Arthritis & Rheumatism, Volume 62,
November 2010 Abstract Supplement
Abstracts of the American College of
Rheumatology/Association of Rheumatology Health Professionals
Annual Scientific Meeting
Atlanta, Georgia November 6-11, 2010.
Human Amniotic Membrane as Scaffold for Human Articular Cartilage Repair.
Muinos-Lopez4, Emma, Diaz-Prado3, Silvia M., Hermida-Gomez2, Tamara, Rendal-Vazquez1, Esther, Fuentes-Boquete3, Isaac, Arufe-Gonda3, María C., De Toro3, Francisco J.
Cryobiology Unit. Complejo Hosp. Univ. A Coruña, A Coruña, Spain
Osteoarticular and Aging Res. Lab. CIBER-BBN, Rheumatology Div, INIBIC-Complejo Hosp, Univ. A Coruña, A Coruña, Spain
Osteoarticular and Aging Res. Lab. CIBER-BBN, Rheumatology Div. INIBIC-Complejo Hosp, Univ. A Coruña, A Coruña, Spain. INIBIC-University of A Coruña, A Coruña, Spain
Osteoarticular and Aging Res. Lab. CIBER-BBN. Rheumatology Div. INIBIC-Complejo Hosp. Univ. A Coruña, Coruña, Spain
Human amniotic membrane (HAM) is a highly abundant and easily accessible tissue that may potentially be an important chondrocyte carrier for cartilage regeneration in vivo.
Develop an in vitro repair model of focal injuries of human articular cartilage, based on the usefulness of cryopreserved HAM as support for chondrocyte proliferation.
Materials and Methods:
Four in vitro repair models of focal injuries of human articular cartilage were developed. For this purpose chondrocytes, isolated from cartilage slices obtained from femoral heads, were grown on the basement layer of HAM for 34 weeks. Then a chondrocyte pellet was implanted into 2 mm focal defects of human articular cartilage and the HAM providing chondrocytes was placed in direct contact with the cartilage surface to be repaired. These implants were cultured in DMEM supplemented with 10% FBS for 8 weeks. The repair tissues were analyzed by histochemistry (hematoxylin-eosin, Masson's trichromic, toluidine blue and safranin O) and immunohistochemistry for type I and II collagen and aggrecan.
Human chondrocytes cultured on HAM and transplanted onto focal injuries of human articular cartilage penetrated into the nearby surface of the chondral defect producing a more regular area and contributed on the closing of the chondral defect. The chondrocyte pellet implanted in the lesion filled the chondral defect and showed good integration between the repair tissue and native cartilage. Stainings were done to detect specific major components of the extracellular matrix to obtain more detailed information about the structure and composition of the repair tissue. Type II collagen, safranin O, toluidine blue and Masson's trichromic stainings of repair tissue were positive, whereas aggrecan and type I collagen stainings were weak (Table 1 and Figure 1). The morphology of the newly-formed tissue exhibited a fibrocartilaginous appearance and high cellularity.
Table 1. Semi-quantitative histochemical and immunohistochemistry analysis of the in vitro human articular cartilage repair model.
|TYPE STAIN||Native Cartilage||Focal Injure|
|Modified Masson's trichromic||+++||-||-||-|
|Type I Collagen||+||++||+||++|
|Type II Collagen||+++||+++||+||+++|
Figure 1. Histochemical and immunohistochemistry analysis of the in vitro human articular cartilage repair model. A. Toluidine blue. B. Safranin O. C. Col I. D. Col II.
These results indicated that cryopreserved amniotic membrane could be used as support for chondrocytes proliferation in cell therapy to repair focal injuries of human articular cartilage.
Project funded by Servizo Galego de Saúde (PS07/84), Catedra Bioiberica de la Universidade da Coruña and Instituto de Salud Carlos III CIBER BBN CB06-01-0040. Silvia Diaz Prado is beneficiary of an Isidro Parga Pondal contract from Xunta de Galicia, A Coruna, Spain.
To cite this abstract, please use the following information:
Muinos-Lopez, Emma, Diaz-Prado, Silvia M., Hermida-Gomez, Tamara, Rendal-Vazquez, Esther, Fuentes-Boquete, Isaac, Arufe-Gonda, María C., et al; Human Amniotic Membrane as Scaffold for Human Articular Cartilage Repair. [abstract]. Arthritis Rheum 2010;62 Suppl 10 :1483