In vitro engineering of fibrocartilage using CDMP1 induced dermal fibroblasts and polyglycolide.
This study was designed to explore the feasibility of using cartilage-derived morphogenetic protein-1 (CDMP1) induced dermal fibroblasts (DFs) as seed cells and polyglycolide (PGA) as scaffold for fibrocartilage engineering. DFs isolated from canine were expanded and seeded on PGA scaffold to fabricate cell/scaffold constructs which were cultured with or without CDMP1. Proliferation and differentiation of DFs in different constructs were determined by DNA assay and glycosaminoglycan (GAG) production. Histological and immunohistochemical staining of the constructs after being in vitro cultured for 4 and 6 weeks were carried out to observe the fibrocartilage formation condition. The fibrocartilage-specific gene expression by cells in the constructs was analyzed by real-time PCR. It was shown that in the presence of CDMP1 the proliferation and GAG synthesis of DFs were significantly enhanced compared to those without CDMP1. Fibrocartilage-like tissue was formed in the CDMP1 induced construct after being cultured for 4 weeks, and it became more matured at 6 weeks as stronger staining for GAG and higher gene expression of collagen type II was observed. Since only weak staining for GAG and collagen type II was observed for the construct engineered without CDMP1, the induction effect on the fibrocartilage engineering can be ascertained when using DFs as seed cells. Furthermore, the potential of using DFs as seed cells to engineer fibrocartilage is substantiated and further study on using the engineered tissue to repair fibrocartilage defects is currently ongoing in our group.
Zhao
Guiqing
G
Department of Plastic and Reconstructive Surgery, Shanghai 9th People's Hospital, Shanghai Jiao Tong University School of Medicine, 639 Zhi Zao Ju Road, Shanghai 200011, PR China.
Yin
Shuo
S
Liu
Guangpeng
G
Cen
Lian
L
Sun
Jian
J
Zhou
Heng
H
Liu
Wei
W
Cui
Lei
L
Cao
Yilin
Y
D053674:Aggrecans / Q000235:genetics / Q000378:metabolism; D000469:Alkaline Phosphatase / Q000378:metabolism; D000818:Animals; D049109:Cell Proliferation; D048430:Cell Shape; D002478:Cells, Cultured; D024042:Collagen Type I / Q000235:genetics / Q000378:metabolism; D000090602:Collagen Type I, alpha 1 Chain; D024043:Collagen Type II / Q000235:genetics / Q000378:metabolism; D020405:Dermis / Q000166:cytology*; D004285:Dogs; D005109:Extracellular Matrix / Q000378:metabolism; D005347:Fibroblasts / Q000166:cytology / Q000378:metabolism*; D051445:Fibrocartilage / Q000033:anatomy & histology / Q000473:pathology / Q000502:physiology*; D020869:Gene Expression Profiling; D006025:Glycosaminoglycans / Q000378:metabolism; D055428:Growth Differentiation Factor 5 / Q000378:metabolism*; D011091:Polyesters / Q000737:chemistry / Q000378:metabolism; D011100:Polyglycolic Acid / Q000737:chemistry / Q000378:metabolism*; D055758:SOX9 Transcription Factor / Q000235:genetics / Q000378:metabolism; D023822:Tissue Engineering / Q000379:methods*; D054457:Tissue Scaffolds
D053674:Aggrecans; D024042:Collagen Type I; D000090602:Collagen Type I, alpha 1 Chain; D024043:Collagen Type II; D006025:Glycosaminoglycans; D055428:Growth Differentiation Factor 5; D011091:Polyesters; D055758:SOX9 Transcription Factor; D011100:Polyglycolic Acid; C033616:poly(lactide); D000469:Alkaline Phosphatase