Hyperelastic properties of human meniscal attachments

Meniscal attachments are ligamentous tissues anchoring the menisci to the underlying subchondral bone. Currently little is known about the behavior of meniscal attachments, with only a few studies quantitatively documenting their properties. The objective of this study was to quantify and compare the tensile mechanical properties of human meniscal attachments in the transverse direction, curve fit experimental Cauchy stress-stretch data to evaluate the hyperelastic behavior, and couple these results with previously obtained longitudinal data to generate a more complete constitutive model. Meniscal attachment specimens were tested using a uniaxial tension test with the collagen fibers oriented perpendicular to the loading axis. Tests were run until failure and load-optical displacement data was recorded for each test. The medial posterior attachment was shown to have a significantly greater elastic modulus (6.42±0.78 MPa) and ultimate stress (1.73±0.32 MPa) when compared to the other three attachments. The Mooney-Rivlin material model was selected as the best fit for the transverse data and used in conjunction with the longitudinal data. A novel computational approach to determining the transition point between the toe and linear regions is presented for the hyperelastic stress-stretch curves. Results from piece-wise non-linear longitudinal curve fitting correlate well with previous linear elastic and SEM findings. These data can be used to advance the design of meniscal replacements and improve knee joint finite element models.     

Geometry,time-dependent and failure properties of human meniscal attachments

Meniscectomies have been shown to lead to osteoarthritis and the success of meniscal replacements remains questionable. It has been suggested that the success of a meniscal replacement is dependent on several factors, one of which is the secure fixation and firm attachment of the replacement to the tibial plateau at the horn locations. To aid in the development of meniscal replacements, the objectives of the current study were to determine the time-dependent and failure properties of human meniscal attachments. In contrast to the time-dependent tests, during uniaxial failure testing a charge-coupled video camera was used to document the local strain and linear modulus distribution across the surface of the attachments. The lateral attachments were statistically smaller in cross-sectional area and longer than the medial attachments. The anterior attachments were statistically longer and had a smaller cross-sectional area than the posterior attachments. From the stress relaxation tests, the load and stress relaxation rates of the medial anterior attachment were statistically greater than the medial posterior attachment. There were no significant differences in the creep, structural properties or the ultimate stress between the different attachments. Ultimate strain varied between attachments, as well as along the length of the attachment. Ultimate strain in the meniscus region (10.4±6.9%) and mid-substance region (12.7±16.4%) was smaller than the bony insertion region (32.2±21.5%). The lateral and anterior attachments were also found to have statistically greater strain than the medial and posterior attachments, respectively. The linear modulus was statistically weaker in the bony insertion region (69.7±33.7 MPa) compared to the meniscus region (153±123 MPa) and mid-substance region (195±121 MPa). Overall the anterior attachments (169±130 MPa) were also found to be statistically stronger than the posterior attachments (90.8±64.9 MPa). These results can be used to help design tissue-engineered replacement menisci and their insertions and show the differences in material properties between attachments, as well as within an attachment.     

Sensitivities of medial meniscal motion and deformation to material properties of articular cartilage, meniscus and meniscal attachments using design of experiments methods

This study investigated the role of the material properties assumed for articular cartilage, meniscus and meniscal attachments on the fit of a finite element model (FEM) to experimental data for meniscal motion and deformation due to an anterior tibial loading of 45 N in the anterior cruciate ligament-deficient knee. Taguchi style L18 orthogonal arrays were used to identify the most significant factors for further examination. A central composite design was then employed to develop a mathematical model for predicting the fit of the FEM to the experimental data as a function of the material properties and to identify the material property selections that optimize the fit. The cartilage was modeled as isotropic elastic material, the meniscus was modeled as transversely isotropic elastic material, and meniscal horn and the peripheral attachments were modeled as noncompressive and nonlinear in tension spring elements. The ability of the FEM to reproduce the experimentally measured meniscal motion and deformation was most strongly dependent on the initial strain of the meniscal horn attachments (epsilon(1H)), the linear modulus of the meniscal peripheral attachments (E(P)) and the ratio of meniscal moduli in the circumferential and transverse directions (E(theta)E(R)). Our study also successfully identified values for these critical material properties (epsilon(1H) = -5%, E(P) = 5.6 MPa, E(theta)E(R) = 20) to minimize the error in the FEM analysis of experimental results. This study illustrates the most important material properties for future experimental studies, and suggests that modeling work of meniscus, while retaining transverse isotropy, should also focus on the potential influence of nonlinear properties and inhomogeneity.     

How the stiffness of meniscal attachments and meniscal material properties affect tibio-femoral contact pressure computed using a validated finite element model of the human knee joint

In an effort to prevent degeneration of articular cartilage associated with meniscectomies, both meniscal allografts and synthetic replacements are subjects of current interest and investigation. The objectives of the current study were to (1) determine whether a transversely isotropic, linearly elastic, homogeneous material model of the meniscal tissue is necessary to achieve a normal contact pressure distribution on the tibial plateau, (2) determine which material and boundary condition (attachments) parameters affect the contact pressure distribution most strongly, and (3) set tolerances on these parameters to restore the contact pressure distribution to within a specified error. To satisfy these objectives, a finite element model of the tibio-femoral joint of a human cadaveric knee (including both menisci) was used to study the contact pressure distribution on the tibial plateau. To validate the model, the contact pressure distribution on the tibial plateau was measured experimentally in the same knee used to create the model. Within physiologically reasonable bounds on five material parameters and four attachment parameters associated with a meniscal replacement, an optimization was performed under 1200 N of compressive load on the set of nine parameters to minimize the difference between the experimental and model results. The error between the experimental and model contact variables was minimized to 5.4%. The contact pressure distribution of the tibial plateau was sensitive to the circumferential modulus, axial/radial modulus, and horn stiffness, but relatively insensitive to the remaining six parameters. Consequently, both the circumferential and axial/radial moduli are important determinants of the contact pressure distribution, and hence should be matched in the design and/or selection of meniscal replacements. In addition, during surgical implantation of a meniscal replacement, the horns should be attached with high stiffness bone plugs, and the attachments of the transverse ligament and deep medial collateral ligament should be restored to minimize changes in the contact pressure distribution, and thereby possibly prevent the degradation of articular cartilage.     

L-精氨酸高产菌的选育及基于代谢流量分布的育种机制

从分析钝齿棒杆菌(Corynebacterium crenatum)的精氨酸合成途径入手,提出了一种通过选育脯氨酸结构类似物抗性突变株以提高其精氨酸合成能力的育种思路。采用亚硝基胍(NTG)诱变处理出发菌株YD8(His-,SGr1.2 mg/mL,D-Argr15 mg/mL),经含15 mg/mL的脯氨酸结构类似物S-甲基半胱氨酸(S-MC)的抗性筛选获得精氨酸高产突变株YDM403(His-,SGr1.2 mg/mL,D-Argr15 mg/mL,S-MCr15 mg/mL),产酸水平可达29.4 g/L,较出发菌株YD8的产酸高出55.0%。通过代谢流量分布分析了菌株YD8和YDM403代谢网络的变化,结果表明,菌株YD8可能存在顺序反馈抑制作用,出发株YD8解除了Arg对Glu到Arg的反馈抑制,而YDM403又解除了Pro对Glu到Pro的反馈抑制,从而使中间物Glu累积量下降而对-αKG到Glu不再有反馈抑制,其通量提高,与此同时从Glu向Arg的代谢通量也相应增加。     

Circumferential measurement and analysis of strain distribution in the human ACL using a photoelastic coating method

Large variable deformations of the ligament cannot be adequately quantified by one-dimensional and/or localized measurements. To obtain accurate measurement of non-uniform strains over the entire surface of anterior cruciate ligament (ACL), we used a photoelastic coating technique and a method that allowed us to photograph an ACL around its longitudinal axis. A cadaver knee was modified to expose its ACL for observation, and the ligament was then coated with a photoelastic material. The knee was locked in a jig that allowed simulation of natural knee motion. The jig containing the knee was then mounted on a stand, which allowed the exposed ACL to be photographed from any angle around its longitudinal axis while set at a chosen degree of knee flexion. The jig itself was rotated on its stand so as to obtain a panoramic view of the ACL at a given knee angle. The obtained images of the photoelastic fringe patterns yielded significant information for understanding how the strain distributions along the fiber bundles change in association with knee motion. From the results we obtained using the photoelastic measuring method, we reached the following conclusions. Reciprocal functioning between the anterior and the posterior bundles from extension to flexion of the knee does occur. Strain distribution is not uniform even along the same bundle. The strain behavior of the ACL under uniaxial tensile test does not duplicate the conditions in which the ACL is damaged during knee motion. The differences in strains on the ACL under active and passive knee motions may not be as large as those reported previously in the literature.     

Regional distribution of axial strain and circumferential residual strain in the layered rabbit oesophagus

The oesophagus is subjected to large axial strains in vivo and the zero-stress state is not a closed cylinder but an open circular cylindrical sector. The closed cylinder with no external loads applied is called the no-load state and residual strain is the difference in strain between the no-load state and zero-stress state. To understand oesophageal physiology and pathophysiology, it is necessary to know the distribution of axial strain, the zero-stress state, the stress-strain relations of oesophageal tissue, and the changes of these states and relationships due to biological remodeling of the tissue under stress. This study is addressed to such biomechanical properties in normal rabbits. The oesophagi were marked on the surface in vivo, photographed, excised (in vitro state), photographed again, and sectioned into rings (no-load state) in an organ bath containing calcium-free Kreb's solution with dextran and EGTA added. The rings were cut radially to obtain the zero-stress state for the non-separated wall and further dissected to separate the muscle and submucosa layers. Equilibrium was awaited for 30min in each state and the specimens were photographed in no-load and the zero-stress states. The oesophageal length, circumferences, layer thicknesses and areas, and openings angle were measured from the digitised images. The oesophagus shortened axially by 35% after excision. The in vivo axial strain showed a significant variation with the highest values in the mid-oesophagus (p<0.001). Luminal area, circumferences, and wall and layer thicknesses and areas varied in axial direction (in all tests p<0.05). The residual strain was compressive at the mucosal surface and tensile at the serosal surface. The dissection studies demonstrated shear forces between the two layers in the non-separated wall in the no-load and zero-stress states. In conclusion, our data show significant axial variation in passive morphometric and biomechanical properties of the oesophagus. The oesophagus is a layered composite structure with nonlinear and anisotropic mechanical behaviour.     

Idiotypic analysis of anti-GL phi antibodies. I. Identification and strain distribution of GL-1 idiotypes

We utilized both the inhibition of antigen binding and direct idiotype binding methods to identify a new set of common idiotype determinants on anti-GL antibodies of various mouse strains. Three anti-idiotypic antisera, each prepared against individually purified B10.WB anti-GL phi antibodies, were able to detect antibody-combining site-associated common idiotypic determinants, designated GL-1 idiotype(s), in antisera with GLT-binding activity obtained from all mouse strains except strains bearing Igh-1e allotype. Anti-GL phi antisera obtained from rabbits, guinea pigs, and rats did not express detectable levels of GL-1 idiotypes. Nonresponder mice to GL phi, upon immunization with GL-F gamma G or GL phi-F gamma G produced anti-GL antibodies expressing GL-1 idiotypes. Although the magnitude of the immune response to various GL-containing polymers is controlled by distinct Ir genes, the common GL-related antigenic determinants on these polymers are able to induce anti-GL antibodies with GL-1 idiotypic specificities.     

Immunocytochemical analysis of prolactin cells in the adult rat adenohypophysis: distribution and quantitation relative to sex and strain

Prolactin immunocytochemistry was conducted 1) in serial sections of whole fixed pituitaries, for cell distribution analysis; and 2) on preparations of enzymatically dispersed anterior pituitary cells maintained for 1-4 days in vitro, for quantitation of the relative population of prolactin (PRL) cells. Both types of analysis were conducted on glands from young adult rats of both sexes in Long-Evans, Sprague-Dawley, and Fischer 344 strains. Cell quantitation results were compared with serum levels, intracellular content, and in vitro release of prolactin, for individual cell preparations. The results show that both PRL cell distribution and relative population numbers are similar between sexes in all three rat strains. Average percentages ranged from 25.9% to 32.1% in all young males and diestrous females. Prolactin-positive numbers of cells correlated positively with intracellular hormone content but not with serum or medium PRL levels. The prolactin cell population was significantly larger in aged Fischer 344 males but was not correlated with intracellular PRL levels. The prolactin cell population was heterogeneous in staining intensity and distribution in both sexes of all three strains.     

产壳聚糖酶菌株的筛选、鉴定及酶学特性分析

【目的】利用筛选培养基,从福建沿海潮间带泥样中分离筛选产壳聚糖酶的菌株,并研究菌株的产酶特性。【方法】通过形态学观察,结合26S rDNA序列进行分类鉴定,采用DNS法测定酶活力。【结果】筛选得到产壳聚糖酶的菌株KQ-1002与草酸青霉(Penicillium oxalicum)的同源性为99%,并初步鉴定为青霉属的一种。发酵培养的最适温度为30°C,最适碳源为1.0%水溶性壳聚糖,最适氮源为1.87%(NH4)2SO4,最适pH为6.0。该菌株液体发酵培养72 h产壳聚糖酶活性最高,经优化后最高产酶量为18 U/mL。纯化后的壳聚糖酶经SDS-PAGE分析其分子量约40 kD。酶促反应最适pH为5.0,最适反应温度为55°C,Km值为1.293 g/L。在离子浓度为1.0×10 3mol/L时,金属离子Cu2+、Hg2+、Ag+对酶的活性均有强烈的抑制作用。壳聚糖酶对不同底物及脱乙酰度的壳聚糖具有不同的降解作用。【结论】筛选获得产壳聚糖酶的真菌菌株KQ-1002的壳聚糖酶活力经优化后提高了约7倍,是一株具有研究和应用潜力的产壳聚糖酶菌株。     

Nanoindentation of human meniscal surfaces

Menisci are crescent shaped fibrocartilaginous structures which support load distribution of the knee. The menisci are specifically designed to fit the contour of the femoral condyles, aiding to disperse the stresses on the tibial plateau and in turn safeguarding the underlying articular cartilage. The importance of the meniscal superficial layer has not been fully revealed and it is suspected that this layer plays a pivotal role for meniscal function. In this study, both femoral (proximal) and tibial (distal) contacting meniscal surfaces were mechanically examined on the nano-level among three distinct regions (anterior, central and posterior) of the lateral and medial menisci. Nanoindentation testing showed no significant differences among regions, surfaces or anatomical locations, possibly elucidating on the homogeneity of the meniscal superficial zone structure (E(instantaneous): 3.17-4.12MPa, E(steady-state): 1.47-1.69MPa). Nanomechanical moduli values were approximately an order of magnitude greater than micro-scale testing derived moduli values. These findings validate the structural homogeneity of the meniscal superficial zone, showing that material properties are statistically similar regardless of meniscal surface and region. Understanding the mechanical behavior of meniscal surfaces is imperative to properly design an effective meniscal replacement.     

A novel methodology for analysis of cell distribution in chimeric mouse organs using a strain specific antibody

Chimeric animals are very useful for analysis of cell lineage, homeostasis in tissue architecture, and cell-cell interactions during both organogenesis and carcinogenesis. However, there is not a generally effective means for marking cells of chimeric mice. We have therefore developed a polyclonal antibody that is useful for this purpose. This antibody specifically recognizes those cells derived from C3H strain mice. The specificity of this antibody was checked by both immunoblotting and immunoadsorption methods. The antigens were immunohistochemically detected in cytoplasm of both epithelial and mesenchymal cells of C3H/HeN strain mouse in many different organs, but not the corresponding cell types from BALB/c or C57BL/10 or several other mouse strains. The validity of these antibodies as markers for C3H cells was further checked by tissue recombination experiments and in mixed cultures of mouse and rat cells. In each case the antibody recognized only the C3H mouse cells. Next, chimeric mice were prepared between strains C3H/HeN and BALB/c, and C3H/HeN and C57BL/10 mice. Chimeras 2-mo old were examined for antigen distribution using the indirect immunofluorescence method. Many tissues in chimeric mice were composed of cells that were both stained and unstained by the anti-C3H specific antigen. The chimeric patterns were classified into four types, A-D. In well-defined structural units such as intestinal crypts, small intestinal villi, kidney convoluted tubules, exocrine gland acini, ovarian follicles, thyroid gland follicles, stomach glands, adrenal cortex, lingual papillae, etc., (A) each unit was composed entirely of either positive or negative cells, or else (B) in some organs each unit was composed of both types of cells. In the uniform tissues without such distinguishable units, such as stratified squamous epithelium, mesenchymal tissue, corpora lutea, pituitary gland, Islets of Langerhans, adrenal medulla etc., (C) the tissue was composed of definite small cell groups made entirely of either positive or negative cells, or else (D) the tissue was composed of both types of cells which were intermingled with one another. These findings strongly suggest that the chimeric patterns demonstrated here reflect the cell proliferative unit in each tissue. This cell marker system has proven useful for analysis of cell lineage and cell renewal systems in many organs of chimeric mice.     

Numerical study of tension and strain distribution around rat molars]

A knowledge of the mechanical processes triggered in the bone and periodontal ligament (PDL) by orthodontic forces applied to a tooth is of decisive importance for an understanding of the subsequent remodelling around the tooth. To investigate these mechanical relationships, three-dimensional finite element (FE) models of the first lower molar in the rat were established. On the basis of digitized serial histological sections, these FE models were generated semi-automatically. Using various simplified geometrical variations, an appropriate FE model for the analysis of the stress and strain distributions was established. The numerical analyses were carried out under a mesially directed force of 0.1 N. Stress distributions in the bone and PDL showed a similar pattern, while strains in the bone were lower than in the PDL by a factor of 10-5. The data confirm the assumption that strain patterns in the PDL may be the key stimulus of bone remodelling.     

Nestling development and the timing of tick attachments

Parasites exposed to fast-developing hosts experience a variety of conditions over a short time period. Only few studies in vertebrate-ectoparasite systems have integrated the timing of ectoparasite infestations in the host's development into the search for factors explaining ectoparasite burden. In this study we examined the temporal pattern of attachment in a nidicolous tick (Ixodes arboricola) throughout the development of a songbird (Parus major). In the first experiment, we exposed bird clutches at hatching to a mix of the 3 tick instars (larvae, nymphs and adults), and monitored the ticks that attached in relation to the average broods' age. In a complementary experiment we focused on the attachment in adult female ticks--the largest and most significant instar for the species' reproduction--after releasing them at different moments in the nestlings' development. Our observations revealed a positive association between the size of the attached instar and the broods' age. Particularly, adult females were less likely to be found attached to recently hatched nestlings, which contrasts with the smaller-sized larvae and nymphs. These differences suggest either an infestation strategy that is adapted to host physiology and development, or a result of selection by the hosts' anti-tick resistance mechanisms. We discuss the implications of our results in terms of tick life-history strategies.