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Biomedical subjects

D G Chang

Publications and source records attributed to D G Chang.

13 recordsLinked to original sources

Quantitation and localization of cartilage degeneration following the induction of osteoarthritis in the rabbit knee.

OBJECTIVE: To develop and apply a new video imaging technique to quantify and localize Indian ink staining of cartilage of the rabbit femorotibial joint after the induction of osteoarthritis by unilateral transection of the anterior cruciate ligament (ACLT). METHODS: Nine weeks after surgery, femora and tibiae from 11 ACLT and contralateral control knees were harvested and positioned to obtain calibrated gray-scale images of the ink-painted articular cartilage surfaces that are opposed with the knee in 90 degrees flexion. Images were processed so that areas of normal cartilage gave a relatively high reflectance score, whereas ink-stained fibrillated cartilage and exposed bone gave low scores. RESULTS: Comparison of the medial and lateral femoral condyles and tibial plateaus (MFC, LFC, MTP, LTP) of control and ACLT knees showed that the area of the MTP not covered by the meniscus had a significantly lower reflectance score (P < 0.001) than other areas. ACLT led to an 11% decrease (P < 0.001) in the overall reflectance score. The reflectance score decreased as a traditional morphological grading of degeneration increased. ACLT-induced degeneration had a predilection for the posteromedial aspects of the joint, and to a lesser extent, the anterolateral aspects. In the tibial plateaus, ACLT caused significant degeneration in the covered, but not the uncovered, areas. Image scores of opposing cartilage surfaces (i.e., MFC vs MTP and LFC vs LTP) were significantly (R = 0.56-0.70, P < 0.001) correlated in ACLT and control knees. DISCUSSION: Identification and characterization of cartilage areas prone to degeneration may be particularly useful for further analysis of biochemical and biomechanical mechanisms in osteoarthritis, as well as the efficacy of therapeutic interventions.

Animals↗

Specificity and functional effects of antibodies to human stem cell factor.

Three monoclonal antibodies (Mabs), 7H6, 4B10 and Genzyme Mab, and a commercially-available polyclonal antiserum (Genzyme) to human Stem Cell Factor (SCF) were compared for their ability to detect native and recombinant SCF in a variety of assays, and for blocking of SCF function. All antibodies were found to bind to the membrane bound isoform as well as soluble SCF and to bind to both glycosylated (yeast MGF) and unglycosylated (E. coli SCF) recombinant factor. Mabs 7H6 and 4B10, as well as the polyclonal antiserum could immunoprecipitate membrane-associated SCF and all the antibodies could detect recombinant soluble SCF on western blots, although the binding of all except 7H6 was partially sensitive to reduction. Titration of the antibodies on CHO cells expressing membrane-associated human SCF showed similar dose-dependence for all Mabs with 70% of maximum binding seen at 3, 5 and 8 micrograms/ml for 7H6, 4B10 and Genzyme Mab respectively, however the maximum binding seen with 7H6 was approximately 2-fold greater than with 4B10 and 7-fold greater than Genzyme Mab. Competitive binding experiments of the Mabs on cells expressing membrane SCF gave non-reciprocal blocking in all cases with 7H6 completely blocking 4B10 and Genzyme Mab binding. All antibodies except the Genzyme Mab effectively blocked SCF binding to c-Kit-expressing cells, and were strongly inhibitory in an assay of in vitro haemopoiesis which is believed to depend on adhesive interactions, as well as the "classical' cytokine-receptor interaction, mediated by SCF binding to c-Kit.

Animals↗

Epitope mapping and immunoneutralization of recombinant human stem-cell factor.

The epitope regions of three anti-[stem-cell factor (SCF)]g have been mapped by characterization of immunoreactivities against truncated forms of SCF in immunoblots and against synthetic peptides in solution-phase competition ELISA. Two of the antibodies, mAb 7H6 and mAb 8H7A, were raised against Escherichia coli-derived human SCF-(1-164) while the third, polyclonal antibody (pAb) 1337, was raised against a peptide corresponding to residues 3-31 of human SCF. The epitopes of mAbs 7H6 and 8H7A have been mapped to residues 61-95 and 95-110, respectively. The epitope of pAb 1337 has been mapped to residues 21-31. The ability of the anti-SCF Ig to recognize E. coli-derived human SCF presented in various formats, i.e. partially denatured (fixed in standard ELISA or on a western blot) or native (in solution), was studied, mAb 7H6 recognized its epitope in partially denatured or native SCF with equally high affinity, while mAb 8H7A and pAb 1337 recognized their epitopes only when SCF was at least partially denatured, mAb 7H6 was found to neutralize in vitro SCF-mediated cell proliferation and SCF binding to its receptor, when present in equimolar concentrations relative to the ligand, suggesting that the epitope region is functionally significant. Evidence that the mAb 7H6 epitope is represented by discontinuous regions (residues within sequences 61-65 and 91-95 are critically involved) is presented. The observation that the mAb 7H6 epitope is discontinuous has implications for the structure of SCF.

Animals↗

Geometric changes in the cervical spinal canal during impact.

SUMMARY OF BACKGROUND DATA: Although the extent of injury after cervical spine fracture can be visualized by imaging, the deformations that occur in the spinal canal during injury are unknown. STUDY DESIGN: This study compared spinal canal occlusion and axial length changes occurring during a simulated compressive burst fracture with the residual deformations after the injury. METHODS: Canal occlusion was measured from changes in pressure in a flexible tube with fluid flowing through it, placed in the canal space after removal of the cord in cadaver specimens. To measure canal axial length, cables were fixed in C1 and led through the foramen transversarium from C2-T1, then out through the base, where they were connected to the core rods of linearly variable differential transformers (LVDT). Axial compressive burst fractures were created in each of ten cadaveric cervical spine specimens using a drop-weight, while force, distraction, and occlusion were monitored throughout the injury event. Pre- and post-injury radiographs and computed tomography scans compared transient and post-injury spinal canal geometry changes. RESULTS: In all cases, severe compressive injuries were produced. Three had an extension component in addition to compression of the vertebra and retropulsion of bone into the canal. The mean post-injury axial height loss measured from radiographs was only 35% of that measured transiently (3.1 mm post-injury, compared with 8.9 mm measured transiently), indicating significant recovery of axial height after impact. Post-injury and transient height loss were not significantly correlated (r2 = 0.230, P = 0.16) demonstrating that it is not a good measure of the extent of injury. Similarly, mean post injury canal area was 139% of the minimum area measured during impact, indicating recovery of canal space, and post-injury and transient values were not significantly correlated (r2 = 0.272, P = 0.12). Mean post-injury midsagittal diameter was 269% of the minimum transient diameter and showed a weak but significant correlation (r2 = 0.481, P = 0.03). CONCLUSIONS: Two potential spinal cord injury-causing mechanisms in axial bursting injuries of the cervical spine are occlusion and shortening of the canal. Post-injury radiographic measurements significantly underestimate the actual transient injury that occurs during impact.

Biomechanical Phenomena↗

Soluble stem cell factor in human serum.

Stem cell factor (SCF) is a recently described factor active in the early stages of hematopoiesis. It can exist in membrane-bound form and in proteolytically released soluble form. The levels and nature of SCF in human serum are described. As determined by an enzyme-linked immunosorbent assay performed for 257 samples, SCF level in serum averaged 3.3 +/- 1.1 ng/mL. The serum SCF was partially purified by immunoaffinity chromatography and analyzed by glycosidase treatments in conjunction with sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunoblotting. The results show that the SCF has N-linked and O-linked carbohydrate and corresponds to the soluble form, at or about 165 amino acids in length. The findings suggest functional importance for soluble SCF in humans.

Adult↗

Functional radiographic diagnosis of the lumbar spine. Flexion-extension and lateral bending.

Several attempts have been made to measure the segmental range of motion in the lumbar spine during flexion-extension with the purpose of gathering additional data for the diagnosis of instability. The previous studies were performed in vitro or in vivo during active motion. The aim of this study was to obtain normal values of passively performed segmental motions. Forty-one healthy adults were examined by means of functional radiographs during flexion-extension and lateral bending. A graphic construction method and a computer-assisted method were used to measure rotations. Comparing with recent in vivo studies, the values obtained for normal angles of rotation were predominately larger. This might be due to the passive examination used in the study. The graphic construction method and computer-assisted method techniques are equally reliable, but the computer-assisted method method yields other important kinematic data, such as translations. It is proposed that passive motion be applied during functional examination of patients with suspected instabilities. However, the large variation of rotational values between individuals in the normal population may limit the clinical usefulness of functional lumbar analysis using this parameter. Future studies should explore the clinical relevance of determining altered segmental mobility in low-back pain patients.

Adult↗

Clinical validation of functional flexion-extension roentgenograms of the lumbar spine.

The purpose of this study was to determine the clinical validity of functional flexion-extension roentgenograms of the lumbar spine in a defined patient population. One hundred and one adults with low-back pain or functional disorders underwent passive functional flexion-extension examinations. Their roentgenograms were analyzed using a computer-assisted method to determine segmental motion parameters such as rotation and translation of the lumbar vertebrae. The patient population was broken down into five groups with similar pathologies or physical conditions, and their motion parameters compared to a normal population and to each other. It was found that all of the patient groups exhibited significantly hypomobile motion, spread equally among all levels, in comparison to the normal population, except for the group of high-performance athletes, who had significant hypermobility. The uniform spread of hypomobility limits the ability to distinguish with any confidence between the four pathologic groups by their motion. Thus, we believe that the analysis of the segmental motion of the lumbar spine using passive flexion-extension roentgenograms does not aid in differentiating the underlying pathologic condition of patients with low-back pain, and that no useful information can be derived form this procedure, especially in relation to the need for surgical intervention.

Adult↗

A radioimmunoassay for 6-ketoprostaglandin E1.

A radioimmunoassay for 6-ketoprostaglandin E1 has been developed. 6-keto-prostaglandin E1 antibodies were produced in rabbits by repeated immunization with 6-ketoprostaglandin E1 coupled to bovine serum albumin. [125]-labeled hapten with high specific radioactivity was prepared by radioiodination of 6-ketoprostaglandin in E1-tyrosine methyl ester conjugate followed by purification with thin layer chromatography. The antibodies showed good specificity toward 6-ketoprostaglandin E1 and crossreacted only significantly with prostaglandin E1. The sensitivity of the assay was 10 pg per assay tube. Application of the radioimmunoassay was demonstrated by the detection of immunoreactive 6-ketoprostaglandin E1 from 6-ketoprostaglandin F1 alpha catalyzed by swine renal NADP+-linked 9-hydroxyprostaglandin dehydrogenase.

Alprostadil↗

The effects of hyaluronan on tissue healing after meniscus injury and repair in a rabbit model.

To assess the effect of hyaluronan on meniscus injury and repair, we had 35 mature New Zealand White rabbits undergo bilateral meniscus injury and repair (19 in the peripheral region, and 16 in the inner region). A longitudinal tear was created in the medial meniscus and repaired with horizontally placed nylon sutures. The left knee joint received intraarticular injections of hyaluronan 1 week after surgery and once a week for 5 weeks. The right knees were injected with phosphate-buffered saline (the carrier vehicle of the hyaluronan). Twelve weeks after repair, tears in the peripheral region showed gross and histologic evidence of healing, with no difference between the vehicle- and hyaluronan-treated menisci. Biochemically, the ratio of reducible collagen cross-links in the hyaluronan-treated menisci was significantly higher than in the vehicle-treated menisci, indicating greater level of collagen remodeling. Biomechanically the vehicle- and hyaluronan-treated menisci demonstrated similarly high tearing load and fracture toughness. In the inner region, poor healing response was observed grossly and histologically in both treatment groups. Water content in the hyaluronan-treated menisci was significantly lower than in the vehicle-treated menisci, indicating a lower level of swelling. Hyaluronan treatment stimulated collagen remodeling in the peripheral region and inhibited swelling of the meniscus repaired in the inner region.

Adjuvants, Immunologic↗