RESUMO
BACKGROUND: Periprosthetic joint infection is a major complication of total joint arthroplasty, with treatment requiring a two-stage exchange procedure and 6 weeks of systemic antibiotics. However, depending on the infection site, intravenous delivery of antibiotics like vancomycin (VCM) can have poor tissue transferability, thus reducing their therapeutic effect. OBJECTIVE: This study demonstrates the 24-week in vivo release profile and antibacterial activity of VCM from calcium phosphate cement impregnated with VCM (CPC/VCM) and compares them with those from polymethylmethacrylate impregnated with VCM (PMMA/VCM). METHODS: Rats were implanted with the test specimens between the fascia and quadriceps. After implantation for 24 weeks, the test specimens were removed and residual VCM was extracted to calculate the concentration of VCM released into rat tissues. We also examined the antibacterial activity of releasable VCM from the removed test specimens by placing them directly onto the surface of agar. RESULTS: CPC/VCM released greater concentrations of VCM for a longer period of time within the 24 weeks than PMMA/VCM. Moreover, CPC/VCM released 1.4 to 26.1-fold more VCM than PMMA/VCM. Using Staphylococcus aureus, antibacterial activity was logarithmically correlated with VCM concentration across the entire concentration range tested (12.5-800 µg/mL). While the area within which inhibition was observed-the inhibition zone-for both CPC/VCM and PMMA/VCM formed and gradually shrank with time after implantation, that for CPC/VCM was significantly larger than that for PMMA/VCM in each week after implantation. CONCLUSION: CPC/VCM releases greater amounts of VCM with antibacterial activity for longer periods of time than PMMA/VCM, suggesting that CPC is effective for facilitating the release of antibiotics for local action in patients with established postoperative infection.
Assuntos
Cimentos Ósseos , Vancomicina , Animais , Antibacterianos/farmacologia , Antibacterianos/uso terapêutico , Fosfatos de Cálcio , Humanos , Polimetil Metacrilato , Ratos , Vancomicina/farmacologiaRESUMO
Bone banks are necessary for providing biological allografts for a series of orthopedic procedures. As nations cope with new realities driven by the 2019 coronavirus disease (COVID-19) pandemic, health-care providers, institutions, and patients share a particular concern about the effect of COVID-19 on organ donation and transplantation. Here, we describe the management of the Kitasato University Bone Bank during the state of emergency declared in response to COVID-19. Living donors received pre-operative screening by PCR, and allograft bone from COVID-19-negative donors was cryopreserved as transplantable tissues. The weekly rate of infection gradually increased from February 2-9 to April 5-11 in the dead donor-derived allograft bone-harvesting region covered by the Bank. It is becoming clear that the virus can be transmitted by asymptomatic patients, and that this route may have facilitated the spread of COVID-19. Therefore, the Bank stopped dead donor donation to consider the safety of medical staff. Three recipients received bone allografts following pre-operative COVID-19 screening by PCR. All patients were asymptomatic after bone allograft. Our experience may provide helpful information for the management of tissue banks.
Assuntos
Bancos de Ossos , COVID-19 , Humanos , Japão , Doadores Vivos , SARS-CoV-2RESUMO
Calcium phosphate cement (CPC) has good release efficiency and has therefore been used as a drug delivery system for postoperative infection. The release profile of CPC has mainly been evaluated by in vitro studies, which are carried out by immersing test specimens in a relatively large amount of solvent. However, it remains unclear whether antibiotic-impregnated CPC has sufficient clinical effects and release in vivo. We examined the in vivo release profile of CPC impregnated with vancomycin (VCM) and compared this with that of polymethylmethacrylate (PMMA) cement. To evaluate the release profile in vitro, the test specimens were immersed in 10 mL sterile phosphate-buffered saline per gram of test specimen and incubated at 37°C for 56 days in triplicate. For in vivo experiments, the test specimens were implanted between the fascia and muscle of the femur of rats. Residual VCM was extracted from the removed test specimens to determine the amount of VCM released into rat tissues. CPC released more VCM over a longer duration than PMMA in vitro. Released levels of VCM from CPC/VCM in vivo were 3.4-fold, 5.0-fold, and 8.6-fold greater on days 1, 7, and 28, respectively, than those released on the corresponding days from PMMA/VCM and were drastically greater on day 56 due to inefficient release from PMMA/VCM. The amount of VCM released from CPC and PMMA was much higher than the minimum inhibitory concentration (1.56 µg) and lower than the detection limit, respectively. Our findings suggest that CPC is a suitable material for releasing antibiotics for local action against established postoperative infection.
Assuntos
Cimentos Ósseos , Polimetil Metacrilato , Vancomicina , Animais , Cimentos Ósseos/química , Cimentos Ósseos/farmacocinética , Cimentos Ósseos/farmacologia , Preparações de Ação Retardada/química , Preparações de Ação Retardada/farmacocinética , Preparações de Ação Retardada/farmacologia , Polimetil Metacrilato/química , Polimetil Metacrilato/farmacocinética , Polimetil Metacrilato/farmacologia , Ratos , Ratos Wistar , Vancomicina/química , Vancomicina/farmacocinética , Vancomicina/farmacologiaRESUMO
Low-Intensity Pulsed Ultrasound (LIPUS) provided a mechanical stimulus, and was thought to promote fracture healing by signal transduction through integrin, a cytoskeletal protein. Meanwhile, teriparatide, a drug for osteoporosis treatment, showed efficacy in promoting bone metabolism. This drug also appeared to prevent fractures in patients with serious osteoporosis by improving bone mineral density and bone quality, which in turn resulted from promoting action for bone metabolism. Further, clinical trials and fundamental research reported that teriparatide demonstrated the effect of promoting fracture healing. Mechanical stimulus by LIPUS had a topical effect on fractures; on the other hand, teriparatide (peptide hormone) had both topical and systemic effects. Both LIPUS and teriparatide had the effect of fracture healing, but it was supposed that the characteristics of each effect were different because of the different mechanism of action. Moreover, the combination therapy of LIPUS and teriparatide was expected to produce synergies. We used elderly rats as models for the femoral fracture to examine the effects of LIPUS and teriparatide on promoting fracture healing for treatment delay by aging. We observed the fracture healing process in 40-week-old rats as an elderly model using simple radiographs, and recognized a delay in fracture healing compared with that of 8-week-old rats. As discussed in histomorphology, it was demonstrated that the period of endochondral ossification, from chondrogenesis to teleost cross-linked callus, was prolonged and the fracture healing process was delayed by aging. Next, we treated the elderly fracture models with LIPUS for 20 minutes a day from the first day after the fracture, and compared them with non-treated models. The bone unions of the treated models were observed earlier than those of non-treated models in the simple radiographs. LIPUS shortened the period of endochondral ossification. Further, we gave the elderly fracture models teriparatide subcutaneously 5 µg/kg three times a week from the first day after the fracture. Bone unions of the treated models were observed earlier than those of non-treated models in simple radiographs as well. In micro CT analysis, it was demonstrated that lamellar bone transforming and bone remodeling of the trabecular structure of external callus were especially accelerated. The results of these trials showed that both LIPUS and teriparatide demonstrated the effect of promoting fracture healing, and each had unique characteristics.
RESUMO
We have studied the mechanism of fracture healing, and the effect of LIPUS, bone graft and growth factor on accelerating fracture healing. We present here the results of our research. To examine callus formation cells in fracture healing, we made marrow GFP chimera mice and a fracture model of marrow mesenchymal stem cell GFP chimera mice. It was demonstrated that periosteal cells were essential for callus formation. We focused on periosteal cells and examined the effect of LIPUS. In an in vitro experiment using a cultured part of the femur, LIPUS promoted ossification of the periosteal tissue. Further, LIPUS accelerated VEGF expression in the experiment using the femoral fracture model of mice. From these results, it was suggested that activation of periosteal cells might play a role in the fracture healing mechanism of LIPUS. Next, we discussed the possibility of combined therapy of LIPUS, bone graft and growth factor. Therapy involving the topical administration of bFGF using a controlled release system and bone graft could promote callus formation. In addition, LIPUS was able to promote membranaceous ossification after the bone graft. It was suggested that combined therapy of LIPUS, bone graft and bFGF could be a new option for treating fractures.
RESUMO
OBJECTIVE: We have conducted a basic study on the influences on ultrasonic properties when LIPUS is applied through wound dressing. According to the results of ex vivo experiments conducted to date, LIPUS showed ultrasonic properties such as transmittance, coefficient of transmission, and a non-uniformity ratio through film wound dressing better than other wound dressing, and it was considered that LIPUS's effect for fracture healing was not influenced by film wound dressing. Then, we discussed the influence on the effect of LIPUS through film wound dressing. METHODS: Thirty male 8-week-old Sprague-Dawley rats were used for the trial. After creating close transverse femoral fractures on the right legs of these 30 rats, they were divided into 3 groups of 10; LIPUS through wound dressing (Group A), LIPUS without wound dressing (Group B), and No LIPUS treatment (Group C). OPSITE Wound, which was thought to have the least influence on ultrasound properties, was used for this trial. Group A and B received LIPUS for 20 minutes a day from the first day after the fractures. LIPUS was generated from Teijin Pharma's device for a basic experiment. When treating Group A, the wound dressing was pasted on the ultrasound terminal in order to apply LIPUS through the dressing. We assessed the time-oriented morphological change of each group in anesthetized condition using simple radiographs on the 8th, 16th, and 24th day after the fractures. RESULTS: Six rats in Group A, 2 in Group B, and 1 in Group C died in anesthesia, and we discussed the remaining 4 rats in Group A, 8 in Group B, and 9 in Group C. We defined more than one teleost callus bridging as bone-union. We also counted a bone remodeling when we recognized the absorption of existing cortical bone and the transformation of new bone to cortical bone in simple radiographs. As a result, compared with Group C, we recognized that both bone union and remodeling accelerated remarkably in Group B, but not in Group A. DISCUSSION: It suggested that LIPUS through wound dressing had negative influences on both period shorting of fracture healing and bone remodeling. When LIPUS was conducted through film wound dressing, transmittance and coefficient of transmission were unchanged; however, the non-uniformity ratio changed slightly. The non-uniformity ratio of the ultrasound transducer had a significant influence on the effect of LIPUS on fracture healing.
RESUMO
Although recent studies suggest that hyperlipidemia is a risk factor for osteoarthritis (OA), the link between OA and hyperlipidemia is not fully understood. As the number of activated, circulating myeloid cells is increased during hyperlipidemia, we speculate that myeloid cells contribute to the pathology of OA. Here, we characterized myeloid cells in STR/Ort mice, a murine osteoarthritis model, under hyperlipidemic conditions. Ratios of myeloid cells in bone marrow, the spleen, and peripheral blood were determined by flow cytometry. To examine the influence of the hematopoietic environment, including abnormal stem cells, on the hematopoietic profile of STR/Ort mice, bone marrow transplantations were performed. The relationship between hyperlipidemia and abnormal hematopoiesis was examined by evaluating biochemical parameters and spleen weight of F2 animals (STR/Ort x C57BL/6J). In STR/Ort mice, the ratio of CD11b(+)Gr1(+) cells in spleens and peripheral blood was increased, and CD11b(+)Gr1(+) cells were also present in synovial tissue. Splenomegaly was observed and correlated with the ratio of CD11b(+)Gr1(+) cells. When bone marrow from GFP-expressing mice was transplanted into STR/Ort mice, no difference in the percentage of CD11b(+)Gr1(+) cells was observed between transplanted and age-matched STR/Ort mice. Analysis of biochemical parameters in F2 mice showed that spleen weight correlated with serum total cholesterol. These results suggest that the increase in circulating and splenic CD11b(+)Gr1(+) cells in STR/Ort mice originates from hypercholesterolemia. Further investigation of the function of CD11b(+)Gr1(+) cells in synovial tissue may reveal the pathology of OA in STR/Ort mice.
Assuntos
Hiperlipidemias/patologia , Células Mieloides/imunologia , Células Mieloides/patologia , Osteoartrite/patologia , Membrana Sinovial/citologia , Membrana Sinovial/patologia , Animais , Antígenos Ly , Antígeno CD11b , Movimento Celular , Hiperlipidemias/complicações , Hiperlipidemias/imunologia , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Endogâmicos , Osteoartrite/complicações , Osteoartrite/imunologiaRESUMO
As the in vivo function of bone marrow-engrafted umbilical cord blood (UCB)-derived mesenchymal cells (UCBCs) after UCB transplantation is unknown, we examined in vivo osteoblastic differentiation using mouse UCB transplantation and fracture models. UCBCs obtained from GFP transgenic mice were intravenously injected into irradiated C57BL/6 mice. After three months, the in vivo osteoblastic differentiation potential of bone marrow-engrafted UCBCs was examined histologically using a mouse fracture model. GFP-positive UCBCs were detected in the bone marrow of recipient mice. On day 7, UCBCs were observed in the fracture gap and surrounding the titanium screws of the fixation device. The UCBCs were also positive for alkaline phosphatase and von Kossa staining. By day 14, UCBCs were observed around and within a formed intramedullary callus. The newly formed woven bone consisted of ALP- and von Kossa-positive cells. Our findings suggest that UCBCs contribute to the fracture healing process after bone marrow engraftment and that UCBC transplantation can fully reconstruct not only hematopoietic cells but also mesenchymal cell lineages.
Assuntos
Células da Medula Óssea , Calo Ósseo/citologia , Calo Ósseo/fisiologia , Fraturas do Fêmur/fisiopatologia , Células-Tronco Fetais/citologia , Consolidação da Fratura , Osteogênese , Animais , Placas Ósseas , Parafusos Ósseos , Diferenciação Celular , Linhagem da Célula , Transplante de Células-Tronco de Sangue do Cordão Umbilical , Modelos Animais de Doenças , Feminino , Células-Tronco Fetais/química , Citometria de Fluxo , Proteínas de Fluorescência Verde/química , Humanos , Injeções Intravenosas , Camundongos , Camundongos Endogâmicos C57BL , Osteoblastos/citologia , Titânio , Quimeras de TransplanteRESUMO
The incidence of spontaneous osteoarthritis (OA) in female STR/Ort mice is much lower than that observed in male STR/Ort mice; however, the reason for the differential incidence of OA between sexes has not been elucidated. Here, we investigated and compared age- and sex-related bone mineral density and architectural changes in male and female STR/Ort mice. Bone architecture and bone mineral density (BMD) of femurs were examined in 5-, 10-, 15-, 20-, and 35-week-old male and female STR/Ort mice by microscopic computed tomography (µCT). Angular degrees of internal tibial torsion (ADITT) were also measured in mice at 5, 15, and 35 weeks of age. Earlier decreases of cancellous volume and BMD were found in male STR/Ort mice. Using µCT, an age-related decline of bone marrow space in femoral diaphysis was observed in both males and females but was more dramatic in females. In addition, an earlier increase of ADITT was observed in male STR/Ort mice, suggesting that internal rotation of the tibia may contribute to OA. Age- and sex-related bone architectural changes clearly differ between male and female STR/Ort mice. These differences in bone structure, particularly ADITT, may explain the differential incidence of OA in STR/Ort mice.
Assuntos
Artrite Experimental/patologia , Densidade Óssea , Osteoartrite do Joelho/patologia , Ovariectomia/efeitos adversos , Fatores Etários , Animais , Artrite Experimental/diagnóstico por imagem , Artrite Experimental/epidemiologia , Modelos Animais de Doenças , Feminino , Humanos , Articulação do Joelho/citologia , Masculino , Camundongos , Camundongos Endogâmicos , Osteoartrite do Joelho/diagnóstico por imagem , Osteoartrite do Joelho/epidemiologia , Radiografia , Fatores Sexuais , Tíbia/diagnóstico por imagem , Tíbia/patologia , Tomografia Computadorizada de Emissão , Anormalidade Torcional/patologiaAssuntos
Artroplastia de Quadril/métodos , Cimentos Ósseos , Prótese de Quadril , Idoso , Feminino , Humanos , Desenho de Prótese , ReoperaçãoRESUMO
Frozen bone-patellar tendon bone allografts are useful in anterior cruciate ligament reconstruction as the freezing procedure kills tissue cells, thereby reducing immunogenicity of the grafts. However, a small portion of cells in human femoral heads treated by standard bone-bank freezing procedures survive, thus limiting the effectiveness of allografts. Here, we characterized the survival rates and mechanisms of cells isolated from rat bones and tendons that were subjected to freeze-thaw treatments, and evaluated the influence of these treatments on the mechanical properties of tendons. After a single freeze-thaw cycle, most cells isolated from frozen bone appeared morphologically as osteocytes and expressed both osteoblast- and osteocyte-related genes. Transmission electron microscopic observation of frozen cells using freeze-substitution revealed that a small number of osteocytes maintained large nuclei with intact double membranes, indicating that these osteocytes in bone matrix were resistant to ice crystal formation. We found that tendon cells were completely killed by a single freeze-thaw cycle, whereas bone cells exhibited a relatively high survival rate, although survival was significantly reduced after three freeze-thaw cycles. In patella tendons, the ultimate stress, Young's modulus, and strain at failure showed no significant differences between untreated tendons and those subjected to five freeze-thaw cycles. In conclusion, we identified that cells surviving after freeze-thaw treatment of rat bones were predominantly osteocytes. We propose that repeated freeze-thaw cycles could be applied for processing bone-tendon constructs prior to grafting as the treatment did not affect the mechanical property of tendons and drastically reduced surviving osteocytes, thereby potentially decreasing allograft immunogenecity.
Assuntos
Transplante Ósseo , Osso e Ossos/citologia , Congelamento , Osteócitos/citologia , Tendões/citologia , Tendões/fisiologia , Animais , Biomarcadores/metabolismo , Fenômenos Biomecânicos/fisiologia , Separação Celular , Forma Celular , Sobrevivência Celular , Fêmur/citologia , Regulação da Expressão Gênica , Humanos , Masculino , Osteoblastos/citologia , Osteoblastos/ultraestrutura , Osteócitos/ultraestrutura , Ratos , Ratos Wistar , Tendões/transplante , Tíbia/citologiaRESUMO
To eliminate the potential for infection, many tissue banks routinely process and terminally sterilize allografts prior to transplantation. A number of techniques, including the use of scanning electron microscopy, bone graft models, and mechanical property tests, are used to evaluate the properties of allograft bone. However, as these methods are time consuming and often destroy the bone sample, the quality assessment of allograft bones are not routinely performed after processing and sterilization procedures. Raman spectroscopy is a non-destructive, rapid analysis technique that requires only small sample volumes and has recently been used to evaluate the mineral content, mineral crystallinity, acid phosphate and carbonate contents, and collagen maturity in human and animal bones. Here, to establish a quality assessment method of allograft bones using Raman spectroscopy, the effect of several common sterilization and preservation procedures on rat femoral bones were investigated. We found that freeze-thawing had no detectable effects on the composition of bone minerals or matrix, although heat treatment and gamma irradiation resulted in altered Raman spectra. Our findings suggest Raman spectroscopy may facilitate the quality control of allograft bone after processing and sterilization procedures.
Assuntos
Densidade Óssea , Matriz Óssea , Transplante Ósseo , Raios gama/efeitos adversos , Análise Espectral Raman , Animais , Densidade Óssea/efeitos da radiação , Matriz Óssea/efeitos da radiação , Fêmur/química , Fêmur/efeitos da radiação , Congelamento/efeitos adversos , Temperatura Alta/efeitos adversos , Masculino , Controle de Qualidade , Ratos , Ratos Wistar , Esterilização/métodosRESUMO
BACKGROUND: Although several types of culture medium have been used for preservation of osteochondral allografts, the viability of chondrocytes decreases with increasing storage duration. We previously showed the University of Wisconsin solution is more suitable for graft preservation than culture medium. QUESTIONS/PURPOSES: We determined whether the addition of allogenic serum to University of Wisconsin solution increases chondrocyte survival during prolonged storage of osteochondral allografts. METHODS: Osteochondral tissue samples harvested from the distal femora of rats were preserved in University of Wisconsin solution supplemented with 0%, 1%, 10%, and 50% allogenic serum at 4 °C for 14 days. Cell viability and chondrocyte degenerative changes of the samples then were assessed using a tetrazolium assay and histologic methods. We also evaluated time-dependent changes in cell viability and histologic findings of samples preserved for 7, 14, and 21 days in University of Wisconsin solution supplemented with or without 10% allogenic serum. RESULTS: After 14 days of preservation, osteochondral tissue samples maintained in University of Wisconsin solution containing 10% or greater allogenic serum exhibited the highest cell viability and lowest degenerative changes in chondrocytes. In the evaluation of time-dependent changes, we found the chondrocyte degenerative changes were greater in cartilage preserved in University of Wisconsin solution alone than in University of Wisconsin solution containing 10% allogenic serum after day 7 or later. CONCLUSIONS: Our results suggest the addition of 10% allogenic serum to University of Wisconsin solution enhances viability of osteochondral tissue samples. CLINICAL RELEVANCE: The use of allogenic serum-supplemented University of Wisconsin solution is expected to prolong the duration of osteochondral allograft storage and result in higher-quality grafts.
Assuntos
Cartilagem/transplante , Condrócitos , Temperatura Baixa , Soluções para Preservação de Órgãos , Soro , Preservação de Tecido/métodos , Adenosina , Alopurinol , Animais , Sobrevivência Celular , Glutationa , Insulina , Masculino , Rafinose , Ratos , Ratos Sprague-DawleyRESUMO
The aim of this study is to clarify the effect of low intensity pulsed ultrasound (LIPUS) on shortening of the fracture healing period and endochondral ossification during the fracture healing process. We first established a model of aging-related delayed union fractures consisting of aged mouse (C57BL/6J; 40 weeks old) with closed femur fractures. We compared the healing process of 40-week-old mice to the healing process of 8-week-old (young) mice using radiological and histological analysis. In aged mice, some cartilage formation was observed 10 days after the fracture; however, endochondral ossification and hard callus bridging were observed 21 and 28 days after the fracture, respectively, whereas cartilage remained in the callus on day 28, suggesting delayed endochondral ossification following bone remodeling. Meanwhile, in aged mice with LIPUS treatment, cartilage formation was similar to that in aged mice without LIPUS; however, hard callus bridging and bone remodeling were observed 21 and 28 days after fracture, respectively, suggesting that LIPUS shortened the healing period due to promotion of endochondral ossification. Immunohistochemical analysis showed marked expression of vascular endothelial growth factor and neovascularization in the fibrous tissue comprising the periosteum that surrounded the whole callus. A cell migration test involving primary cultured human endothelial cells also showed promotion of cell migration by LIPUS. These results suggested that endothelial cell migration and neovascularization, which were observed around fracture sites, played a part in the mechanism of promotion of endochondral ossification by LIPUS.
Assuntos
Fraturas do Fêmur/terapia , Fêmur/patologia , Consolidação da Fratura , Fraturas Fechadas/terapia , Fraturas não Consolidadas/terapia , Osteogênese , Terapia por Ultrassom/métodos , Animais , Remodelação Óssea , Calo Ósseo/citologia , Calo Ósseo/diagnóstico por imagem , Movimento Celular , Fraturas do Fêmur/diagnóstico por imagem , Fraturas do Fêmur/patologia , Fêmur/diagnóstico por imagem , Fêmur/fisiopatologia , Fraturas Fechadas/diagnóstico por imagem , Fraturas Fechadas/patologia , Fraturas não Consolidadas/diagnóstico por imagem , Fraturas não Consolidadas/patologia , Humanos , Imageamento Tridimensional , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Modelos Animais , Neovascularização Fisiológica , Osteogênese/efeitos da radiação , Tomografia Computadorizada por Raios X , Fator A de Crescimento do Endotélio Vascular/metabolismoRESUMO
This study aimed to clarify the relationship between the mechanical environment at the fracture site and endogenous fibroblast growth factor-2 (FGF-2). We compared two types of fracture healing with different callus formations and cellular events using MouseFix(TM) plate fixation systems for murine fracture models. Left femoral fractures were induced in 72 ten-week-old mice and then fixed with a flexible (Group F) or rigid (Group R) Mouse Fix(TM) plate. Mice were sacrificed on days 3, 5, 7, 10, 14, and 21. The callus volumes were measured by 3D micro-CT and tissues were histologically stained with hematoxylin & eosin or safranin-O. Sections from days 3, 5, and 7 were immunostained for FGF-2 and Proliferating Cell Nuclear Antigen (PCNA). The callus in Group F was significantly larger than that in Group R. The rigid plate allowed bone union without a marked external callus or chondrogenesis. The flexible plate formed a large external callus as a result of endochondral ossification. Fibroblastic cells in the granulation tissue on days 5 and 7 in Group F showed marked FGF-2 expression compared with Group R. Fibroblastic cells showed ongoing proliferation in granulation tissue in group F, as indicated by PCNA expression, which explained the relative granulation tissue increase in group F. There were major differences in early phase endogenous FGF-2 expression between these two fracture healing processes, due to different mechanical environments.
Assuntos
Calo Ósseo/metabolismo , Calo Ósseo/fisiologia , Fraturas do Fêmur/metabolismo , Fraturas do Fêmur/fisiopatologia , Fator 2 de Crescimento de Fibroblastos/metabolismo , Consolidação da Fratura/fisiologia , Fixadores Internos , Estresse Mecânico , Animais , Proliferação de Células , Modelos Animais de Doenças , Fraturas do Fêmur/patologia , Fibroblastos/citologia , Camundongos , Antígeno Nuclear de Célula em Proliferação/metabolismoRESUMO
Many investigators are currently studying the use of decellularized tissue allografts from human cadavers as scaffolds onto which patients' cells could be seeded, or as carriers for genetically engineered cells to aid cell transplantation. However, it is difficult to seed cells onto very dense regular connective tissue which has few interstitial spaces. Here, we discuss the development of a chemotactic cell seeding technique using solvent-preserved human meniscus. A chemokinetic response to recombinant human bone morphogenetic protein-2 (rhBMP-2) was observed in a monolayer culture of primary chondrocytes derived from femoral epiphyseal cartilage of 2-day-old rats. The rhBMP-2 significantly increased their migration upto 10 ng/ml in a dose-dependent manner. When tested with solvent-preserved human meniscus as a scaffold, which has few interstitial spaces, rhBMP-2 was able to induce chondrocytes to migrate into the meniscus. After a 3-week incubation, newly-formed cartilaginous extracellular matrix was synthesized by migrated chondrocytes throughout the meniscus, down to a depth of 3 mm. These findings demonstrate that rhBMP-2 may be a natural chemokinetic factor in vivo, which induces migration of proliferative chondrocytes into the narrow interfibrous spaces. Our results suggest a potential application of rhBMP-2 for the designed distribution of chondrocytes into a scaffold to be used for tissue engineering.
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Proteína Morfogenética Óssea 2/metabolismo , Condrócitos/citologia , Meniscos Tibiais/citologia , Engenharia Tecidual/métodos , Fator de Crescimento Transformador beta/metabolismo , Animais , Movimento Celular , Células Cultivadas , Fêmur/citologia , Lâmina de Crescimento/citologia , Humanos , Meniscos Tibiais/ultraestrutura , Ratos , Ratos Wistar , Proteínas Recombinantes/metabolismo , SolventesAssuntos
Sangue Fetal/citologia , Sangue Fetal/transplante , Células-Tronco Mesenquimais/citologia , Células-Tronco Pluripotentes/citologia , Animais , Diferenciação Celular , Proteínas de Fluorescência Verde/genética , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Transgênicos , Músculo Esquelético/citologia , Miocárdio/citologia , Neurônios/citologia , Osteoblastos/citologiaRESUMO
To test whether mechanical loading produces faster healing in aged mice, fractured femurs of aged 1-year-old mice were subjected to low-intensity pulsed ultrasound (LIPUS), a treatment that is routinely used to help heal fractures in humans. Cyclooxygenase-2 knockout mice (COX-2(-/-)), which lack an immediate early mediator of mechanical stimulation, were also studied by histochemistry, microcomputed tomography and quantitative polymerase chain reaction to determine the role of COX-2. The healing in the aged COX-2(-/-) mice is slow during the endochondral bone remodeling (>30 d), a period generally prolonged in senescence. For aged wild-type mice, LIPUS halved the endochondral phase to about 10 d, whereas that was not the case for aged COX-2(-/-) mice, which showed no apparent shortening of the prolonged endochondral-phase healing time. Injecting prostaglandin E(2) receptor agonists, however, rescued the COX-2(-/-) callus from insensitivity to LIPUS. In conclusion, COX-2 is a limiting factor in the delayed endochondral bone healing and is induced by LIPUS, which normalizes healing rate to the wild-type level.
Assuntos
Ciclo-Oxigenase 1/metabolismo , Fraturas do Fêmur/fisiopatologia , Fraturas do Fêmur/terapia , Fêmur/fisiopatologia , Fêmur/efeitos da radiação , Consolidação da Fratura/efeitos da radiação , Proteínas de Membrana/metabolismo , Terapia por Ultrassom/métodos , Envelhecimento/efeitos da radiação , Animais , Camundongos , Camundongos Knockout , Doses de Radiação , Transdução de Sinais/efeitos da radiaçãoRESUMO
PURPOSE: To evaluate the discrepancy between the anatomical axis of the distal femur of Japanese patients and the stem position of 5 types of femoral components. METHODS: Lateral radiographs of 12 men and 88 women aged 31 to 83 (mean, 59) years with rheumatoid arthritis were evaluated. The discrepancy between the anatomical axis of the distal femur and the stem position of 5 types of femoral components (Nexgen LCCK, Press-Fit Condylar, Scorpio, Total Stabilizer, and Rotating Hinge) was determined by superimposing the template of each model over each lateral radiograph. RESULTS: The anatomical axis varied widely among our patients, as did the stem position of the 5 femoral components. Stems of all 5 femoral components tended to be more posterior than the anatomical axis. The discrepancy was smallest in the Nexgen LCCK, followed by the Press-Fit Condylar components. It was >3 mm in the other 3 models. In 35% of the patients, none of the prosthesis could be placed in an appropriate position. Smaller-size prostheses appear necessary for the Japanese. CONCLUSION: The stem position should be an important factor guiding selection of the appropriate model. The currently available femoral components may not be appropriate for the Japanese. Prostheses with appropriately positioned stems for Japanese patients with rheumatoid arthritis should be developed.
Assuntos
Artrite Reumatoide/patologia , Povo Asiático , Fêmur/patologia , Prótese do Joelho , Desenho de Prótese , Adulto , Idoso , Idoso de 80 Anos ou mais , Artrite Reumatoide/etnologia , Artrite Reumatoide/cirurgia , Artroplastia do Joelho , Estudos de Coortes , Feminino , Fêmur/diagnóstico por imagem , Humanos , Japão , Masculino , Pessoa de Meia-Idade , Ajuste de Prótese , RadiografiaRESUMO
PURPOSE: To compare quality of life, knee function, and physical activity in 33 elderly women with or without early-stage knee osteoarthritis (OA). METHODS: 33 Japanese elderly women (mean age, 66 years) with (n=18) or without (n=15) early-stage knee OA symptoms (knee pain and decreased range of motion [ROM]) were recruited. The height, weight, and body mass index, disease severity, quality of life (according to the Japanese Knee Osteoarthritis Measure [JKOM]), knee function (knee extension strength, ROM, 10-m gait time), and the amount of physical activity (net energy expenditure and step count) of the 2 groups were compared. RESULTS: The 2 patient groups did not differ significantly with respect to mean patient age, height, and body mass index, except for weight. Regarding knee function, mean knee extension strength, ROM (extension but not flexion), and 10-m gait speed (comfortable and maximum) were significantly inferior in patients with knee OA than in controls. Regarding the mean amount of physical activity undertaken, patients with knee OA did not differ significantly from controls with respect to net energy expenditure (179 vs. 212 Kcal/day) and step count (8016 vs. 9729 steps/day). Net energy expenditure (r= -0.65, p=0.04) and step count (r= -0.62, p=0.02) correlated negatively with JKOM scores in patients with knee OA but not in the controls. CONCLUSION: In Japanese elderly women with knee OA, quality of life (JKOM scores) correlated negatively with physical activity (net energy expenditure and step count). The 2 groups undertook similar amounts of physical activity, although those with knee OA exhibited less knee extension strength. Decreased knee extension strength coupled with high levels of physical activity may exacerbate the development of knee OA.