Superficial Zone Protein Expression Research Articles

Tubular perfusion system for chondrocyte culture and superficial zone protein expression.

Tissue engineering is an alternative method for articular cartilage repair. Mechanical stimulus has been found to be an important element to the healthy development of chondrocytes and maintenance of their native phenotype. To enhance nutrient transport and apply mechanical stress, we have developed a novel bioreactor, the tubular perfusion system (TPS), to culture chondrocytes in three-dimensional scaffolds. In our design, chondrocytes are encapsulated in alginate scaffolds and placed into a tubular growth chamber, which is perfused with media to enhance nutrient transfer and expose cells to fluid flow. Results demonstrate that TPS culture promotes the proliferation of chondrocytes compared to static culture as shown by DNA content and histochemical staining. After 14 days of culture, low messenger RNA expression of proinflammatory and apoptotic markers in TPS bioreactor culture confirmed that a flow rate of 3 mL/min does not damage the chondrocytes embedded in alginate scaffolds. Additionally, cells cultured in the TPS bioreactor showed increased gene expression levels of aggrecan, type II collagen, and superficial zone protein compared to the static group, indicative of the emergence of the superficial zone specific phenotype. Therefore, the TPS bioreactor is an effective means to enhance the proliferation and phenotype maintenance of chondrocytes in vitro.

Induction of Chondrogenesis and Expression of Superficial Zone Protein in Synovial Explants with TGF-β1 and BMP-7

Superficial zone protein (SZP) functions as a boundary lubricant in articular cartilage and decreases the coefficient of friction. As lubrication of articular cartilage is critical for normal joint function, the ability to secrete SZP at the surface of tissue-engineered cartilage is a prerequisite for optimal lubrication. Synovium-derived mesenchymal stem cells (MSCs) are thought to be an attractive cell source for cartilage regeneration. However, optimization of a three-dimensional environment is necessary for tissue engineering. In this study, we investigated whether synovial explants, which would preserve the physiologic microenvironment for MSCs therein, have the potential of SZP secretion after chondrogenic differentiation by treatment with transforming growth factor-β1 (TGF-β1) and bone morphogenetic protein-7 (BMP-7). Immunostaining and enzyme-linked immunosorbent assay analysis demonstrated that synovial explants can synthesize and secrete SZP following chondrogenic differentiation in response to TGF-β1 and BMP-7. Interestingly, the combined treatment with TGF-β1 and BMP-7 or treatment first with TGF-β1 followed by BMP-7 was more effective than other treatment groups in both chondrogenic differentiation and SZP secretion. In conclusion, synovial explants represent not only a superb source of progenitors/stem cells for the regeneration of the surface zone of articular cartilage, but also a useful model system for the in vitro differentiation into mature articular cartilage phenotypes in response to morphogens for tissue engineering of articular cartilage.

Probing the microenvironmental conditions for induction of superficial zone protein expression

To determine the in vitro conditions which promote expression of superficial zone protein (SZP). Chondrocytes from 6-month-old calves were expanded in monolayer culture and the expression of SZP in alginate bead and monolayer culture was quantified with quantitative real time-polymerase chain reaction (qRT-PCR) and immunostaining. The effect of oxygen tension on SZP expression was determined by qRT-PRC analysis of cells cultured in two dimension (2D) and three dimension (3D) under hypoxic (1% pO2) or normoxic (21% pO2) conditions. Finally, to examine the effect of cyclic tensile strain on expression of SZP in 2D and 3D cultures, chondrocytes encapsulated in alginate beams or seeded on type I collagen coated polydimethylsiloxane (PDMS) chambers were subjected to 5% strain at 1 Hz, 2 h/day for 4 days or 2 h at the fourth day of culture and mRNA levels were quantified. Bovine chondrocytes in monolayer showed a drastic decrease in SZP expression, similar in trend to the commonly reported downregulation of type II collagen (Col2). Chondrocytes embedded in alginate beads for 4 days re-expressed SZP but not Col2. SZP expression was higher under normoxic conditions whereas Col2 was upregulated only in alginate beads under hypoxic conditions. Cyclic mechanical strain showed a tendency to upregulate mRNA levels of SZP. A microenvironment encompassing a soft encapsulation material and 21% oxygen is sufficient for fibroblastic chondrocytes to re-express SZP. These results serve as a guideline for the design of stratified engineered articular cartilage and suggest that microenvironmental cues (oxygen tension level) strongly influence the pattern of SZP expression in vivo.

Dedifferentiation and Redifferentiation of Articular Chondrocytes from Surface and Middle Zones: Changes in MicroRNAs-221/-222, -140, and -143/145 Expression

Articular cartilage contains three functional zones (superficial, middle, and deep) characterized by distinct structure, composition, and biomechanical properties. One of the unsolved major challenges in cartilage tissue engineering is to produce tissue that mimics the zonal organization of the native articular cartilage. An increasing number of studies aim to design zonal organization into tissue-engineered cartilage by forming a stratified construct using zonal cell subpopulations. However, in vitro monolayer expansion of chondrocytes, which is generally required to obtain high cell numbers necessary for tissue engineering and autologous chondrocyte implantation, leads to dedifferentiation of chondrocytes into fibroblast-like cells, resulting in loss of zonal markers, such as the superficial zone protein (SZP) of the superficial zone as well as chondrocytic phenotype markers, such as type II collagen and aggrecan. Several microRNAs (miRNAs), including miR-221, miR-222, miR-143, and miR-145, have been identified from bovine articular cartilage as superficial zone-enriched miRNAs. miR-140 has been known as a cartilage-specific miRNA whose expression is implicated in chondrocyte differentiation and cartilage tissue homeostasis. As miRNAs play an important role in regulating gene expression during cell differentiation and maintaining tissue homeostasis, we determined the expression of the miRNAs with zonal differentiation and homeostasis. We investigated how chondrocyte dedifferentiation during multiple passages and redifferentiation in a three-dimensional (3D) agarose culture regulates the expression of these miRNAs by quantitative reverse transcription-polymerase chain reaction. Additionally, the effect of transforming growth factor beta 1 (TGF-β1), which is known to enhance chondrocytic differentiation and SZP expression, on these miRNAs was evaluated. The expression of miR-221 and miR-222 increased during dedifferentiation and during redifferentiation in a 3D culture and TGF-β1 restored them to normalcy. miR-140 dramatically decreased during dedifferentiation and its expression is partially recovered in a 3D culture with TGF-β1. miR-143 and miR-145 in the superficial chondrocytes decreased during dedifferentiation and further decreased in a 3D culture, but TGF-β1 partially recovered their expression in a 3D culture. In conclusion, the expression patterns of the miRNAs will be of functional utility for strategies and approaches to tissue engineering of the articular cartilage.

Inducing articular cartilage phenotype in costochondral cells

IntroductionCostochondral cells may be isolated with minimal donor site morbidity and are unaffected by pathologies of the diarthrodial joints. Identification of optimal exogenous stimuli will allow abundant and robust hyaline articular cartilage to be formed from this cell source.MethodsIn a three factor, two level full factorial design, the effects of hydrostatic pressure (HP), transforming growth factor β1 (TGF-β1), and chondroitinase ABC (C-ABC), and all resulting combinations, were assessed in third passage expanded, redifferentiated costochondral cells. After 4 wks, the new cartilage was assessed for matrix content, superficial zone protein (SZP), and mechanical properties.ResultsHyaline articular cartilage was generated, demonstrating the presence of type II collagen and SZP, and the absence of type I collagen. TGF-β1 upregulated collagen synthesis by 175% and glycosaminoglycan synthesis by 75%, resulting in a nearly 200% increase in tensile and compressive moduli. C-ABC significantly increased collagen content, and fibril density and diameter, leading to a 125% increase in tensile modulus. Hydrostatic pressure increased fibril diameter by 30% and tensile modulus by 45%. Combining TGF-β1 with C-ABC synergistically increased collagen content by 300% and tensile strength by 320%, over control. No significant differences were observed between C-ABC/TGF-β1 dual treatment and HP/C-ABC/TGF-β1.ConclusionsEmploying biochemical, biophysical, and mechanical stimuli generated robust hyaline articular cartilage with a tensile modulus of 2 MPa and a compressive instantaneous modulus of 650 kPa. Using expanded, redifferentiated costochondral cells in the self-assembling process allows for recapitulation of robust mechanical properties, and induced SZP expression, key characteristics of functional articular cartilage.

Induction of superficial zone protein (SZP)/lubricin/PRG 4 in muscle-derived mesenchymal stem/progenitor cells by transforming growth factor-β1 and bone morphogenetic protein-7

IntroductionArticular cartilage (AC) is an avascular tissue with precise polarity and organization. The three distinct zones are: surface, middle and deep. The production and accumulation of the superficial zone protein (SZP), also known as lubricin, by the surface zone is a characteristic feature of AC. To date, there is a wealth of evidence showing differentiation of AC from mesenchymal stem cells. Most studies that described chondrogenic differentiation did not focus on AC with characteristic surface marker SZP/lubricin. The present investigation was initiated to determine the induction of SZP/lubricin in skeletal muscle-derived mesenchymal stem/progenitor cells (MDMSCs) by transforming growth factor-β1 (TGF-β1) and bone morphogenetic protein-7 (BMP-7).MethodsMDMSCs were cultured as a monolayer at a density of 1 × 105 cells/well in 12-well tissue culture plates. Cell cultures were treated for 3, 7 and 10 days with TGF-β1 and BMP-7. The medium was analyzed for SZP. The cells were used to isolate RNA for RT-PCR assays for SZP expression.ResultsThe SZP/lubricin increased in a time-dependent manner on Days 3, 7 and 10 in the medium. As early as Day 3, there was a three-fold increase in response to 3 ng/ml of TGF-β1 and 300 ng/ml of BMP-7. This was confirmed by immunochemical localization of SZP as early as Day 3 after treatment with TGF-β1. The expression of SZP mRNA was enhanced by TGF-β1.ConclusionsThe present investigation demonstrated the efficient and reproducible induction of SZP/lubricin accumulation by TGF-β1 and BMP-7 in skeletal MDMSCs. Optimization of the experimental conditions may permit the utility of MDMSCs in generating surface zone-like cells with phenotypic markers of AC and, therefore, constitute a promising cell source for tissue engineering approaches of superficial zone cartilage.

Increased friction coefficient and superficial zone protein expression in patients with advanced osteoarthritis

To quantify the concentration of superficial zone protein (SZP) in the articular cartilage and synovial fluid of patients with advanced osteoarthritis (OA) and to further correlate the SZP content with the friction coefficient, OA severity, and levels of proinflammatory cytokines. Samples of articular cartilage and synovial fluid were obtained from patients undergoing elective total knee replacement surgery. Additional normal samples were obtained from donated body program and tissue bank sources. Regional SZP expression in cartilage obtained from the femoral condyles was quantified by enzyme-linked immunosorbent assay (ELISA) and visualized by immunohistochemistry. Friction coefficient measurements of cartilage plugs slid in the boundary lubrication system were obtained. OA severity was graded using histochemical analyses. The concentrations of SZP and proinflammatory cytokines in synovial fluid were determined by ELISA. A pattern of SZP localization in knee cartilage was identified, with load-bearing regions exhibiting high SZP expression. SZP expression patterns were correlated with friction coefficient and OA severity; however, SZP expression was observed in all samples at the articular surface, regardless of OA severity. SZP expression and aspirate volume of synovial fluid were higher in OA patients than in normal controls. Expression of cytokines was elevated in the synovial fluid of some patients. Our findings indicate a mechanochemical coupling in which physical forces regulate OA severity and joint lubrication. The findings of this study also suggest that SZP may be ineffective in reducing joint friction in the boundary lubrication mode at an advanced stage of OA, where other mechanisms may dominate the observed tribological behavior.

Effects of mechanical stimuli on the synthesis of superficial zone protein in chondrocytes

Superficial zone protein (SZP) has been demonstrated to contribute to the boundary lubrication in synovial joints. This study was designed to clarify the modulation of SZP expression by mechanical stress in articular chondrocytes. Cyclic tensile strains of 7 and 21% cell elongation were applied to cultured chondrocytes obtained from porcine mandibular condylar cartilage. The mRNA levels of SZP, IL-1 beta, and TGF-beta1 were examined by a quantitative real-time PCR analysis. Protein level of SZP was examined by Western blotting. The SZP mRNA level was significantly upregulated after 12, 24, and 48 h by 7% elongation. Although SZP mRNA level was upregulated by 21% elongation after 12 h, it decreased to a lower level than the control after 48 h. The TGF-beta1 mRNA level exhibited an almost similar change to SZP. The IL-1 beta mRNA level was not changed markedly by 7% elongation. However, the IL-1 beta mRNA level was significantly increased by a 12-h application of 21% elongation. Western blot analysis revealed that the SZP expression was increased by 7% elongation, but decreased remarkably by 21% elongation. It is suggested from these findings that the SZP expression level in the chondrocytes is enhanced by optimal mechanical stimuli, but inhibited by excessive loading partly affected by TGF-beta1 and IL-1 beta, leading to the deterioration of joint lubrication.

Superficial Zone Protein (Lubricin) in the Different Tissue Compartments of the Knee Joint: Modulation by Transforming Growth Factor Beta 1 and Interleukin-1 Beta

Superficial-zone protein (SZP), also known as lubricin, is a key mediator of boundary lubrication and plays an important role in the functional integrity of the diarthrodial joint. The aim of this investigation was to examine the role of transforming growth factor beta (TGF-beta) and interleukin-1 beta (IL-1beta) on the expression of SZP in various compartments of the bovine knee joint: the superficial zone of articular cartilage, synovium, meniscus, and anterior and posterior cruciate ligaments. The effects of TGF-beta1 and IL-1beta on SZP expression were examined in explants and cells from the different tissue compartments. TGF-beta1 up-regulated the expression of SZP in cultured explants, but IL-1beta down-regulated it. Quantitative analysis of secreted proteins in the medium of the cells demonstrated significant stimulation by TGF-beta1 and inhibition by IL1-beta of the accumulation of SZP protein in all four tissues. Real-time polymerase chain reaction analysis revealed that TGF-beta1 significantly up-regulated SZP expression and that IL-1beta down-regulated it. These results revealed the modulation of SZP expression in various compartments of the knee joint by TGF-beta1 and IL-1beta. In addition, SZP was found to be immunolocalized at the surface layer of cells in histological sections of all four tissue compartments. Collectively, results of the current study on regulation of SZP expression by TGF-beta and IL-1 help provide new insights, into tissue engineering strategies to repair and regenerate the different tissue compartments in the articular joint with optimal lubrication.

Induction of chondrogenesis and expression of superficial zone protein (SZP)/lubricin by mesenchymal progenitors in the infrapatellar fat pad of the knee joint treated with TGF-β1 and BMP-7

Superficial zone protein (SZP) is a key mediator of boundary lubrication of articular cartilage in joints. In this investigation, we made the unexpected discovery that SZP was expressed in infrapatellar fat pad (IFP) from bovine knee. Quantitative analysis of secreted proteins in the medium of the IFP stromal cells demonstrated a significant stimulation by TGF-β1 and BMP-7. Real-time PCR analysis revealed the SZP expression was up-regulated by TGF-β1 and BMP-7. Chondrogenically differentiated IFP progenitor cells were stimulated by TGF-β1 and BMP-7 to synthesize and secrete SZP. SZP mRNA was significantly up-regulated by chondrogenic induction for 21 days. These findings indicate that the stimulation of SZP expression by TGF-β and BMP-7 may lead to functional improvement of damaged intraarticular tissues and that IFP progenitor cells may be a potential useful source for inducing superficial zone of articular cartilage by tissue engineering for regeneration of damaged articular cartilage due to osteoarthritis

Engineered cartilage generated by nasal chondrocytes is responsive to physical forces resembling joint loading

To determine whether engineered cartilage generated by nasal chondrocytes (ECN) is responsive to different regimens of loading associated with joint kinematics and previously shown to be stimulatory of engineered cartilage generated by articular chondrocytes (ECA). Human nasal and articular chondrocytes, harvested from 5 individuals, were expanded and cultured for 2 weeks into porous polymeric scaffolds. The resulting ECN and ECA were then maintained under static conditions or exposed to the following loading regimens: regimen 1, single application of cyclic deformation for 30 minutes; regimen 2, intermittent application of cyclic deformation for a total of 10 days, followed by static culture for 2 weeks; regimen 3, application of surface motion for a total of 10 days. Prior to loading, ECN constructs contained significantly higher amounts of glycosaminoglycan (GAG) and type II collagen compared with ECA constructs. ECN responded to regimen 1 by increasing collagen and proteoglycan synthesis, to regimen 2 by increasing the accumulation of GAG and type II collagen as well as the dynamic modulus, and to regimen 3 by increasing the expression of superficial zone protein, at the messenger RNA level and the protein level, as well as the release of hyaluronan. ECA constructs were overall less responsive to all loading regimens, likely due to the lower extracellular matrix content. Human ECN is responsive to physical forces resembling joint loading and can up-regulate molecules typically involved in joint lubrication. These findings should prompt future in vivo studies exploring the possibility of using nasal chondrocytes as a cell source for articular cartilage repair.

Mechanotransduction of bovine articular cartilage superficial zone protein by transforming growth factor β signaling

Mechanical signals are key determinants in tissue morphogenesis, maintenance, and restoration strategies in regenerative medicine, although molecular mechanisms of mechanotransduction remain to be elucidated. This study was undertaken to investigate the mechanotransduction process of expression of superficial zone protein (SZP), a critical joint lubricant. Regional expression of SZP was first quantified in cartilage obtained from the femoral condyles of immature bovines, using immunoblotting, and visualized by immunohistochemistry. Contact pressure mapping in whole joints was accomplished using pressure-sensitive film and a load application system for joint testing. Friction measurements on cartilage plugs were acquired under boundary lubrication conditions using a pin-on-disk tribometer modified for reciprocating sliding. Direct mechanical stimulation by shear loading of articular cartilage explants was performed with and without inhibition of transforming growth factor beta (TGFbeta) signaling, and SZP content in media was quantified by enzyme-linked immunosorbent assay. An unexpected pattern of SZP localization in knee cartilage was initially identified, with anterior regions exhibiting high levels of SZP expression. Regional SZP patterns were regulated by mechanical signals and correlated with tribological behavior. Direct relationships were demonstrated between high levels of SZP expression, maximum contact pressures, and low friction coefficients. Levels of SZP expression and accumulation were increased by applying shear stress, depending on location within the knee, and were decreased to control levels with the use of a specific inhibitor of TGFbeta receptor type I kinase and subsequent phospho-Smad2/3 activity. These findings indicate a new role for TGFbeta signaling in the mechanism of cellular mechanotransduction that is especially significant for joint lubrication.

Expression of superficial zone protein in mandibular condyle cartilage

Superficial zone protein (SZP) has been shown to function in the boundary lubrication of articular cartilages of the extremities. However, the expression of SZP has not been clarified in mandibular cartilage which is a tissue that includes a thick fibrous layer on the surface. This study was conducted to clarify the distribution of SZP on the mandibular condyle and the regulatory effects of humoral factors on the expression in both explants and fibroblasts derived from mandibular condyle. The distribution of SZP was determined in bovine mandibular condyle cartilage, and the effects of interleukin-1beta (IL-1beta) and transforming growth factor-beta (TGF-beta) on SZP expression were examined in condyle explants and fibroblasts derived from the fibrous zone of condyle cartilage. SZP was highly distributed in the superficial zone of intact condyle cartilage. The SZP expression was up-regulated by TGF-beta in both explants and cultured fibroblasts, whereas the expression was slightly down-regulated by IL-1beta. A significant increase in accumulation of SZP protein was also observed in the culture medium of the fibroblasts treated with TGF-beta. These results suggest that SZP plays an important role in boundary lubrication of mandible condylar cartilage, is synthesized locally within the condyle itself, and exhibits differential regulation by cell mediators relevant to mandibular condyle repairing and pathologies.

Growth factor impact on articular cartilage subpopulations

We have examined the effects of growth factor stimulation on superficial and growth zone chondrocyte populations. Zonal articular chondrocytes from 8-month-old Spanish goat distal femurs were plated in monolayer cultures and stimulated by using insulin-like growth factor I (IGF-I), basic fibroblast growth factor (bFGF), and transforming growth factor-beta1 (TGF-beta1). Gene expression for collagen I and II, aggrecan, and superficial zone protein were evaluated every week for 3 weeks. Finally, proteoglycan and collagen deposition were measured for each experimental group. Major differences existed in the behavior of superficial and growth zone chondrocytes, the most apparent being the higher capacity for protein synthesis by the growth zone population. Variations also existed regarding growth factor treatment. TGF-beta1 had the greatest effect on proliferation over 8 days. With respect to differentiation, IGF-I increased average collagen II gene expression in the growth zone populations in comparison with growth zone controls. IGF-I increased aggrecan gene expression for the same groups. Superficial zone populations exhibited lower collagen II, collagen I, and aggrecan gene expression than the growth zone populations under all conditions. However, superficial zone protein expression was dramatically elevated in superficial zone populations by TGF-beta1. Collagen I expression showed a general increase under all conditions compared with initial values. Combined biosynthesis results showed that the superficial populations secreted little to no collagen, especially collagen II, in comparison with their growth zone counterparts. Glycosaminoglycan production was also much lower than for the growth zone groups. TGF-beta1 and IGF-I increased collagen II production in the growth zone populations. TGF-beta1 increased glycosaminoglycan secretions in the superficial zone populations and in the growth zone populations, whereas IGF-I produced an increase in glycosaminoglycan secretion only in the growth zone populations. Thus, growth factors elicit different proliferation, gene expression, and biosynthesis responses from zonal chondrocyte subpopulations.

Surface Motion Upregulates Superficial Zone Protein and Hyaluronan Production in Chondrocyte-Seeded Three-Dimensional Scaffolds

A cartilage engineering bioreactor has been developed that provides joint-specific kinematics. This study investigated the effect of articular motion on the gene expression of superficial zone protein (SZP) and hyaluronan synthases (HASs) and on the release of SZP and hyaluronan of chondrocytes seeded onto biodegradable scaffolds. Cylindrical (8 x 4 mm) porous polyurethane scaffolds were seeded with bovine articular chondrocytes and subjected to static or dynamic compression, with and without articulation against a ceramic hip ball. After loading, the mRNA expression of SZP and HASs was analyzed, and SZP immunoreactivity and hyaluronan concentration of conditioned media were determined. Surface motion significantly upregulated the mRNA expression of SZP and HASs. Axial compression alone had no effect on SZP and increased HAS mRNA only at high strain amplitude. SZP was immunodetected only in the media of constructs exposed to surface motion. The release of hyaluronan into the culture medium was significantly enhanced by surface motion. These results indicate that specific stimuli that mimic the kinematics of natural joints, such as articular motion, may promote the development of a functional articular surface-synovial interface.

Tissue engineering of stratified articular cartilage from chondrocyte subpopulations

Objective: To test if subpopulations of chondrocytes from different cartilage zones could be used to engineer cartilage constructs with features of normal stratification. Design: Chondrocytes from the superficial and middle zones of immature bovine cartilage were cultured in alginate, released, and seeded either separately or sequentially to form cartilage constructs. Constructs were cultured for 1 or 2 weeks and were assessed for growth, compressive properties, and deposition, and localization of matrix molecules and superficial zone protein (SZP). Results: The cartilaginous constructs formed from superficial zone chondrocytes exhibited less matrix growth and lower compressive properties than constructs from middle zone chondrocytes, with the stratified superficial-middle constructs exhibiting intermediate properties. Expression of SZP was highest at the construct surfaces, with the localization of SZP in superficial-middle constructs being concentrated at the superficial surface. Conclusions: Manipulation of subpopulations of chondrocytes can be useful in engineering cartilage tissue with a biomimetic approach, and in fabricating constructs that exhibit stratified features of normal articular cartilage.

Articular Cartilage Superficial Zone Protein (SZP) Is Homologous to Megakaryocyte Stimulating Factor Precursor and Is a Multifunctional Proteoglycan with Potential Growth-Promoting, Cytoprotective, and Lubricating Properties in Cartilage Metabolism

We have performed cDNA sequencing and homology analyses to elucidate the complete amino acid composition for a superficial zone protein (SZP) from human and bovine cartilage which has previously been shown to be a proteoglycan specifically synthesized by chondrocytes located at the surface of bovine articular cartilage and also some synovial lining cells. The results of this study indicate that cartilage SZP is homologous with a glycoprotein first described as the precursor protein of a megakaryocyte stimulating factor (MSF). Sequence comparisons and analyses indicate that (i) the amino acid composition of SZP is highly conserved between bovine and human species, (ii) SZP contains structural motifs at the N- and C-termini which are similar to those found in vitronectin and which may impart cell-proliferative and matrix-binding properties to the molecule, and (iii) SZP contains large and small mucin-like repeat domains composed of the sequences KEPAPTTT/P (76–78 repeats) andXXTTTX(6–8 repeats), respectively, which occur within a large central region of ∼940 amino acids. The mucin-like domains are likely to be substituted with O-linked oligosaccharides which would impart lubricating properties to SZP which in part accumulates at the articular cartilage–synovial fluid interface. Additionally, we have shown that interleukin-1 inhibits the biosynthesis of chondrocyte SZP, while TGF-β and IGF-1 increase its biosynthesis, and that in pathological (osteoarthritic) human articular cartilage SZP mRNA can be expressed as an alternatively spliced variant lacking exons 4 and 5 which encode a potential heparin binding domain. The occurrence of different SZP alternative splice variants and the differential expression of SZP in the presence of cytokines and growth factors suggest that SZP may play an important cytoprotective role by preventing cellular adhesion to the articular cartilage surface in normal cartilage metabolism. Modifications to the structure of SZP, coupled with inhibition of SZP synthesis during inflammation, may account for the attachment and invasion of pannus observed in inflammatory joint diseases.