RESUMO
Many studies focused on the annulus fibrosus (AF) injury in rodent tail model for the intervertebral disk degeneration (IDD) research. However, previous studies caused tremendous injury of intervertebral disk (IVD) by penetrating whole disk. This study aimed to build a progressive IDD rodent tail model by a novel device for precise and minimally invasive puncture in AF. A precise puncture device was customized by 3D Printing Technique. 40 rodent tail IVDs were randomly grouped as follows: group A, non-puncture; group B, annulus needle puncture (ANP) for 4 week; group C, ANP for 8 week; and group D, ANP for 12 week. Pre- and post-puncture IVD height on radiographs and IVD signal intensity on T2 magnetic resonance imaging (MRI) were measured. Average bone density (ABD) on the end of coccygeal vertebrae between punctured disk was measured on the radiographs. Hematoxylin and eosin, TUNEL staining methods, immunofluorescence for cleaved-caspas3 and immunohistochemistry for aggrecan and collagen II were performed. Progressively and significantly increasing IVD height loss and degenerative grade were observed following the time points. The ABD was respectively, 81.20 ± 4.63 in group A, 83.93±3.18 in group B, 92.65 ± 4.32 in group C, 98.87 ± 6.69 in group D. In both group C and group D, there were significant differences with group A. In histology, increasing number of AF cells was noted in group B. In both group C and D, the fissures in AF were obviously observed, and a marked reduction of AF cells were also observed. In all ANP groups, there were significant decrease in number of NP cells, as well as aggrecan and collagen II contents. TUNEL assay showed cellular apoptosis were stimulated in all puncture group, especially in group D. A progressive IDD rat model could be standardly established by the micro-injury IVD puncture using a novel 3D printing device. This animal model provided a potential application for research of progressive hyperosteogeny following IDD development.
RESUMO
Notch signaling is one of the evolutionarily conserved signaling pathways in multicellular organisms. It plays an important role in embryonic development. During skeletal development of vertebrates, it regulates bone homeostasis by manipulating both osteoblastogenesis and osteoclastogenesis through different mechanisms. However, due to the different nature of Notch signaling in mesenchymal stem cell and osteoblast, regulation of Notch signaling in bone-related diseases remains unsettled. Previous studies by cell culture and mouse models showed contradictory results regarding the role of Notch signaling in bone homeostasis. To clarify the role of Notch signaling in osteogenesis, we established a zebrafish model, in which Notch1a intracellular domain (N1aICD) was specifically expressed in the osteoblasts. We found that overexpression of N1aICD in osteoblasts caused hyperosteogeny in the column region of zebrafish with the morphology of narrowed neural/hemal canals. Moreover, increased metabolic activity of osteoblasts instead of augmenting osteoblast number led to hyperosteogeny in N1aICD-overexpressed zebrafish. In summary, we successfully established a transgenic zebrafish line overexpressing N1aICD to clarify the in-vivo function of Notch signaling during osteoblastogenesis. In the future, this fish line can serve as a valuable tool to test the therapeutic drugs for hyperosteogeny.
Assuntos
Desenvolvimento Embrionário/genética , Proteínas de Homeodomínio/genética , Proteínas do Tecido Nervoso/genética , Osteogênese/genética , Receptor Notch1/genética , Proteínas de Peixe-Zebra/genética , Peixe-Zebra/genética , Animais , Animais Geneticamente Modificados , Doenças Ósseas/genética , Doenças Ósseas/patologia , Diferenciação Celular/genética , Regulação da Expressão Gênica no Desenvolvimento/genética , Humanos , Células-Tronco Mesenquimais/metabolismo , Camundongos , Osteoblastos/metabolismo , Receptores Notch/genética , Transdução de Sinais/genética , Peixe-Zebra/crescimento & desenvolvimentoRESUMO
OBJECTIVE:To establish the quality standard of Patch for hyperosteogeny.METHODS:Mastic and Myrrh in Patch for hyperosteogeny were qualitatively identified by TLC,and the content of paeoniflorin in preparation were determined by HPLC.RESULTS:The identification of TLC has a good resolution and specificity.The linear range of paeoniflorin were 0.204～1.428 ?g(r=0.999 2) with an average recovery of 99.41%(RSD=0.64%,n=6).CONCLUSION:Established standard can be used for the quality control of Patch for hyperosteogeny.
RESUMO
@#ObjectiveTo study the histological change of cervical body osteophytosis. Methods7 specimens of osteophytosis were sectioned from the cadavers'cervical vertebrae for histological observation.Results and ConclusionThere are five phases of the progress of the osteophytosis, i.e.,ossified in cartilage, precalcified, calcified, preossified and ossified.
RESUMO
Objective: To study the feasibility of percutaneous absorption of Compound Patch of Hyperosteogeny(CPH). Methods: The content of ferulic acid,an index composition in percutaneous receptor fluid and release receptor fluid were determined by HPLC.Results: The results of in vitro transdermal delivery experiment and in vitro release experiment showed that ferulic acid permeated at the constant speed of 0.2142?g?cm -2 ?h -1 in 24 hours and its release coincided with Higuchi Equation.Futhermore,the release rate was 14.53?g?cm -2 ?h -1/2 . Conclusion: CPH is a skeleton controlledtransdermal delivery system whose permeation speed is limited by skin.
