Matrigel scaffold combined with Ad-hBMP7-transfected chondrocytes improves the repair of rabbit cartilage defect

نویسندگان

  • Xiaopeng Xia
  • Jing Li
  • Bo Xia
  • Huilin Yang
  • Dongmei Zhang
  • Bin Zhou
  • Jie Zhang
  • Man Zhou
  • Fan Liu
چکیده

The aim of this study was to explore an effective method for the repair of cartilage defects using chitosan/glycerophosphate (C/GP) gel- and Matrigel-engineered human bone morphogenetic protein 7 (hBMP7)-expressing chondrocytes. Rabbit chondrocytes were obtained, cultured in vitro and transfected with an adenovirus containing hBMP7 and green fluorescent protein (Ad-hBMP7-GFP). The expression of hBMP7 in the transfected cells was tested by reverse transcription-polymerase chain reaction (RT-PCR) and western blotting. The phenotype of the transfected cells was evaluated by detecting the yields of collagen II and hyaluronic acid using RT-PCR and enzyme-linked immunosorbent assay (ELISA). The growth of chondrocytes in the C/GP gel and Matrigel was accessed by measuring the cell growth rate, hematoxylin and eosin (H&E) staining and observation under a scanning microscope. Twelve adult male New Zealand white rabbits were randomly divided into three groups. Two cartilage defects were created in the rabbits' knees by aseptic surgery. Group A (n=4) did not receive any treatment, group B (n=4) were treated with C/GP gel and Matrigel-engineered Ad-mock-GFP-transfected chondrocytes, and group C (n=4) were treated with C/GP gel and Matrigel-engineered Ad-hBMP7-GFP-transfected chondrocytes. Rabbits were sacrificed at 4 weeks after transplantation, and the repair effect was measured by the Wakitani scoring method. On the basis of the RT-PCR and western blot results, hBMP7 was efficiently overexpressed in the Ad-hBMP7-GFP-transfected chondrocytes. The ELISA results showed that the yields of collagen II and hyaluronic acid in Ad-hBMP7-GFP-transfected chondrocytes were significantly higher than those in Ad-mock-GFP-transfected chondrocytes. Chondrocytes have a better morphology and arrangement in a Matrigel scaffold than in C/GP, as assessed by H&E staining and scanning microscopy. According to the Wakitani score, Matrigel combined with Ad-hBMP7-GFP-transfected chondrocytes successfully promoted the repair of cartilage defects in rabbit knees.

برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید

ثبت نام

اگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید

منابع مشابه

Partial Resurfacing of the Distal Femoral Cartilage Defect with Stem Cell- Seeded Poly-Vinyl-Alcohol (PVA) Scaffold

Objective- To evaluate the biological compatibility of differentiated stem cells embedded in poly-vinyl-alcohol (PVA) scaffolds for repair of distal femoral cartilage defect.   Design- Experimental in vivo study. Animals- Twelve adult male New Zealand white rabbits were used which were divided into two groups (I, II) six rabbits each.   Procedures- Mesenchymal stem cells were isolated from h...

متن کامل

Preconditioning of Rabbit Mesenchymal Stem Cells in Polyglycolic Acid (PGA) Scaffold using Low-Intensity Ultrasound Improved Regeneration of Cartilage in Rabbit Articular Cartilage Defect Model

This study investigated the effect of low intensity ultrasound (LIUS) stimulation of mesenchymal stem cells (MSCs) in vitro on the repair of cartilage defect after implantation of the construct in vivo. Rabbit MSCs were cultured in the polyglycolic acid (PGA) scaffold and preconditioned with (MSCs/US+) or without (MSCs/US-) LIUS stimulation during the chondrogenic differentiation for 1 week in ...

متن کامل

Co-Culture of Mesenchymal Stem Cells with Mature Chondrocytes: Producing Cartilage Construct for Application in Cartilage Regeneration

Background: Cell-based treatment approach using differentiated mesenchymal stem cells (MSCs) and mature chondrocytes has been considered as an advanced treatment for cartilage repair. We investigated the differentiated level of these two cell types that is crucial for their repair capacity for cartilage defect at a co-culture micro mass system. Methods: Passaged-2 MSCs isolated from the mouse b...

متن کامل

Comparison of Autogenic Costal Cartilage with Chitosan Scaffold in Canine Humeral Defect Healing

Objective- Current trends emphasize the acceleration of fracture healing on the ground that in doing so, the limitation of mobility and complications associated with recovery period are reduced. The present study aims to compare autogenic costal cartilage with Chitosan scaffold in canine humeral defect healing. Design- Experimental study Animal-15 adult male dogs Procedures-...

متن کامل

Sonic Hedgehog Improves Redifferentiation of Dedifferentiated Chondrocytes for Articular Cartilage Repair

Sonic hedgehog (Shh) is involved in the induction of early cartilaginous differentiation of mesenchymal cells in the limb. We investigated whether Shh could promote redifferentiation of dedifferentiated chondrocytes and have a favorable effect on the regeneration of cartilage. Articular chondrocytes of rats were separated and cultured. The redifferentiation of dedifferentiated chondrocytes tran...

متن کامل

ذخیره در منابع من


  با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید

برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید

ثبت نام

اگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید

عنوان ژورنال:

دوره 13  شماره 

صفحات  -

تاریخ انتشار 2017