Biomedical subjects

G J van Osch

Publications and source records attributed to G J van Osch.

At least 19 recordsLinked to original sources

PubMed2001

Monoclonal antibody 11-fibrau: a useful marker to characterize chondrocyte differentiation stage.

The aim of this study was to determine the feasibility of discriminating between differentiated and dedifferentiated chondrocytes by using the Mab 11-fibrau. Mab 11-fibrau did not bind to differentiated chondrocytes in cartilage of human knee joint, auricle, or nasal septum. During monolayer culture, when cells dedifferentiate, the number of 11-fibrau positive cells gradually increased and reached up to 100% after 4 passages. When differentiated chondrocytes were cultured in alginate, most (90--95%) of the cells remained 11-fibrau negative, in accordance with previous studies demonstrating that differentiated chondrocytes cultured in alginate keep their phenotype. Dedifferentiated (11-fibrau positive) cells were subjected to different redifferentiation regimes. As a well-known fact, cultures in alginate in medium where FCS was replaced by IGF1 and TGF beta 2 results in increased collagen type II formation, indicative for redifferentiation. However, the cells remained 11-fibrau positive, suggesting they are not (yet) fully redifferentiated. On the other hand, when dedifferentiated cells (after 4 passages in monolayer culture) were seeded in a biomaterial and implanted subcutaneously in a nude mouse, the newly formed cartilage matrix contained collagen type II and the 11-fibrau staining on the cells had disappeared. Our results indicate that 11-fibrau may be a reliable and sensitive marker of chondrocyte phenotype.

Animals↗

PubMed2001

A new in vivo model for testing cartilage grafts and biomaterials: the 'rabbit pinna punch-hole' model.

In this study an animal model was developed for evaluation of the feasibility of cartilage grafts. In the cartilage of the external ear of the rabbit multiple holes, 6 mm in diameter, were punched, leaving the adherent skin intact. Different experimental groups were evaluated. First, the punch-hole model was validated under various conditions to study spontaneous or perichondrial initiated regeneration of the cartilage defect. When both cartilage and perichondrium was excised no spontaneous repair of the cartilage defect was observed. When perichondrium is present, variable patch-like closure of the punch hole was found. As 'golden standard' a punched out piece of cartilage was reimplanted directly. This condition showed adequate closure of the punch hole, however, no perfect integration of graft and surrounding cartilage was observed. Secondly, to evaluate the 'punch-hole model' a biomaterial, trabecular demineralized bovine bone matrix (DBM), was implanted and tested as a scaffold for tissue engineering techniques in vivo and in vitro. Direct implantation of DBM did not lead to any cartilage formation to close the defect. In vivo engineered cartilage, generated by enveloping DBM in perichondrium for 3 weeks, could adequately close the punch hole. When DBM was seeded with isolated chondrocytes in vitro before implantation in the defect, a highly fragmented graft, with some islets of viable cells was seen. To promote an efficient and reliable evaluation of cartilage grafts a semi-quantitative grading system was developed. Items such as quality, quantity and integrity of the cartilage graft were included in a histomorphological grading system to provide information about the properties of a specific cartilage graft. To validate the grading system, all conditions were scored by two independent observers. An excellent reliability (R = 0.96) was seen between the observers. In summary, the rabbit pinna punch-hole model is a reliable and efficient method for first evaluation of cartilage grafts. The results can be easily analyzed using a semi-quantitative grading system.

Animals↗

PubMed2001

Growth factor expression in cartilage wound healing: temporal and spatial immunolocalization in a rabbit auricular cartilage wound model.

OBJECTIVE: The ability of cartilage to regenerate following injury is limited, potentially leading to osteoarthritis. Integrative cartilage repair, necessary for durable restoration of cartilage lesions, can be regarded as a wound healing process. Little is known about the effects of growth factors regulating acute cartilage wound healing in vivo. In this study the temporal expression patterns of growth factors and proteoglycan content in cartilage wound edges in vivo were studied. DESIGN: Cartilage wounds were created in rabbit ear cartilage using a 6 mm biopsy punch. Specimens were subsequently harvested 1, 3, 7, 14 and 28 days after surgery. Paraffin sections were thionin stained to visualize proteoglycan loss and replacement. Immunohistochemical staining of TGFbeta1, TGFbeta3, IGF-1, IGF-II and FGF-2 was used to define growth factor expression at the cartilage wound sites. RESULTS: Almost no effect of cartilage wounding was observed one day after surgery. A decrease of proteoglycan content, with a maximal loss at day 7, and a subsequent restoration was observed at the wound edges. Growth factor expression increased simultaneously. Maximal immunostaining for IGF1, IGFII, FGF2 and TGF-beta3 was observed at day 7, followed by a gradual decrease. Increased expression of TGFbeta1 lasted from day 3 until day 14. CONCLUSION: We have demonstrated the ability of chondrocytes to increase growth factor expression and to restore the rapid decrease in proteoglycan content in the initial phase following acute wounding. A temporal increase in intracellular growth factor expression suggests an autocrine and/or paracrine metabolic stimulation, which can be regarded a sign of chondrocytes repair capacity.

Animals↗

PubMed2001

In vitro redifferentiation of culture-expanded rabbit and human auricular chondrocytes for cartilage reconstruction.

To construct an autologous cartilage graft using tissue engineering, cells must be multiplied in vitro; they then lose their cartilage-specific phenotype. The objective of this study was to assess the capacity of multiplied ear chondrocytes to re-express their cartilage phenotype using various culture conditions. Cells were isolated from the cartilage of the ears of three young and three adult rabbits and, after multiplication in monolayer culture, they were seeded in alginate and cultured for 3 weeks in serum-free medium with insulin-like growth factor 1 (IGF-1) and transforming growth factor-beta2 (TGF-beta2) in three different dose combinations. As a control, cells were cultured in 10% fetal calf serum, which was demonstrated in previous experiments to be unable to induce redifferentiation. Chondrocytes from the ears of young, but not adult, rabbits, synthesized significantly more glycosaminoglycan when serum was replaced by insulin-like growth factor-1 and transforming growth factor-beta2. The number of collagen type II-positive cells was increased from 10 percent to 97 percent in young cells and to 33 percent in adult cells. Using human ear cells from 12 patients (aged 7 to 60 years), glycosaminoglycan synthesis could also be stimulated by replacing serum with insulin-like growth factor and transforming growth factor-beta. Although the number of collagen type II-positive cells could be increased under these conditions, it never reached above 10 percent. Data from five patients showed that further optimization of the culture conditions by adding ITS+ and cortisol significantly increased (doubled or tripled) both glycosaminoglycan synthesis and collagen type II expression. In conclusion, this study demonstrates a method to regain cartilage phenotype in multiplied ear cartilage cells. This improves the chances of generating human cartilage grafts for the reconstruction of external ears or the repair of defects of the nasal septum.

Adolescent↗

PubMed2000

Tissue-engineered cartilage using serially passaged articular chondrocytes. Chondrocytes in alginate, combined in vivo with a synthetic (E210) or biologic biodegradable carrier (DBM).

In vitro multiplication of isolated autologous chondrocytes is required to obtain an adequate number of cells to generate neo-cartilage, but is known to induce cell-dedifferentiation. The aim of this study was to investigate whether multiplied chondrocytes can be used to generate neo-cartilage in vivo. Adult bovine articular chondrocytes, of various differentiation stages, were suspended in alginate at densities of 10 or 50 million/ml, either directly after isolation (P0) or after multiplication in monolayer for one (P1) or three passages (P3). Alginate with cells was seeded in demineralized bovine bone matrix (DBM) or a fleece of polylactic/polyglycolic acid (E210) and implanted in nude mice for 8 weeks. The newly formed tissue was evaluated by Alcian Blue and immunohistochemical staining for collagen type-II and type-I. Structural homogeneity of the tissue, composed of freshly isolated as well as serially passaged cells, was found to be enhanced by high-density seeding (50 million/ml) and the use of E210 as a carrier. The percentage of collagen type-II positive staining P3-cells was generally higher when E210 was used as a carrier. Furthermore, seeding P3-chondrocytes at the highest density (50 million/ml) enhanced collagen type-II expression. This study shows promising possibilities to generate structurally regular neo-cartilage using multiplied chondrocytes in alginate in combination with a fleece of polylactic/polyglycolic acid.

Alcian Blue↗

PubMed2000

Fixation-dependent immunolocalization shift and immunoreactivity of intracellular growth factors in cartilage.

The effects of fixation on immunolocalization and immunoreactivity in cartilage tissues were studied using monoclonal antibodies against peptides that can effectively stimulate chondrocytes in vitro and have been shown to play a role in musculoskeletal tissue regeneration: transforming growth factor beta1, transforming growth factor beta3, insulin-like growth factor I, insulin-like growth factor II and fibroblast growth factor 2. Paraffin sections fixed in buffered formalin, buffered paraformaldehyde, Carnoy and methacarn, as well as cryosections, were tested. A strong immunoreaction was observed in tissue fixed in formaldehyde-based fixatives, with a resemblance to that in cryopreserved tissues. Immunoreactivity was reduced in alcohol-fixed tissues. Furthermore, a striking intracellular immunolocalization shift from cytoplasm to nucleus was observed using alcohol-based fixatives as compared to cryopreserved or formaldehyde-based fixatives. We concluded that, for the detection and localization of growth factors in cartilage tissues, fixation in buffered formalin or paraformaldehyde is optimal.

Animals↗

PubMed1999

The role of trabecular demineralized bone in combination with perichondrium in the generation of cartilage grafts.

The use of a composite graft of bovine trabecular demineralized bone matrix (DBM) and perichondrium has been found a reliable method for in vivo generation of cartilage. In the present study, the mechanism whereby this commercially available matrix increases cartilage formation was investigated. First, the time course of cartilage formation in vivo, in the combined implant of perichondrium and DBM in the rabbit ear was studied, with special focus on tissue reactions to DBM. DBM was colonized by macrophages from day 3 post-operatively, reaching a maximum after 2 weeks. Only a minimal number of neutrophils was found. After 3 weeks the DBM appeared to be resorbed. In the first week the DBM was invaded with chondroblasts, and chondrogenesis occurred between the first and second week of implantation. After 3 weeks, the initially formed islets of cartilage had fused. Next, the chondrogenic capacity of DBM itself was investigated by implantation of DBM without perichondrium. This never resulted in cartilage formation. Immunohistochemistry showed only a faint staining of the DBM for growth factors. This indicates a minimal chondrogenic effect of DBM alone and the requirement of perichondrium as cell provider. In order to define the conditions which cause chondrogenesis in composites of perichondrium and DBM, a series of in vitro culture experiments was performed in which the in vivo situation was mimicked step by step. The basic condition was perichondrium cultured in medium with 10% FCS. In this condition, cartilage formation was variable. Because in the in vivo situation both DBM and macrophages can release growth factors, the effect of IGF1, TGFbeta2 or OP1 added to the culture medium was tested. Neither the incidence nor the amount of cartilage formation was stimulated by addition of growth factors. Perichondrium wrapped around DBM in vitro gave cartilage formation in the perichondrium but the incidence and amount were not significantly stimulated compared to cultures of perichondrium without DBM. However, cartilage-like cells were found in the DBM suggesting an effect of DBM on perichondrium-derived cells. Finally, macrophages and/or blood were added to the composite DBM-perichondrium to mimic the in vivo situation as close as possible. However, no effect of this treatment was found. In conclusion, this study indicates that DBM itself has few chondrogenic qualities but functions merely as a spacer for cell ingrowth. The fast resorption of DBM by macrophages in vivo seems of importance for the cartilage forming process, but in vitro the presence of macrophages (in combination with blood) could not enhance chondrogenesis.

Animals↗

PubMed1999

Optimization of chondrocyte expansion in culture. Effect of TGF beta-2, bFGF and L-ascorbic acid on bovine articular chondrocytes.

In vitro multiplication of isolated chondrocytes is needed to repair articular cartilage defects with autologous material. In this study we used monolayer cultures of bovine articular chondrocytes. The effect of transforming growth factor beta-2, basic fibroblast growth factor or L-ascorbic acid on cell multiplication, in the presence of 10% fetal calf serum, was measured in primary culture, the third and tenth passage. TGF beta-2 stimulated the proliferation of chondrocytes in the primary culture and L-ascorbic acid stimulated in the third passage. On the basis of these results, we chose an optimal addition scheme in which TGF beta-2 was added in primary culture and first passage, followed by addition of L-ascorbic acid in the second and third passage; this resulted in a 7-fold increase in cell number compared to the control group, in about 4 weeks. Our findings stress the importance of adding the right growth factor at the right moment. Collagen type II expression was lost after the third passage, in the control as well as in the experimental condition. The ability to produce hyaline cartilage specific matrix components is essential, if multiplied cells are to be used to repair cartilage defects.

Animals↗

PubMed1998

Wound healing of cartilage structures in the head and neck region.

This study was performed to determine the various processes involved in the behaviour of hyaline cartilage during the wound healing period after trauma or surgery of vulnerable structures like the nasal septal cartilage and the cricoid. The results of different procedures (perpendicular and parallel to the cartilage surface) in young and young-adult animals were analyzed: septal incision at different locations (young-old), cricoid split (young-old), suturing cartilage, closing defects with autologous cartilage (young), biomaterials (young) and newly engineered cartilage in 4- and 24-week-old rabbits (series of ten animals). Cartilage of the young rabbit and child have similar hyaline cartilage with a varying distribution in thickness. Thinner areas are more susceptible to malformations. Incisions through younger cartilage give rise to some new cartilage formation covered by a new layer of perichondrium: through older, differentiated cartilage the incision causes superficial but permanent necrosis. Edges of cut cartilage mostly do heal by formation of fibrous junctions. This forms a weak spot, sensitive to deviations. The same fate goes for the healing between the autologous graft and the surrounding pre-existent cartilage. Trauma parallel to the surface, leads to inconsistent quantity of neocartilage. With ageing the wound healing and regenerative capacities decrease. In general, biomaterials are less accepted by the surrounding tissues and would impede further growth. Only newly engineered, and thus less differentiated (younger) cartilage of hyaline nature, appeared to be well accepted at the interface with the edges of a cartilage defect. There are indications that the release of growth factors might play a role in cartilage wound healing. In the child as well as the adult, wound healing of hyaline cartilage structures is incomplete, and surgery remains 'experimental' surgery. The clinical implications of gradual loss of the regenerative capacity of hyaline cartilage should be further investigated.

Animals↗

PubMed1998

The immediate effects of local trauma on the shape of the cricoid cartilage.

Injury-induced abnormal development of the cricoid ring has been demonstrated in previous growth studies. In this study we focused on the immediate effects of various types of lesions to the cricoid, eliminating the influence of inserting muscles. In isolated, vital cricoids (cricoid explants) the anterior arch was split, creating a small gap between the cut ends. Previous injury to the internal surface of the cricoid ring resulted in a three to four fold increase of the diameter of the gap, actually widening the interrupted cricoid. On the contrary, injuring the external surface of the cricoid cartilage prior to anterior cricoid split, leads to an overlap of the cut edges, and a narrowing of the ring. These injury-specific changes in shape of the cricoid ring are ascribed to the release of interlocked stresses, present in the cartilage. It is suggested that the demonstrated methods to change the shape of the cricoid ring in a predictable way, are relevant for the treatment of patients with cricoid malformation.

Analysis of Variance↗

PubMed1998

Effect of transforming growth factor-beta on proteoglycan synthesis by chondrocytes in relation to differentiation stage and the presence of pericellular matrix.

The effects of transforming growth factor-beta (TGF-beta) on proteoglycan synthesis of chondrocytes are controversial. The hypothesis that the differential effect of TGF-beta is related to the differentiation stage of the chondrocytes is investigated in this study. Rabbit auricular chondrocytes were cultured in alginate. When seeded in alginate immediately after isolation, cells keep their cartilaginous phenotype. When cells are first cultured in monolayer, they lose their cartilaginous phenotype and become dedifferentiated. We used three different cell populations: (1) Differentiated cells (P0: immediately after isolation); (2) partially (de)differentiated cells (P1: after one passage in monolayer); (3) dedifferentiated cells (P4: after four passages in monolayer). Cells were characterized by morphology using electron microscopy, amount of proteoglycans using the Farndale assay and type of collagen produced using immunohistochemistry. The effects of addition of 10 ng/ml TGF-beta2 for 7 days to P0, P1 and P4 cells were compared. TGF-beta was added either directly from the start of the alginate culture, or after a preculture period of three weeks in alginate. The amount of proteoglycans was increased in all chondrocyte populations when TGF-beta was added immediately after seeding in alginate, indicating that the effect of TGF-beta on proteoglycan synthesis does not depend on the differentiation stage of cells. After preculture in alginate, stimulation of proteoglycan synthesis (as measured by amount of proteoglycans and 35S-sulfate incorporation) had vanished. This effect was independent of differentiation stage . A dose-response experiment with TGF-beta (1, 10, 50 ng/ml) confirmed this differentiation-stage-independent effect of TGF-beta on proteoglycan synthesis. Stimulation by TGF-beta can be retained after enzymatic digestion of the pericellular matrix and reseeding of the cells in alginate, indicating the importance of pericellular matrix for the effect of TGF-beta on matrix synthesis. Alkaline phosphatase (ALP) activity was largely inhibited by TGF-beta in P0 chondrocytes, either with or without preculture in alginate. After culturing in monolayer, ALP activity was not substantially changed by TGF-beta. This indicates that the effect of TGF-beta on ALP activity, in contrast to the effect on proteoglycan synthesis, does depend on the differentiation stage of the cells. Furthermore, the fact that ALP synthesis in P0 cells is still inhibited by TGF-beta after preculture indicates that these cells remain responsive to TGF-beta. This provides additional evidence for the importance of the pericellular matrix for regulation of the effect of TGF-beta on proteoglycan synthesis. The results indicate that, in pathological cartilage, matrix depletion might be the trigger for increased matrix synthesis in reaction to TGF-beta, suggesting an important role for TGF-beta in cartilage repair.

Animals↗

PubMed1998

Differential effects of IGF-1 and TGF beta-2 on the assembly of proteoglycans in pericellular and territorial matrix by cultured bovine articular chondrocytes.

OBJECTIVES: Knowledge of matrix assembly is necessary to understand the pathogenesis of disease processes and to find solutions for repair of articular cartilage lesions. The influence of growth factors on matrix assembly is largely unknown. We investigated whether, and to what degree, insulin-like growth factor (IGF-1) and transforming growth factor beta-2 (TGF beta-2) influence proteoglycan synthesis and accumulation in the cell-associated matrix compartment (consisting of pericellular and territorial matrix) compared to the further-removed matrix compartment (consisting of the interterritorial matrix). DESIGN: Bovine articular chondrocytes were cultured in alginate beads for day 13. The effect of addition of 25 ng/ml IGF-1 or 25 ng/ml TGF beta-2 during the last 7 days in culture was determined. Cell-associated and further-removed matrix compartments were separated by centrifugation after sodium citrate/EDTA treatment. The amount of DNA, the total amount of proteoglycans and the amount of newly synthesized proteoglycans were analyzed biochemically. Morphometric analysis on electron micrographs was used to calculate the volumes of the cell-associated and further-removed matrix components. RESULTS: It was demonstrated in control beads that 25 +/- 8% of the proteoglycans were laid down in the cell-associated matrix compartment compared with 75 +/- 8% in the further-removed matrix compartment. The cell-associated matrix compartment in intact beads could be recognized in electron microscopy by a delineation of dense amorph material. Morphometric evaluation showed a relative volume of the cell-associated matrix compartment of 5.2 +/- 0.6% compared with 91.3 +/- 0.8% of the further-removed matrix compartment and 3.5 +/- 0.3% of the area occupied by cells. Combination of biochemical and morphometric results showed that the concentration of proteoglycans in the cell-associated matrix compartment was 3.63 +/- 0.32 mg/ml. By adding IGF-1 or TGF beta-2, the amount of both total accumulated proteoglycans and newly synthesized [35S]proteoglycans at day 13 in culture increased. In addition to an overall rise in proteoglycan content, IGF-1 significantly increased (24%) the percentage of proteoglycans laid down in the cell-associated matrix compartment while not changing the relative volume of this compartment (5.2 +/- 0.8%). This leads to a 82% (P < 0.05) increase in the proteoglycan concentration in the cell-associated matrix compartment compared to control beads. In contrast, TGF beta-2 significantly decreased (24%) the relative amount of proteoglycans in the cell-associated matrix compartment which was paralleled by a reduction of the relative volume from 5.2 +/- 0.6 to 3.6 +/- 1.4%. This leads to a significant increase of 87% of the proteoglycan concentration in the cell-associated matrix compartment. CONCLUSIONS: This study demonstrates that both IGF-1 and TGF beta-2 significantly but differently influence proteoglycan synthesis and accumulation in the cell-associated matrix compartment of cultured bovine chondrocytes in alginate. Both growth factors increase the concentration of proteoglycans in the cell-associated matrix compartment. However, addition of TGF beta-2 to bovine articular chondrocytes cultured in alginate beads for 13 days results in a significant reduction of the relative volume of the pericellular matrix compartment compared to controls, indicating differences in assembly of the matrix.

Animals↗

PubMed1998

Efficacy of perichondrium and a trabecular demineralized bone matrix for generating cartilage.

A pedicled auricular perichondrial flap wrapped around trabecular demineralized bovine bone matrix can generate an autologous cartilage graft. In earlier experimental studies, it was demonstrated that this graft could be used for nasal and cricoid reconstruction. It was assumed that the vascularization of the perichondrial flap was obligatory, but it was never proven that the flap should be pedicled. Moreover, for clinical use, the dimensions of the auricle would set restrictions to the size of the graft generated. Therefore, the possibility to generate cartilage with a composite graft of a free perichondrial flap wrapped around demineralized bovine bone matrix, by using young New Zealand White rabbits, was studied. This composite graft was implanted at poorly (subcutaneously in the abdominal wall; n = 12), fairly (subcutaneously in the pinna; n = 12), and well-vascularized sites (quadriceps muscle; n = 12). As a control, trabecular demineralized bovine bone matrix was implanted without perichondrial cover. Half of these grafts (n = 6) were harvested after 3 weeks, and the remaining grafts (n = 6) after 6 weeks of implantation. In histologic sections of these grafts, the incidence of cartilage formation was scored. Furthermore, the amount of newly formed cartilage was calculated by computerized histomorphometry. Trabecular demineralized bovine bone matrix without perichondrial cover demonstrated early resorption; no cartilage or bone was formed. In demineralized bovine bone matrix wrapped in perichondrium, early cartilage formed after 3 weeks at well- and fairly vascularized sites. No cartilage could be detected in grafts placed at a poorly vascularized site after 3 weeks; minimal cartilage formed after 6 weeks. In summary, the highest incidence of cartilage formed when trabecular demineralized bovine bone matrix was wrapped either in a pedicled auricular perichondrial flap or in a free perichondrial flap, which was placed at a well-vascularized site. Second, a significantly higher percentage of the total area of the graft was cartilaginized at well-vascularized sites after 3 weeks. The newly generated cartilage contained collagen type II and proteoglycans with hyaluronic acid binding regions, whereas collagen type I was absent, indicating the presence of hyaline cartilage. This study demonstrates that new cartilage suitable for a graft can be generated by free perichondrial flaps, provided that the site of implantation is well vascularized. Consequently, the size of such a graft is no longer limited to the dimensions of the auricle.

Animals↗

PubMed1997

Growth dynamics of the cricoid cartilage and subglottic injury. An autoradiographic and histometric study in the rabbit.

Earlier studies in young growing rabbits have suggested that post-traumatic developmental deformities of the cricoid might play a role in failing treatment in children with similar injuries. Specific patterns of malformations in the animals occurred preferentially at the anterior side of the cricoid ring. In this study, the growth dynamics of the cricoid were studied by histometry and autoradiography. Results indicated that (1) the cell/matrix ratio remains unchanged from 4 to 24 weeks of age, with both tissue components thus contributing equally to growth of the cricoid; (2) mitoses occur in cartilage and perichondrium, indicating interstitial as well as appositional growth; (3) mitotic activity is mainly restricted to the first 4 weeks of life, while cell hypertrophy is thereafter the dominant feature; (4) the highest degree of mitotic activity and cell hypertrophy is found in the anterior half of the cricoid ring which therefore can be considered as a growth center. This center might play a role in the development of trauma-induced deformation in the anterior part of the cricoid ring.

Aging↗

PubMed1996

Quantification of morphometric changes in murine experimental osteoarthritis using image analysis.

The aim of this study was to evaluate a method for the quantification of cartilage erosions and osteophyte sizes in a murine model of osteoarthritis (OA). Mice in which OA was induced in the knee joint by intra-articular injection of bacterial collagenase were used. With an interactive image analysis system, the areas occupied by osteophytes and the areas of erosions of the articular cartilage were measured on histological sections by two independent observers at two time points. Measurements of osteophyte areas and cartilage loss at the tibial plateau showed good reproducibility, whereas measurement of cartilage loss at the femoral condyles was less reproducible. Measurement of three frontal total knee joint sections from the middle part of the joint provided a reliable measure for cartilage damage and osteophyte size in the total joint. A cumulative score was developed, composed of both cartilage loss and osteophyte size, which can be used as a general measure for OA of the knee joint. The presented method of quantitative scoring makes it possible to perform correlation studies and to investigate the effect of therapeutic interventions on the osteoarthritis process.

Animals↗

PubMed1996

The relation between cartilage damage and osteophyte size in a murine model for osteoarthritis in the knee.

The aim of this study was to investigate the relationship between location and size of osteophytes and cartilage loss in an instability-induced experimental model for osteoarthritis. Osteoarthritis was induced in murine knee joints by injection of highly purified bacterial collagenase, causing joint instability. The size of the osteophytes and the cartilage loss were measured at different locations in the joint using image analysis on histological sections of total kees. Cartilage damage did not occur without osteophytes. Osteophytes were located on both medial and lateral sides, independent of the location of cartilage damage, but the size of the osteophytes was related to the amount of cartilage damage on the corresponding side. Cartilage loss on the lateral tibial plateau correlated well with the size of lateral osteophytes, in particular with the osteophyte at the margin of the lateral tibial plateau. Cartilage loss on the medial tibial plateau appeared to have a good correlation with the size of medial osteophytes, which was most pronounced for the osteophyte on the medial margin of the tibial plateau. This side-specific correlation between cartilage damage and osteophyte formation suggests compartmentalization of the osteoarthritic process.

Animals↗

PubMed1996

In vitro laxity-testers for knee joints of mice.

The knee joints of mice can be used as a model for studying the effects of interventions on knee laxity. The goal of this study was to quantify knee joint laxity in vitro. Three devices were developed: a positioning- and cementing device, an anterior-posterior (AP) laxity tester and a varus-valgus (VV) laxity tester. The positioning and cementing device was used to position the joint in a reproducible way and to attach clamping pins to the proximal femur and distal tibia using PM MA. The clamping pins were used to fix the joint to the AP- and VV-testers. In both testers the load was applied by means of a spindle-actuated spring while load and displacements were measured simultaneously. The load--displacement data were used to calculate displacement and compliance parameters. The performance of the testers was evaluated by testing 5 normal knee joints of 5 mice. Total AP-translation at + or - 0.8 N was 0.43 (+ or - 0.16 S.D.) mm with compliances of 0.14 (+ or - 0.05 S.D.) mm N(1) and 0.12 ( + or - 10.05 S.D.) mm N(-1) at 0.8 N posterior and anterior force, respectively. Total VV-rotation at + or - 4 Nmm was 17.2 (+ or - 2.6 S.D.) degrees with compliances of 0.9 degrees Nmm(-1) (+ or - 0.2 degrees Nmm(-1) S.D.) and 1.0 Nmm(-1) (+ or - 0.4 degrees Nmm(-1) S.D.) at 4 Nmm valgus and varus moment, respectively. The contributions of the deformations of the bones and the fixtures to the rotations were negligible in the VV-test. In the AP-test they account for approximately 0.07 ( + or - 0.03 S.D.)mm of the total AP-translation. This will not affect the utilization of the device for comparative analysis. It is concluded that in in vitro evaluation of AP- and VV-laxity in knees of mice is feasible with sufficient accuracy for evaluation of changes after ligament damage.

Animals↗

PubMed1996

Relation of ligament damage with site specific cartilage loss and osteophyte formation in collagenase induced osteoarthritis in mice.

OBJECTIVE: To investigate the correlation between initial ligament damage and development of subsequent osteoarthritic changes. METHODS: Collagenase was injected intraarticularly into the knee joint of mice of strain C57B16 or C57B110. After 3 days, ligament damage was evaluated by measurements of knee laxity in the anterior-posterior direction as a measure of cruciate ligament function, and in the varus-valgus direction as a measure of collateral ligament function. The amount and location of cartilage loss and osteophyte formation were determined at Day 42. RESULTS: Significant correlations between the amount of laxity changes and the severity of cartilage loss (r = 0.78), the amount of laxity changes and the size of osteophytes (r = 0.87), and between the severity of cartilage loss and osteophyte size (r = 0.94) were demonstrated. The amount of cartilage loss and the degree of osteophyte formation at the medial side of the joint depended mainly on the severity of cruciate ligament damage. This is contrast to changes at the lateral side of the joint, which appeared not to be associated with the severity of ligament damage. CONCLUSION: A strong relationship exists between the severity of cruciate ligament damage and the severity of osteoarthritic changes on the medial side of the joint. In the lateral joint compartment, prone to spontaneous osteoarthritis in the mouse strain studied, this relation is absent.

Animals↗