Effects of alginate/collagen scaffold on cell proliferation and differentiation of human adipose-derived mesenchymal stem cells
10.3760/cma.j.issn.1002-0098.2017.04.013
- VernacularTitle: 海藻酸钠-胶原支架材料对人脂肪间充质干细胞增殖与分化的影响
- Author:
Wei CHENG
1
;
Xiaopeng HAN
1
;
Suli MOU
2
;
Fan YANG
3
;
Liping LIU
1
Author Information
1. Department of Prosthodontic, Yantai Stomatological Hospital Affiliated to Binzhou Medical College, Yantai Shandong 264001, China
2. Department of Endodontics, Yantai Stomatological Hospital Affiliated to Binzhou Medical College, Yantai Shandong 264001, China
3. Department of Operative Dentistry & Endodontics, School of Stomatology, The Fourth Military Medical University & State Key Laboratory of Military Stomatology & National Clinical Research Center for Oral Diseases & Shaanxi Key Laboratory of Stomatology, Xi'an 710032, China
- Publication Type:Journal Article
- Keywords:
Hydrogels;
Mesenchymal stem cells;
Cell proliferation;
Cell differentiation
- From:
Chinese Journal of Stomatology
2017;52(4):259-264
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To build scaffold materials with different concentrations of alginate and collagen, and to observe the effects of alginate/collagen ratio on the proliferation of human adipose-derived mesenchymal stem cells (hAMSC) and osteogenic differentiation. The optimal concentration of alginate/collagen will be chosen for constructing hydrogel that will be used for bone tissue engineering.
Methods:Soluble hydrogel scaffold materials containing alginate/collagen were prepared, and the following groups were established based on different alginate/collagen ratio: 4∶1 (group A), 2∶1 (group B), and 1∶1 (group C). Cell proliferation on the material surface was observed using the cell counting kit-8 (CCK-8) assay, while cell viability in each material group were observed using live/dead staining. Quantitative real-time PCR(qPCR) was used to measure the differential expression of osteogenesis-related genes on and in the materials. Immunofluorescence staining was used to measure the differential gene expression of osteogenesis-related proteins in each group.
Results:The results from the CCK-8 assay showed increasing cell proliferation rate on the lyophilized hydrogel material surface as the collagen concentration increased, and the highest cell proliferation was observed in group C. Live/dead staining assay indicated that cells were able to proliferate in all three types of hydrogel materials, and the highest cell viability was found in material from group B ([87.50±2.65]%). qPCR showed that the expression of osteogenesis-related genes in group C was the highest, among the three groups, while the expression of osteocalcin in group B was significantly higher than those in the other two groups (P<0.05). Immunofluorescence staining was carried out for osteocalcin on and in the hydrogel material and the results were consistent with that of qPCR.
Conclusions:The alginate/collagen scaffold materials did not show adverse effects on the cell proliferation of hAMSC and osteogenenic differentiation. Bone tissue engineering can use 10% hydrogel material, and when the sodium alginate and collagen have a ratio of 2∶1, the hydrogel can be conducive to cell differentiation and proliferation.
