Biosynthesis of proteoglycan in vitro by cartilage from human osteochondrophytic spurs.
نویسندگان
چکیده
Proteoglycan biosynthesis by human osteochondrophytic spurs (osteophytes) obtained from osteoarthritic femoral heads at the time of surgical joint replacement was studied under defined culture conditions in vitro. Osteophytes were primarily present in two anatomic locations, marginal and epi-articular. Minced tissue slices were incubated in the presence of [(35)S]sulphate or [(14)C]glucosamine. Osteophytes incorporated both labelled precursors into proteoglycan, which was subsequently characterized by CsCl-isopycnic-density-gradient ultracentrifugation and chromatography on Sepharose CL-2B. The material extracted with 0.5m-guanidinium chloride showed 78.1% of [(35)S]sulphate in the A1 fraction after centrifugation. Only 23.0% of the [(35)S]sulphate in this A1 fraction was eluted in the void volume of Sepharose CL-2B under associative conditions. About 60-80% of the [(35)S]sulphate in the tissue 4m-guanidinium chloride extract was associated with monomeric proteoglycan (fraction D1). The average partition coefficient (K(av.)) of the proteoglycan monomer on Sepharose CL-2B was 0.28-0.33. Approx. 12.4% of this monomer formed stable aggregates with high-molecular-weight hyaluronic acid in vitro. Sepharose CL-2B chromatography of fractions with lower buoyant densities (fractions D2-D4) demonstrated elution profiles on Sepharose CL-2B substantially different than that of fraction D1, indicative of the polydisperse nature of the newly synthesized proteoglycan. Analysis of the composition and chain size of the glycosaminoglycans showed the following: (1) preferential elution of both [(35)S]sulphate and [(14)C]glucosamine in the 0.5m-LiCl fraction on DEAE-cellulose; (2) the predominant sulphated glycosaminoglycan was chondroitin 6-sulphate (60-70%), with 9-11% keratan sulphate in the monomer proteoglycan; (3) K(av.) values of 0.38 on Sephadex G-200 and 0.48 on Sepharose CL-6B were obtained with papain-digested and NaBH(4)-treated D1 monomer respectively. A comparison of the synthetic with endogenous glycosaminoglycans indicated similar types. These studies indicated that human osteophytes synthesized in vitro sulphated proteoglycans with some characteristics similar to those of mature human articular cartilage, notably in the size of their proteoglycan monomer and predominance of chondroitin 6-sulphate. They differed from articular cartilage primarily in the lack of substantial quantities of keratan sulphate and aggregation properties associated with monomer interaction with hyaluronic acid.
منابع مشابه
Surgam - Product Information
Class Tiaprofenic acid is a non-steroidal, anti-inflammatory, analgesic compound, developed by Roussel Uclaf, and belongs to the propionic acid class. Site and Mode of Action Tiaprofenic acid displays analgesic, anti-inflammatory and anti-pyretic activity. Classical pharmacological studies in animals both in vitro and in vivo have shown that tiaprofenic acid inhibits the biosynthesis of prostag...
متن کاملStudy of Human Chondrocyte Redifferntiation Capacity in Three-Dimensional Hydrogel Culture
Objective(s) Articular cartilage tissue defects cannot be repaired by the proliferation of resident chondrocytes. Autologous chondrocyte transplantation (ACT) is a relatively new therapeutic approach to cover full thickness articular cartilage defects by in vitro grown chondrocytes from the joint of a patient. Therefore, we investigated the redifferentiation capability of human chondrocytes ma...
متن کاملInduction of Chondrogenic Differentiation of Human Adipose-Derived Stem Cells with TGF-β3 in Pellet Culture System
Objective Adult stem cells which are derived from different tissues, with their unique abilities to self-renew and differentiate into various phenotypes have the potential for cell therapy and tissue engineering. Human adipose tissue is an appropriate source of mesenchymal stem cells with wide differentiation potential for tissue engineering research. In this study isolated stem cells from hum...
متن کاملTransforming Growth Factor-β1 Preserves Bovine Nasal Cartilage against Degradation Induced by Interleukin-1α in Explant Culture
Background and Aims: Chondrocytes and their differentiation play a central role in joint diseases. Effect of the transforming growth factor (TGF)-β1 on chondrocyte characteristics and differentiation is not clearly understood. This study was undertaken to investigate the effects of TGF-β1 on tissue characteristics and morphology of chondrocytes against degradation induced by interleuk...
متن کاملLong-term effects of osteogenic protein-1 on biosynthesis and proliferation of human articular chondrocytes.
OBJECTIVE To study the long-term effects of OP-1 on the biosynthesis and proliferation of human articular chondrocytes. METHODS Chondrocytes were released from human cartilage from 10 organ donors of different ages and cultured in alginate. They were exposed to OP-1 (0-200 ng/ml) for 3 to 60 days. Proteoglycan (35S-sulfate) and collagen (3H-proline) synthesis were measured by radiolabeling. P...
متن کاملذخیره 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 ناماگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید
ورودعنوان ژورنال:
- The Biochemical journal
دوره 206 2 شماره
صفحات -
تاریخ انتشار 1982