Method for inducing mesenchymal stem cells to be directionally differentiated into osteoblasts

Abstract

The invention belongs to the field of biomedical engineering, and particularly relates to a method for inducing mesenchymal stem cells to be directionally differentiated into osteoblasts. The method for inducing the mesenchymal stem cells to be directionally differentiated into the osteoblasts has the advantages that the problem of deficiency of a simple, convenient, economical, safe and effectivemethod for inducing mesenchymal stem cells to be differentiated into osteoblasts in the prior art can be solved; macromolecular three-dimensional structures are constructed, appropriate micro-environments are provided, collagen composition and mediation intercellular signal cascade amplification are adopted, culture is carried out on the mesenchymal stem cells in osteogenic induction culture media, accordingly, expression of osteogenesis-related transcription factors can be strengthened, and the mesenchymal stem cells can be promoted to be differentiated into the osteoblasts.

Description

technical field

[0001] The invention belongs to the field of biomedical engineering, and in particular relates to a method for inducing directional differentiation of bone marrow mesenchymal stem cells into osteoblasts. Background technique

[0002] Bone marrow mesenchymal stem cells (BMSCs) are a kind of adult stem cells that can self-renew and have multi-directional differentiation potential. Under appropriate induction conditions, they can differentiate into cartilage, bone, muscle, fat, nerve, etc. Various tissue cells. MSCs have become a research hotspot in the field of basic medicine and clinical tissue and organ damage repair and regeneration, but there are still some technical problems, such as low differentiation efficiency or non-directional differentiation to form heterogeneous cells or even abnormal differentiation into tumor cells. In the application of bone tissue engineering, directional induction of MSCs to differentiate into osteoblasts is a key step in the...

Examples

Embodiment 1

[0050] Example 1 Method for preparing three-dimensional polymer structure and inducing osteogenesis

[0051] Weigh 1g PLGA, 10% chitosan, put into trifluoroethanol solution, stir magnetically, dissolve evenly. Under the parameters of positive pressure 18.5kv, negative pressure 2.8kv, humidity 35%, and temperature 19°C, a film with a three-dimensional structure was prepared by electrospinning, and the obtained film was dried in a vacuum oven for 48 hours to remove the solvent; The thin film material with a three-dimensional structure was subjected to surface functionalization treatment with dopamine, in a Tris-HCl solution with a pH value of 8.5 and a concentration of 0.3 wt% dopamine, reacted for 24 hours, and washed three times with deionized water. After pretreatment in 2% glutaraldehyde for 1 h, they were washed 3 times with deionized water. In a 5wt% collagen solution, react for 24 hours, control the temperature at 37° C., wash with deionized water three times, and dry ...

Embodiment 2

[0052] Example 2 Method for preparing three-dimensional polymer structure and inducing osteogenesis

[0053] Weigh 1g PLGA, 10% chitosan, put into trifluoroethanol solution, stir magnetically, dissolve evenly. Under the parameters of positive pressure 18.5kv, negative pressure 2.8kv, humidity 35%, and temperature 19°C, a film with a three-dimensional structure was prepared by electrospinning, and the obtained film was dried in a vacuum oven for 48 hours to remove the solvent; The thin film material with a three-dimensional structure was subjected to surface functionalization treatment with dopamine, in a Tris-HCl solution with a pH value of 8.5 and a concentration of 0.3 wt% dopamine, reacted for 24 hours, and washed three times with deionized water. After pretreatment in 2% glutaraldehyde for 1 h, they were washed 3 times with deionized water. In a 5wt% collagen solution, react for 24 hours, control the temperature at 37° C., wash with deionized water three times, and dry ...

technical field

[0001] The invention belongs to the field of biomedical engineering, and in particular relates to a method for inducing directional differentiation of bone marrow mesenchymal stem cells into osteoblasts. Background technique

[0002] Bone marrow mesenchymal stem cells (BMSCs) are a kind of adult stem cells that can self-renew and have multi-directional differentiation potential. Under appropriate induction conditions, they can differentiate into cartilage, bone, muscle, fat, nerve, etc. Various tissue cells. MSCs have become a research hotspot in the field of basic medicine and clinical tissue and organ damage repair and regeneration, but there are still some technical problems, such as low differentiation efficiency or non-directional differentiation to form heterogeneous cells or even abnormal differentiation into tumor cells. In the application of bone tissue engineering, directional induction of MSCs to differentiate into osteoblasts is a key step in the...

Examples

Embodiment 1

[0050] Example 1 Method for preparing three-dimensional polymer structure and inducing osteogenesis

[0051] Weigh 1g PLGA, 10% chitosan, put into trifluoroethanol solution, stir magnetically, dissolve evenly. Under the parameters of positive pressure 18.5kv, negative pressure 2.8kv, humidity 35%, and temperature 19°C, a film with a three-dimensional structure was prepared by electrospinning, and the obtained film was dried in a vacuum oven for 48 hours to remove the solvent; The thin film material with a three-dimensional structure was subjected to surface functionalization treatment with dopamine, in a Tris-HCl solution with a pH value of 8.5 and a concentration of 0.3 wt% dopamine, reacted for 24 hours, and washed three times with deionized water. After pretreatment in 2% glutaraldehyde for 1 h, they were washed 3 times with deionized water. In a 5wt% collagen solution, react for 24 hours, control the temperature at 37° C., wash with deionized water three times, and dry ...

Embodiment 2

[0052] Example 2 Method for preparing three-dimensional polymer structure and inducing osteogenesis

[0053] Weigh 1g PLGA, 10% chitosan, put into trifluoroethanol solution, stir magnetically, dissolve evenly. Under the parameters of positive pressure 18.5kv, negative pressure 2.8kv, humidity 35%, and temperature 19°C, a film with a three-dimensional structure was prepared by electrospinning, and the obtained film was dried in a vacuum oven for 48 hours to remove the solvent; The thin film material with a three-dimensional structure was subjected to surface functionalization treatment with dopamine, in a Tris-HCl solution with a pH value of 8.5 and a concentration of 0.3 wt% dopamine, reacted for 24 hours, and washed three times with deionized water. After pretreatment in 2% glutaraldehyde for 1 h, they were washed 3 times with deionized water. In a 5wt% collagen solution, react for 24 hours, control the temperature at 37° C., wash with deionized water three times, and dry ...

Images