RESUMEN
Excessive bone formation occurring in such conditions as paravertebral ligamentous ossification, hallux osteophytes or some neoplastic tumors, presents a significant problem, both epidemiological and clinical. Since pathogenesis of this disorder is still unclear, we studied its mechanism in experimental model utilizing inducible orthotopic osteogenesis. Periosteal bone apposition stimulated by Moloney sarcoma is characterized by unusually high volume of new bone tissue appearing subperiosteally in the bone adjacent to the tumor. Genes engaged in this growth have not been characterized so far. Here we show the results of mRNA Representation Difference Analysis in Moloney sarcoma, which reveal high expression of four genes coding extracellular matrix proteins: osteopontin, fibronectin, stromelysin-1 and tenascin. These findings suggest that the uncommon dynamics of the Moloney sarcoma-induced osteogenesis depends on high expression of these extracellular matrix proteins.
Asunto(s)
Fibronectinas/metabolismo , Metaloproteinasa 3 de la Matriz/metabolismo , Virus del Sarcoma Murino de Moloney/fisiología , Osteogénesis/fisiología , Osteopontina/metabolismo , Periostio/virología , Sarcoma Experimental/etiología , Tenascina/metabolismo , Animales , Northern Blotting , Ratones , Ratones Endogámicos BALB CRESUMEN
Several studies have demonstrated the pleiotropic effects of statins. Since Wang and associates reported that in rabbits lovastatin reduced steroid-induced bone loss, numerous authors have confirmed these data, however, others have reported conflicting results. In this study, the effects of fluvastatin on bone formation were investigated in early and late phase of osteogenesis. In the first set of experiments (early phase of osteogenesis) CFW/Ll mice were randomly divided into three groups. Two groups were injected with Moloney-murine sarcoma virus (Mo-MSV) into right thighs to induce orthotopic bone formation. Mice in the experimental group received fluvastatin for 11 consecutive days. Thirty days after Mo-MSV inoculation, total serum cholesterol, triglycerides, high- and low-density lipoprotein cholesterol, alkaline phosphatase (AP) were measured and bone mineral increase was calculated. In the second set of experiments (late phase of osteogenesis), fluvastatin was administered from day 11 after Mo-MSV inoculation for 20 consecutive days. Fluvastatin administration in the early phase of osteogenesis made no significant difference in average bone increase compared with mice receiving placebo. Lipid profile and AP were not significantly affected. During late phase of osteogenesis, the average increase in femural dry mass was significantly lower in the group of mice receiving fluvastatin than in the control group. Also, Mo-MSV-initiated tumors disappeared earlier in mice treated with fluvastatin. This may be attributed to the antioncogenic potential of fluvastatin. These results also point out that orthotopic bone formation at the sites of Mo-MSV inoculation in mice seems to be a useful model to examine the pleiotropic effects of statins.
Asunto(s)
Ácidos Grasos Monoinsaturados/farmacología , Indoles/farmacología , Osteogénesis/efectos de los fármacos , Fosfatasa Alcalina/sangre , Animales , Ácidos Grasos Monoinsaturados/administración & dosificación , Fluvastatina , Indoles/administración & dosificación , Lípidos/sangre , Ratones , Ratones Endogámicos , Virus del Sarcoma Murino de Moloney , Neoplasias/tratamiento farmacológico , Periostio/efectos de los fármacos , Periostio/virologíaRESUMEN
An increase in femoral and tibio/fibular bone mass following periosteal membrane stimulation by Moloney sarcoma virus inoculation into thigh muscles of mice was measured in situ on formalin fixed excised hind limbs using a Hologic 4500A Fan Beam X-ray bone densitometer adapted for small bone samples. These results were verified by measurements of constant dry bone mass of the same bones liberated from soft limb tissues by NaOH hydrolysis. There was no consistent data correlation found between the DEXA scan and dry bone mass evaluations. It is concluded that the sensitivity of the DEXA measurement is unsuitable when assessing very small bone samples, weighing merely 20-30 mg.
Asunto(s)
Densidad Ósea/fisiología , Huesos/diagnóstico por imagen , Miembro Posterior/anatomía & histología , Ratones/anatomía & histología , Absorciometría de Fotón , Animales , Miembro Posterior/diagnóstico por imagen , Masculino , Virus del Sarcoma Murino de Moloney , Periostio/fisiología , Periostio/virologíaRESUMEN
OBJECTIVE: To evaluate both the potential for transferring genes to periosteal cells using an adeno-associated virus (AAV) vector and the potential for gene expression after transplantation of those cells to a cartilage defect in vivo. METHODS: Periosteum was obtained from the tibia of 6-week-old rabbits and enzymatically digested. The isolated periosteum derived cells were cultured and the subconfluence cells were infected with a recombinant AAV expressing the LacZ gene (AAV-LacZ). Collagen gel containing the LacZ transferred, periosteum derived cells was transplanted into a full thickness articular cartilage defect in 10 rabbits. RESULTS: Infected cells still growing on the plate continued to express LacZ at least 12 weeks after AAV infection, with the highest percentage of LacZ positive cells reaching 74.4%. The LacZ positive cells were recognized at the transplant sites in 8 out of 10 knees. CONCLUSION: Gene expression in periosteum derived cells was sustained in vitro for at least 12 weeks using the AAV vector, and for 2 weeks ex vivo after transplantation into a cartilage defect.
Asunto(s)
Adenovirus Humanos/genética , Cartílago Articular/patología , Vectores Genéticos/genética , Periostio/citología , Transducción Genética , Cicatrización de Heridas , Animales , Cartílago Articular/enzimología , Cartílago Articular/lesiones , Trasplante de Células , Células Cultivadas , Marcadores Genéticos , Humanos , Articulaciones/cirugía , Operón Lac/genética , Periostio/enzimología , Periostio/virología , Conejos , beta-Galactosidasa/metabolismoRESUMEN
Several alphaviruses, including the Sindbis-group viruses, Ross River virus, O'nyong-nyong virus, and Chikungunya virus, are associated with outbreaks of acute and persistent arthralgia and arthritis in humans. Mechanisms underlying alphavirus-induced arthralgia and arthritis are not clearly understood, though direct viral replication within or around the affected joints is thought to contribute to disease. S.A.AR86 is a Sindbis-group alphavirus closely related to the arthralgia-associated Ockelbo and Girdwood S.A viruses. Following inoculation with S.A.AR86 derived from a molecular clone, infectious virus was isolated from bone and joint tissue 1 to 6 days postinfection. Studies using either in situ hybridization or S.A.AR86-derived double promoter viruses and replicons expressing green fluorescent protein localized sites of viral replication to the periosteum, tendons, and endosteum within the epiphyses of the long bones adjacent to articular joints. These results demonstrate that alphaviruses associated with arthralgia in humans replicate within bone-associated connective tissue adjacent to articular joints in an adult mouse model.