The local concentration of Ca2+ correlates with BMP7 expression and osseointegration in patients with total hip arthroplasty.

BACKGROUND: A successful osseointegration of total hip arthroplasty (THA) relies on the interplay of implant surface and bone marrow microenvironment. This study was undertaken to investigate the impact of perioperative biochemical molecules (Ca2+, Mg2+, Zn2+, VD, PTH) on the bone marrow osteogenetic factors (BMP2, BMP7, Stro-1+ cells) in the metaphyseal region of the femoral head, and further on the bone mineral density (BMD) of Gruen R3. METHODS: Bone marrow aspirates were obtained from the discarded metaphysis region of the femoral head in 51 patients with THA. Flow cytometry was used to measure the Stro-1+ expressing cells. ELISA was used to measure the concentrations of bone morphologic proteins (BMP2 and BMP7) and the content of TRACP5b in serum. TRAP staining was used to detect the osteoclast activity in the hip joint. The perioperative concentrations of the biochemical molecules above were measured by radioimmunoassay. The BMD of Gruen zone R3 was examined at 6 months after THA, using dual-energy X-ray absorptiometry (DEXA). RESULTS: Our data demonstrated that the concentration of Ca2+ was positively correlated with BMP7 expression, and with the postoperative BMD of Gruen zone R3. However, the concentration of Mg2+ had little impact on the R3 BMD, although it was negatively correlated with the expression of BMP7. Osteoclast activity in hip joint tissue of patients with femoral neck fractures was increased. Compared with the patients before THA, the levels of TRACP5b in serum of patients after THA were decreased. The data also suggested that the other biochemical molecules, such as Zn2+, VD, and PTH, were not significantly correlated with any bone marrow osteogenetic factors (BMP2, BMP7, Stro-1+ cells). The postoperative R3 BMD of patients of different gender and age had no significant difference. CONCLUSIONS: These results indicate the local concentration of Ca2+ may be an indicator for the prognosis of THA patients.

Multiple mesenchymal progenitor cell subtypes with distinct functional potential are present within the intimal layer of the hip synovium.

BACKGROUND: The synovial membrane adjacent to the articular cartilage is home to synovial mesenchymal progenitor cell (sMPC) populations that have the ability to undergo chondrogenesis. While it has been hypothesized that multiple subtypes of stem and progenitor cells exist in vivo, there is little evidence supporting this hypothesis in human tissues. Furthermore, in most of the published literature on this topic, the cells are cultured before derivation of clonal populations. This gap in the literature makes it difficult to determine if there are distinct MPC subtypes in human synovial tissues, and if so, if these sMPCs express any markers in vivo/in situ that provide information in regards to the function of specific MPC subtypes (e.g. cells with increased chondrogenic capacity)? Therefore, the current study was undertaken to determine if any of the classical MPC cell surface markers provide insight into the differentiation capacity of sMPCs. METHODS: Clonal populations of sMPCs were derived from a cohort of patients with hip osteoarthritis (OA) and patients at high risk to develop OA using indexed cell sorting. Tri-differentiation potential and cell surface receptor expression of the resultant clones was determined. RESULTS: A number of clones with distinct differentiation potential were derived from this cohort, yet the most common cell surface marker profile on MPCs (in situ) that demonstrated chondrogenic potential was determined to be CD90+/CD44+/CD73+. A validation cohort was employed to isolate cells with only this cell surface profile. Isolating cells directly from human synovial tissue with these three markers alone, did not enrich for cells with chondrogenic capacity. CONCLUSIONS: Therefore, additional markers are required to further discriminate the heterogeneous subtypes of MPCs and identify sMPCs with functional properties that are believed to be advantageous for clinical application.

Mechanical properties of human bone-implant interface tissue in aseptically loose hip implants.

The main cause of failure in total hip replacement is aseptic loosening which is associated with the formation of a periprosthetic fibrous (interface) tissue. Despite important applications for finite element modeling of loose implants, the mechanical properties of the bone-implant interface tissue have never been measured in humans. In this study, we performed unconfined compression tests to characterize the mechanical properties of the interface tissue and to determine the parameters of various hyperelastic material models which were fitted to the measurements. Human interface tissues were retrieved during 21 elective revision surgeries from aseptically loosened cemented (N=10) and uncemented hip implants (N=11). Specimens were tested at a fixed deformation rate of 0.1mm/min up to a maximum force of 10N. Elastic moduli for low and high strain regions of the stress-strain curves were determined. Interface tissue from aseptically loose cemented prostheses shows higher elastic moduli (mean=1.85MPa, 95% C.I.=1.76-1.95MPa) in the high strain region as compared to that of the interface tissue from the cementless group (mean=1.65MPa, 95% C.I.=1.43-1.88MPa). The 5-terms Mooney-Rivlin model ( [Formula: see text] ) described the stress-strain behavior the best. Large variations in the mechanical behavior were observed both between specimens from the same patient as between those of different patients. The material model parameters were therefore estimated for the mean data as well as for the curves with the highest and lowest strain at the maximum load. The model parameters found for the mean data were C1=-0.0074MPa, C2=0.0019MPa, C3=0MPa, C4=-0.0032MPa and C5=0MPa in the cemented group and C1=-0.0137MPa, C2=0.0069MPa, C3=0.0026MPa, C4=-0.0094MPa and C5=0MPa in the cementless group. The results of this study can be used in finite element computer.

Scaffold-assisted cartilage tissue engineering using infant chondrocytes from human hip cartilage.

OBJECTIVE: Studies about cartilage repair in the hip and infant chondrocytes are rare. The aim of our study was to evaluate the use of infant articular hip chondrocytes for tissue engineering of scaffold-assisted cartilage grafts. METHOD: Hip cartilage was obtained from five human donors (age 1-10 years). Expanded chondrocytes were cultured in polyglycolic acid (PGA)-fibrin scaffolds. De- and re-differentiation of chondrocytes were assessed by histological staining and gene expression analysis of typical chondrocytic marker genes. In vivo, cartilage matrix formation was assessed by histology after subcutaneous transplantation of chondrocyte-seeded PGA-fibrin scaffolds in immunocompromised mice. RESULTS: The donor tissue was heterogenous showing differentiated articular cartilage and non-differentiated tissue and considerable expression of type I and II collagens. Gene expression analysis showed repression of typical chondrocyte and/or mesenchymal marker genes during cell expansion, while markers were re-induced when expanded cells were cultured in PGA-fibrin scaffolds. Cartilage formation after subcutaneous transplantation of chondrocyte loaded PGA-fibrin scaffolds in nude mice was variable, with grafts showing resorption and host cell infiltration or formation of hyaline cartilage rich in type II collagen. Addition of human platelet rich plasma (PRP) to cartilage grafts resulted robustly in formation of hyaline-like cartilage that showed type II collagen and regions with type X collagen. CONCLUSION: These results suggest that culture of expanded and/or de-differentiated infant hip cartilage cells in PGA-fibrin scaffolds initiates chondrocyte re-differentiation. The heterogenous donor tissue containing immature chondrocytes bears the risk of cartilage repair failure in vivo, which may be possibly overcome by the addition of PRP.

Microscopic analysis of the iliofemoral and ischiofemoral ligaments in the hip joint: collagen fiber direction and crimp distribution.

Since no previous studies have described the functional significance of the iliofemoral and ischiofemoral ligaments on the basis of microscopic analyses, we examined the direction of collagen fiber alignment and crimp distribution of the collagen fibers in sections cut in different directions. Polarized microscopic images of sections along the longitudinal (L) and transverse (T) planes of each ligament were obtained from 20 cadavers (8 males and 12 females, age at death 81.7 ± 9.4 years old). Results showed that the microscopic direction of collagen fibers in the iliofemoral ligament was parallel to the macroscopic direction, suggesting that this ligament may play a part in restricting extension of the hip joint. In contrast, the microscopic direction of collagen fibers in the ischiofemoral ligament was not parallel to the macroscopic direction, suggesting that this ligament may contribute not only to the restriction of medial rotation but also retstriction of flexion of the hip joint. From the low density of the crimp distribution in the L plane, the iliofemoral ligament may contribute to stability of the hip joint in the standing position in the living body. In conclusion, the microscopic observations of the direction of collagen fibers as well as the crimp distribution shown in the present study provide a better understanding of the functional significance of the iliofemoral and ischiofemoral ligaments.

In vitro evaluation of stiffness graded artificial hip joint femur head in terms of joint stresses distributions and dimensions: finite element study.

The aim of the present work is to evaluate the artificial hip joint femur head that is made of Stiffness Graded (SG) material in terms of joint stresses distributions and dimensions. In this study, 3D finite element models of femur head that is made of SG material and traditional femur heads made of Stainless Steel (SS), Cobalt Chromium alloy (Co Cr Mo) and Titanium alloy (Ti) have been developed using the ANSYS Code. The effects on the total artificial hip joint system stresses due to using the proposed SG material femur head (with low stiffness at the outer surface and high stiffness at its core) have been investigated. Also, the effects on the polymeric cup contact stresses due to the use of different sizes of femur heads, presence of metal backing shell and presence of radial clearance (gap) between cup and femur head have been investigated. The finite element results showed that using SG femur head resulted in a significant reduction in the cup contact stresses even for small femur heads compared with Ti alloy, SS and Co Cr Mo femur heads. The presence of radial clearance resulted in significant increase in the cup stresses especially for small femur heads. Finally, the presence of SS metal backing shell resulted in slight increase in the hip joint stresses especially for small femur head joints. This work analyzes successfully the usage of proposed SG material as femur head in order to reduce the predicted stresses at the total hip joint replacement due to the redistribution of strain energy in the hip prostheses. Therefore, the present results suggest that minor changes in design and geometrical parameters of the hip joint have significant consequences on the long term use of the joint and should be taken into consideration during the design of the hip joint.

Vascularity of the hip labrum during the foetal period.

Tears of the hip labrum have been recognized as a cause of hip pain and clicking. It has been reported that labrum tears are associated with increased microvessel formation. The purpose of this study was to identify the regional vascularity of the acetabular labrum during late foetal development. The acetabular labrum was examined from 21 formalinized foetuses of the age 5th to 10th months of gestation (mean 6.4±0.99). The acetabulum of each specimen was anatomically prepared and divided into four quadrants. The number of blood vessels in labrum quadrants was counted during microscopic examination. A total of 599 of blood vessels were found in all specimens: 159 in quadrant I, 150 in quadrant II, 127 in quadrant III and 163 in quadrant IV. The capsular part of the labrum contained 357 vessels and the articular part contained 242. The total number of blood vessels within the capsular parts of all specimens (357) was significantly greater than the number within the articular parts (242) (p<0.028). There was some evidence to suggest that with increasing foetal age, the number of blood vessels in the labrum decreased. However, taking into consideration the number of vessels in particular quadrants of the labrum, the great frequency of labral tears in the anterosuperior part of the labrum could not be explained.

ERCC1 expression in aseptic loosening after total hip replacement.

Particle-induced osteolysis is a major cause of aseptic loosening after total joint replacement. The purpose of the current study was to evaluate the DNA damage repair capacity of macrophages in patients with aseptic hip loosening by determination of ERCC1. Moreover, we wanted to elucidate if the potency of the DNA-repair mechanisms correlates with the survival of joint implants. For this purpose we compared the immunohistochemical ERCC1 expression in capsules and interface membranes of patients with loosening of a hip replacement in the first 10 years after implantation with those in patients with late loosening. In analogy with ERCC1 studies on cancer in humans we calculated the semi-quantitative H-score by multiplying the staining intensity with the proportion score of positive stained macrophages. The level of ERCC1 reaction in the specimens taken from patients with early aseptic loosening (mean H-score 0.57) was clearly lower in comparison with those from patients undergoing exchange hip arthroplasty later than 10 years after surgery (mean H-score 2.24). We determined an H-score for ERCC1 expression of 1 as a cutoff point giving a sensitivity and specificity of 100% for identification of early aseptic loosening after less than 10 years. In summary, lower levels of ERCC1 were found in patients with early aseptic loosening compared to patients with aseptic loosening later than 10 years.

Mithramycin downregulates proinflammatory cytokine-induced matrix metalloproteinase gene expression in articular chondrocytes.

Interleukin-1 (IL-1), IL-17 and tumor necrosis factor alpha (TNF-alpha) are the main proinflammatory cytokines implicated in cartilage breakdown by matrix metalloproteinase (MMPs) in arthritic joints. We studied the impact of an anti-neoplastic antibiotic, mithramycin, on the induction of MMPs in chondrocytes. MMP-3 and MMP-13 gene expression induced by IL-1beta, TNF-alpha and IL-17 was downregulated by mithramycin in human chondrosarcoma SW1353 cells and in primary human and bovine femoral head chondrocytes. Constitutive and IL-1-stimulated MMP-13 levels in bovine and human cartilage explants were also suppressed. Mithramycin did not significantly affect the phosphorylation of the mitogen-activated protein kinases, extracellular signal-regulated kinase, p38 and c-Jun N-terminal kinase. Despite effective inhibition of MMP expression by mithramycin and its potential to reduce cartilage degeneration, the agent might work through multiple unidentified mechanisms.

Influence of proteoglycan contents and of tissue hydration on the frictional characteristics of articular cartilage.

In this work, the hypothesis that water content and substances present on the articular surface play an important role in lubrication through the formation of a layer with a high content of water on the articular surface is analysed. The hydrophilic properties of proteoglycans exposed at the articular surface and hydration of tissue are the main responsible factors for the formation of this layer. The role of the articular surface in the frictional characteristics of articular cartilage was examined using specimens (femoral condyles of pigs) with intact and wiped surfaces tested in intermittent friction tests. Results indicated that the intact condition presented low friction in comparison with the wiped condition. The measured water loss of the articular cartilage after sliding and loading indicated a gradual decrease in the water content as the time evolved, and rehydration was observed after the submersion of unloaded specimens in the saline bath solution. Micrographic analyses indicated the presence of a layer covering the articular surface, and histological analyses indicated the presence of proteoglycans in this superficial layer. The hydration of the cartilage surface layer and proteoglycan in this layer influence lubrication.

The origin of osteoprogenitor cells responsible for heterotopic ossification following hip surgery: an animal model in the rabbit.

PURPOSE: To investigate the source of osteoprogenitor cells responsible for heterotopic ossification (HO) following total hip arthroplasty in an animal model. METHODS: New Zealand White (NZW) rabbits (n = 20) received a radiation treatment 24 h preoperatively to the hip joint of one hindquarter and to the femoral shaft of the contralateral side. Subjects underwent bilateral hip surgery 24 h after treatment. Subjects were euthanized and radiographed 4 months postoperatively. Heterotopic ossification was graded according to a modified Brooker scale. Mean grade, intra-observer reliability, and statistical significance (p < 0.05) were evaluated to compare the severity of heterotopic ossification between hindquarters treated with hip irradiation versus those treated with femoral shaft irradiation. RESULTS: The Fleiss Weighted Kappa Statistic indicated "almost perfect" (0.872) intra-rater reliability of radiographic heterotopic ossification grading. The average heterotopic ossification grade for the group receiving radiation to the hip was significantly greater than that for the group receiving radiation to the femoral shaft (2.575 versus 2.0, p < 0.02). CONCLUSION: Although both have some beneficial effect, our results demonstrate that irradiation of the femoral canal is significantly more effective than irradiation of the hip joint and abductor musculature for heterotopic ossification prophylaxis. This suggests that osteoprogenitor cells responsible for heterotopic ossification originate from both the hip abductors and the femoral canal, but the data provide indirect evidence that the femoral canal may be a more dominant source of these cells in the rabbit model.

Age-related changes in parathyroid hormone-related protein and vascular endothelial growth factor in human osteoblastic cells.

Osteogenesis and angiogenesis occur in a coordinated manner in skeletal tissue, so that impaired angiogenesis is associated with decreased bone formation in aged subjects. However, the interaction between bone endothelium and osteoblastic cells is poorly understood. Parathyroid hormone-related protein (PTHrP), a bone factor which modulates osteoblastic cell growth and/or differentiation, stimulates vascular endothelial growth factor (VEGF), a potent angiogenic factor, in primary cultures of human osteoblastic (hOB) cells. In the present study, we examined the age-related changes of both factors in these cells. Human OB cells were isolated from trabecular bone samples from knee or hip explants obtained from 45 osteoarthritic patients: 12 <60 years (21-59 years), 5 women and 7 men, and 33 >60 years (61-82 years), 20 women and 13 men. Cell total RNA was isolated, and mRNA analysis was performed by reverse transcription-polymerase chain reaction. Relative ratios of amplified products with respect to glyceraldehyde-3-phosphate dehydrogenase were then calculated. PTHrP and VEGF were measured in the cell-conditioned medium, after stimulation with (or without) 10 nM 1,25(OH)(2)D(3) for 72 h, using specific immunoradiometric assay and a competitive immunoassay, respectively. A positive correlation was found between PTHrP and VEGF (both mRNA and secreted protein), and also between PTHrP mRNA and the secreted protein levels, in these cells. PTHrP, both mRNA and protein secretion levels, and VEGF secreted values were higher in knee hOB cells than in hip hOB cells only in the younger group. In addition, a decrease in the secreted levels of these factors occurs with aging only in hOB cells from knee. Treatment with 10 nM 1,25(OH)(2)D(3) induced a lower inhibitory response of PTHrP secretion, and a higher stimulatory response of secreted VEGF, in hOB cells with age. These findings indicate that age-related bone loss in humans is associated with a decrease in the osteoblastic secretion of both PTHrP and VEGF in the knee, a predominantly trabecular bone. These data might provide a rationale to explain the impaired angiogenesis associated with trabecular bone loss in aging.

Patterns of osteocytic endothelial nitric oxide synthase expression in the femoral neck cortex: differences between cases of intracapsular hip fracture and controls.

Evidence indicates that extensive amalgamation of adjacent resorbing osteons is responsible for destroying the microstructural integrity of the femoral neck's inferior cortex in osteoporotic hip fracture. Such osteonal amalgamation is likely to involve a failure to limit excessive resorption, but its mechanistic basis remains enigmatic. Nitric oxide (NO) inhibits osteoclastic bone destruction, and in normal bone cells its generation by endothelial nitric oxide synthase (eNOS, the predominant bone isoform) is enhanced by mechanical stimuli and estrogen, which both protect against fracture. To determine whether eNOS expression in osteocytes reflects their proposed role in regulating remodeling, we have examined patterns of osteocyte eNOS immunolabeling in the femoral neck cortex of seven cases of hip fracture and seven controls (females aged 68-96 years). The density of eNOS+ cells (mm(-2)) was 53% lower in the inferior cortex of the fracture cases (p < 0.0004), but was similar in the superior cortex. eNOS+ osteocytes were, on average, 22% further from their nearest blood supply, than osteocytes in general (p < 0.0001) and the nearest eNOS+ osteocyte was 57% further from its nearest canal surface (p < 0.0001). This differential distribution of eNOS+ osteocytes was significantly more pronounced in the cortices of fracture cases (p < 0.0001). We conclude that the normal regional and osteonal pattern of eNOS expression by osteocytes is disrupted in hip fracture, particularly at sites that are loaded most by physical activity. These results suggest that eNOS+ osteocytes may normally act as sentinels confining resorption within single osteons. A reduction in their number, coupled to an increase in their remoteness from canal surfaces, may thus permit the irreversible merging of resorbing osteons, and thus contribute to the marked increase in the fragility of osteoporotic bone.

Biosynthetic response of cultured articular chondrocytes to mechanical vibration.

The objective of this study was to determine the effects of mechanical vibration loading on DNA and proteoglycan syntheses in cultured rabbit articular chondrocytes. Chondrocyte culture plates were placed in a vibratory apparatus and subjected to a mechanical vibratory load at various frequencies and periods in culture. Mechanical vibration was applied at a sinusoidal waveform of 1.4 g acceleration with frequencies of 200, 300, 400, 800, and 1600 Hz. 3H-Thymidine and H2(35)SO4 incorporation were used to detect radiolabeled DNA and proteoglycan syntheses, respectively. A frequency of 300 Hz showed a time-dependent augmentation of DNA synthesis and gave a maximal increase at day 3 with periodic vibration (8 h per day) and at 72 h or longer with continuous vibration. It also promoted proteoglycan synthesis in long-term culture (from 3 to 15 days) by periodic vibration. However. frequencies above 400 Hz suppressed biosynthesis. These results suggest that mechanical vibration at certain frequencies may modulate biosynthetic response of articular chondrocytes.

Cartilaginous differentiation in the joint capsule.

The proliferation and differentiation of cells are greatly influenced by their environment. Many growth factors and cytokines are reported to be environmental factors that affect the proliferation and differentiation of cells. Mechanical stress is also considered to influence these physiological reactions. The joint capsule, which is a part of the joint tissue, plays a very important role in the stability of the joint and in maintaining the intracapsular phenomenon. In patients with dislocated hip arthropathy, this capsule is involved in the weightbearing function by forming a sliding surface between the capsule and the femoral head articular cartilage. The surface of the tissue macroscopically shows cartilaginous change, which indicates cartilaginous differentiation caused by mechanical stress. We examined the cartilage-specific proteoglycan component, which is composed of cartilaginou matrix at the differentiation site. We investigated proteoglycan production, molecular size, and the gene expression of cartilaginous substrate. At the inner layer of the weightbearing area of the joint capsule, proteoglycan production was significantly higher than that of other noncartilaginous tissue. We also identified the gene expression of cartilaginous proteoglycan using the reverse transcription polymerase chain reaction (RT-PCR) method.

Wear debris and cytokine production in the interface membrane of loosened prostheses.

In this study, thirty-nine patients were examined. All of them suffered from hip joint prostheses loosening and underwent revision surgery. Bioptic samples were collected at the interface between bone and implant either at the stem or cotyle level. Immunohistochemistry was performed to detect IL-1alpha, IL-1beta, IL-6 and TNF, cytokines that directly cause bone resorption and indirectly induce synthesis of other bone resorbing cytokines. Quantitative analysis of the positive cells and correlation with clinical data was performed. It resulted that there is a great variability in positive cells for cytokines according to the harvest site; anyway, cytokines tend to be higher in patients carrying a joint prosthesis with polyethylene acetabular component and it is associated with plastic wear particles, even though there is no direct correlation between wear amount and cytokine levels. There is a statistically significant negative correlation between metal wear and a cytokine (IL-6); cytokines levels do not depend on the implant time to failure and do not correlate with pain score. As expected, cytokines levels tend to be lower in subjects being treated with non-steroidal antiinflammatory drugs. It can be concluded that plastic wear is the factor inducing the highest cytokine levels in the tissues around the prosthesis at the interface; cytokines that are an indicator of osteolysis risk.

Influence of skeletal site of origin and donor age on 1,25(OH)2D3-induced response of various osteoblastic markers in human osteoblastic cells.

Age-related bone loss may be a consequence of a lack of osteoblastic formation and/or function. In vitro, the osteoblastic response to 1,25(OH)2D3, an important regulator of osteoblastic function, appears to depend on the stage of osteoblastic maturation. In this study, we examined the response to 1,25(OH)2D3 of C-terminal type I procollagen (PICP), alkaline phosphatase (ALP), and osteocalcin (OC) secretion in primary cultures of osteoblastic cells from human trabecular bone (hOB). Forty-four bone samples were obtained from subjects undergoing knee arthroplastia, 20 aged 50-70 (64 +/- 5), and 24 >70 (73 +/- 2) years. Another 33 bone samples were obtained from subjects undergoing hip arthroplastia, 21 were aged 50-70 (64 +/- 4) and 12 >70 (75 +/- 5) years. Pooling knee and hip hOB cell cultures, we found that PICP secretion decreased after 1,25(OH)2D3 in hOB cells from the older group (>70 years). Treatment with 1,25(OH)2D3 increased ALP secretion in these cells only in the younger group (50-70 years), whereas it increased OC secretion in hOB cells in both age groups. By pooling hOB cell cultures from both age groups we found that knee hOB cells increased OC secretion, and decreased PICP secretion, after 1,25(OH)2D3. This metabolite also increased OC secretion in hip hOB cells. Considering the influence of donor age at the same skeletal site, 1,25(OH)2D3 was found to stimulate ALP secretion only in knee hOB cells in the younger group. In contrast, this metabolite decreased ALP secretion in hip hOB cells in the older group. PICP secretion decreased after 1,25(OH)2D3 only in hOB cells in the older group, at both skeletal sites. In age-matched cultures, OC secretion was lower in hip hOB cells compared with those from the knee in the older group, but was similar in these cell cultures from both skeletal sites in the younger group. OC secretion after 1,25(OH)2D3 stimulation did not show age differences in knee hOB cells, but was lower in hip hOB in the older group. In summary, our results demonstrate that the response of various osteoblastic markers to 1,25(OH)2D3 in primary cultures of hOB cells depends on the donor age and skeletal site of origin.

Oxidative stress induced by humic acid solvent extraction fraction in cultured rabbit articular chondrocytes.

Kashin-Beck disease (KBD), an endemic, chronic osteoarthritic disorder with necrosis of chondrocytes, commonly occurs in China. The humic substance present in the drinking water of endemic areas has been proposed as one of the causative factors. In this study an in vitro cell culture system was used to investigate the damaging effects of humic acid (HA), a constituent of humic substance, on cultured rabbit articular chondrocytes. The commercial Aldrich humic acid (AHA) was fractionated with a series of organic solvents including n-hexane, benzene, ethyl acetate, and methanol. Among the several fractions of AHA, the ethyl acetate fraction (AHA-[EA]) displayed the most potent inhibitory effect on the survival of chondrocytes in clonogenic assays. Cellular injury induced by AHA-[EA] was evaluated by measuring cell viability with methylthiazol tetrazolium (MTT) and by determining the release of intracellular lactate dehydrogenase (LDH). Incubation of chondrocytes with AHA-[EA] (100-500 microg/ml) for 12 h produced a concentration-dependent decrease in cell viability and increase in LDH release. In addition, AHA-[EA] triggered lipid peroxidation manifested by elevated malondialdehyde (MDA) formation. In chemiluminescence assay, AHA-[EA] at the concentrations of 150-600 microg/ml caused 6- to 15-fold increases of luminol-amplified chemiluminescence responses, which are considered to reflect the production of hydrogen peroxide (H2O2). Moreover, pretreating the cells with 500-750 U/ml of catalase significantly prevented the loss of cell viability, while superoxide dismutase (SOD) enhanced the adverse effect of 300 microg/ml AHA-[EA]. Data suggest that the injury to chondrocytes induced by AHA-[EA] may be first through O2.- production, which is then converted into H2O2, thus initiating lipid peroxidation and leading to chondronecrosis observed in KBD.

Structural peculiarities of the tissue formed in modelling of the articular surface.

A model of articular cavity with smooth cover was produced by influence of shape-forming element on regenerating bone tissue. Morphological, ultrastructural, histochemical, and biochemical peculiarities of the tissue of the formed articular surface are described.

Histologic, biochemical, and ion analysis of tissue and fluids retrieved during total hip arthroplasty.

Large amounts of metal and polyethylene debris and high ion readings are found in capsule and fibrous membranes of both loose titanium and cobalt-chromium stems. Prostaglandin E2, interleukin-1, and collagenase levels are elevated when compared to control values with collagenase having the highest and most consistent elevations. Synovial fluid and blood ion readings were elevated in loose cemented and cementless stems made from both materials. Blood ion readings were not elevated in fixed stems. Fixed stems had much less particulate debris in soft tissues. The data showed that failure of most metal hip stems was initially due to a mechanical cause, with high debris and ion counts occurring secondarily in capsule and fibrous membranes. Particulate debris and high ion readings are primarily a focal problem contained by the periprosthetic fibrous connective-tissue encapsulation within the femoral canal and joint capsules. No systemic problems were manifest in any of the patients examined and followed in this study.