title: Characterization of chondrocyte scaffold carriers for cell-based gene therapy in articular cartilage repair.
abstract: Articular cartilage lesions in the knee are common injuries. Chondrocyte transplant represents a promising therapeutic modality for articular cartilage injuries. Here, we characterize the viability and transgene expression of articular chondrocytes cultured in three-dimensional scaffolds provided by four types of carriers. Articular chondrocytes are isolated from rabbit knees and cultured in four types of scaffolds: type I collagen sponge, fibrin glue, hyaluronan, and open-cell polylactic acid (OPLA). The cultured cells are transduced with adenovirus expressing green fluorescence protein (AdGFP) and luciferase (AdGL3-Luc). The viability and gene expression in the chondrocytes are determined with fluorescence microscopy and luciferase assay. Cartilage matrix production is assessed by Alcian blue staining. Rabbit articular chondrocytes are effectively infected by AdGFP and exhibited sustained GFP expression. All tested scaffolds support the survival and gene expression of the infected chondrocytes. However, the highest transgene expression is observed in the OPLA carrier. At 4 weeks, Alcian blue-positive matrix materials are readily detected in OPLA cultures. Thus, our results indicate that, while all tested carriers can support the survival of chondrocytes, OPLA supports the highest transgene expression and is the most conductive scaffold for matrix production, suggesting that OPLA may be a suitable scaffold for cell-based gene therapy of articular cartilage repairs.
authors: Department of Orthopaedic Surgery of the Affiliated Hospitals, Chongqing Medical University, Chongqing 400016, China; Molecular Oncology Laboratory, Department of Orthopaedic Surgery and Rehabilitation Medicine, The University of Chicago Medical Center, Chicago, Illinois 60637.
authors: Wei
authors: W
authors: Shui
authors: Liangjun
authors: L
authors: Yin
authors: Jeffrey
authors: J
authors: Luo
authors: Ruidong
authors: R
authors: Li
authors: Wenwen
authors: W
authors: Zhang
authors: Jiye
authors: J
authors: Zhang
authors: Wei
authors: W
authors: Huang
authors: Ning
authors: N
authors: Hu
authors: Xi
authors: X
authors: Liang
authors: Zhong-Liang
authors: ZL
authors: Deng
authors: Zhenming
authors: Z
authors: Hu
authors: Lewis L
authors: LL
authors: Shi
authors: Hue H
authors: HH
authors: Luu
authors: Rex C
authors: RC
authors: Haydon
authors: Tong-Chuan
authors: TC
authors: He
authors: Sherwin H
authors: SH
authors: Ho
mesh_terms: D000256:Adenoviridae / Q000378:metabolism; D000818:Animals; D002358:Cartilage, Articular / Q000187:drug effects / Q000473:pathology*; D002469:Cell Separation; D002470:Cell Survival / Q000187:drug effects / Q000235:genetics; D019902:Chondrocytes / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism*; D005109:Extracellular Matrix / Q000187:drug effects / Q000378:metabolism; D005786:Gene Expression Regulation / Q000187:drug effects; D018014:Gene Transfer Techniques; D015316:Genetic Therapy; D005822:Genetic Vectors / Q000378:metabolism; D057809:HEK293 Cells; D006801:Humans; D019344:Lactic Acid / Q000494:pharmacology; D008297:Male; D011091:Polyesters; D011108:Polymers / Q000494:pharmacology; D011817:Rabbits; D011995:Recombination, Genetic; D054457:Tissue Scaffolds / Q000737:chemistry*; D019076:Transgenes / Q000235:genetics; D014945:Wound Healing / Q000187:drug effects
chemical_list: D011091:Polyesters; D011108:Polymers; D019344:Lactic Acid; C033616:poly(lactide)
keywords: articular cartilage; autologous chondrocyte implantation; chondrocyte; ex vivo gene therapy; open-cell poly lactic acid (OPLA); scaffold carrier
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