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Regulation and control research method of cell cilium transportprotein 80 in rapid maxillary expansion mechanical biological signal transduction

A transporter protein, rapidly expanding technology, applied in biochemical equipment and methods, microbial measurement/testing, animal cells, etc., can solve problems such as unclear mechanisms

Inactive Publication Date: 2018-11-06
AFFILIATED STOMATOLOGICAL HOSPITAL OF NANJING MEDICAL UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the mechanism of how osteosynthetic tissue cells perceive mechanical stimulation signals and convert them into biochemical signals is still unclear

Method used

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0049] The research method of the regulation mechanism of intraciliary transporter 80 in the rapid expansion of maxillary mechanical biological signal transduction includes the following steps:

[0050] a. Construct an animal model of rapid maxillary enlargement in mice. The mice are no less than 6 weeks old and their first and second molars have erupted;

[0051] b. Construction of IFT80 gene-silencing osteoblasts; fetal mouse skull parietal bones were taken to culture osteoblasts, and negative shRNA interference sequences targeting mouse IFT80 genes were constructed, packaged into lentiviral particles to infect osteoblasts, and the best inhibition efficiency was screened out ShRNA interference sequence;

[0052] c. Construct an in vitro stretch culture osteoblast model, the mechanical stretch stress parameters are: 2% deformation, 0.5Hz; divided into static culture blank group, static culture negative sequence interference group, static culture IFT80 gene interference group, stretc...

Embodiment 2

[0084] To clarify the process of rapid maxillary expansion of the palatal suture bone reconstruction, continuous time point Micro-CT to observe the shape and bone density of the palatal suture; HE, total collagen, ALP and TRAP staining to observe the formation and absorption of new bone in the palatal suture tissue, Real-timeRT -PCR and Western-blot to evaluate the expression changes of osteogenic differentiation related indicators, such as bone sialine protein (BSP), osteopontin (OPN), osteocalcin (OCN), ALP and type I collagen (Col I); Calcein double-labeling experiment to evaluate the deposition rate of new bone in the palatal suture 14 days after arch expansion.

[0085] To clarify the cell proliferation in the rapid expansion of the palatal suture tissue in the upper jaw, and perform immunohistochemical staining of the nuclear proliferation antigen PCNA and Ki67.

[0086] To clarify the temporal and spatial expression of IFT80 in the process of distracting bone formation in th...

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Abstract

The invention discloses a regulation and control research method of cell cilium transportprotein 80 in rapid maxillary expansion mechanical biological signal transduction. The method comprises the following steps: a, establishing a rapid maxillary expansion mouse animal model; b, establishing IFT80 gene silence osteoblast; c, establishing an in-vitro stretching culture osteoblast model; d, establishing a co-culture model of osteoblast and osteoclast; e, carrying out in-vivo transfection experiment; f, confirming a relationship of IFT80 spatio-temporal expression and rapid maxillary expansion midline palatine suture bone remolding; g, confirming regulation and control mechanisms of IFT80 upon proliferation and differentiation of in-vitro stretching cultured osteoblast, primary cilium formation and mechanical biological signal transduction; h, confirming influence of IFT80 gene silence upon in-vitro stretching cultured osteoblast in regulating functions of osteoclast. The method has great instruction significances for treating such malocclusion.

Description

Technical field [0001] The present invention relates to the technical field of biological genes, and in particular to a method for studying the regulation mechanism of the intraciliary transporter 80 in the upper jaw rapid expansion mechanical biological signal transduction. Background technique [0002] Underdevelopment of the maxillary width is a common malocclusion in orthodontics. Rapid Maxillary Expansion (RME) is the most commonly used method to treat such malocclusion. The principle is to guide the upper jaw to move left and right by expanding the palatal suture, and then the expanded palatal suture tissue proliferates, repairs and deposits bone. The top of the upper jaw is widened, and the width of the upper dental arch is increased, finally achieving the purpose of treatment. However, after rapid expansion of the palatal suture, the mineralization and regeneration of new bone takes more time. The new bone that has not been calcified at the edge of the palatal suture can ...

Claims

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Application Information

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IPC IPC(8): C12N5/077C12Q1/6883G01N15/00G01N15/10G01N15/14G01N33/50G01N33/569
CPCC12N5/0654C12Q1/6883C12Q2600/158G01N15/00G01N15/10G01N15/1434G01N33/5044G01N33/56966
Inventor 王华孙雯张卫兵潘永初
Owner AFFILIATED STOMATOLOGICAL HOSPITAL OF NANJING MEDICAL UNIV
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