RESUMO
Recent investigations suggest that physeal morphologic features have a major role in the capital femoral epiphysis stability and slipped capital femoral epiphysis (SCFE) pathology, with a smaller epiphyseal tubercle and larger peripheral cupping of the femoral epiphysis being present in hips with progressive SCFE compared to healthy controls. Yet, little is known on the causal versus remodeling nature of these associations. This study aimed to use preoperative magnetic resonance imaging (MRI) of patients with unilateral SCFE to perform a comparison of the morphology of the epiphyseal tubercle, metaphyseal fossa, and peripheral cupping in hips with SCFE versus the contralateral uninvolved hips. Preoperative MRIs from 22 unilateral SCFE patients were used to quantify the morphological features of the epiphyseal tubercle (height, width, and length), metaphyseal fossa (depth, width, and length), and peripheral cupping height in three dimension. The quantified anatomical features were compared between hips with SCFE and the contralateral uninvolved side across the whole cohort and within SCFE severity subgroups using paired t-test. We found significantly smaller epiphyseal tubercle heights (p < 0.001) across all severities of SCFE when compared to their uninvolved contralateral side. There was a marginally smaller metaphyseal fossa length (p = 0.05) in SCFE hips compared to their contralateral uninvolved hips, with mild SCFE hips specifically having smaller fossa and epiphyseal lengths (p < 0.05) than their contralateral uninvolved side. There were no side-to-side differences in any other features of the epiphyseal tubercle, metaphyseal fossa and peripheral cupping across all severities (p > 0.05). These findings suggest a potential causal role of epiphyseal tubercle in SCFE pathogenesis.
Assuntos
Articulação do Quadril , Escorregamento das Epífises Proximais do Fêmur , Humanos , Articulação do Quadril/diagnóstico por imagem , Articulação do Quadril/patologia , Escorregamento das Epífises Proximais do Fêmur/diagnóstico por imagem , Escorregamento das Epífises Proximais do Fêmur/patologia , Fêmur/diagnóstico por imagem , Fêmur/patologia , Epífises/diagnóstico por imagem , Epífises/patologia , Lâmina de Crescimento/patologia , Estudos RetrospectivosRESUMO
INTRODUCTION: Disuse-induced bone loss is caused by a suppression of osteoblastic bone formation and an increase in osteoclastic bone resorption. There are few data available for the effects of environmental conditions, i.e., atmospheric pressure and/or oxygen concentration, on osteoporosis. This study examined the effects of mild hyperbaric oxygen at 1317 hPa with 40% oxygen on unloading-induced osteoporosis. MATERIALS AND METHODS: Eighteen 8-week old male Wistar rats were randomly divided into three groups: the control for 21 days without unloading and mild hyperbaric oxygen (NOR, n = 6), the unloading for 21 days and recovery for 10 days without mild hyperbaric oxygen (HU + NOR, n = 6), and the unloading for 21 days and recovery for 10 days with mild hyperbaric oxygen (HU + MHO, n = 6). RESULTS: The cortical thickness and trabecular bone surface area were decreased in the HU + NOR group compared to the NOR group. There were no differences between the NOR and HU + MHO groups. Osteoclast surface area and Sclerostin (Sost) mRNA expression levels were decreased in the HU + MHO group compared to the HU + NOR group. These results suggested that the loss of the cortical and trabecular bone is inhibited by mild hyperbaric oxygen, because of an inhibition of osteoclasts and enhancement of bone formation with decreased Sost expression. CONCLUSIONS: We conclude that exposure to mild hyperbaric oxygen partially protects from the osteoporosis induced by hindlimb unloading.
Assuntos
Elevação dos Membros Posteriores/fisiologia , Oxigenoterapia Hiperbárica , Osteoporose/fisiopatologia , Osteoporose/terapia , Animais , Peso Corporal , Proteínas Morfogenéticas Ósseas/genética , Proteínas Morfogenéticas Ósseas/metabolismo , Osso Esponjoso/patologia , Osso Esponjoso/fisiopatologia , Osso Cortical/patologia , Osso Cortical/fisiopatologia , Marcadores Genéticos/genética , Lâmina de Crescimento/patologia , Masculino , Osteocalcina/genética , Osteocalcina/metabolismo , Osteoclastos/patologia , Osteoporose/genética , Coativador 1-alfa do Receptor gama Ativado por Proliferador de Peroxissomo/genética , Coativador 1-alfa do Receptor gama Ativado por Proliferador de Peroxissomo/metabolismo , Ligante RANK/genética , Ligante RANK/metabolismo , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Ratos WistarRESUMO
Dexamethasone (Dex) is known to cause significant bone growth impairment in childhood. Although previous studies have suggested roles of osteocyte apoptosis in the enhanced osteoclastic recruitment and local bone loss, whether it is so in the growing bone following Dex treatment requires to be established. The current study addressed the potential roles of chemokine CXCL12 in chondroclast/osteoclast recruitment and bone defects following Dex treatment. Significant apoptosis was observed in cultured mature ATDC5 chondrocytes and IDG-SW3 osteocytes after 48 hours of 10-6 M Dex treatment, and CXCL12 was identified to exhibit the most prominent induction in Dex-treated cells. Conditioned medium from the treated chondrocytes/osteocytes enhanced migration of RAW264.7 osteoclast precursor cells, which was significantly inhibited by the presence of the anti-CXCL12 neutralizing antibody. To investigate the roles of the induced CXCL12 in bone defects caused by Dex treatment, young rats were orally gavaged daily with saline or Dex at 1 mg/kg/day for 2 weeks, and received an intraperitoneal injection of anti-CXCL12 antibody or control IgG (1 mg/kg, three times per week). Aside from oxidative stress induction systemically, Dex treatment caused reductions in growth plate thickness, primary spongiosa height, and metaphysis trabecular bone volume, which are associated with induced chondrocyte/osteocyte apoptosis and enhanced chondroclast/osteoclast recruitment and osteoclastogenic differentiation potential. CXCL12 was induced in apoptotic growth plate chondrocytes and metaphyseal bone osteocytes. Anti-CXCL12 antibody supplementation considerably attenuated Dex-induced chondroclast/osteoclast recruitment and loss of growth plate cartilage and trabecular bone. CXCL12 neutralization did not affect bone marrow osteogenic potential, adiposity, and microvasculature. Thus, CXCL12 was identified as a potential molecular linker between Dex-induced skeletal cell apoptosis and chondroclastic/osteoclastic recruitment, as well as growth plate cartilage/bone loss, revealing a therapeutic potential of CXCL12 functional blockade in preventing bone growth defects during/after Dex treatment. © 2018 American Society for Bone and Mineral Research.
Assuntos
Apoptose/efeitos dos fármacos , Osso Esponjoso , Quimiocina CXCL12/metabolismo , Dexametasona/efeitos adversos , Lâmina de Crescimento , Músculo Esquelético/metabolismo , Animais , Anticorpos Neutralizantes/farmacologia , Osso Esponjoso/crescimento & desenvolvimento , Osso Esponjoso/patologia , Linhagem Celular , Quimiocina CXCL12/antagonistas & inibidores , Dexametasona/farmacologia , Lâmina de Crescimento/crescimento & desenvolvimento , Lâmina de Crescimento/patologia , Masculino , Camundongos , Músculo Esquelético/patologia , Células RAW 264.7 , Ratos , Ratos Sprague-DawleyRESUMO
BACKGROUND: Whilst hypocalcemic complications from vitamin D deficiency are considered rare in high-income countries, they are highly prevalent among Black, Asian and Minority Ethnic (BAME) group with darker skin. To date, the extent of osteomalacia in such infants and their family members is unknown. Our aim was to investigate clinical, cardiac and bone histomorphometric characteristics, bone matrix mineralization in affected infants and to test family members for biochemical evidence of osteomalacia. CASE PRESENTATION: Three infants of BAME origin (aged 5-6 months) presented acutely in early-spring with cardiac arrest, respiratory arrest following seizure or severe respiratory distress, with profound hypocalcemia (serum calcium 1.22-1.96 mmol/L). All infants had dark skin and vitamin D supplementation had not been addressed during child surveillance visits. All three had severely dilated left ventricles (z-scores + 4.6 to + 6.5) with reduced ejection fraction (25-30%; normal 55-70), fractional shortening (7 to 15%; normal 29-40) and global hypokinesia, confirming hypocalcemic dilated cardiomyopathy. They all had low serum levels of 25 hydroxyvitamin D (25OHD < 15 nmol/L), and elevated parathyroid hormone (PTH; 219-482 ng/L) and alkaline phosphatase (ALP; 802-1123 IU/L), with undiagnosed rickets on radiographs. One infant died from cardiac arrest. At post-mortem examination, his growth plate showed a widened, irregular zone of hypertrophic chondrocytes. Histomorphometry and backscattered electron microscopy of a trans-iliac bone biopsy sample revealed increased osteoid thickness (+ 262% of normal) and osteoid volume/bone volume (+ 1573%), and extremely low bone mineralization density. Five of the nine tested family members had vitamin D deficiency (25OHD < 30 nmol/L), three had insufficiency (< 50 nmol/L) and 6/9 members had elevated PTH and ALP levels. CONCLUSIONS: The severe, hidden, cardiac and bone pathology described here exposes a failure of public health prevention programs, as complications from vitamin D deficiency are entirely preventable by routine supplementation. The family investigations demonstrate widespread deficiency and undiagnosed osteomalacia in ethnic risk groups and call for protective legislation.
Assuntos
Cardiomiopatia Dilatada/etiologia , Parada Cardíaca/etiologia , Hipocalcemia/complicações , Grupos Minoritários , Osteomalacia/etiologia , Insuficiência Respiratória/etiologia , Raquitismo/complicações , Densidade Óssea , Inglaterra , Feminino , Lâmina de Crescimento/patologia , Humanos , Hipocalcemia/etnologia , Hipocalcemia/patologia , Ílio/patologia , Lactente , Masculino , Raquitismo/etnologia , Raquitismo/patologiaRESUMO
Cadmium (Cd) and lead (Pb) are toxic elements that accumulate to the largest extent in bones. Rats were used to investigate whether tannic acid (TA; 0.5%, 1.0%, 1.5%. 2.0%, or 2.5%) would have a protective effect on the structure and properties of bones in the case of exposure to Cd and Pb (diet: 7 mg Cd/kg and 50 mg Pb/kg) for 6 weeks. The effects of administration of TA in Cd- and Pb-poisoned rats on bone characteristics and the morphology of articular and growth cartilages were determined. All the rats administered Cd and Pb had an enhanced Cd and Pb concentration in blood plasma and bone and reduced bone Ca content irrespective of the TA administration. Cd and Pb alone reduced the mechanical endurance and histomorphometric parameters of trabecular bone and the thickness of the growth plate and articular cartilage. Tannic acid improved cancellous bone parameters in the rat exposed to Cd and Pb. A diet rich in TA improved articular cartilage constituents in heavy metal-poisoned rats. These results suggest that alimentary TA supplementation can counteract in a dose-dependent manner some of the destructive changes evoked by Cd and Pb possibly by reducing the exposure.
Assuntos
Osso e Ossos/efeitos dos fármacos , Intoxicação por Cádmio/prevenção & controle , Cartilagem Articular/efeitos dos fármacos , Lâmina de Crescimento/efeitos dos fármacos , Intoxicação por Chumbo/prevenção & controle , Substâncias Protetoras/uso terapêutico , Taninos/uso terapêutico , Animais , Fenômenos Biomecânicos/efeitos dos fármacos , Densidade Óssea/efeitos dos fármacos , Conservadores da Densidade Óssea/administração & dosagem , Conservadores da Densidade Óssea/uso terapêutico , Osso e Ossos/química , Osso e Ossos/patologia , Cádmio/sangue , Cádmio/toxicidade , Intoxicação por Cádmio/patologia , Intoxicação por Cádmio/fisiopatologia , Cálcio/sangue , Osso Esponjoso/química , Osso Esponjoso/efeitos dos fármacos , Osso Esponjoso/patologia , Cartilagem Articular/química , Cartilagem Articular/patologia , Relação Dose-Resposta a Droga , Lâmina de Crescimento/química , Lâmina de Crescimento/patologia , Chumbo/sangue , Chumbo/toxicidade , Intoxicação por Chumbo/patologia , Intoxicação por Chumbo/fisiopatologia , Masculino , Osteomalacia/etiologia , Osteomalacia/prevenção & controle , Osteoporose/etiologia , Osteoporose/prevenção & controle , Substâncias Protetoras/administração & dosagem , Distribuição Aleatória , Ratos Wistar , Taninos/administração & dosagem , ToxicocinéticaRESUMO
Most toxic physeal changes are characterized microscopically by altered chondrocyte development, proliferation, or maturation in the growth plate and eventually result in disordered appositional bone growth. Many therapeutic drugs directly or indirectly target proteins involved in chondrocytic differentiation and maturation pathways, so toxic physeal injury has become increasingly common in preclinical toxicologic pathology. While physeal dysplasia has been associated with several different drug classes including bisphosphonates, vascular endothelial growth factor receptor inhibitors, fibroblast growth factor receptor inhibitors, transforming growth factor beta receptor inhibitors, and vascular targeting agents, physeal changes often share similar morphologic features including thickening and disorganization of the hypertrophic layer, increased numbers of hypertrophic chondrocytes, altered mineralization of endochondral ossification, and/or increased thickness of subphyseal bone. Knowledge of genetic and nutritional diseases affecting bone growth has been important in helping to determine which specific target drugs may be affecting that could result in toxic physeal lesions. A pathophysiologic mechanism for most physeal toxicants has been determined in detail using a variety of investigative techniques. However, due to the signaling cross talk and the tight regulation required for chondrocyte maturation in the physis, several growth factor pathways are likely to be affected simultaneously with pharmacologic disruption of physeal homeostasis and inhibition of one factor necessary for chondrocyte function often affects others.
Assuntos
Desenvolvimento Ósseo/efeitos dos fármacos , Doenças Ósseas/induzido quimicamente , Avaliação Pré-Clínica de Medicamentos , Efeitos Colaterais e Reações Adversas Relacionados a Medicamentos , Lâmina de Crescimento/efeitos dos fármacos , Animais , Doenças Ósseas/fisiopatologia , Osso e Ossos/efeitos dos fármacos , Osso e Ossos/fisiopatologia , Condrócitos/efeitos dos fármacos , Condrócitos/metabolismo , Difosfonatos/efeitos adversos , Modelos Animais de Doenças , Lâmina de Crescimento/patologia , Humanos , Receptores de Fatores de Crescimento de Fibroblastos/antagonistas & inibidores , Receptores de Fatores de Crescimento Transformadores beta/antagonistas & inibidores , Fator A de Crescimento do Endotélio Vascular/antagonistas & inibidoresRESUMO
The aim of the study was to investigate the effect of 2-oxoglutaric acid (2-Ox) supplementation (a precursor of glutamine and hydroxyproline, the most abundant amino acid of collagen) on cartilage and bone in pigs after fundectomy. Pigs at the age of forty days were subjected to fundectomy and divided into two groups depending on 2-Ox supplementation (at the daily dosage of 0.4 g/kg of body weight). Other pigs were sham operated. Pigs were euthanized at the age of eight months. An analysis of the morphometry of trabeculae, growth plate and articular cartilage in fundectomy-induced osteopenic bone was performed. Moreover, the levels of expression of osteocalcin, osteopontin and osteoprotegerin in trabecular bone and osteocalcin in articular cartilage were evaluated. Articular cartilage was thinnest in fundectomized pigs and thickest in 2-Ox-supplemented animals after fundectomy. Moreover, 2-Ox supplementation after fundectomy enhanced the total thickness of the growth plate and trabeculae in fundectomized pigs. The most evident signal for osteocalcin and osteoprotegerin in trabecular bone was in sham-operated and 2-Ox-supplemented pigs; a low reaction was observed in the fundectomized group. Additionally, as a long-term postoperative consequence, a change was observed in the expression of osteocalcin in articular cartilage. It seems that 2-Ox is suitable for use in preventing the negative effects of fundectomy on cancellous bone and cartilage.
Assuntos
Doenças Ósseas Metabólicas/dietoterapia , Osso Esponjoso , Cartilagem Articular , Lâmina de Crescimento , Ácidos Cetoglutáricos/uso terapêutico , Animais , Peso Corporal , Doenças Ósseas Metabólicas/etiologia , Doenças Ósseas Metabólicas/patologia , Osso Esponjoso/patologia , Cartilagem Articular/metabolismo , Cartilagem Articular/patologia , Colágeno/metabolismo , Fundo Gástrico/cirurgia , Lâmina de Crescimento/patologia , Masculino , Osteocalcina/biossíntese , Proteoglicanas/metabolismo , SuínosRESUMO
BACKGROUND: We previously demonstrated the benefits of daily, oral pentosan polysulfate (PPS) treatment in a rat model of mucopolysaccharidosis (MPS) type VI. Herein we compare these effects to once weekly, subcutaneous (s.c.) injection. The bioavailability of injected PPS is greater than oral, suggesting better delivery to difficult tissues such as bone and cartilage. Injected PPS also effectively treats osteoarthritis in animals, and has shown success in osteoarthritis patients. METHODOLOGY/PRINCIPAL FINDINGS: One-month-old MPS VI rats were given once weekly s.c. injections of PPS (1, 2 and 4 mg/kg, human equivalent dose (HED)), or daily oral PPS (4 mg/kg HED) for 6 months. Serum inflammatory markers and total glycosaminoglycans (GAGs) were measured, as were several histological, morphological and functional endpoints. Overall, weekly s.c. PPS injections led to similar or greater therapeutic effects as daily oral administration. Common findings between the two treatment approaches included reduced serum inflammatory markers, improved dentition and skull lengths, reduced tracheal deformities, and improved mobility. Enhanced effects of s.c. treatment included GAG reduction in urine and tissues, greater endurance on a rotarod, and better improvements in articular cartilage and bone in some dose groups. Optimal therapeutic effects were observed at 2 mg/kg, s.c.. No drug-related increases in liver enzymes, coagulation factor abnormalities or other adverse effects were identified following 6 months of s.c. PPS administration. CONCLUSIONS: Once weekly s.c. administration of PPS in MPS VI rats led to equal or better therapeutic effects than daily oral administration, including a surprising reduction in urine and tissue GAGs. No adverse effects from s.c. PPS administration were observed over the 6-month study period.
Assuntos
Mucopolissacaridose VI/tratamento farmacológico , Poliéster Sulfúrico de Pentosana/administração & dosagem , Poliéster Sulfúrico de Pentosana/farmacologia , Administração Oral , Animais , Disponibilidade Biológica , Fenômenos Biomecânicos , Cartilagem Articular/efeitos dos fármacos , Cartilagem Articular/patologia , Relação Dose-Resposta a Droga , Esquema de Medicação , Feminino , Fêmur/diagnóstico por imagem , Fêmur/efeitos dos fármacos , Glicosaminoglicanos/metabolismo , Lâmina de Crescimento/efeitos dos fármacos , Lâmina de Crescimento/patologia , Injeções Subcutâneas , Masculino , Movimento/efeitos dos fármacos , Mucopolissacaridose VI/metabolismo , Mucopolissacaridose VI/patologia , Mucopolissacaridose VI/fisiopatologia , Poliéster Sulfúrico de Pentosana/farmacocinética , Poliéster Sulfúrico de Pentosana/uso terapêutico , Ratos , Coluna Vertebral/diagnóstico por imagem , Coluna Vertebral/efeitos dos fármacos , Tomografia Computadorizada por Raios XRESUMO
OBJECTIVE: We investigated the combined roles of a low-nutrition diet (low levels of protein, iodine, and selenium) and T-2 toxin in bone development and to establish an experimental animal model of Kashin-Beck disease (KBD) that reliably mimics the disease's pathological changes for further study of the pathogenesis and prevention of the disease. METHODS: Sprague-Dawley rats were randomly divided among four groups: group A, normal diet; group B, normal diet plus T-2 toxin; group C, low-nutrition diet; and group D, low-nutrition diet plus T-2 toxin exposure. The radiographic and histopathological changes in the tibial growth zone, plate cartilage and metaphysis were examined. RESULTS: In group D, all epiphyseal plates were blurred, thin, and irregular. Tibias were significantly shorter in group D than in groups A and B. After 4 weeks, epiphyseal plates showed chondrocyte necrosis, with the more obvious necrosis appearing in groups C and D. The positive rate of lamellar necrosis was significantly higher in group D than in groups B and A (P < 0.01). In group D, metaphyseal trabecular bone was sparse, disordered, and disrupted, and massive transverse trabecular bone appeared in the metaphysis at 12 weeks. CONCLUSIONS: A rat model of KBD induced by a low-nutrition diet and T-2 toxin exposure demonstrated radiographic and histopathological abnormalities of the proximal epiphyseal plate and the tibial metaphysis that are very similar to the bone changes found in patients with KBD. This animal model will be helpful for further study of the pathogenesis and prevention of KBD.
Assuntos
Modelos Animais de Doenças , Doença de Kashin-Bek/etiologia , Desnutrição/complicações , Toxina T-2/toxicidade , Fenômenos Fisiológicos da Nutrição Animal/fisiologia , Animais , Cartilagem Articular/patologia , Condrócitos/efeitos dos fármacos , Condrócitos/patologia , Proteínas Alimentares/administração & dosagem , Feminino , Lâmina de Crescimento/efeitos dos fármacos , Lâmina de Crescimento/crescimento & desenvolvimento , Lâmina de Crescimento/patologia , Iodo/administração & dosagem , Doença de Kashin-Bek/sangue , Doença de Kashin-Bek/patologia , Doença de Kashin-Bek/fisiopatologia , Masculino , Desnutrição/sangue , Desnutrição/fisiopatologia , Necrose/etiologia , Ratos , Ratos Sprague-Dawley , Ratos Wistar , Selênio/administração & dosagem , Selênio/sangue , Tiroxina/sangue , Tíbia/crescimento & desenvolvimento , Tíbia/patologia , Tri-Iodotironina/sangue , Aumento de Peso/efeitos dos fármacos , Aumento de Peso/fisiologiaRESUMO
The objective of this study is to observe pathogenic lesions of joint cartilages in rats fed with T-2 toxin under a selenium deficiency nutrition status in order to determine possible etiological factors causing Kashin-Beck disease (KBD). Sprague-Dawley rats were fed selenium-deficient or control diets for 4 weeks prior to their being exposed to T-2 toxin. Six dietary groups were formed and studied 4 weeks later, i.e., controls, selenium-deficient, low T-2 toxin, high T-2 toxin, selenium-deficient diet plus low T-2 toxin, and selenium-deficient diet plus high T-2 toxin. Selenium deficiencies were confirmed by the determination of glutathione peroxidase activity and selenium levels in serum. The morphology and pathology (chondronecrosis) of knee joint cartilage of experimental rats were observed using light microscopy and the expression of proteoglycans was determined by histochemical staining. Chondronecrosis in deep zone of articular cartilage of knee joints was seen in both the low and high T-2 toxin plus selenium-deficient diet groups, these chondronecrotic lesions being very similar to chondronecrosis observed in human KBD. However, the chondronecrosis observed in the rat epiphyseal growth plates of animals treated with T-2 toxin alone or T-2 toxin plus selenium-deficient diets were not similar to that found in human KBD. Our results indicate that the rat can be used as a suitable animal model for studying etiological factors contributing to the pathogenesis (chondronecrosis) observed in human KBD. However, those changes seen in epiphyseal growth plate differ from those seen in human KBD probably because of the absence of growth plate closure in the rat.
Assuntos
Doenças das Cartilagens/patologia , Cartilagem Articular/patologia , Doença de Kashin-Bek/patologia , Selênio/deficiência , Joelho de Quadrúpedes/patologia , Toxina T-2/toxicidade , Ração Animal/efeitos adversos , Ração Animal/análise , Animais , Peso Corporal/efeitos dos fármacos , Doenças das Cartilagens/induzido quimicamente , Cartilagem Articular/efeitos dos fármacos , Cartilagem Articular/metabolismo , Condrócitos/efeitos dos fármacos , Condrócitos/metabolismo , Condrócitos/patologia , Modelos Animais de Doenças , Relação Dose-Resposta a Droga , Glutationa Peroxidase/sangue , Lâmina de Crescimento/efeitos dos fármacos , Lâmina de Crescimento/patologia , Doença de Kashin-Bek/fisiopatologia , Masculino , Necrose/induzido quimicamente , Necrose/patologia , Proteoglicanas/metabolismo , Ratos , Ratos Sprague-Dawley , Selênio/sangue , Selênio/farmacocinética , Joelho de Quadrúpedes/metabolismoRESUMO
Fibroblast growth factor 23 (FGF23) is a circulating factor secreted by osteocytes that is essential for phosphate homeostasis. In kidney proximal tubular cells FGF23 inhibits phosphate reabsorption and leads to decreased synthesis and enhanced catabolism of 1,25-dihydroxyvitamin D3 (1,25[OH]2 D3 ). Excess levels of FGF23 cause renal phosphate wasting and suppression of circulating 1,25(OH)2 D3 levels and are associated with several hereditary hypophosphatemic disorders with skeletal abnormalities, including X-linked hypophosphatemic rickets (XLH) and autosomal recessive hypophosphatemic rickets (ARHR). Currently, therapeutic approaches to these diseases are limited to treatment with activated vitamin D analogues and phosphate supplementation, often merely resulting in partial correction of the skeletal aberrations. In this study, we evaluate the use of FGFR inhibitors for the treatment of FGF23-mediated hypophosphatemic disorders using NVP-BGJ398, a novel selective, pan-specific FGFR inhibitor currently in Phase I clinical trials for cancer therapy. In two different hypophosphatemic mouse models, Hyp and Dmp1-null mice, resembling the human diseases XLH and ARHR, we find that pharmacological inhibition of FGFRs efficiently abrogates aberrant FGF23 signaling and normalizes the hypophosphatemic and hypocalcemic conditions of these mice. Correspondingly, long-term FGFR inhibition in Hyp mice leads to enhanced bone growth, increased mineralization, and reorganization of the disturbed growth plate structure. We therefore propose NVP-BGJ398 treatment as a novel approach for the therapy of FGF23-mediated hypophosphatemic diseases.
Assuntos
Fatores de Crescimento de Fibroblastos/metabolismo , Compostos de Fenilureia/farmacologia , Pirimidinas/farmacologia , Receptores de Fatores de Crescimento de Fibroblastos/antagonistas & inibidores , Raquitismo Hipofosfatêmico/metabolismo , Transdução de Sinais/efeitos dos fármacos , Animais , Peso Corporal/efeitos dos fármacos , Desenvolvimento Ósseo/efeitos dos fármacos , Proteínas da Matriz Extracelular/deficiência , Proteínas da Matriz Extracelular/metabolismo , Fêmur/efeitos dos fármacos , Fêmur/patologia , Fator de Crescimento de Fibroblastos 23 , Lâmina de Crescimento/efeitos dos fármacos , Lâmina de Crescimento/patologia , Homeostase/efeitos dos fármacos , Íons , Rim/efeitos dos fármacos , Rim/metabolismo , Camundongos Endogâmicos C57BL , Minerais/metabolismo , Compostos de Fenilureia/uso terapêutico , Pirimidinas/uso terapêutico , Receptores de Fatores de Crescimento de Fibroblastos/metabolismo , Raquitismo Hipofosfatêmico/tratamento farmacológico , Raquitismo Hipofosfatêmico/patologia , Cauda/anatomia & histologia , Vitamina D/análogos & derivados , Vitamina D/biossínteseRESUMO
Vitamin A (VA) and its active form, retinoic acid (RA), are regulators of skeletal development. In the present study, we investigated if maternal VA intake during pregnancy and lactation, as well as direct oral supplementation of neonates with VA + RA (VARA) in early life, alters neonatal bone formation and chondrocyte gene expression. Offspring of dams fed 3 levels of VA (marginal, adequate, and supplemented) for 10 wk were studied at birth (P0) and postnatal day 7 (P7). One-half of the newborns received an oral supplement of VARA on P1, P4, and P7. Tissues were collected on P0 and 6 h after the last dose on P7. Pup plasma and liver retinol concentrations were increased by both maternal VA intake and VARA (P < 0.01). Although maternal VA did not affect bone mineralization as assessed by von Kossa staining, newborn femur length was increased with maternal VA (P < 0.05). VARA supplementation of neonates increased the length of the hypertrophic zone only in VA-marginal pups, close to that in neonates from VA-adequate dams, suggesting VARA caused a catching up of growth that was limited by low maternal VA intake. Maternal diet did not alter type X nor type II collagen mRNA. However, VARA-treated pups from VA-supplemented dams had reduced mRNA for aggrecan, a major component of cartilage matrix, and increased mRNA for matrix metalloproteinase (MMP)13, which catalyzes the degradation of aggrecan and collagens. These results suggest that moderately high maternal VA intake combined with neonatal VARA supplementation can reduce the ratio of aggrecan:MMP, which may unfavorably alter early bone development.
Assuntos
Agrecanas/metabolismo , Fêmur/metabolismo , Regulação da Expressão Gênica no Desenvolvimento , Metaloproteinase 13 da Matriz/metabolismo , Osteogênese , Deficiência de Vitamina A/dietoterapia , Vitamina A/efeitos adversos , Agrecanas/genética , Animais , Animais Recém-Nascidos , Condrócitos/metabolismo , Suplementos Nutricionais/efeitos adversos , Feminino , Fêmur/patologia , Lâmina de Crescimento/patologia , Lactação , Fígado/metabolismo , Fenômenos Fisiológicos da Nutrição Materna , Metaloproteinase 13 da Matriz/genética , Gravidez , Ratos , Ratos Sprague-Dawley , Tretinoína/efeitos adversos , Tretinoína/uso terapêutico , Vitamina A/sangue , Vitamina A/metabolismo , Vitamina A/uso terapêutico , Deficiência de Vitamina A/congênito , Deficiência de Vitamina A/metabolismo , Deficiência de Vitamina A/patologia , DesmameRESUMO
Estrogens enhance skeletal growth during early sexual maturation, whereas high estradiol levels during late puberty result in growth plate fusion in humans. Although the growth plates do not fuse directly after sexual maturation in rodents, a reduction in growth plate height is seen by treatment with a high dose of estradiol. It is unknown whether the effects of estrogens on skeletal growth are mediated directly via estrogen receptors (ERs) in growth plate cartilage and/or indirectly via other mechanisms such as the growth hormone/insulin-like growth factor 1 (GH/IGF-1) axis. To determine the role of ERα in growth plate cartilage for skeletal growth, we developed a mouse model with cartilage-specific inactivation of ERα. Although mice with total ERα inactivation displayed affected longitudinal bone growth associated with alterations in the GH/IGF-1 axis, the skeletal growth was normal during sexual maturation in mice with cartilage-specific ERα inactivation. High-dose estradiol treatment of adult mice reduced the growth plate height as a consequence of attenuated proliferation of growth plate chondrocytes in control mice but not in cartilage-specific ERα(-/-) mice. Adult cartilage-specific ERα(-/-) mice continued to grow after 4 months of age, whereas growth was limited in control mice, resulting in increased femur length in 1-year-old cartilage-specific ERα(-/-) mice compared with control mice. We conclude that during early sexual maturation, ERα in growth plate cartilage is not important for skeletal growth. In contrast, it is essential for high-dose estradiol to reduce the growth plate height in adult mice and for reduction of longitudinal bone growth in elderly mice.
Assuntos
Desenvolvimento Ósseo , Cartilagem/metabolismo , Receptor alfa de Estrogênio/metabolismo , Lâmina de Crescimento/metabolismo , Animais , Densidade Óssea/efeitos dos fármacos , Desenvolvimento Ósseo/efeitos dos fármacos , Cartilagem/efeitos dos fármacos , Cartilagem/patologia , Estradiol/farmacologia , Receptor alfa de Estrogênio/deficiência , Feminino , Fêmur/efeitos dos fármacos , Fêmur/patologia , Inativação Gênica/efeitos dos fármacos , Lâmina de Crescimento/efeitos dos fármacos , Lâmina de Crescimento/patologia , Camundongos , Modelos Biológicos , Tamanho do Órgão/efeitos dos fármacos , Especificidade de Órgãos/efeitos dos fármacos , Ovariectomia , Maturidade Sexual/efeitos dos fármacosRESUMO
Severe hyperhomocysteinemia, as seen in classic homocystinuria, is associated with several skeletal malformations and osteopenia. Moreover, hyperhomocysteinemia during pregnancy has been associated with multiple developmental defects in the fetus. This study was undertaken to determine whether offspring of hyperhomocysteinemic mothers have demonstrable changes in bone volume and the epiphyseal growth plate. Ten adult female Sprague-Dawley rats were randomly assigned to the control or experimental group. The experimental group received 100 mg/kg/day of homocysteine in their drinking water for 3 weeks before mating and for the total duration of pregnancy. In each group, three pups per mother were randomly selected. The histomorphometric properties of tibial, radial and vertebral growth plates of newborn rats and the volume fraction of bone were compared between groups. The plasma homocysteine concentration at the end of study was significantly higher in dams in the experimental group (16.42+/-1.5 vs. 4.7+/-1.7 mumol/L, P<0.05). In offspring born to dams given the homocysteine supplement, the volume fraction of bone in the tibia (30.7+/-1.5% vs. 36.8+/-1.9%, P<0.05), radius (29.6+/-1.1% vs. 37.4+/-2%, P<0.05) and vertebra (34.4+/-1.8% vs. 41+/-1.9%, P<0.05) were significantly decreased whereas vertical heights of proliferative (423+/-25.1 vs. 301.8+/-28.1 microm for radius and 131.9+/-5.9 vs. 107.8+/-3.5 microm for vertebra) and hypertrophic zones (213.1+/-12 vs. 163.3+/-7.5 microm for tibia, 153.2+/-7.7 vs. 121.1+/-7.9 microm for radius and 112+/-9.9 vs. 88.4+/-10.1 microm for the vertebra) were increased (P<0.05). The results showed that the administration of homocysteine caused osteopenia in newborn rats. In addition, these data suggest that hyperhomocysteinemia may induce disruption of normal development of epiphyseal cartilage in the rat embryo.
Assuntos
Doenças Ósseas Metabólicas/induzido quimicamente , Idade Gestacional , Hiper-Homocisteinemia/fisiopatologia , Osteogênese/fisiologia , Complicações na Gravidez/fisiopatologia , Animais , Animais Recém-Nascidos , Feminino , Lâmina de Crescimento/anatomia & histologia , Lâmina de Crescimento/patologia , Homocisteína/sangue , Homocisteína/toxicidade , Hiper-Homocisteinemia/sangue , Hiper-Homocisteinemia/induzido quimicamente , Masculino , Microscopia , Gravidez , Complicações na Gravidez/sangue , Complicações na Gravidez/induzido quimicamente , Rádio (Anatomia)/anatomia & histologia , Rádio (Anatomia)/patologia , Ratos , Ratos Sprague-Dawley , Coluna Vertebral/anatomia & histologia , Coluna Vertebral/patologia , Tíbia/anatomia & histologia , Tíbia/patologiaRESUMO
An episode of ischemia followed by reperfusion of the femoral head (FH) is thought to be a common pathway in the pathogenesis of femoral head necrosis (FHN). Femoral head histology was investigated after short-term high-dose steroid treatment and femoral head ischemia and reperfusion in a large animal model. Twenty-two pigs were randomized to receive methylprednisolone 20mg/day/kg bodyweight intamuscularly for 3 days followed by methylprednisolone 10mg/day/kg bodyweight for 11 days (n=11), whereas the control group (n=11) received no treatment. After 6h of unilateral hip-joint pressure increase to 250mmHg, the pressure was discontinued and reperfusion was allowed for 4h. Undecalcified histology was performed for the femoral head subchondral region, the mid-region, and the region adjacent to the growth plate. Congestion phenomena were predominantly discerned in femoral head sections of the tamponaded hips. Histomorphometry revealed fat cell hyperthrophy and reduced hemopoetic marrow cells in the femoral heads of the steroid-treated group of animals. The number of blood vessels and bone trabeculae remained unchanged. These characteristics may correlate with early-stage femoral head necrosis.
Assuntos
Cabeça do Fêmur/citologia , Articulação do Quadril/fisiologia , Metilprednisolona/farmacologia , Tecido Adiposo/patologia , Animais , Células da Medula Óssea/efeitos dos fármacos , Células da Medula Óssea/patologia , Células da Medula Óssea/fisiologia , Cateterismo/métodos , Feminino , Cabeça do Fêmur/irrigação sanguínea , Cabeça do Fêmur/efeitos dos fármacos , Cabeça do Fêmur/patologia , Lâmina de Crescimento/citologia , Lâmina de Crescimento/efeitos dos fármacos , Lâmina de Crescimento/patologia , Lâmina de Crescimento/fisiologia , Articulação do Quadril/efeitos dos fármacos , Injeções Intramusculares , Isquemia/patologia , Isquemia/fisiopatologia , Masculino , Metilprednisolona/administração & dosagem , Necrose , Cloreto de Potássio/farmacologia , Pressão , Reperfusão , SuínosRESUMO
Vitamin D plays a major role in mineral and skeletal homeostasis through interaction with the nuclear vitamin D receptor (VDR) of target cells. Recent reports have indicated that some cellular effects of vitamin D may occur via alternative signaling pathways, but concrete evidence for mineral homeostasis has not been shown in vivo. To investigate this issue, the actions of calcitriol (1,25D) and maxacalcitol (OCT), which were developed for treatment of uremia-induced secondary hyperparathyroidism, were analyzed in VDR knockout (VDR(-/-)) mice. The VDR(-/-) mice were fed a rescue diet immediately after weaning. 1,25D, OCT or a control solution was administered intraperitoneally to these mice three times a week for eight weeks. Biological markers and bone growth were measured and bone histomorphometric analysis of the calcein-labeled tibia was performed 24 h after the final administration. Significantly higher levels of serum Ca(2+) were observed in 1,25D- and OCT-treated mice, but the serum parathyroid hormone level was unchanged by both agents. Impaired bone growth, enlarged and distorted cartilaginous growth plates, morphological abnormalities of cancellous and cortical bones; a morbid osteoid increase, lack of calcein labeling, and thinning of cortical bone, were all significantly improved by 1,25D and OCT. The significance of these effects was confirmed by bone histomorphometrical analysis. Upregulation of the calbindin D(9k) mRNA expression level in the duodenum may explain these findings, since this protein is a major modulator of Ca transport in the small intestine. We conclude that 1,25D and OCT both at a high dose exert significant effects on Ca and skeletal homeostasis with the principal improvement of Ca status in VDR(-/-) mice, and some of these effects may occur through an alternative vitamin D signaling pathway.
Assuntos
Osso e Ossos/efeitos dos fármacos , Osso e Ossos/metabolismo , Calcitriol/análogos & derivados , Cálcio/metabolismo , Homeostase/efeitos dos fármacos , Receptores de Calcitriol/deficiência , Animais , Transporte Biológico/efeitos dos fármacos , Osso e Ossos/anormalidades , Osso e Ossos/patologia , Calbindinas , Calcitriol/farmacologia , Canais de Cálcio/genética , Canais de Cálcio/metabolismo , Relação Dose-Resposta a Droga , Duodeno/efeitos dos fármacos , Duodeno/metabolismo , Regulação da Expressão Gênica/efeitos dos fármacos , Lâmina de Crescimento/efeitos dos fármacos , Lâmina de Crescimento/patologia , Camundongos , Camundongos Knockout , Osteogênese/efeitos dos fármacos , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Receptores de Calcitriol/metabolismo , Proteína G de Ligação ao Cálcio S100/genética , Proteína G de Ligação ao Cálcio S100/metabolismo , Canais de Cátion TRPV/genética , Canais de Cátion TRPV/metabolismoRESUMO
OBJECTIVE: To non-invasively investigate the changes to epiphyseal bone occurring in a longitudinal pre-clinical model of osteoarthritis (OA) using in vivo micro-computed tomography (micro-CT). DESIGN: In vivo micro-CT images were acquired using a bench-top micro-CT scanner, which produces three-dimensional data with isotropic voxel spacing of 0.046 mm. Male rodents were scanned prior to surgical destabilization, consisting of anterior cruciate ligament transection and partial medial menisectomy (ACLX). Subsequent scans were performed every 4 weeks post-ACLX, for up to 5 months. Volumetric bone mineral density (vBMD) was measured in specific, anatomically segmented regions within each image. The ACLX rodent data were compared with the contralateral non-operated hind limb of the same animal, as well as a sham-operated group (SHAM) of animals, for each time point. End-point histology compared changes to cartilage and bone between the ACLX and control animals. RESULTS: The micro-CT protocol produced sufficient spatial resolution and signal-to-noise ratio (SNR=19) to quantify subchondral bone pathology, with an acceptable entrance exposure to radiation (0.36 Gy). Significantly lower vBMD was measured in the ACLX group, vs SHAM rodents, at 1, 4, and 5 months post-surgery (P<0.05). Qualitative observations of ACLX joints revealed significant loss of cartilage, subchondral bone cysts, and calcification of tendon similar to changes found in humans. CONCLUSIONS: This study demonstrates in vivo micro-CT as an effective method for investigating the development of rodent knee OA longitudinally. This method can be applied, in future pre-clinical trials, to non-destructively monitor the efficacy of pharmacological interventions.
Assuntos
Osso e Ossos/diagnóstico por imagem , Osso e Ossos/patologia , Microrradiografia/métodos , Osteoartrite/diagnóstico por imagem , Animais , Ligamento Cruzado Anterior/cirurgia , Artrite Experimental , Cistos Ósseos/diagnóstico por imagem , Cistos Ósseos/patologia , Densidade Óssea/fisiologia , Progressão da Doença , Lâmina de Crescimento/diagnóstico por imagem , Lâmina de Crescimento/patologia , Masculino , Osteoartrite/patologia , Ratos , Ratos Sprague-Dawley , Índice de Gravidade de Doença , Joelho de Quadrúpedes , Tomografia Computadorizada por Raios X/métodosRESUMO
Ovariectomy of immature female rats, results in significant decrease of trabecular bone volume and in cortical bone thickness. Previously, we found that estradiol-17beta (E(2)) restored bone structure of ovariectomized (Ovx) female rats to values obtained in intact sham-operated female rats. E(2) also selectively stimulated creatine kinase (CK) specific activity a hormonal-genomic activity marker. In the present study, we compared the effects of E(2) and the phytoestrogens: daidzein (D), biochainin A (BA), genistein (G), carboxy-derivative of BA (cBA), and the SERM raloxifene (Ral) in Ovx, on both histological changes of bones and CK, when administered in multiple daily injections for 2.5 months. Bone from Ovx rats, showed significant disrupted architecture of the growth plate, with fewer proliferative cells and less chondroblasts. The metaphysis underneath the growth plate, contained less trabeculae but a significant increased number of adipocytes in the bone marrow. D like E(2) and Ral but not G, BA, or cBA, restored the morphology of the tibiae, similar to that of control sham-operated animals; the bony trabeculeae observed in the primary spongiosa was thicker, with almost no adipocytes in bone marrow. Ovariectomy resulted also in reduced CK, which in both epiphysis and diaphysis was stimulated by all estrogenic compounds tested. In summary, only D stimulated skeletal tissues growth and differentiation as effectively as E(2) or Ral, suggesting that under our experimental conditions, D is more effective in reversing menopausal changes than any of the other isolated phytoestrogens which cannot be considered as one entity.
Assuntos
Osso e Ossos/efeitos dos fármacos , Creatina Quinase/metabolismo , Isoflavonas/farmacologia , Fitoestrógenos/farmacologia , Adipócitos/patologia , Animais , Medula Óssea/patologia , Osso e Ossos/enzimologia , Osso e Ossos/patologia , Estradiol/administração & dosagem , Estradiol/farmacologia , Feminino , Genisteína/administração & dosagem , Genisteína/farmacologia , Lâmina de Crescimento/efeitos dos fármacos , Lâmina de Crescimento/patologia , Isoflavonas/administração & dosagem , Ovariectomia , Fitoestrógenos/administração & dosagem , Cloridrato de Raloxifeno/administração & dosagem , Cloridrato de Raloxifeno/farmacologia , Ratos , Ratos Wistar , Tíbia/efeitos dos fármacos , Tíbia/enzimologia , Tíbia/patologia , Malha Trabecular/efeitos dos fármacos , Malha Trabecular/enzimologia , Malha Trabecular/patologiaRESUMO
Chemotherapy often induces bone growth defects in pediatric cancer patients; yet the underlying cellular mechanisms remain unclear and currently no preventative treatments are available. Using an acute chemotherapy model in young rats with the commonly used antimetabolite methotrexate (MTX), this study investigated damaging effects of five once-daily MTX injections and potential protective effects of supplementary treatment with antidote folinic acid (FA) on cellular activities in the tibial growth plate, metaphysis, and bone marrow. MTX suppressed proliferation and induced apoptosis of chondrocytes, and reduced collagen-II expression and growth plate thickness. It reduced production of primary spongiosa bone, volume of secondary spongiosa bone, and proliferation of metaphyseal osteoblasts, preosteoblasts and bone marrow stromal cells, with the cellular activities being most severely damaged on day 9 and returning to or towards near normal levels by day 14. On the other hand, proliferation of marrow pericytes was increased early after MTX treatment and during repair. FA supplementation significantly suppressed chondrocyte apoptosis, preserved chondrocyte proliferation and expression of collagen-II, and attenuated damaging effects on production of calcified cartilage and primary bone. The supplementation also significantly reduced MTX effects on proliferation of metaphyseal osteoblastic cells and of bone marrow stromal cells, and enhanced pericyte proliferation. These observations suggest that FA supplementation effectively attenuates MTX damage on cellular activities in producing calcified cartilage and primary trabecular bone and on pools of osteoblastic cells and marrow stromal cells, and that it enhances proliferation of mesenchymal progenitor cells during bone/bone marrow recovery.
Assuntos
Desenvolvimento Ósseo/efeitos dos fármacos , Células da Medula Óssea/efeitos dos fármacos , Células da Medula Óssea/patologia , Leucovorina/farmacologia , Metotrexato/efeitos adversos , Células Estromais/efeitos dos fármacos , Células Estromais/patologia , Animais , Apoptose/efeitos dos fármacos , Proteínas Reguladoras de Apoptose/genética , Osso e Ossos/citologia , Osso e Ossos/efeitos dos fármacos , Bromodesoxiuridina/metabolismo , Proliferação de Células/efeitos dos fármacos , Condrócitos/citologia , Condrócitos/efeitos dos fármacos , Colágeno Tipo II/genética , Colágeno Tipo II/metabolismo , Regulação da Expressão Gênica/efeitos dos fármacos , Lâmina de Crescimento/efeitos dos fármacos , Lâmina de Crescimento/patologia , Masculino , Tamanho do Órgão/efeitos dos fármacos , Osteoblastos/citologia , Osteoblastos/efeitos dos fármacos , Pericitos/citologia , Pericitos/efeitos dos fármacos , Ratos , Ratos Sprague-DawleyRESUMO
We demonstrated previously that receiving long-term phototherapy was associated with early impairment of growth plate structure in neonatal rats, and oxidative stress may be the main risk factor for growth plate injury. The purpose of this study was to examine the histomorphometric effects of pentoxifylline treatment on the growth plate. Sixty weanling Sprague-Dawley rats were randomly separated into three equal groups. Group A, the control group, did not receive phototherapy and pentoxifylline. Groups B and C were exposed to phototherapy for 7 d. In addition to phototherapy, group C was also given pentoxifylline during the study period. Compared with zonal lengths on d 7 after initiation of phototherapy, group B had significantly lower values than group A for all zonal lengths (p < 0.001). Zonal lengths of growth plate were increased significantly with pentoxifylline treatment in group C for 7 d compared with group B (p < 0.001). After phototherapy, group B had significantly higher values than groups A and C for plasma malondialdehyde levels (p < 0.001). The pentoxifylline was found here to have some potential to reduce the effects of phototherapy on growth plate in neonatal rats at a relatively low dose.