluate fusion. Allograft incorporation was assessed using a modified Bridwell-Lenke scale, and changes in bone density within the adjacent vertebral bodies were monitored. At 12 months, all investigational patients had radiographic evidence of new bone formation and incorporation of the allografts into the adjacent vertebral endplates; there were no changes in fusion status at 24 months. Fusions were documented in 89% of patients in the control group at 12 months. The percentage declined to 81.5% at 24 months with 10% of the patients in the autograft group showing incomplete healing and 11% having no healing of the allograft dowels. On CT scans, 14 of the investigational patients (14 of 79; 18%) developed a transient, localized area of bone remodeling within the vertebral body adjacent to the allograft dowel. All were healed by 24 months. The transient appearance of these zones had no effect on clinical outcomes. No patient in the control group developed bone remodeling zones. We think that the use of rhBMP-2 is a promising method of facilitating allograft dowel incorporation and new bone formation in patients undergoing ALIF with allograft dowels.

Evaluation of different grafting materials in three-wall intra-bony defects around dental implants in beagle dogs

The purpose of this study was to evaluate histologically and compare the healing of three-wall defects using xenogeneic demineralized bone matrix putty (xDBM putty), porous β-tricalcium phosphate (β-TCP), or newly developed non-crystalline calcium phosphate glass around submerged SLA (Sand blasted with Large-grit and Acid Etched) surface dental implants in dogs. All of the treatment groups in this study showed no histologic evidence of new bone formation around implants or of bone to implant contact originating from grafted material. No beneficial effects on osseointegration in intrabony defects around implants was attributed to the use of these grafting materials.

Growth and transplantation of a custom vascularised bone graft in a man

A major goal of research in bone transplantation is the ability to avoid creation of secondary bone defects. We aimed to repair an extended mandibular discontinuity defect by growth of a custom bone transplant inside the latissimus dorsi muscle of an adult male patient. Three-dimensional computed tomography (CT) scanning and computer-aided design techniques were used to produce an ideal virtual replacement for the mandibular defect. These data were used to create a titanium mesh cage that was filled with bone mineral blocks and infiltrated with 7 mg recombinant human bone morphogenetic protein 7 and 20 mL of the patient's bone marrow. Thus prepared, the transplant was implanted into the latissimus dorsi muscle and 7 weeks later transplanted as a free bone-muscle flap to repair the mandibular defect. In-vivo skeletal scintigraphy showed bone remodelling and mineralisation inside the mandibular transplant both before and after transplantation. CT provided radiological evidence of new bone formation. Postoperatively, the patient had an improved degree of mastication and was satisfied with the aesthetic outcome of the procedure. Heterotopic bone induction to form a mandibular replacement inside the latissimus dorsi muscle in a human being is possible. This technique allows for a lower operative burden compared with conventional techniques by avoiding creation of a secondary bone defect. It also provides a good three-dimensional outcome.

Frontal ostium neo-osteogenesis and restenosis after modified endoscopic Lothrop procedure in an animal model.

All patients who undergo a modified endoscopic Lothrop procedure have postoperative narrowing of the enlarged frontal ostium. The aim of this study is to evaluate neo-osteogenesis and restenosis of the frontal ostium and its effect on mucociliary clearance. Fourteen sheep underwent an endoscopic modified Lothrop procedure. Pre- and postoperative nuclear medicine gamma scintigraphy of the frontal sinuses was performed. The sizes of the frontal ostia were measured and biopsies taken from the bone of the frontal ostium. Histological evidence of new bone formation was found in 56% of biopsies. The average preoperative mucociliary clearance half times (T1/2) at 15 and 30 min were 70 and 74 min, respectively, and postoperatively were 50 and 67 min. There was a non-significant trend towards poorer clearance in sinuses with neo-osteogenesis. The average size of the frontal ostium decreased by 28%. There was no relationship between the size of the ostium and neo-osteogenesis. Neo-osteogenesis was seen in 56% of biopsies with a 28% reduction in size of the frontal ostium after 224 days. Mucociliary clearance did not alter significantly.

Anterior thoracic spinal fusion in dogs by injection of recombinant human bone morphogenetic protein-2 and a synthetic polymer.

The purpose of this study was to achieve spinal fusion in the absence of bone graft material using a new, injectable, and semi-liquid synthetic polymer (polylactic acid polyethylene glycol [PLA-PEG] block copolymer) containing recombinant human bone morphogenetic protein-2 (rhBMP-2). Twenty-seven skeletally mature beagles underwent anterior thoracic spinal fusion at T9-T10. Group I (n = 9) was injected with 1 mL of PLA-PEG block copolymer carrier alone into space under the vertebral pleura and the anterior longitudinal ligament. Group II (n = 9) was injected with 1 mL of PLA-PEG carrier containing 500 microgram of rhBMP-2. Group III (n = 9) was injected with 1 mL of PLA-PEG carrier containing 1000 microgram of rhBMP-2. In the Group I animals, no evidence of new bone formation was noted at the implanted sites both radiographically and histologically. In contrast, all of the nine animals in Group III showed new bone formation in 12 weeks, and four of the nine animals in Group II showed bony mass at the injected sites. However, vertebral bony fusion was incomplete despite the significant amount of new bone formation in both groups that showed new bone formation. In addition to resulting in improvements in the surgical procedure, injection of rhBMP-2 and a synthetic polymer is useful for bone formation for spinal fusion.

The Anabolic Effects of Vitamin D-Binding Protein- Macrophage Activating Factor (DBP-MAF) and a Novel Small Peptide on Bone

Vitamin D-binding protein-macrophage activating factor (DBP-MAF) has previously been shown to stimulate bone resorption and correct the skeletal defects associated with osteopetrosis in two nonallelic mutations in rats. This same protein and a small fragment of the protein have now been shown to demonstrate an anabolic effect on the skeleton of both newborn and young adult, intact rats. The novel peptide fragment was synthetically produced based on the human amino acid sequence at the site of glycosylation in the third domain of the native protein (DBP). The peptide tested is 14 amino acids in length and demonstrates no homologies other than to that region of DBP. Newborn rats were injected i.p. with saline, peptide (0.4 ng/g body wt.) or DBP-MAF (2 ng/g body wt.) every other day from birth to 14 days of age. On day 16 the rats were euthanized and the long bones collected for bone densitometry by pQCT. After 2 weeks of treatment with either the whole protein (DBP-MAF) or the small peptide, bone density was significantly increased in the treated animals compared to the saline controls. Young adult female rats (180 grams) were given s.c. injections of saline or peptide (0.4 ng/g body wt. or 5 ng/g body wt.) every other day for 2 weeks; 2 days after the final injections, the rats were euthanized and the femurs and tibias collected for bone densitometry. Both doses of the peptide resulted in significant increases in bone density as determined by pQCT. Young adult rats were injected locally with a single dose of the peptide (1 microg) or saline into the marrow cavity of the distal femur. One week after the single injection, the bones were collected for radiographic and histological evaluation. The saline controls showed no evidence of new bone formation, whereas the peptide-treated animals demonstrated osteoinduction in the marrow cavity and osteogenesis of surrounding cortical and metaphyseal bone. These data suggest that DBP-MAF and the synthetic peptide represent therapeutic opportunities for the treatment of a number of bone diseases and skeletal disorders. Systemic administration could be used to treat osteoporosis and a number of other osteopenias, and local administration could be effective in fractures, bony defect repairs, spinal surgery, and joint replacement.

Autoclaving the ossicles provides safe autografts in cholesteatoma

Objective: The choice of the graft in ossicular chain reconstruction during middle ear surgery for cholesteatoma is a subject still discussed on. In order to clarify the discussion of reuse of the autologous ossicles obtained during middle ear surgery for cholesteatoma, we evaluated the probability of residual disease histologically and the safety of the ossicles after autoclavization, the most promoting alternative method to eradicate residual cholesteatoma and infection on them. Methods: The specimens used in this study were eroded twenty-seven ossicles (22 incuses, 5 malleoli) which were removed from the 27 consecutive patients operated because of cholesteatomatous middle ear disease. They were grouped as follows: Group 1, Fifteen ossicles examined histopathologically directly. Group 2, Five ossicles autoclaved for 20 min at 134 °C and then examined histopathologically. Group 3, Five ossicles autoclaved for 20 min at 134 °C after mechanical surface cleaning by a fine diamond drill, examined histopathologically. Group 4, Two ossicles removed from two different patients were placed in their mastoid cavities in order to be examined after access in the second-look operation. While one ossicle was only autoclaved, the other was mechanically cleaned by a drill before autoclavization (for 20 min at 134 °C). The ossicles were examined histopathologically after the removal at the second stage operation performed 12 months later. Results: In Group 1, all ossicles showed evidence of periosteal thickening. Additional findings were surface cholesteatoma or epithelia in 13 ossicles, surface inflammation in 12 ossicles, granulation tissue in 10 ossicles, osteitis in six ossicles. In Group 2, all five ossicles had preserved their lamellar structure but, no vital cells were seen. The lacunes that had the osteocytes was almost completely empty. The inflammatory cells were eliminated from the ossicles. In Group 3, ossicles were found well preserved with their lamellar structures and contours, with empty lacunes and eliminated inflammatory cells. In Group 4, in two ossicles of this group the lacunes were replaced by the new migrated viable osteocytes with evidence of new bone formation and neovascularisation. No new inflammatory focus or epithelia were found on the surfaces of the ossicles. The shape and the contour of the ossicles remained unchanged. Conclusion: In cholesteatoma surgery, ossicles with minimal erosion and adequate thickness can be used after autoclavization. In this study, it was observed histopathologically that the autoclaving autologous ossicles before ossiculoplasty in cholesteatomatous middle ear is a safe and reliable method.

Behavior of natural coral in a human osteoporotic bone

The research concerning a preventive treatment of osteoporotic femoral neck started in 1990 because the surgical procedure of unstable femoral neck fractures is difficult, after effects frequent and there number will increase in the next decade. The goal is to reinforce the femur with a biomaterial. Natural coral is bioresorbable and biocompatible. It acts as an autologus bone graft in the reconstruction of either cortical or cancellous bone and increases the mechanical resistance. This work shows evidence of newbone formation in osteoporotic unbroken femur thanks to the biomaterial; consequently, there preventive surgical treatment should be taken in consideration.

The posterior fat pad sign in association with occult fracture of the elbow in children.

An elevated posterior fat pad visible on a lateral radiograph of a child's elbow following trauma is generally considered to be suggestive of an intracapsular fracture about the elbow. However, in previous studies, the prevalence of fracture in elbows with an elevated posterior fat pad and no other radiographic evidence of fracture has ranged from only 6 percent (two of thirty-one) to 29 percent (nine of thirty-one). We are not aware of any prospective studies, limited to children, on the value of an elevated posterior fat pad as an indicator of an occult fracture about the elbow. While it is common practice to manage children who have radiographic evidence of an elevated posterior fat pad as if they have a fracture, scientific evidence for this approach is lacking. Forty-five consecutive children who had an average age of four and a half years, a history of trauma to the elbow, and an elevated posterior fat pad without other radiographic evidence of a fracture were enrolled in the study. At an average of three weeks after the injury, anteroposterior, lateral, and two oblique radiographs were made and evaluated for evidence of fracture-healing. If there was evidence of new-bone formation on any of these four radiographs, it was considered to indicate a fracture of the elbow. Thirty-four (76 percent) of the forty-five patients had evidence of a fracture. Eighteen (53 percent) of the thirty-four had a supracondylar fracture of the humerus; nine (26 percent), a fracture of the proximal part of the ulna; four (12 percent), a fracture of the lateral condyle; and three (9 percent), a fracture of the radial neck. This prospective study demonstrated that the posterior fat pad sign was predictive of an occult fracture of the elbow following trauma in thirty-four (76 percent) of forty-five children who had no other evidence of fracture on anteroposterior, lateral, and oblique radiographs after the injury. This finding is in contrast to those of previous studies, in which the highest prevalence of fracture in elbows with an elevated posterior fat pad and no other radiographic evidence of fracture was 29 percent (nine of thirty-one elbows). Our results support the practice of managing children who have a history of trauma to the elbow, an elevated posterior fat pad, and no other radiographic evidence of fracture as if they have a nondisplaced fracture about the elbow.