Gérard Ville

Hypertrophic repair of articular cartilage in experimental osteoarthrosis.

Section of the anterior cruciate ligament has been performed in the knee of 11 mature dogs. The macroscopically normal cartilage from patella and femoral trochlea of animals killed from 2 to 32 weeks after operation was used for histological, histomorphometrical, and biochemical analysis. Previously undescribed degenerative lesions of the superficial matrix were observed, and there was evidence for secondary healing of these lesions. An early and progressive decrease in superficial cell density and a later progressive increase in cartilage thickness without any change in the cell density of the middle and deep cartilage layers was found. A slight increase in water content with no reduction in glycosaminoglycan content was observed. The results suggest that joint laxity results initially in superficial degenerative changes and later in hypertrophic regenerative changes due to cell proliferation and increased matrix synthesis. Hypertrophic remodeling of articular cartilage in response to abnormal mechanical stresses is postulated.

Reactions to a bovine collagen implant: Clinical and immunologic study in 705 patients

A small percentage of patients treated with bovine collagen implants have adverse reactions involving both the cellular and humoral types of immune response. We report a clinical follow-up of 705 subjects treated with a new bovine collagen implant, Atelocollagen (Koken Co. Ltd., Tokyo, Japan). Sensitization to the implant was evaluated in all subjects by skin testing, and humoral response was monitored in 166 subjects by measuring the level of circulating antibodies directed against bovine collagen. Twenty-seven patients (3.8%) exhibited a positive response to a skin test, and of the remaining 656 patients, an adverse reaction to the implant developed in 2.3%. We found a strong correlation between the presence of antibodies to collagen and a positive response to skin testing (92%) or an adverse reaction (100%). In the case of a borderline clinical response to bovine collagen implantation, anticollagen serologic tests appeared to be a useful tool for the identification of clinically reactive patients.

Biochemical properties and immunolocalization of minor collagens in foetal calf cartilage

Type II collagen is the major collagen of cartilage. However, new collagenous chains have been described in different cartilaginous tissues, recently. The la, 2a and 3a chains were extracted from human and bovine hyaline cartilage [1]. Other minor collagenous components were extracted from neonatal pig and human cartilage noted M, CFI, CF2 [2-4], from bovine nasal cartilage and human intervertebral disc noted CPS1, CPS2 [5,6] and from chicken sternal cartilage noted HMW, LMW [7,8] and M 1, M 2 [9]. Here, we report the partial characterization of 3 collagenous fractions obtained after limited pepsin treatment of foetal calf cartilage and isolated according to their solubility properties. The 1.2 M NaC1 fraction contains the la, 2t~ and 3a chains. The 2.0 M NaC1 and 3.0 M NaCl fractions contain at least 7 collagenous chains, which are related to the disulfide-bonded new chains enumerated above, but show some major differences, Antibodies against the 1.2 M and 2.0 M NaC1 fractions were raised in rabbits and their specificity tested by radioimmunoprecipitation. The localization of the corresponding collagenous chains was achieved by indirect immunofluorescence in epiphyseal proper and growth cartilage from foetal calf cartilage. 2. MATERIALS AND METHODS

A new radioimmunoassay using a commercially available solid support for the detection of IgE antibodies against muscle relaxants

It is well established that muscle-relaxant drugs may be responsible for anaphylactoid reactions during anesthesia. In this study, we developed an in vitro test with a commercially available solid phase for the detection of specific IgE directed to the tertiary or quaternary ammonium groups of neuromuscular-blocking drugs. The solid-phase complex was P-aminophenylphosphoryl-choline (PAPPC) immobilized on agarose, and an RIA was performed with an antihuman IgE labeled with 125I. The results, expressed as the percentage of 125I-labeled anti-IgE linked to the solid phase, were at 0.41 +/- 0.19 for 34 control sera from nonallergic healthy adults, with an upper limit estimated at 1%. The values obtained with the sera of 31 allergic patients ranged from 0.6% to 41% with a sensitivity of 97%. The specificity and the positive predictive value of the PAPPC RIA were 97% and 94%, respectively. These results were compared with results of other RIAs with morphine, trimethylamine, triethylamine immobilized on epoxy-activated Sepharose, and choline hydrochloride immobilized on Sepharose (quaternary ammonium Sepharose RIA) and with Phadebas RAST succinylcholine and Phadebas RAST alcuronium. The PAPPC RIA appears to be the most efficient test to screen sera for the presence of IgE antibodies directed to neuromuscular-blocking drugs. One major advantage is that this solid phase is commercially available and ready to use. This advantage will improve the accuracy in the comparison of the results with results from different laboratories.

Serum type I collagen and N-terminal peptide of type III procollagen in chronic hepatitis: Relationship to liver histology and conventional liver tests

The aim of this study was to compare serum N-terminal peptide of type III procollagen to aminotransferases and gamma-globulins as a marker for histological activity in patients with chronic hepatitis and to assess the role of type I collagen, a new serum marker, as a marker of fibrosis in these patients. Sixty patients with biopsy-proven chronic hepatitis were included in this study. Liver disease was virus B-related in 29, autoimmune in five, drug-induced in five, and of unknown etiology in 21. Each biopsy was independently assessed by two liver pathologists. Two histological scores, a score of activity and a score of fibrosis, were established. Serum N-terminal peptide of type III procollagen and type I collagen were assayed by liquid phase RIA. Significant correlations were noted between serum N-terminal peptide of type III procollagen and scores of activity (r = 0.70, p less than 10(-4)) and fibrosis (r = 0.45, p = 0.0005), and between serum type I collagen and scores of activity (r = 0.46, p = 0.0004) and fibrosis (r = 0.67, p less than 10(-4)). When the correlation between scores of activity and fibrosis (r = 0.52, p = 10(-4)) was considered by partial correlation, serum N-terminal peptide of type III procollagen was correlated with the score of activity (r = 0.63, p less than 10(-3)) but not with the score of fibrosis, and serum type I collagen was correlated with the score of fibrosis (r = 0.58, p less than 10(-3)), but not with the score of activity.(ABSTRACT TRUNCATED AT 250 WORDS)

Wound healing of human skin transplanted onto the nude mouse: II. An immunohistological and ultrastructural study of the epidermal basement membrane zone reconstruction and connective tissue reorganization

The reconstruction of human epidermis during healing of human skin wounded after grafting onto the nude mouse was described in a previous paper (M. Démarchez, P. Sengel, and M. Pruniéras, 1986, Dev. Biol. 113, 90-96). The regeneration of the epidermal basement membrane zone (BMZ) and the reorganization of the connective tissue are the subjects of the present study. They were investigated by two complementary methods: electron microscopy to analyze the BMZ reorganization, and indirect immunofluorescence with species-specific and cross-reacting antibodies directed against laminin, bullous pemphigoid antigen, mouse or human collagens of types I or IV, human elastic fibers, fibronectin, fibrin, actin, and human vimentin, to examine the species origin and distribution of BMZ and connective tissue components during the regeneration process. It is reported that grafted human skin preserves its own immunological markers not only in the epidermis but also in the BMZ and dermis as well, and that, after injury, its regeneration proceeds according to the following sequence of overlapping events: production of a mouse granulation tissue; reepidermization by human cells; reconstruction of a BMZ with human characteristics; formation of a human neodermis. It is concluded that human skin grafted onto the nude mouse is able to regenerate its three structural compartments, namely, the epidermis, BMZ, and dermis. Interestingly, it appeared, also, that the connective tissue regeneration would be a two-step mechanism including the sequential formation of two tissues of distinct sources, namely, a granulation tissue and a neodermis.

Enzymatic isotopic assay for human plasma histamine.

Abstract We propose a modified enzymatic isotopic method for human plasma histamine determination. The procedure is simple, rapid, and needs only 20 μl of plasma without any extraction or sample pretreatment. The sensitivity is 0.1 μg/l and the histamine content of plasma from 50 healthy subjects was found to be 0.77 ± 0.61 μ mg/l.

Histamine release assay and radioimmunoassay for the detection of IgE antibodies against neuromuscular blocking drugs.

It is well established that muscle-relaxant drugs may be responsible for anaphylactoid reactions during anaesthesia. In the present work we evaluated in 41 patients who had experienced anaphylactoid reactions during general anesthesia, the value of different radioimmunoassays (RIAs) and of an histamine release assay (HRA). The RIA was performed with different solid phases as PAPPC (para amino phenyl phosphoryl choline), morphine, TMA (trimethylamine) and TEA (triethylamine). The results were expressed as the percentage of 125I anti-human IgE adsorbed onto the solid phase. The sensitivity was estimated respectively at 95, 82, 93 and 64%. The results were significantly in accordance with those obtained with QAS-RIA (from Guéant) but a weak correlation was seen with Phadebas Rast succinylcholine (r = 0.85 and 0.53 respectively). The HRA gave with the NMBD incriminated a sensitivity of 88%. The correlation between HRA and IDR for the drug involved was 89% and 43% for the other NMBDs. The PAPPC RIA was apparently the most efficient test to screen sera for the presence of IgE antibodies whatever the NMBDs involved in anaphylaxis. An advantage is that this solid phase is commercially available and therefore this RIA can be used routinely with a high sensitivity, allowing the comparison of the results obtained by different laboratories.

Immunolocalization of cathepsin D in dental tissues

Cathepsin D antigenicity was localized at the light and electron microscopic levels within dental cells, but not in extracellular matrix. Different intracellular sites for cathepsin D were found depending on the cell type: the enzyme was detected in secretory vesicles of the odontoblasts and in the lysosome-like structures of the ameloblasts. Otherwise, these results suggest that the secretory vesicles of the odontoblasts may contain both cathepsin D and type I collagen. These data might implicate cathepsin D in the enamel and the dentin formations.

The Effects of PEG on Second Antibody Immunoprecipitation and Its Use in Immunoassay

The effects of PEG on the second antibody immunoprecipitation have been studied in several radioimmunological systems. The following parameters have been studied in the assays: the average molecular weight and concentration of PEG, the species of the animal producing the first and second antibody, the nature of the antigen, the concentration of carrier immunoglobulins, and the rate of the immunoprecipitation. Thus, optimal conditions for the use of PEG with a second antibody have been defined in order to be used in any liquid phase immunoassay.