RESUMO
Serum albumin-coated bone allografts (BoneAlbumin) have successfully supported bone regeneration in various experimental models by activating endogenous progenitors. However, the effect of tissue aging, linked to declining stem cell function, has yet to be explicitly examined within the context of BoneAlbumin's regenerative capacity. Stem cell function was tested with an in vitro attachment assay, which showed that albumin coating increases stem cell attachment on demineralized bone surfaces in an aging cell population. Bone regeneration was investigated in vivo by creating critical size bone defects on the parietal bones of aging female rats. Demineralized bone matrices with and without serum albumin coating were used to fill the defects. Bone regeneration was determined by measuring the density and the size of the remaining bone defect with computed tomography (CT). Microcomputed tomography (MicroCT) and mechanical testing were performed on the parietal bone explants. In vivo CT and ex vivo microCT measurements showed better regeneration with albumin-coated grafts. Additionally, the albumin-coated group showed a twofold increase in peak fracture force compared with uncoated allografts. In the present study, serum albumin-coated demineralized bone matrices successfully supported faster and functionally superior bone regeneration in aging rats. Because stem cell function, a key contributor of bone remodelling, decreases with age and serum albumin is an effective activator of endogenous progenitor cells, this method could be an effective and safe adjuvant in bone regeneration of aging adult and osteo-compromised populations.
Assuntos
Envelhecimento/fisiologia , Aloenxertos/fisiologia , Transplante Ósseo , Osso e Ossos/fisiologia , Materiais Revestidos Biocompatíveis/farmacologia , Osteogênese/efeitos dos fármacos , Albumina Sérica/farmacologia , Aloenxertos/efeitos dos fármacos , Animais , Fenômenos Biomecânicos , Osso e Ossos/efeitos dos fármacos , Adesão Celular/efeitos dos fármacos , Feminino , RatosRESUMO
INTRODUCTION: The aim of this study was to compare the paranasal sinus volumes obtained by manual and semiautomatic imaging software programs using both CT and CBCT imaging. METHODS: 121 computed tomography (CT) and 119 cone beam computed tomography (CBCT) examinations were selected from the databases of the authors' institutes. The Digital Imaging and Communications in Medicine (DICOM) images were imported into 3-dimensonal imaging software, in which hand mode and semiautomatic tracing methods were used to measure the volumes of both maxillary sinuses and the sphenoid sinus. The determined volumetric means were compared to previously published averages. RESULTS: Isometric CBCT-based volume determination results were closer to the real volume conditions, whereas the non-isometric CT-based volume measurements defined coherently lower volumes. By comparing the 2 volume measurement modes, the values gained from hand mode were closer to the literature data. Furthermore, CBCT-based image measurement results corresponded to the known averages. CONCLUSIONS: Our results suggest that CBCT images provide reliable volumetric information that can be depended on for artificial organ construction, and which may aid the guidance of the operator prior to or during the intervention.
Assuntos
Tomografia Computadorizada de Feixe Cônico/métodos , Processamento de Imagem Assistida por Computador/métodos , Prótese Maxilofacial , Seios Paranasais , Órgãos Artificiais/normas , Humanos , Tamanho do Órgão , Seios Paranasais/diagnóstico por imagem , Seios Paranasais/patologia , Seios Paranasais/cirurgiaRESUMO
Blood serum fractions are hotly debated adjuvants in bone replacement therapies. In the present experiment, we coated demineralized bone matrices (DBM) with serum albumin and investigated stem cell attachment in vitro and bone formation in a rat calvaria defect model. In the in vitro experiments, we observed that significantly more cells adhere to the serum albumin coated DBMs at every time point. In vivo bone formation with albumin coated and uncoated DBM was monitored biweekly by computed tomography until 11 weeks postoperatively while empty defects served as controls. By the seventh week, the bone defect in the albumin group was almost completely closed (remaining defect 3.0 ± 2.3%), while uncoated DBM and unfilled control groups still had significant defects (uncoated: 40.2 ± 9.1%, control: 52.4 ± 8.9%). Higher density values were also observed in the albumin coated DBM group. In addition, the serum albumin enhanced group showed significantly higher volume of newly formed bone in the microCT analysis and produced significantly higher breaking force and stiffness compared to the uncoated grafts (peak breaking force: uncoated: 15.7 ± 4 N, albumin 46.1 ± 11 N). In conclusion, this investigation shows that implanting serum albumin coated DBM significantly reduces healing period in nonhealing defects and results in mechanically stronger bone. These results also support the idea that serum albumin coating provides a convenient milieu for stem cell function, and a much improved bone grafting success can be achieved without the use of exogenous stem cells.
Assuntos
Materiais Revestidos Biocompatíveis , Matriz Extracelular/química , Osteogênese/efeitos dos fármacos , Crânio/lesões , Células-Tronco/metabolismo , Animais , Técnica de Desmineralização Óssea , Adesão Celular , Materiais Revestidos Biocompatíveis/química , Materiais Revestidos Biocompatíveis/farmacologia , Masculino , Ratos , Ratos Wistar , Albumina Sérica , Crânio/metabolismo , Crânio/patologiaRESUMO
The key drawback of using demineralized bone matrix (DBM) is its low initial mechanical stability due to the severe depletion of mineral content. In the present study, we investigated the long-term regeneration of DBM in a critical size bone defect model and investigated the remineralization after 6 months. Bone defects were created in the cranium of male Wistar rats which were filled with DBM or left empty as negative control. In vivo bone formation was monitored with computed tomography after 11, 19, and 26 weeks postoperatively. After 6 months, parietal bones were subjected to micro-CT. Mineral content was determined with spectrophotometric analysis. After 11 weeks the DBM-filled bone defects were completely closed, while empty defects were still open. Density of the DBM-treated group increased significantly while the controls remained unchanged. Quantitative analysis by micro-CT confirmed the in vivo results, bone volume/tissue volume was significantly lower in the controls than in the DBM group. The demineralization procedure depleted the key minerals of the bone to a very low level. Six months after implantation Ca, P, Na, Mg, Zn, and Cr contents were completely restored to the normal level, while K, Sr, and Mn were only partially restored. The remineralization process of DBM is largely complete by the 6th month after implantation in terms of bone density, structure, and key mineral levels. Although DBM does not provide sufficient sources for any of these minerals, it induces a faster and more complete regeneration process. © 2015 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 104B: 1336-1342, 2016.
Assuntos
Calcificação Fisiológica , Matriz Extracelular/transplante , Osteogênese , Crânio , Microtomografia por Raio-X , Animais , Seguimentos , Masculino , Ratos , Ratos Wistar , Crânio/diagnóstico por imagem , Crânio/lesões , Crânio/metabolismo , Crânio/patologiaRESUMO
Partitioning of polypeptides between protein folding and amyloid formation is of outstanding pathophysiological importance. Using yeast phosphoglycerate kinase as model, here we identify the features of the energy landscape that decide the fate of the protein: folding or amyloidogenesis. Structure formation was initiated from the acid-unfolded state, and monitored by fluorescence from 10 ms to 20 days. Solvent conditions were gradually shifted between folding and amyloidogenesis, and the properties of the energy landscape governing structure formation were reconstructed. A gradual transition of the energy landscape between folding and amyloid formation was observed. In the early steps of both folding and misfolding, the protein searches through a hierarchically structured energy landscape to form a molten globule in a few seconds. Depending on the conditions, this intermediate either folds to the native state in a few minutes, or forms amyloid fibers in several days. As conditions are changed from folding to misfolding, the barrier separating the molten globule and native states increases, although the barrier to the amyloid does not change. In the meantime, the native state also becomes more unstable and the amyloid more stable. We conclude that the lower region of the energy landscape determines the final protein structure.