Cartilage oligomeric matrix protein gene multilayers inhibit osteogenic differentiation and promote chondrogenic differentiation of mesenchymal stem cells.
There are still many challenges to acquire the optimal integration of biomedical materials with the surrounding tissues. Gene coatings on the surface of biomaterials may offer an effective approach to solve the problem. In order to investigate the gene multilayers mediated differentiation of mesenchymal stem cells (MSCs), gene functionalized films of hyaluronic acid (HA) and lipid-DNA complex (LDc) encoding cartilage oligomeric matrix protein (COMP) were constructed in this study via the layer-by-layer self-assembly technique. Characterizations of the HA/DNA multilayered films indicated the successful build-up process. Cells could be directly transfected by gene films and a higher expression could be obtained with the increasing bilayer number. The multilayered films were stable for a long period and DNA could be easily released in an enzymatic condition. Real-time polymerase chain reaction (RT-PCR) assay presented significantly higher (p<0.01) COMP expression of MSCs cultured with HA/COMP multilayered films. Compared with control groups, the osteogenic gene expression levels of MSCs with HA/COMP multilayered films were down-regulated while the chondrogenic gene expression levels were up-regulated. Similarly, the alkaline phosphatase (ALP) staining and Alizarin red S staining of MSCs with HA/COMP films were weakened while the alcian blue staining was enhanced. These results demonstrated that HA/COMP multilayered films could inhibit osteogenic differentiation and promote chondrogenic differentiation of MSCs, which might provide new insight for physiological ligament-bone healing.
Department of Orthopaedic Surgery, the Second Affiliated Hospital, Medical College of Zhejiang University, NO. 88 Jiefang Road, Hangzhou 310009, China. 21218130@zju.edu.cn.
Peng
P
Guo
Department of Orthopaedic Surgery, the Second Affiliated Hospital, Medical College of Zhejiang University, NO. 88 Jiefang Road, Hangzhou 310009, China. zlshi78@gmail.com.
Zhong-Li
ZL
Shi
Department of Orthopaedic Surgery, the Second Affiliated Hospital, Medical College of Zhejiang University, NO. 88 Jiefang Road, Hangzhou 310009, China. liuan2013zju@gmail.com.
An
A
Liu
Department of Orthopaedic Surgery, the Second Affiliated Hospital, Medical College of Zhejiang University, NO. 88 Jiefang Road, Hangzhou 310009, China. lintiao112009@gmail.com.
Tiao
T
Lin
Department of Orthopaedic Surgery, the Second Affiliated Hospital, Medical College of Zhejiang University, NO. 88 Jiefang Road, Hangzhou 310009, China. fanggangbi@gmail.com.
Fang-Gang
FG
Bi
Department of Orthopaedic Surgery, the Second Affiliated Hospital, Medical College of Zhejiang University, NO. 88 Jiefang Road, Hangzhou 310009, China. shimingmin@gmail.com.
Ming-Min
MM
Shi
Department of Orthopaedic Surgery, the Second Affiliated Hospital, Medical College of Zhejiang University, NO. 88 Jiefang Road, Hangzhou 310009, China. zrjwsj@zju.edu.cn.
Shi-Gui
SG
Yan
D064236:Cartilage Oligomeric Matrix Protein / Q000235:genetics* / Q000378:metabolism; D002454:Cell Differentiation / Q000235:genetics*; D020219:Chondrogenesis / Q000235:genetics*; D005786:Gene Expression Regulation; D049452:Green Fluorescent Proteins / Q000378:metabolism; D057809:HEK293 Cells; D006801:Humans; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000378:metabolism*; D010012:Osteogenesis / Q000235:genetics*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D060888:Real-Time Polymerase Chain Reaction; D013056:Spectrophotometry, Ultraviolet; D013194:Staining and Labeling; D013499:Surface Properties; D014162:Transfection; D014867:Water
D064236:Cartilage Oligomeric Matrix Protein; D012333:RNA, Messenger; D014867:Water; D049452:Green Fluorescent Proteins