Indentation Stiffness Research Articles

A study of global vs. local properties for maleic anhydride modified polypropylene nanocomposites

Nanoindentation of organomodified clay filled maleated polypropylene (MAPP) was investigated. The study aims to identify the relative increase in local stiffness in comparison to the increase in mechanical properties of the bulk in polypropylene-based nanocomposites. Such a study allows one to assess confined material property in addition to increased filler volume at the local scale. A mixture of highly intercalated and well exfoliated clay structures, when dispersed in MAPP matrix, was observed under transmission electron microscopy. The degree of exfoliation was found to increase with clay loading, which was attributed to the higher viscosity and mechanical shear forces during melt compounding. Instrumented indentation was performed on (1) clay aggregate supported by MAPP matrix, (2) clay–matrix boundary, and (3) the MAPP matrix. The clay aggregated region generally showed higher stiffness as compared to the matrix. And, the relative increase in indentation stiffness is substantially higher than the relative increase in tensile and compressive stiffnesses for clay reinforced systems. Polymer chain confinement and topological constraint appeared to be operative to enhance local stiffness in the clay aggregated region. Good correlation was, however, obtained between the change in macroscopic stiffness and the change in highly local indentation stiffness as a result of clay reinforcement.

Small Indentations of Rubber Blocks: Effect of Size and Shape of Block and of Lateral Compression

Abstract Many gaskets and seals consist of a long rubber strip or thin-walled ring, placed on a flat rigid surface and indented by a flat-ended rigid indenter. We have examined their resistance to small indentations by FEA. They are treated as infinitely-long elastic blocks of rectangular cross-section, resting on a rigid frictionless base. The indentation stiffness is calculated for various ratios of indenter tip width to block width and to block thickness, using two restraint conditions on the outer surfaces: frictionless walls (zero outwards displacement), as for a gasket placed in a recess; or stress-free, as for a gasket with no lateral restraint. For an infinitely-wide and infinitely-thick block, the theoretical resistance to indentation is zero. For comparison, the indentation stiffness is calculated for cylindrical rubber blocks of varied radius and thickness, indented by a flat-ended cylindrical indenter. In this case the result for an infinitely-large block is finite. A second study treats indentation of a rubber block, pre-compressed in the surface plane. Biot showed that the indentation stiffness of a half-space becomes zero at a critical compression, about 33% for equi-biaxial compression and 44 % for plane strain compression, for both a neo-Hookean and a Mooney-Rivlin elastic solid. FEA calculations were made of the indentation stiffness of neo-Hookean blocks of various sizes, pre-compressed to various degrees. The results are compared with Biot's result. In an Appendix, the critical degree of compression is calculated for a more realistic strain energy function than either the neo-Hookean or the Mooney-Rivlin approximation.

Protection of Sandwich Plates from Low-velocity Impact

Sandwich plates used in marine structures are made of laminated face sheets bonded to a structural foam core. Low-velocity impact by a stiff indenter may cause local deflection of the face sheet and permanent deformation or crushing of an underlying volume of the foam core. Upon unloading, the face sheet tends to spring back, while the permanent strain in the core gives rise to residual stresses that may nucleate local interfacial cracks. Extension of such interfacial cracks by applied loads can compromise structural integrity. This article examines the protective effect of thin, ductile interlayers, inserted between the face sheet and the foam core, that absorb the face sheet deflection and thus prevent or reduce the extent of damage to the foam core. Finite element solutions are obtained for a stiff, incompressible, and very ductile polyurethane (PUR) interlayer, and also for a compliant and compressible elastomeric foam (EF) layer, in a sandwich plate made of quasi-isotropic AS4/3501-6 laminated face sheets and H100 PVC core. Loading is applied by a concentrated force acting on a rigid cylindrical indenter. Results show that the PUR interlayer better supports the impacted face sheet than does the foam core or the EF interlayer. The PUR layer also reduces the extent of permanent deformation of the foam core. At a given magnitude of applied load, the energy release rate for cracks extended by residual stresses at the interface between the foam core and either interlayer is much lower than that at the interface between the face sheet and the foam core in the conventional design.

Anisotropic nanomechanical properties ofNephila clavipesdragline silk

Spider silk is a material with unique mechanical properties under tension. In this study, we explore the anisotropic mechanical properties of spider silk using instrumented indentation. Both quasistatic indentation and dynamic stiffness imaging techniques were used to measure the mechanical properties in transverse and longitudinal sections of silk fibers. Quasistatic indentation yielded moduli of 10 ± 2 GPa in transverse sections and moduli of 6.4 ± 0.5 GPa in longitudinal sections, demonstrating mechanical anisotropy in the fiber. This result was supported by dynamic stiffness imaging, which also showed the average reduced modulus measured in the transverse section to be slightly higher than that of the longitudinal section. Stiffness imaging further revealed an oriented microstructure in the fiber, showing microfibrils aligned with the drawing axis of the fiber. No spatial distribution of modulus across the silk sections was observed by either quasistatic or stiffness imaging mechanics.

Pulsatile dynamic stiffness of cartilage‐like materials and use of agarose gels to validate mechanical methods and models

Stiffness is a fundamental indicator of the functional state of articular cartilage. Reported test modes include compressive incremental strain to determine the equilibrium modulus, and sinusoidal strain to determine the dynamic modulus and stress/strain loss angle. Here, initial development is described for a method recognizing that gait is pulsatile. Agarose gels have been used by others for validation or comparison of mechanical test methods and models for cartilage and proteoglycan aggregate. Accordingly, gels ranging from 0.5 to 20% agarose were prepared. Pulsatile stiffness in both indentation and unconfined compression were closely reproducible. Stiffness as a function of agarose concentration rose exponentially, as found using other methods. Indentation stiffness was higher than for unconfined compression and ranged from approximately 2.0 kPa for 0.5% gel to approximately 3,800 kPa for 20% gel. Pulsatile dynamic stiffness appears to be a useful method, although further development is needed. Agarose gel stiffness values obtained by other methods were reviewed for comparison. Unfortunately, reported values for a given agarose concentration ranged widely (e.g. fourfold) even when test methods were similar. Causes appear to include differences in molecular weight and gel preparation time-temperature regimens. Also, agarose is hygroscopic, leading to unintended variations in gel composition. Agarose gels are problematic materials for validation or comparison of cartilage mechanical test methods and models.

Indentation Stiffness of Repair Tissue After Autologous Chondrocyte Transplantation

Indentation Stiffness of Repair Tissue After Autologous Chondrocyte Transplantation

Microtubules Modulate the Stiffness of Cardiomyocytes Against Shear Stress

Although microtubules are involved in various pathological conditions of the heart including hypertrophy and congestive heart failure, the mechanical role of microtubules in cardiomyocytes under such conditions is not well understood. In the present study, we measured multiple aspects of the mechanical properties of single cardiomyocytes, including tensile stiffness, transverse (indentation) stiffness, and shear stiffness in both transverse and longitudinal planes using carbon fiber-based systems and compared these parameters under control, microtubule depolymerized (colchicine treated), and microtubule hyperpolymerized (paclitaxel treated) conditions. From all of these measurements, we found that only the stiffness against shear in the longitudinal plane was modulated by the microtubule cytoskeleton. A simulation model of the myocyte in which microtubules serve as compression-resistant elements successfully reproduced the experimental results. In the complex strain field that living myocytes experience in the body, observed changes in shear stiffness may have a significant influence on the diastolic property of the diseased heart.

Analysis of Stored Osteochondral Allografts at the Time of Surgical Implantation

Background To date, the morphological, biochemical, and biomechanical characteristics of articular cartilage in osteochondral allografts that have been stored have not been fully described. Hypothesis Osteochondral allografts procured and stored commercially for a standard period as determined by tissue banking protocol will have compromised chondrocyte viability but preserved extracellular matrix quality. Study Design Controlled laboratory study. Methods Unused cartilage from 16 consecutive osteochondral allografts was sampled during surgery after tissue bank processing and storage. Ten grafts were examined for cell viability and viable cell density using confocal microscopy, proteoglycan synthesis via 35SO4 uptake, and glycosaminoglycan content and compared with fresh cadaveric articular cartilage. Biomechanical assessment was performed on the 6 remaining grafts by measuring the indentation stiffness of the cartilage. Results The mean storage time for the transplanted specimens was 20.3 ± 2.9 days. Chondrocyte viability, viable cell density, and 35SO4 uptake were significantly lower in allografts at implantation when compared to fresh, unstored controls, whereas matrix characteristics, specifically glycosaminoglycan content and biomechanical measures, were unchanged. In addition, chondrocyte viability in the stored allografts was preferentially decreased in the superficial zone of cartilage. Conclusion Human osteochondral allografts stored for a standard period (approximately 3 weeks) before implantation undergo decreases in cell viability, especially in the critically important superficial zone, as well as in cell density and metabolic activity, whereas matrix and biomechanical characteristics appear conserved. The exact clinical significance of these findings, however, is unknown, as there are no prospective studies examining clinical outcomes using grafts stored for extended periods. Clinical Relevance Surgeons who perform this procedure should understand the cartilage characteristics of the graft after 21 days of commercial storage in serum-free media.

Simultaneous elastic and electromechanical imaging by scanning probe microscopy: Theory and applications to ferroelectric and biological materials

An approach for combined imaging of elastic and electromechanical properties of materials, referred to as piezoacoustic scanning probe microscopy (PA-SPM), is presented. Applicability of this technique for elastic and electromechanical imaging with nanoscale resolution in such dissimilar materials as ferroelectrics and biological tissues is demonstrated. The PA-SPM signal formation is analyzed based on the theory of nanoelectromechanics of piezoelectric indentation and signal sensitivity to materials properties and imaging conditions. It is shown that simultaneous measurements of local indentation stiffness and indentation piezocoefficient provide the most complete description of the local electroelastic properties for transversally isotropic materials, thus making piezoacoustic SPM a comprehensive imaging and analysis tool. The contrast formation mechanism in the low frequency regime is described in terms of tip-surface contact mechanics. Signal generation volumes for electromechanical and elastic signals are determined and relative sensitivity of piezoresponse force microscopy (PFM) and atomic force acoustic microscopy (AFAM) for topographic cross-talk is established.

Meniscal regeneration using tissue engineering with a scaffold derived from a rat meniscus and mesenchymal stromal cells derived from rat bone marrow

The purpose of this study was to regenerate a meniscus using a scaffold from a normal meniscus and mesenchymal stromal cells derived from bone marrow (BM-MSCs). Thirty Sprague-Dawley rat menisci were excised and freeze-thawed three times with liquid nitrogen to kill the original meniscal cells. Bone marrow was aspirated from enhanced green fluorescent protein transgenic Sprague-Dawley rats. BM-MSCs were isolated, cultured for 2 weeks, and 2 x 10(5) cells were then seeded onto the meniscal scaffolds. Using a fluorescent microscope and immunohistochemical staining, repopulation of enhanced green fluorescent protein positive cells was observed in the superficial zone of the scaffold after 1 week of culture, and then in the deep zone after 2 weeks. At 4 weeks, expression of extracellular matrices was detected histologically and expression of mRNA for aggrecan and type X collagen was detected. Stiffness of the cultured tissue, assessed by the indentation stiffness test, had increased significantly after 2 weeks in culture, and approximated the stiffness of a normal meniscus. From this study, we conclude that a scaffold derived from a normal meniscus seeded with BM-MSCs can form a meniscus approximating a normal meniscus.

Indentation Stiffness of Repair Tissue after Autologous Chondrocyte Transplantation

Our main hypothesis was that indentation stiffness of the repair tissue approaches the values of adjacent cartilage 1 year after autologous chondrocyte transplantation. We also wanted to investigate the differences between osteochondritic lesions and full-thickness lesions. Thirty patients with cartilage lesions were operated on with autologous chondrocyte transplantation. The repair was evaluated arthroscopically, indentation stiffness was measured, and clinical evaluations were done. The stiffness of the repair tissue improved to 62% (mean 2.04 +/- 0.83 N, mean +/- SD) of adjacent cartilage (3.58 +/- 1.04 N). Fifty-three percent of the patients graded their knee as excellent or good and 47% of the patients graded their knee as fair at the followup. In six patients the normalized stiffness was at least 80%, suggesting hyaline-like repair. The indentation stiffness of the osteochondritis dissecans lesion repairs (1.45 +/- 0.46 N; n = 7) was less than that of the nonosteochondritis dissecans lesion repair sites (2.37 +/- 0.72 N; n = 19). Gadolinium-enhanced magnetic resonance imaging of the cartilage (dGEMRIC) during followup of four patients suggested proteoglycan replenishment, although all grafts showed low indentation values. Low stiffness values may indicate incomplete maturation or predominantly fibrous repair. The indentation analysis showed that the repair tissue stiffness could, in some cases, reach the same level as the adjacent cartilage, but there was a large variation among the grafts.

Indentation Micromechanics of Fibroblast-Populated Fibrin Constructs

AbstractDuring normal wound healing, a fibrin clot is formed within the first few minutes and is replaced, over several days, by collagen and other extracellular matrix components which are populated with fibroblasts [Clark, 1996]. This process leads to the rebuilding of dermal tissue on which the epidermal tissue is slowly rebuilt. In severe acute and chronic wounds dermal replacement materials that mimic these processes are needed. Current trends are to use scaffolds as structural substitutes as well as carriers for growth factors and cells for wound treatment. Fibrin-based sealants have been used over the last 30 years in hemostasis and tissue sealing applications and in the last 5 years as a scaffold. Fibrin sealant consists of two primary components, fibrinogen and thrombin which form a fibrin clot when mixed. However, a basic understanding of the microstructure/property relationships of fibrin/collagen constructs is not yet fully established. Furthermore, while previous studies showed that cells proliferate and differentiate within 3D fibrin clots, their effect on structural mechanics of the fibrin clot has not been examined.Here, an indentation protocol was established to determine the effects of fibrin/collagen biochemistry and time-dependent cellular response on the elastic parameters of the constructs. Specifically, 4 ml fibrin/collagen constructs were prepared with varying compositions of human-derived fibrinogen (5 - 33 mg/ml), thrombin (1 or 2 U/ml) and bovine collagen (0, 25, 50, 75, 100 weight % of 2.4 mg/ml collagen). Constructs were prepared with/without the presence of human foreskin fibroblasts (ATCC NIH3T3) seeded at a density of 100K cells/ml. Using a 3-mm diameter punch indenter, the indentation load-displacement response was measured after 1, 5 and 10 days of incubation at 37°C in a humidified air/5% CO2 atmosphere. Four replicates per experimental condition were prepared, and three indentations per replicate were performed.For the unpopulated fibrin, there was a linear (R=0.983) relationship between the indentation stiffness and fibrinogen concentration. Also, there was a nonlinear relationship between indentation stiffness and concentration of collagen in the fibrin/collagen constructs. Finally, although in most cases there was no measurable change in the stiffness of the cell-populated tissue constructs with incubation time, in one of the cell-populated formulations, however, the indentation stiffness decreased steadily with increasing incubation time. These results are analyzed within the context of existing analytical micromechanics models for the relationships between scaffold structure, e.g., porosity / fibril diameter, and stiffness.

Biomechanical characteristics of the normal medial and lateral porcine knee menisci

The purpose of this investigation was to examine the compressive properties of the porcine meniscus at a variety of topographical locations using a creep indentation experiment. Three different solution techniques were used to analyse the creep response of the tissue. Specifically, the indentation stiffness, aggregate modulus, permeability, Poisson's ratio, and shear modulus were determined at six different testing locations (anterior, central, and posterior regions; femoral and tibial sides) of both the medial and lateral porcine menisci. Results indicate topographical variations among the testing locations, with the femoral-anterior portion of the medial meniscus having the highest indentation stiffness (350+/-110 kPa), aggregate modulus (270+/-90 kPa), and shear modulus (140+/-40 kPa). The tibial-posterior region of the medial meniscus exhibited the lowest indentation stiffness (170+/-40 kPa), aggregate modulus (130+/-30 kPa), and shear modulus (60+/-20 kPa). No statistical differences were found at the six tested locations of the lateral meniscus.

Low-Intensity Ultrasound Enhances Maturation of Callus after Segmental Transport

The purpose of this study was to determine whether low-intensity ultrasound can be used to enhance callus maturation. Fifteen-millimeter bone defects at the metatarsal bones of sheep were treated with a segmental bone transport for 16 days. The callus formations in the bone defects were allowed to mature for 63 days before the animals were sacrificed. Eighteen sheep were operated on and divided into two groups. One group was treated with low-intensity ultrasound for 20 minutes per day, whereas the other group served as an untreated control group. Biomechanical tests after removal of the metatarsals showed significantly higher axial compression stiffness and significantly higher indentation stiffness of callus tissue in the healing zone in the group treated with ultrasound. Also, histologic analysis of the cortical defect zone showed significantly more callus formation and more active zones of endochondral ossification in the group treated with ultrasound. Stimulation of callus maturation by ultrasound is possible, similar to stimulation of fresh fracture healing, and may be used to shorten clinical treatment times.

Follow-Up of Osteochondral Plug Transfers in a Goat Model

Background Osteochondral transfer procedures are increasingly used to resurface full-thickness articular cartilage defects. There has not been long-term assessment/description of autogenous donor and recipient sites. Hypothesis The healing process occurs at the donor/host cartilage and bone interfaces. Study Design Histologic, biochemical, and biomechanical changes were assessed 6 months after an osteochondral transfer in a goat model. Methods Eight adult goats were studied. In the 6 osteochondral transfer goats, 2 autogenous plugs were transferred from the femoral trochlea to defects in the weightbearing portion of the medial femoral condyle. The goats were allowed free range for 6 months. Randomly assigned plugs were assessed. Results Knees of the sacrificed animals had preservation of the joint space with mild chondromalacic changes in both transfer and contralateral control groups. Histologically, no evidence of cartilage (host/donor) healing was seen. Subchondral bone of the plug was contiguous with the surrounding recipient bone. Cellular viability in the autogenous osteochondral plug was seen, and 35SO4 uptake of the articular cartilage was not statistically different from the contralateral control condyle. The indentation stiffness of the transfer plug (mosaicplasty) and the contralateral donor site were similar—much stiffer than normal cartilage including surrounding condylar cartilage. Large structural stiffness of transferred cores and donor sites appeared to be related to their thinner cartilage layer. Conclusions At 6-month follow-up, a cleft between host and transferred articular regions remained, with no integration between the two. Clinical Relevance Autogenous transplantation of osteochondral plugs is possible with integration of subchondral bone and preservation of chondral viability.

Site- and exercise-related variation in structure and function of cartilage from equine distal metacarpal condyle

Determine (1) the site-associated response of articular cartilage of the equine distal metacarpal condyle to training at a young age as assessed by changes in indentation stiffness and alterations in cartilage structure and composition, and (2) relationships between indentation stiffness and indices of cartilage structure and composition. Experimental animals (n=6) were trained on a track (increasing exercise to 1km/day by 5 months); controls (n=6) were pasture-reared. Animals were euthanized at 18 months and four osteochondral samples were harvested per metacarpal condyle from dorsal-medial, dorsal-lateral, palmar-medial, and palmar-lateral aspects. Cartilage was analyzed for India ink staining (quantified as reflectance score (RS)), short-term indentation stiffness (sphere-ended, 0.4mm diameter), thickness, and biochemical composition. Cartilage structural, biochemical and biomechanical properties varied markedly with site in the joint. Sites just medial and just lateral to the sagittal ridge showed signs of early degeneration, with relatively low RS, indentation stiffness, and collagen content, and relatively high water content. Effects of exercise and side (left vs right) were not detected for any measure. Overall, indentation stiffness correlated positively with RS and collagen content, and inversely with thickness and water content. Gentle exercise-imposed mechanical stimulation did not markedly affect articular cartilage function or structure. However, the marked site-associated variation suggests that biomechanical environment can initiate degenerative changes in immature cartilage during joint growth and maturation.

Spatially-localized correlation of dGEMRIC-measured GAG distribution and mechanical stiffness in the human tibial plateau

The concentration of glycosaminoglycan (GAG) in articular cartilage is known to be an important determinant of tissue mechanical properties based on numerous studies relating bulk GAG and mechanical properties. To date limited information exists regarding the relationship between GAG and mechanical properties on a spatially-localized basis in intact samples of native tissue. This relation can now be explored by using delayed gadolinium-enhanced MRI of cartilage (dGEMRIC––a recently available non-destructive magnetic resonance imaging method for measuring glycosaminoglycan concentration) combined with non-destructive mechanical indentation testing. In this study, three tibial plateaus from patients undergoing total knee arthroplasty were imaged by dGEMRIC. At 33–44 test locations for each tibial plateau, the load response to focal indentation was measured as an index of cartilage stiffness. Overall, a high correlation was found between the dGEMRIC index ( T 1 Gd ) and local stiffness (Pearson correlation coefficients r=0.90, 0.64, 0.81; p<0.0001) when the GAG at each test location was averaged over a depth of tissue comparable to that affected by the indentation. When GAG was averaged over larger depths, the correlations were generally lower. In addition, the correlations improved when the central and peripheral (submeniscal) areas of the tibial plateau were analyzed separately, suggesting that a factor other than GAG concentration is also contributing to indentation stiffness. The results demonstrate the importance of MRI in yielding spatial localization of GAG concentration in the evaluation of cartilage mechanical properties when heterogeneous samples are involved and suggest the possibility that the evaluation of mechanical properties may be improved further by adding other MRI parameters sensitive to the collagen component of cartilage.

INVESTIGATION OF INDENTATION METHODS FOR PROPERTIES DETERMINATION IN HARD FILM – SOFT SUBSTRATE SYSTEMS

ABSTRACTElastic modulus values of the thin films utilized in three different hard film- soft substrate systems were measured using a combination of traditional and continuous stiffness indentation. These systems were chosen to represent typical systems currently utilized in MEMS and included Si/SU8/W, Si/SiO2/Ti/Pt/PZT, and Si/SiO2/Ti/Pt. The last system was coated with either a compressive or nonstressed Cr film. By taking into account ratio between the creep due to the polymer substrate, SU8, to the unloading rate, the modulus of W was measured to be 400 GPa. The modulus of the SU8 was also determined to be 6 GPa. Comparing both the CSM and traditional indentation for the Si/SiO2/Ti/Pt/PZT system showed that the dynamic motion of the indenter caused pile-up in the PZT and resulting in the overestimation of the PZT modulus. This pile-up is a function of the sinusoidal loading frequency. Instead, the modulus of the PZT was measured by shallow depths of traditional indentation that resulted in the PZT modulus of 100 GPa. The hard film-soft substrate systems were show to follow the same trend as stressed soft-film hard-substrate systems with residual stress. The modulus was over estimated for compressively stressed Cr films.

Indentation testing of human cartilage: sensitivity to articular surface degeneration.

To determine, for clinical indentation testing of human articular cartilage, the effects of aging and degeneration on indentation stiffness and traditional indices of cartilage degeneration; the relationship between indentation stiffness and indices of degeneration; and the sensitivity and specificity of indentation stiffness to cartilage degeneration. Osteochondral cores from femoral condyles of cadaveric human donors were harvested. Samples were distributed into experimental groups based on donor age (young [20-39 years], middle [40-59 years], and old [>/=60 years]), and a macroscopic articular surface appearance that was either normal or mildly degenerate, without deep erosion. Samples were analyzed for indentation stiffness, cartilage thickness, India ink staining (quantitated as the reflected light score), and Mankin-Shapiro histopathology score. Indentation stiffness, India ink staining, and the histopathology score each varied markedly between normal-sample and degenerate-sample groups but varied relatively little between normal samples obtained from different age groups. A decrease in indentation stiffness (softening) correlated with a decrease in the reflectance score and an increase in the overall histopathology score, especially the surface irregularity component of the histopathology score. Receiver operating characteristic analysis suggested that the indentation testing could accurately detect cartilage degeneration as indicated by macroscopic appearance, India ink staining, and histopathology score. The indentation stiffness of the normal to mildly degenerate samples tested in this study was sensitive to mild degeneration at the articular surface and was insensitive to changes associated with normal aging or to slight variations in cartilage thickness. This suggests that indentation testing may be a useful clinical tool for the evaluation of early-stage degenerative changes in articular cartilage.

Prolonged storage effects on the articular cartilage of fresh human osteochondral allografts.

Fresh osteochondral allograft transplantation is a well-established technique for the treatment of cartilage defects of the knee. It is believed that the basic paradigm of the technique is that the transplantation of viable chondrocytes maintains the articular cartilage matrix over time. Allograft tissue is typically transplanted up to forty-two days after the death of the donor, but it is unknown how the conditions and duration of storage affect the properties of fresh human osteochondral allografts. This study examined the quality of human allograft cartilage as a function of storage for a duration of one, seven, fourteen, and twenty-eight days. We hypothesized that chondrocyte viability, chondrocyte metabolic activity, and the biochemical and biomechanical properties of articular cartilage would remain unchanged after storage for twenty-eight days. Sixty osteochondral plugs were harvested from ten fresh human femoral condyles within forty-eight hours after the death of the donor and were stored in culture medium at 4 degrees C. At one, seven, fourteen, and twenty-eight days after harvest, the osteochondral plugs were analyzed for (1) viability and viable cell density by confocal microscopy, (2) proteoglycan synthesis by quantification of (35)SO(4) incorporation, (3) glycosaminoglycan content, (4) indentation stiffness, (5) compressive modulus and hydraulic permeability by static and dynamic compression testing, and (6) tensile modulus by equilibrium tensile testing. Chondrocyte viability and viable cell density remained unchanged after storage for seven and fourteen days (p > 0.7) and then declined at twenty-eight days (p < 0.001). Proteoglycan synthesis remained unchanged at seven days (p > 0.1) and then declined at fourteen days (p < 0.01) and twenty-eight days (p < 0.001). No significant differences were detected in glycosaminoglycan content (p > 0.8), indentation stiffness (p > 0.4), compressive modulus (p > 0.05), permeability (p > 0.3), or equilibrium tensile modulus after storage for twenty-eight days (p > 0.9). These data demonstrate that fresh human osteochondral allograft tissue stored for more than fourteen days undergoes significant decreases in chondrocyte viability, viable cell density, and metabolic activity, with preservation of glycosaminoglycan content and biomechanical properties. The cartilage matrix is preserved during storage for twenty-eight days, but the chondrocytes necessary to maintain the matrix after transplantation decreased over that time-period.