Cartilage Repair Capacity within a Single Full-Thickness Chondral Defect in a Porcine Autologous Matrix-Induced Chondrogenesis Model Is Affected by the Location within the Defect.

OBJECTIVE: Large articular cartilage defects are a challenge to regenerative surgery. Biomaterial scaffolds might provide valuable support for restoration of articulating surface. The performance of a composite biomaterial scaffold was evaluated in a large porcine cartilage defect. DESIGN: Cartilage repair capacity of a biomaterial combining recombinant human type III collagen (rhCo) and poly-(l/d)-lactide (PLA) was tested in a porcine model. A full-thickness chondral defect covering the majority of the weightbearing area was inflicted to the medial femoral condyle of the right knee. Spontaneous cartilage repair and nonoperated healthy animals served as controls. The animals were sacrificed after a 4-month follow-up. The repair tissue was evaluated with the International Cartilage Repair Society (ICRS) macroscopic score, ICRS II histological score, and with micro-computed tomography. Additionally, histopathological evaluation of lymph nodes and synovial samples were done for toxicological analyses. RESULTS: The lateral half of the cartilage defect in the operated groups showed better filling than the medial half. The mean overall macroscopic score for the rhCo-PLA, spontaneous, and nonoperated groups were 5.96 ± 0.33, 4.63 ± 0.42, and 10.98 ± 0.35, respectively. The overall histological appearance of the specimens was predominantly hyaline cartilage in 3 of 9 samples of the rhCo-PLA group, 2 of 8 of the spontaneous group, and 9 of 9 of the nonoperated group. CONCLUSIONS: The use of rhCo-PLA scaffold did not differ from spontaneous healing. The repair was affected by the spatial properties within the defect, as the lateral part of the defect showed better repair than the medial part, probably due to different weightbearing conditions.

Changes in spatial collagen content and collagen network architecture in porcine articular cartilage during growth and maturation.

OBJECTIVES: The present study was designed to reveal changes in the collagen network architecture and collagen content in cartilage during growth and maturation of pigs. METHODS: Femoral groove articular cartilage specimens were collected from 4-, 11- and 21-month-old domestic pigs (n=12 in each group). The animal care conditions were kept constant throughout the study. Polarized light microscopy was used to determine the collagen fibril network birefringence, fibril orientation and parallelism. Infrared spectroscopy was used to monitor changes in the spatial collagen content in cartilage tissue. RESULTS: During growth, gradual alterations were recorded in the collagen network properties. At 4 months of age, a major part of the collagen fibrils was oriented parallel to the cartilage surface throughout the tissue. However, the fibril orientation changed considerably as skeletal maturation progressed. At 21 months of age, the fibrils of the deep zone cartilage ran predominantly at right angles to the cartilage surface. The collagen content increased and its depthwise distribution changed during growth and maturation. A significant increase of the collagen network birefringence was observed in the deep tissue at the age of 21 months. CONCLUSIONS: The present study revealed dynamic changes of the collagen network during growth and maturation of the pigs. The structure of the collagen network of young pigs gradually approached a network with the classical Benninghoff architecture. The probable explanation for the alterations is growth of the bone epiphysis with simultaneous adaptation of the cartilage to increased joint loading. The maturation of articular cartilage advances gradually with age and offers, in principle, the possibility to influence the quality of the tissue, especially by habitual joint loading. These observations in porcine cartilage may be of significance with respect to the maturation of human articular cartilage.

Evaluation of cartilage repair in the distal femur after autologous chondrocyte transplantation using T2 relaxation time and dGEMRIC.

OBJECTIVES: To investigate the potential of combining T2 relaxation time and delayed gadolinium-enhanced magnetic resonance imaging of cartilage (dGEMRIC) measurements after autologous chondrocyte transplantation (ACT). MATERIALS AND METHODS: T2 and dGEMRIC maps were measured in the sagittal and coronal directions in 12 patients 10-15 months after ACT surgery. Grafts were assessed for bulk full thickness, superficial and deep tissue T2 and dGEMRIC values, and were compared to control cartilage. RESULTS: All ACT grafts showed filling of the repair area to the level of or above the articular surface. Matrix of the grafts lacked the classical laminar structure and appeared more heterogenous on T2 maps than control cartilage. As compared to control cartilage, ACT grafts showed significantly longer T2 values for bulk tissue as well as for the superficial 50% and deep 50% of tissue except for superficial cartilage in the coronal direction. dGEMRIC assessment in the sagittal and coronal directions did not show a significant difference between bulk, superficial or deep tissue as compared to the control cartilage. Superficial and deep ACT tissue did not differ statistically in terms of their T2 or dGEMRIC values. CONCLUSIONS: These preliminary results suggest that, according to T2 measurements, ACT repair tissue at 10-15 months differs from normal cartilage and probably lacks the preferential collagen arrangement of normal cartilage, while according to dGEMRIC a varying degree of proteoglycan replenishment takes place. Combining these two quantitative magnetic resonance imaging techniques enables a more comprehensive characterization of cartilage repair than before.

Immature porcine knee cartilage lesions show good healing with or without autologous chondrocyte transplantation.

OBJECTIVE: The purpose of this study was to find out how deep chondral lesions heal in growing animals spontaneously and after autologous chondrocyte transplantation. METHODS: A 6mm deep chondral lesion was created in the knee joints of 57 immature pigs and repaired with autologous chondrocyte transplantation covered with periosteum or muscle fascia, with periosteum only, or left untreated. After 3 and 12 months, the repair tissue was evaluated with International Cartilage Repair Society (ICRS) macroscopic grading, modified O'Driscoll histological scoring, and staining for collagen type II and hyaluronan, and with toluidine blue and safranin-O staining for glycosaminoglycans. The repair tissue structure was also examined with quantitative polarized light microscopy and indentation analysis of the cartilage stiffness. RESULTS: The ICRS grading indicated nearly normal repair tissue in 65% (10/17) after the autologous chondrocyte transplantation and 86% (7/8) after no repair at 3 months. At 1 year, the repair tissue was nearly normal in all cases in the spontaneous repair group and in 38% (3/8) in the chondrocyte transplantation group. In most cases, the cartilage repair tissue stained intensely for glycosaminoglycans and collagen type II indicating repair tissue with true constituents of articular cartilage. There was a statistical difference in the total histological scores at 3 months (P=0.028) with the best repair in the spontaneous repair group. A marked subchondral bone reaction, staining with toluidine blue and collagen type II, was seen in 65% of all animals. CONCLUSIONS: The spontaneous repair ability of full thickness cartilage defects of immature pigs is significant and periosteum or autologous chondrocytes do not bring any additional benefits to the repair.

Quantitative ultrasound imaging of spontaneous repair of porcine cartilage.

OBJECTIVE: Arthroscopy offers qualitative means to evaluate the surface of articular cartilage. However, possible degeneration of the deep cartilage and subchondral bone remains undetected. High frequency ultrasound imaging is an advanced cartilage evaluation method which is conceivable to arthroscopic use and brings diagnostic information also from deeper cartilage and subchondral bone. DESIGN: In this study, we characterized spontaneous repair of porcine cartilage in situ with quantitative 2D-ultrasound imaging. At the age of 7-8 months, a cartilage lesion (diameter 6mm, not penetrating into subchondral bone) was created on the lateral facet of the right femoral trochlea (n=8). The animals were sacrificed 3 months after the surgery. The lesion site, adjacent cartilage and the corresponding control area at the contralateral (left) knee were imaged in situ with 20 MHz ultrasound. Ultrasound reflection coefficients were determined from the cartilage surface (R) and from the cartilage-bone interface (R(bone)). Microtopography of the articular surface was quantified by calculating ultrasound roughness index (URI) parameter from the ultrasonically determined surface profile. RESULTS: Lesion site was spontaneously filled with visually cartilage-like soft tissue with smooth surface. However, ultrasonic images and histological analyses revealed erosion of subchondral bone under the lesion site. Ultrasound reflection (R) at the surface of the spontaneously repaired tissue was significantly lower (-73.5+/-7.6%, P<0.05) than at the surface of intact cartilage. R(bone) was lowest at the lesion site. The surface roughness of spontaneously repaired cartilage was significantly higher than that of the intact tissue (44.0+/-26.0 microm vs 7.5+/-2.3 microm, P<0.05). CONCLUSIONS: Quantitative ultrasound parameters offered diagnostic information revealing impaired structural integrity of the spontaneously repaired porcine cartilage and subchondral bone. These changes are not detectable by traditional arthroscopic means.

Subchondral bone reaction associated with chondral defect and attempted cartilage repair in goats.

Repair of cartilage damage with autologous chondrocyte transplantation (ACT) has become popular in clinical use during the past few years. Although clinical results have mostly been successful, several unanswered questions remain regarding the biological mechanism of the repair process. The aim of this study was to develop a goat model for ACT. The repair was not successful due to the graft delamination, but we characterize the subchondral changes seen after the procedure. A chondral lesion was created in 14 goat knees, operated on 1 month later with ACT, and covered with periosteum or a bioabsorbable poly-L/D-lactide scaffold. After 3 months, only two of the five lesions repaired with ACT showed partly hyaline-like repair tissue, and all lesions (n = 4) with the scaffold failed. Even though the lesions did not extend through the calcified cartilage, the bone volume and collagen organization of bone structure were decreased when assessed by quantitative polarized light microscopy. There was a significant loss of bone matrix and distortion of the trabecular structure of subchondral bone, which extended several millimeters into the bone. The subchondral bone demonstrated strong hyaluronan staining in the bone marrow and cartilaginous areas with signs of endochondral ossification, suggesting structural remodeling of the bone. The goat model used here proved not to be an optimal model for ACT. The changes in subchondral bone may alter the biomechanical properties of the subchondral plate and thus the long-term survival of the repair tissue after ACT.

Computer-generated guided-wave holography: application to beam splitting.

The use of guided-wave synthetic holograms in integrated optics is extended beyond the conventional Fresnel lens and Bragg grating technology. As an example, beam splitters based on Fourier-domain holograms are proposed as an alternative to the usual channel waveguide devices. We demonstrate fanout to six with +/-5% (+/-0.2-dB) uniformity error using a binary phase grating and fanout to eight with +/-20% (+/-0.8-dB) uniformity error using a multilevel grating.

Binary surface-relief gratings for array illumination in digital optics.

Separable binary-phase array illuminators for fan-out up to 1024 x 1024 and ~65% two-dimensional efficiency are designed by simulated annealing with constraints for maximizing the minimum feature size. A new nonseparable trapezoidal coding technique is introduced and applied to design high-efficiency (~75%-80%) array generators for fan-out up to 16 x 16. A rigorous electromagnetic diffraction theory is used to evaluate the range of validity of the scalar designs (both grating period and input angle are considered), to analyze fabrication errors (slanted groove walls and undercutting), and to design binary resonance-domain one-dimensional array generators with 90%-100% efficiency. Trapezoidal gratings for low fan-out (8 x 8), separable gratings for high fan-out (up to 128 x 128), and a 1 x 5 resonance domain (100% efficient) reflection grating are demonstrated experimentally.

Hybrid kinoform fanout holograms in dichromated gelatin.

The technique for fabricating hybrid holograms, introduced by Bartelt and Case [Appl. Opt. 21, 2886 (1982)], is applied here to record extremely high-efficiency (>90%) space-invariant fanout holograms in dichromated gelatin. The object wave front corresponding to that produced by a kinoform with a continuous-phase profile is derived by appropriate spatial filtering from a binary computer-generated hologram fabricated using electron-beam lithography.

High-efficiency, wide-bandwidth optical fanout elements in dichromated gelatin.

We have combined the flexibility of computer-generated binary holograms with the high optical quality of dichromated gelatin holography to fabricate a range of efficient, wide-bandwidth, and compact space-invariant fanout elements. Our approach is based on a certain novel coherent spatial filtering technique, termed the inverse central dark ground method, which permits the use of a binary-amplitude hologram corresponding to the desired binaryphase profile in the object arm of the interferometric recording setup.

Fabrication of space invariant fanout components in dichromated gelatin.

Efficient and compact space invariant fanout holographic optical elements have been fabricated by combining the flexibility in design of computer-generated holograms with the versatility of conventional interferometric holography in dichromated gelatin. Effects of K-ratio variation and the phase distortion in the reconstruction fidelity are discussed. In addition, various preprocessing and postprocessing techniques have been employed to increase the optical damage threshold of these components to well over 100 W cm(-2).

Realization of general nondiffracting beams with computer-generated holograms.

A new class of solutions to the scalar wave equation was introduced recently that represents transversely localized but totally nondiffracting fields. We show by the method of stationary phase that any of these wave fields can be realized approximately with a laser and a single computer-generated hologram. We briefly discuss various techniques for coding and fabrication of the required hologram and the associated diffraction efficiencies. Using both binary-amplitude and four-level phase holograms, we demonstrate experimentally the formation of arbitrary-order Bessel beams and rotationally nonsymmetric beams.

Photolithographic fabrication method of computer-generated holographic interferograms.

We consider the fabrication of high-quality interferogram-type diffractive optical elements with conventional photolithographic techniques and compare the results with those achievable with electron-beam lithography. The fringes associated with the phase transfer function of the binary phase holographic interferogram are approximated with rectangles, which can be realized at submicron accuracy using a pattern generator and step-and-repeat camera. The effects of the rectangle quantization are analyzed both numerically and experimentally with the aid of diffraction patterns produced by simple focusing elements. Both resolution and diffraction efficiency of the best holograms approach their theoretical values.