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Method for promoting osteogenic differentiation of mesenchymal stem cells (MSCs) and application

A technology of stem cell and osteogenic differentiation, applied in new species of animal cells, biochemical equipment and methods, applications, etc., can solve the problems of slow fracture healing time, high recurrence rate, increased treatment complexity and risk, etc.

Active Publication Date: 2021-02-19
中山大学附属第八医院
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Among them, the occurrence of osteoporotic fractures is a difficult point in the treatment of osteoporosis, because the treatment of osteoporotic fractures has problems such as slow fracture healing time and high recurrence rate.
Moreover, if the patient has advanced age, general health decline, coexisting multi-system diseases, low immunity, etc., the probability of complications in surgical treatment will be higher, thus increasing the complexity and risk of treatment.

Method used

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  • Method for promoting osteogenic differentiation of mesenchymal stem cells (MSCs) and application
  • Method for promoting osteogenic differentiation of mesenchymal stem cells (MSCs) and application
  • Method for promoting osteogenic differentiation of mesenchymal stem cells (MSCs) and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Example 1 Animal experiment: Effect of overexpression of N-acetyltransferase 10 (N-acetyltransferase 10, NAT10) on bone mass production

[0030] The process of animal experimentation is as follows figure 1 Shown:

[0031] (1) Animal model selection and grouping

[0032] Eight-week-old female C57BL / 6 mice were selected and randomly divided into 4 groups with 6 mice in each group.

[0033] (2) Construction of osteoporosis mouse model

[0034] Two groups of 8-week-old female C57BL / 6 mice underwent bilateral ovariectomy (OVX), named OVX-NC (ovariectomy normal control group) and OVX-NAT10 OE (ovariectomy NAT10 overexpression group) respectively. , and the other two groups performed incision (sham operation) as blank control (Sham), respectively named Sham-NC (sham operation normal control group) and Sham-NAT10OE (sham operation NAT10 overexpression group).

[0035] (2) Construction of overexpression NAT10 adenovirus

[0036] The target shuttle plasmid (pADV-mCMV-Nat10-3...

Embodiment 2

[0044] Embodiment 2 Animal experiment: Scaffold osteogenesis under the skin of nude mice

[0045] (1) Animal model selection and grouping

[0046] Eight-week-old female BALB / c nude mice were selected and randomly divided into 2 groups with 6 mice in each group.

[0047] (2) Construction of NAT10 overexpression lentiviral vector

[0048] The day before transfection, 293T cells were digested and passaged into a 100mm dish, and the density of the cells was controlled to be 70-80% during transfection; the medium was replaced with 10mL Opti-MEM medium 1 hour before transfection; The viral vector plasmid 32ug (backbone plasmid: shuttle plasmid = 1:1) (backbone plasmid is provided by Heyuan Biotechnology Co., Ltd., shuttle plasmid is Ubi-MSC-3FLAG-SV40-EGFP-IRES-puromycib / Ubi-MSC- NAT10-3FLAG-SV40-EGFP-IRES-puromy cibn) was dissolved in Opti-MEM medium with a total volume of 500uL, mixed gently and allowed to stand for 5 minutes; the transfection reagent was dissolved in Ooti-MEM m...

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Abstract

The invention belongs to the technical field of bone tissue engineering, and particularly relates to a method for promoting osteogenic differentiation of mesenchymal stem cells and application. The invention provides the method for promoting osteogenic differentiation of the mesenchymal stem cells (MSCs), that is, N-acetyltransferase 10 (NAT10) is overexpressed in vivo by a biological method, through overexpression of NAT10 in mice by adenovirus according to the method, it is found that NAT10 can reverse bone mass loss of osteoporosis mice, enhance osteogenic differentiation potential of the MSCs, effectively increase local bone mass, increase bone mineral density, enhance bone strength, improve local osteoporosis and improve bone metabolism, and it is indicated that NAT10 can be used forconstructing a novel stem cell osteogenic material. Therefore, the method can provide a new treatment scheme for osteoporosis treatment.

Description

technical field [0001] The invention belongs to the technical field of bone tissue engineering, and in particular relates to a method and application for promoting osteogenic differentiation of mesenchymal stem cells. Background technique [0002] Osteoporosis (OP) is a systemic metabolic bone disease, which occurs due to the decline of bone mass caused by bone resorption, the destruction of bone microstructure, and the increase of bone fragility. A systemic metabolic bone disease (WHO) characterized by recurrent fractures. At the same time, osteoporosis is also one of the main risk factors for jaw bone loss, which can lead to gingival recession, root exposure, and severe cases can lead to abutment teeth loosening and falling off, alveolar bone resorption, jaw atrophy and other oral problems. , which brings great difficulties to clinical orthodontics, denture restoration and implant restoration. At present, this disease has a high incidence rate and great harm in the world...

Claims

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Application Information

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IPC IPC(8): C12N15/861C12N15/867A01K67/027C12N5/0775A61L27/12A61L27/10A61L27/38A61L27/50A61L27/54
CPCC12N15/86A01K67/0278C12N5/0662C12N9/1029A61L27/12A61L27/10A61L27/3834A61L27/3847A61L27/50A61L27/54C12N2740/15043C12N2710/10343A01K2267/03A01K2227/105A01K2217/072C12N2513/00C12N2517/02C12N2533/18A61L2430/02A61L2400/06A61L2300/412Y02A50/30
Inventor 沈慧勇杨文马梦君李鸿宇王鹏吴燕峰米汝佳卢艺萱
Owner 中山大学附属第八医院
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