Bone grafts engineered from human adipose-derived stem cells in dynamic 3D-environments.

Modular tissue engineering (TE) is a promising alternative to overcome the limits in traditional TE. In the present study, adipose tissue derived stem cells (ADSC)-laden microcarriers are used as building blocks (microtissues) that self-assemble into macrotissues in a bottom-up approach. These bone grafts were compared with a classical top-down approach (scaffolds). This concept was compared with bone marrow derived stem cells (BMSC) as cell source. Cells were immunophenotypically analyzed, followed by 2D/3D osteogenic differentiation in static/dynamic conditions. The bone graft quality was evaluated by (immuno)histochemistry and gene expression. After 6 weeks of dynamic culturing, scaffolds were highly colonized although not in the center and the osteogenic gene expression was higher in contrast to static cultures. A cell-to-microcarrier ratio of 5 × 10(6) cells-0.09 g microcarriers leaded to aggregate formation resulting in microtissues with subsequent macrotissue formation. ADSC/BMSC on scaffolds showed a downregulation of Runx2 and collagen I, demonstrating the end-stage, in contrary to microcarriers, where an upregulation of Runx2, collagen I together with BSP and osteocalcin was observed. This paper showed that high quality bone grafts (2 cm³) can be engineered in a bottom-up approach with cell-laden microcarriers.

Efeito da desmineralização ácida da interface enxerto-leito na consolidação de enxertos ósseos autógenos em bloco / Influence of acid demineralization of contacting osseous surfaces on the consolidation of autogenous onlay bone grafts

Para testar a hipótese de que a desmineralização in situ das superfícies de contato enxerto-leito, e a forma como o enxerto é estabilizado ao leito, podem influenciar os mecanismos envolvidos na consolidação do enxerto, fragmentos ósseos de 10 mm de diâmetro foram removidos das metáfises proximais tibiais de 36 coelhos (Oryctolagus Cuniculus) e transplantados para uma área adjacente. Na tíbia esquerda dos animais, as superfícies de contato do enxerto e do leito hospedeiro foram desmineralizadas com ácido cítrico pH 1,0 por 3 minutos antes dos enxertos serem fixados ao leito. Na tíbia direita, o transplante do bloco ósseo não foi precedido de desmineralização. Metade dos enxertos foi imobilizada sobre o leito pela superposição de uma membrana não reabsorvível de politetrafluoretileno colada com cianoacrilato ao leito à distância da interface enxerto-leito. A outra metade dos enxertos foi fixada por um parafuso de titânio no centro do enxerto. Assim, foram formados 4 grupos de estudo: membrana (M), membrana + ácido (MA), parafuso (P) e parafuso + ácido (PA). Três animais de cada grupo forneceram espécimes para análise microscópica quantitativa e qualitativa aos 15, 30 e 45 dias de pós-operatório. A análise qualitativa demonstrou que não houve formação óssea na interface em nenhum espécime aos 15 dias e que nos demais períodos, em todos os grupos, a quantidade de tecido ósseo neoformado na interface e seu estágio de maturação aumentaram com o tempo. Ambos os métodos de fixação empregados foram eficientes em manter os enxertos em posição, porém a membrana promoveu menor reabsorção da estrutura do enxerto. A análise quantitativa computadorizada revelou que, aos 30 dias, os grupos MA e PA apresentaram maior área de formação óssea na interface (71,34 ± 12,03%; 56,74 ± 2,15% respectivamente) em relação aos grupos M e P (51,75 ± 11,02%; 43,95 ± 4,05% respectivamente) e superfícies de consolidação óssea mais extensas (93,41 ± 5,95%; 93,73 ± 4,96%...

In order to test the hypothesis that the demineralization "in situ" of contacting surfaces of bone graft/bone bed and the fixation method used for graft stabilization can influence the mechanisms involved in the consolidation of the graft, bone fragments of 10 mm in diameter were removed from the proximal tibial metaphysis of thirty-six male rabbits (Oryctolagus Cuniculus) and transplanted to an adjacent area. In the left tibia of the animals, the contacting surfaces of the graft and host bed were demineralized with citric acid pH 1.0 for 3 minutes before the grafts were fixed to the receptor bed. In the right tibia, the bone block transplantation was not preceded by demineralization. Half of the grafts were immobilized on the bone bed by a nonresorbable polytetrafluoroethylene membrane glued with cyanoacrylate adhesive to the host bed distant from the bone graft-bone bed interface. The other half of the grafts were fixed by a titanium screw in the center of the graft. Thus, four groups were formed: membrane (M), membrane + acid (MA), screw (P) and screw + acid (PA). Three animals from each group provided specimens for quantitative and qualitative microscopic analysis at 15, 30 and 45 days postoperatively. Qualitative analysis showed no significant bone formation at the interface in any specimen of the groups at 15 days and on the other periods in all groups, the amount of newly formed bone at the interface as well as the stage of bone maturation increased with time. Both fixation methods were effective in maintaining the graft in position, but the membrane resulted in less resorption of the graft. Quantitative analysis, performed by means of a computer program for image analysis, showed that at day 30, groups MA and PA, showed greater area of bone formation at the interface (71.34 ± 12.03%; 56.74 ± 2 15%) than groups M and P (51.75 ± 11.02%, 43.95 ± 4.05%) and more osseointegrated bone surfaces (93.41 ± 5.95%, 93.73 ± 4.96%) than those...

Volume changes of iliac crest autogenous bone grafts after vertical and horizontal alveolar ridge augmentation of atrophic maxillas and mandibles: a 6-year computerized tomographic follow-up.

PURPOSE: To evaluate by computerized tomography the long-term volume resorption of autogenous corticocancellous grafted bone harvested from the ilium and used in an alveolar augmentation procedure followed by endosseous dental implant placement. PATIENTS AND METHODS: Eleven maxillary grafts (8 positioned horizontally) and 13 mandibular grafts (10 positioned vertically) were placed in 16 patients. Using software programs, pre- and postsurgical computerized tomographic scans were used to compare volumes of grafts over time (up to 6 yr) to determine the annual percentage of remaining bone and the overall percentage of bone resorption that could be expected. Yearly measurements of volumes and percentages of remaining bone were then compared statistically. RESULTS: At the 6-year survey for blocks grafted in the mandible, an average resorption rate of 87% was obtained; for maxillary grafts at the same survey, complete resorption of the grafts (mean, 105.5%) was recorded. In general, bone resorption appeared slow, except for that recorded in the first 2 years of healing, the only period in which statistical comparisons among all time points showed significant differences for all variables. CONCLUSIONS: Volumetric measurements of the grafts and their related percentages of remaining bone attested to a progressive and unavoidable bone resorption of almost all the grafted bone in the maxilla and mandible. Although the present data were from a heterogeneous group of defects treated with horizontal and vertical procedures, clinicians, when performing alveolar bone augmentation with an autogenous hip bone, should aim at titanium dental implant osseointegration, not only in the augmented bone but also in the native bone below the graft.

Alendronate enhances osseous healing in a rat calvarial defect model.

AIM: The aim of this study was to evaluate the effect of alendronate on osseous wound healing in an experimental model. METHODS: Critical size defects were created in calvaria of 40 male Wistar rats. The animals were divided into four groups of 10 animals each: autogenous bone graft group; autogenous bone graft with systemic alendronate group (0.01 mg/kg body weight per day for 8 weeks); autogenous bone graft with local alendronate group (1mg/mL for 5 min); non-treatment (control) group. Animals were sacrificed after 8 weeks; osteoblast number, lamellar bone formation, and area of newly formed bone were analysed. RESULTS: The osteoblast number significantly increased in the autogenous bone graft with local alendronate group compared to the autogenous bone graft group (p<0.05). Both systemic and local application of the alendronate significantly increased the new bone formation compared to the autogenous bone graft group (p<0.05) with no significant difference between local or systemic use (p>0.05). Local alendronate and autogenous bone graft use significantly increased the total bone area compared to autogenous bone graft alone (p<0.05). CONCLUSION: Alendronate enhances the new bone formation by autogenous bone graft in the rat calvarial defect model suggesting that the inhibition of the osteoclastic activity allows an increased rate of bone apposition, which could be applicable to the inflammation-induced destruction of the periodontal tissues during disease.

Evaluation of a novel reconstituted bone xenograft using processed bovine cancellous bone in combination with purified bovine bone morphogenetic protein.

BACKGROUND: Xenogeneic grafting represents an alternative to autogenous grafting in osseous reconstruction and exhibits many beneficial properties. However, the usefulness of xenogeneic bone relies on necessary processing procedures for removing antigens and viruses, and preserving biological activities simultaneously. By chemical treatment of bovine cancellous bone to make it an antigen-free scaffold, and extraction of bone morphogenetic protein (BMP) from bovine cortical bone, followed by recombination of the scaffold with the BMP, we developed a new grafting material, reconstituted bone xenograft (RBX). METHODS: In this study, scanning electron microscope and energy dispersive X-ray were first employed to observe the structure and components of RBX. Then the biomechanical property was evaluated by applying compression in a materials testing machine. Subsequently, the immunologic evaluation was performed by measuring galactose-alpha-1,3-galactose (α-gal) epitope in vivo and proinflammatory cytokine (TNF-α) secreted by human monocytic cell line (THP-1) in vitro. Finally, this RBX was implanted into segmental radial defects in a rabbit model, and its ability to treat large bone defects was specifically evaluated. RESULTS: Although the compressive strength of RBX was 10% lower than that of unprocessed bovine cancellous bone (UBCB), the basic porous structure and natural components were still kept in this composite. The α-gal xenoantigen level was significantly lower in RBX (P < 0.05) compared with UBCB. Moreover, the TNF-α level was significantly (P < 0.05) reduced compared with UBCB when THP-1 was exposed to RBX. On the other hand, RBX appeared to induce cartilage formation from immature cell populations and resulted in osteogenesis through endochondral-like ossification from 4 to 12 weeks in repairing segmental bone defects. CONCLUSIONS: These results demonstrate that RBX, with its natural microstructure and components, certain mechanical strength and strong osteoinductivity without evoking immune rejection, has significant potential for the treatment of bone defects.

Autogenous bone graft: basic science and clinical implications.

No single biomaterial is optimum for every craniomaxillofacial application. Instead, surgeons should consider the advantages and disadvantages of each alternative in a given clinical situation, and select the material with lowest overall cost and morbidity, and the highest likelihood of success. Autogenous bone is still considered the gold standard for most applications; it becomes vascularized and osseointegrates with surrounding bone, thus minimizing the risk of infection, dislodgement, or break-down. Limitations include added operative time for graft harvest, donor site morbidity, graft resorption, molding challenges, and limited availability, especially in the pediatric population. Numerous alternatives to bone graft have become available to address these limitations; unfortunately, most of these products are expensive, do not osseointegrate, and have unpredictable biologic activity. Understanding the physiologic behavior of autogenous bone graft can help clarify the indications for its use and provide a conceptual framework for achieving the best possible outcome when this alternative is chosen.

The effect of bisphosphonate and intermittent human parathyroid hormone 1-34 treatments on cortical bone allografts in rabbits.

AIM: This study compares the effect of bisphosphonate and intermittent PTH administration on haversian remodeling in cortical bone allografts in rabbits. MATERIALS AND METHODS: An intercalary heat-treated cortical bone allograft was applied to a segment skeletal defect in the left femur of Japanese white rabbits. The rabbits were randomly assigned to one of three groups: the vehicle control group (CNT); the bisphosphonate group (B group); and the intermittent PTH treatment group (P group). Periodic radiographic evaluation was performed and peripheral quantitative computerized tomography (pQCT) was used to evaluate the total bone area (Area), bone mineral density (BMD), and bone mineral content (BMC). The allografts also underwent histological examination. RESULTS: The P group was radiographically superior in the latter stage, compared with the other groups. pQCT analysis of the allografts showed that the B group had a significantly higher Area and BMC. These parameters in the latter stage were significantly lower in the P group than in the other groups. The allograft of the B group was histologically mostly necrotic bone, whereas allograft of the P group showed abundant newly formed bone. CONCLUSION: In rabbits, bisphosphonate prevents resorption, but suppresses remodeling and incorporation; by contrast, PTH increases resorption and accelerates allograft remodeling and incorporation. Based on our preliminary data, we suggest that further research on the manner of administration of bisphosphonate and PTH - which have contrasting effects - can be beneficial in maintaining bone strength and in regulating remodeling and allograft incorporation.

Bone morphogenic protein: an elixir for bone grafting--a review.

Bone morphogenetic proteins (BMPs) are multifunctional growth factors that belong to the transforming growth factor beta superfamily. This literature review focuses on the molecular biology of BMPs, their mechanism of action, and subsequent applications. It also discusses uses of BMPs in the fields of dentistry and orthopedics, research on methods of delivering BMPs, and their role in tissue regeneration. BMP has positive effects on bone grafts, and their calculated and timely use with other growth factors can provide extraordinary results in fractured or nonhealing bones. Use of BMP introduces new applications in the field of implantology and bone grafting. This review touches on a few unknown facts about BMP and this ever-changing field of research to improve human life.

Anterior cervical interbody constructs: effect of a repetitive compressive force on the endplate.

Graft subsidence following anterior cervical reconstruction can result in the loss of sagittal balance and recurring foraminal stenosis. This study examined the implant-endplate interface using a cyclic fatigue loading protocol in an attempt to model the subsidence seen in vivo. The superior endplate from 30 cervical vertebrae (C3 to T1) were harvested and biomechanically tested in axial compression with one of three implants: Fibular allograft; titanium mesh cage packed with cancellous chips; and trabecular metal. Each construct was cyclically loaded from 50 to 250 N for 10,000 cycles. Nondestructive cyclic loading of the cervical endplate-implant construct resulted in a stiffer construct independent of the type of the interbody implant tested. The trabecular metal construct demonstrated significantly more axial stability and significantly less subsidence in comparison to the titanium mesh construct. Although the allograft construct resulted in more subsidence than the trabecular metal construct, the difference was not significant and no difference was found when comparing axial stability. For all constructs, the majority of the subsidence during the cyclic testing occurred during the first 500 cycles and was followed by a more gradual settling in the remaining 9,500 cycles.

Isolating stromal stem cells from periodontal granulation tissues.

Stem cell therapy is a promising area in regenerative medicine. Periodontal granulation tissues are often discarded during conventional surgery. If stromal stem cells can be isolated from these tissues, they can be used for subsequent surgery on the same patient. Fifteen human periodontal granulation tissue samples were obtained from intrabony defects during surgery. Immunohistochemistry (IHC) was carried out on five of the samples to identify STRO-1, a marker of mesenchymal stem cells. Five samples underwent flow cytometry analysis for the same marker. The remaining five samples were characterized by "colony formation unit-fibroblast" (CFU-f) assay and selected for proliferation assay, flow cytometry of stem cell markers, immunocytochemistry (ICC), multipotent differentiation assays, and repairing critical-size defects in mice. The ratio of STRO-1(+) cells detected by IHC was 5.91 ± 1.50%. The analysis of flow cytometry for STRO-1 was 6.70 ± 0.81%. Approximately two thirds of the CFU-f colonies had a strong reaction to STRO-1 in ICC staining. The cells were multipotent both in vitro and in vivo. Mice given bone grafts and stem cells showed significantly better bone healing than those without stem cells. Multipotent stromal stem cells can be isolated from human periodontal granulation tissues. These cells improve new bone formation when transplanted in mouse calvarial defects. Isolating stem cells from relatively accessible sites without extra procedures is clinically advantageous. This study demonstrated that human periodontal granulation tissues contain isolatable multipotent stem cells. The cells may be a good source for autotransplantation in subsequent treatment.

Orbital stress analysis: part II: design and fixation of autogenous bone graft used to repair orbital blowout fracture.

PURPOSE OF THE STUDY: The purpose of this study was to develop a finite element model (FEM) of a human orbit, who experienced a pure orbital blowout fracture, to study the effect of the geometrical mismatch-induced stresses on the orbital floor/graft interface and how to improve the graft design when restoring the orbital floor. MATERIALS AND METHODS: A FEM of the orbit and the globe of 1 patient who experienced pure orbital blowout fracture and treated with autogenous bone graft was generated based on computed tomographic scans. Simulations were performed with a computer using a commercially available finite element software NISA (EMRC, Troy, MI). The FEM was then used to study the effects of changing the geometry, position, material properties, and method of fixation of the autogenous bone graft on its predictions. RESULTS: The factors that had the biggest impact on the predicted principal strain magnitudes were absence of cancellous bone (up to 60%) and bony support of the graft (up to 50%). Applying rigid fixation reduced stresses by 30% posteriorly and by almost 100% anteriorly. Alterations to the geometry of the bone graft, such as an increase in its thickness, increased principal strain magnitudes (up to 42%). CONCLUSIONS: Applying rigid fixation reduced principal stresses significantly. The role of rigid fixation becomes more prominent when there is no bony support posteriorly and/or medially. This study also highlights the importance of preserving cancellous bone, when harvesting and preparing the autogenous bone graft to reconstruct the orbital floor. The possibility that absence of cancellous bone and the resulting stresses may be a source of graft resorption and/or failure cannot be excluded.

Island osteoperiosteal flaps with interpositional bone grafting in rabbit tibia: preliminary study for development of new bone augmentation technique.

PURPOSE: Loss of alveolar vertical height is one of the most challenging conditions in the field of implantology. A few augmentation techniques have been proposed to address this challenge, including guided bone regeneration, alveolar distraction osteogenesis, sandwich osteotomies and autogenous block grafting. This is a pre-clinical study of lengthening the alveolar bone height using a thin osteoperiosteal bone flap, the "island bone flap" or i-flap, to establish vitality, stability and incorporation of the augmentation material after healing. MATERIALS AND METHODS: A rabbit tibia model was designed for this study and included 8 rabbits. An osteoperiosteal osteotomy was performed through the periosteum cutting 1 mm of the outer cortex of the bone in order to elevate a bone flap. The bone flap was detached, and remained attached to the periosteum as a free floating osteoperiosteal flap, and the resultant defect was filled with xenograft. RESULTS: The histological analysis demonstrates formation of maturing trabeculae in the site of the i-flap. In cases of trauma to the bone marrow during the surgical procedure, a centripetal gradient of bone remodeling from the surgical site toward the periphery was evident, while the bony component of the i-flap maintained vital. CONCLUSIONS: Creating a free floating osteoperiosteal flap, used here with interpositional grafting in a rabbit tibia, appears to heal by both appositional modeling and creeping substitution remodeling. Volumetric augmentation persisted despite exuberant bone turnover in this animal model. This technique holds promise as a possible augmentation bone grafting approach for use in alveolar reconstruction.

The effect of impaction and a bioceramic coating on bone ingrowth in porous titanium particles.

BACKGROUND AND PURPOSE: Porous titanium (Ti) particles can be impacted like cancellous allograft bone particles, and may therefore be used as bone substitute in impaction grafting. We evaluated the effect of impaction and of a thin silicated biphasic calcium phosphate coating on osteoconduction by Ti particles. METHODS: The bone conduction chamber of Aspenberg was used in goats and filled with various groups of coated or uncoated small Ti particles (diameter 1.0-1.4 mm). Impacted allograft bone particles and empty chambers were used in control groups. Fluorochromes were administered at 4, 8, and 12 weeks. Maximum bone ingrowth distance was evaluated by histomorphometric analysis. RESULTS: Histology of Ti particle graft cylinders showed a dense matrix with narrow inter-particle and intra-particle pores (< 100 µm), occluding the lumen of the bone chamber. Bone ingrowth distances gradually increased with time in all groups. Maximum bone ingrowth distance was higher in originally empty chambers than those with allograft bone particles (p = 0.01) and Ti particles (p < 0.001). Maximum bone ingrowth in allograft bone particles was higher than in all Ti groups (p ≤ 0.001). Impaction reduced osteoconduction and the coating partially compensated for the negative effect of impaction, but these differences were not statistically significant. No osteolytic reactions were found. INTERPRETATION: Osteoconduction in the bone conduction chamber was reduced more by the insertion of small Ti particles than by insertion of small allograft bone particles. The osteoconductive potential of porous Ti particles should be studied further with larger-sized particles, which may allow bone ingrowth after impaction through larger inter-particle pores.

Platelet-rich plasma (PRP) enhances bone healing in non-united critical-sized defects: a preliminary study involving rabbit models.

INTRODUCTION: The use of bone grafts in treating non- or delayed unions as the result of large bone loss is well established. However, despite good outcomes, the time to achieve complete union is still considerably long. To overcome this problem, the use of platelet-rich plasma (PRP) has been advocated albeit with varying success. To determine the true effectiveness of PRP in treating non-/delayed unions, a study was conducted using (n=12) rabbit models. METHODS AND MATERIALS: Critical-sized defects measuring 2cm created in the midshaft of the right rabbit tibias were stabilised using 2.7-mm small fragment plates. A spacer placed in the defects to create a delay in bone union was replaced at 3 weeks with artificial bone grafts (Coragraft®), with or without PRP. The operated limbs were radiographed following the defect creation and at 3, 7 and 11 weeks (at sacrifice). Bone healing and histological changes were later assessed and scored using the appropriate grading systems. Four groups were compared for quality of healing: (group-A) control group, that is, no PRP or Coragraft; (group-B) PRP; (group-C) Coragraft; and (group-D) PRP and Coragraft. RESULTS: Group-D demonstrated the best bone healing based on radiological, histological and gross findings (Kruskall-Wallis: p<0.05). Group-C had significantly higher scores than group-B, whilst group-A had significantly lower scores than all other groups (Mann-Whitney U: p<0.05). CONCLUSION: The use of PRP with bone graft significantly improves the quality of bone healing. However, the use of PRP without bone substitute does not provide adequate repair tissue and, therefore, provides little benefit when used independently.

Influence of space-filling materials in subantral bone augmentation: blood clot vs. autogenous bone chips vs. bovine hydroxyapatite.

AIM: The first objective of the present study was to compare the short- and long-term 3D volume stability of sub-sinusal bone regeneration in rabbits using different space fillers. The second objective was to assess qualitatively and quantitatively the early bone formation process and long-term behavior of the regenerated bone. MATERIALS AND METHODS: Fifteen rabbits underwent a double sinus lift procedure using: blood clot (Clot), autogenous bone chips (Auto) and bovine hydroxyapatite (BHA). Animals were euthanized at 1 week, 5 weeks and 6 months. Samples were subjected to X-ray microtomography and histology. Variations in the volume of bone augmentations were calculated at different time points. Qualitative analysis was performed using 7 µm sections and quantitative histomorphometric analyses were carried out using scanning electron microscopy. RESULTS: From baseline (100%) to 5 weeks, the augmented volumes declined to 17.3% (Clot), 57.6% (Auto) and 90.6% (BHA). After 6 months, only 19.4% (Clot) and 31.4% (Auto) of initial volumes were found, while it remained more stable in the BHA group (84%). At 1 week, an initial osteogenesis process could be observed in the three groups along the bone walls. At 5 weeks, despite a significant decline in the volume, newly formed bone density was higher with Clot and Auto than with BHA. At 6 months, bone densities were statistically similar in the three groups. However, after 6 months, the surface invaded by newly formed bone (regenerated area) was significantly higher when BHA was used as space filler. In the BHA group, the biomaterial area slightly decreased from 42.7% (1 week) to 40% (5 weeks) and 34.9% (6 months) and the density of the composite regenerated tissue (bone+BHA) reached >50% at 6 months. CONCLUSIONS AND CLINICAL IMPLICATIONS: The three space fillers allowed bone formation to occur. Nevertheless, augmented volumes declined in the Clot and Auto groups, while they remained stable with BHA. A slowly resorbable biomaterial might be suitable in sub-sinusal bone augmentation for preventing the re-expansion process and for augmenting the density of the regenerated tissues.

Microvascular bone grafting: a new long-term solution for intraosseous arteriovenous malformations of the mandible in children.

BACKGROUND: Intraosseous arteriovenous malformations (AVMs) in the maxillofacial area are rarely reported in the current literature. These malformations have been associated with severe hemorrhage resulting in significant morbidity and mortality. The recommended gold standard treatment of AVMs is an endovascular embolization, combined with surgery. Especially in children, disease management remains a challenge for the surgeon due to the process of bone growth. AIM: In this report, we describe our experience with a microvascular bone graft as another possible surgical technique for the treatment of intraosseous AVMs in children. PATIENT AND METHODS: A 15-year-old boy was admitted to our department with a life-threatening hemorrhage due to an AVM of the left mandible. The attempt of an ordinary tooth extraction had lead to the emergency. Several embolizations and surgical interventions were required. Finally, a bony reconstruction with a microvascular bone graft from the right iliac crest was performed in order to achieve a normal form and function of the mandible. Postoperative recovery of the patient was unremarkable, and no recurrence was reported. Dental rehabilitation and a good esthetic outcome were achieved by insertion of dental implants. RESULTS: The performed interventions resulted in a complete anatomic and clinical cure. CONCLUSION: We suggest microvascular bone grafts from the anterior iliac crest as a valuable alternative in the long-term treatment of intraosseous AVMs, especially for extensive defects and in children.

Treatment of recalcitrant, multiply operated tibial nonunions with the RIA graft and rh-BMP2 using intramedullary nails.

Nonunions of the tibia continue to present some of the most difficult challenges in orthopaedic fracture care. Whether the consequence of the initial presenting injury, co-morbidity or subsequent attempts at fixation, the biological environment is often compromised. Compounding this problem is a lack of consensus on the best approach to addressing nonunited tibia fractures, placing them at risk for multiple, and sometimes ill-informed attempts at nonunion repair. We present nine cases of recalcitrant tibial nonunions which had previously undergone 4 or more attempts at repair treated with a protocol using RIA graft, rh-BMP2 and intramedullary nail fixation.

Reconstrucción nasal con cartílago costal de banco de hueso / Nasal reconstruction with rib cartilage graft

Presentamos una serie de reporte de casos de 10 pacientes con deformidad rinoseptal de distinta etiología; traumáticas, infecciosas y quirúrgicas. Estos pacientes presentaban obstrucción nasal como principal síntoma además de la deformidad estética. Los pacientes fueron tomados de la consulta externa del servicio de Otorrinolaringología del Hospital de San José. A los pacientes se les realizó reconstrucción nasal con cartílago costal de banco de hueso, se les practicó un abordaje por técnica abierta y se utilizó el injerto de banco de hueso. Los pacientes fueron seguidos por 7 meses como mínimo y máximo 16 meses. Los pacientes se evaluaron con registro fotográfico pre y posquirúrgicamente. Dentro del periodo de seguimiento no hubo infección del sitio operatorio o extrusión del injerto. La mejoría funcional y estética fueron evaluadas con grado de satisfacción del paciente y del cirujano como buenas-excelentes. La aplicabilidad de la técnica mostró buenos resultados y aplicación en diferentes grupos etarios y etiológicos.

We are presenting a series of case reports of 10 patients with nasal deformity of a varied etiology:traumatic, infectious, and surgical. All these patients exhibited nasal obstruction as their main symptomadditional to aesthetic deformity. All patients were taken from the external consultation of the Otolaryngology service at Hospital de San José. A nasal reconstruction with costal cartilage frombone bank was performed on the patients. The patients were approached through the open techniqueapproach using the graft from the bone bank. Patients were followed at least 7 months and maximum 16 months. Patients were assessed with photographic record pre and post-operatory. During the follow-up period there was no infection of the operatory site nor was there extrusion of the graft. Functional and aesthetic improvement was assessed with good-excellent as the degree of satisfaction of both the patients and the surgeon. The applicability of the technique showed good results as well as its application to various age and etiologic groups.

Bone regeneration in surgically created defects filled with autogenous bone: an epifluorescence microscopy analysis in rats.

UNLABELLED: Although the search for the ideal bone substitute has been the focus of a large number of studies, autogenous bone is still the gold standard for the filling of defects caused by pathologies and traumas, and mainly, for alveolar ridge reconstruction, allowing the titanium implants installation. OBJECTIVES: The aim of this study was to evaluate the dynamics of autogenous bone graft incorporation process to surgically created defects in rat calvaria, using epifluorescence microscopy. MATERIAL AND METHODS: Five adult male rats weighing 200-300 g were used. The animals received two 5-mm-diameter bone defects bilaterally in each parietal bone with a trephine bur under general anesthesia. Two groups of defects were formed: a control group (n=5), in which the defects were filled with blood clot, and a graft group (n=5), in which the defects were filled with autogenous bone block, removed from the contralateral defect. The fluorochromes calcein and alizarin were applied at the 7th and 30th postoperative days, respectively. The animals were killed at 35 days. RESULTS: The mineralization process was more intense in the graft group (32.09%) and occurred mainly between 7 and 30 days, the period labeled by calcein (24.66%). CONCLUSIONS: The fluorochromes showed to be appropriate to label mineralization areas. The interfacial areas between fluorochrome labels are important sources of information about the bone regeneration dynamics.

Quantitative assessment of the bone morphogenetic protein expression from alternate bone graft harvesting sites.

OBJECTIVE: Bone morphogenetic proteins (BMPs) play important roles in the stimulation of osteogenesis and osteoinduction during bone fracture healing and their expression levels may be important for bone graft efficacy. The objective of this study was to determine if there are variations in the expression of BMPs and their receptors in various bone graft harvesting sites. We analyzed autogenous marrow aspirates obtained from three different graft sites for the mRNA levels of BMPs and their receptors. METHODS: Using real-time polymerase chain reaction, we analyzed the mRNA levels of BMPs and their receptors in autogenous bone marrow aspirates obtained from three different bone graft sites of 10 different human subjects. Collection of autogenous bone marrow from the iliac crest, the proximal humerus, and the proximal tibia was performed using standard sterile techniques in the operating room as part of surgery to treat an established fracture nonunion. RESULTS: The mRNA levels of BMP-2 and BMP-5 were the highest in the bone marrow aspirates from the three different sites, whereas the mRNA levels of the other osteoinductive BMPs (BMP-4, -5, -6, -7, -8, and -9) were lower. The mRNA levels of BMP-3, an inhibitor of osteogenesis, were the lowest in the bone marrow aspirates of all three different sites. There were no statistical significant differences in the mRNA levels of any of the BMPs or their receptors investigated in this study in the bone marrow of the three different sites. CONCLUSION: Because no statistical significant differences in the mRNA levels of the BMPs and their receptors were detected in the bone marrow aspirates from the three different sites, our findings suggest that potential differences of various graft sites in the augmentation of bone healing does not result from different expression levels of BMPs.