The interaction of collagen and acid mucopolysaccarides. A model for connective tissue

1. The interaction of acid mucopolysaccharides of connective tissue with solubilized collagen of native, or near-native, structure was investigated by free solution electrophoresis at pH7.0. 2. Complex-formation was detected by the appearance of a third peak in the ascending limb only, indicating reversible association. 3. Complex-formation was destroyed by prior heating of solubilized collagen, indicating a probable requirement for high molecular weight or internal structure of the protein. 4. Hyaluronate and chondroitin sulphate of mol.wt. 50000 gave complexes with soluble collagen at I 0.4, whereas heparin and chondroitin sulphate of mol.wt. 15000-18000 did not. All mucopolysaccharides yielded complexes at I 0.1. The stability of the complex appears mainly dependent on electrostatic forces and is increased with increase in chain length of the polysaccharide. 5. Solubilized collagen interacted to yield gels with the ;native' chondroitin sulphate-protein macro-molecule from cartilage. 6. A schematic model for the interaction of collagen and chondroitin sulphate-protein macromolecules shows parallel-ordered interaction of collagen fibrils with chondroitin sulphate side chains of the chondroitin sulphate-protein macromolecule. The biological implications of this model are discussed, particularly in relation to the ordered structures and the ionic-network properties of the intercellular components of connective tissue.     

Characteristics of bone tissue metabolism in adolescents with undifferentiated connective tissue dysplasia

Biochemical markers of bone metabolism, including osteocalcin, total aminoterminal propeptide type I collagen (PINP), and the product of degradation of carboxy-terminal telopeptide type I collagen (β-CrossLaps) were studied in 17 adolescents 11–14 years of age with undifferentiated connective tissue dysplasia (UDCT). Decreased serum concentrations of bone formation markers (PINP and osteocalcin) against the background of normal levels of the bone resorption marker (β-CrossLaps) indicated that the processes of bone remodeling were disturbed and characterized by low-intensity bone-tissue formation with the relative predominance of resorption. The detected bone metabolism disturbances contributed to the development of osteopenia, which gives us grounds to include adolescents with UDCT to the risk group for early-onset osteoporosis.     

A rapid connective tissue stain for glycol methacrylate embedded tissue

No reliable connective tissue stains for GMA sections were available until recently. However, the use of toluidine blue in combination with basic fuchsin appeared to be a rapid and reliable connective tissue stain for glycol methacrylate (GMA) embedded tissue.     

Indices of biopolymer metabolism in connective tissue during repeated stress

Experiments on rats have shown that daily strong stress actions (immobilization) brought about the marked increase in the 11-oxycorticosteroid level and indices of the biopolymer exchange in the connective tissue (free oxyproline, glycosaminoglycanes, sialic acids). Under the acupuncture action (weak stresses) all the above indices were changed mainly after the first influence. Preliminary acupuncture stimulations prevent deep changes in the exchange of biopolymers in the connective tissue, developing under strong stresses.     

Mixed connective tissue disease. A clinico-serological study of 17 cases

Mixed connective tissue disease (MCTD) was described as a distinct clinical syndrome in 1972. Since then many cases have been reported in the literature worldwide. In this study we present our experience with a group of 17 Mexican patients with this syndrome, and we analyze their clinical and serological features, as well as the causes of death in these patients. The patients are Mexican mestizos living in Guadalajara and most of them have been followed-up at Hospital General de Occidente for a period of 1–10 years. The female/male ratio was 16:1, and their age ranged from 14–55 years with a mean of 29 years. The disease duration has ranged from 1–17 years, with a mean of 6 years. Among the clinical manifestations we have found a high frequency of lymphadenopathy when compared with published series (13/17 or 76%), and the laboratory findings in our patients included a very high polyclonal increase of gammaglobulins (93%), lymphopenia (76%), direct immunofluorescence speckled nuclear epidermal deposits in skin biopsies (75%) and positive rheumatoid factor (65%). Other clinical and serological features were similar to those reported in other series of patients with MCTD. Six of the 17 patients have died (35%), and in 3 of them (17.5%) the cause of death was due to an infectious disease that suddenly presented, and apparently was not related to a concomitant high dose of steroids or malnutrition in the patients. It seems that in addition to the already well known autoimmune abnormalities that occur in MCTD, there are other features like the presence of lymphadenopathy, the high polyclonal increase of gammaglobulins, and the lymphopenia, that reflect the profound disturbance of the immune system in this syndrome, possibly contributing to the sudden appearance of a severe infectious disease in some of our patients.Abbreviations ANA antinuclear antibody - CIE counterimmunoelectrophoresis - MCTD mixed connective tissue disease - PHA passive hemagglutination - PM polymyositis - RF rheumatoid factor - SLE systemic lupus erythematosus - SS systemic sclerosis (SS)     

Modulation of connective tissue metabolism by partially purified human interleukin 1

We have investigated the relationship between the monokine interleukin 1 (IL-1) and the connective tissue-stimulating activities produced by monocytes such as mononuclear cell factor (MCF). Using almost exclusively human tissue we have monitored a wide range of MCF-like activities through the partial purification of IL-1 by gel filtration and isoelectric focusing. Activities measured include stimulation of chondrocytes to produce prostaglandins, plasminogen activator and proteoglycanase, enhancement of synovial cell proliferation, and stimulation of cartilage resorption, in addition to IL-1 (lymphocyte activating factor) activity. The activities described show the same molecular heterogeneity; the active material has similar potencies in the different systems, and removal of IL-1 activity by pretreatment with phenylglyoxal also results in loss of the connective tissue-stimulating activities. These results show that the factors responsible for this wide range of activities are very closely related to IL-1 and give further evidence in support of the possible involvement of IL-1 in the processes of joint destruction occurring in chronic inflammatory conditions such as rheumatoid arthritis.     

The regulation of connective tissue metabolism by vasoactive intestinal polypeptide.

Immunohistochemical studies have confirmed the innervation of bone with neuropeptidergic neurons containing vasoactive intestinal polypeptide (VIP), substance P (SP) and calcitonin gene-related peptide (CGRP). In this study, we report effects of VIP on connective tissue cell metabolism. VIP stimulated PGE2 production in human articular chondrocytes, human osteoblast-like cells and human synovial cells, however, stromelysin production was unaffected. VIP also stimulated cAMP production in human osteoblast-like cells, but not in human articular chondrocytes or synovial cells. These findings are suggestive of a role of VIP in connective tissue cell metabolism which may contribute to the inflammatory processes of arthritis.     

Principles of polarization-optical analysis in the study of connective tissue

Possibilities of polarization-optic methods for studying macromolecular organization of the intercellular matrix of the connective tissue are demonstrated. Topo-optic reactions, being histochemical, make it possible to reveal not only presence of the molecules studied, but also quantitatively determine degree of their orientational regularity in the structural organization of the intercellular matrix. Determination of changes in the orientational regularity of single molecules contributes to revealing morphogenetic peculiarities of the intercellular matrix of the connective tissue, normal and under development of the destructive-distrophic process. Methodological principles of reactions to collagen, glycosaminoglycanes, glycoproteins, elastin and non-collagenous proteins are presented. Their advantages comparing certain known histochemical reactions, as well as their limitations are demonstrated. Certain illustrative material is presented.     

Physical behavior of gelatin gels: a simple model for connective tissue

The swelling behavior of gelatin gels when immersed in buffer or in osmotically active solutions of dextran 500 has been studied, and the results can be described by means of the Flory-Rehner theory of the thermodynamic properties of three-dimensional network structures. The density of crosslinkages formed in the gel is found to be related to the concentration of gelatin at which gelation occurs. Values of rigidity moduli are estimated from the density of crosslinkages by use of the statistical thermodynamic theory of rubber elasticity. These values are in excellent agreement with direct measurements.     

Microspectrophotometric study of the lipid content in regenerating connective tissue

An experimental quantitative histological investigation was performed to study lipids in cellular elements of the regenerated connective tissue. Flap wounds were cut on the back of white mice and granulation tissue dissected 1, 3, 7, 14 30 and 60 days after the operation was investigated. Lipids and cellular elements were revealed with Sudan III-IV black B and Nile blue. Lipid quantity was estimated by means of a modified scanning integrating microphotometer. The greatest amount of lipids in the cellular elements of the granulation tissue was revealed on the 3d day of the experiment, total optic density (TOD) of lipids in leucocytes was 0.83, TOD in histiocytes--0.6. On the 7th day the amount of lipids decreased, their TOD in leucocytes was 0.6, in histiocytes--0.4. By the 14th day only phospholipids were revealed, in histiocytes their TOD was 0.3. In subsequent days phospholipid contents continued decreasing and by the 30th day their TOD was 0.2. By the 60th day, when a scar was formed in the wound, in fact, there were no lipids in the cellular elements. The microphotometric investigation performed demonstrated qualitative and quantitative characteristics of lipids in the cellular elements of the granulation tissue in dynamics and their close dependence on the time of wound healing. It was also revealed that the regenerative process was accompanied by decreasing lipid amount in the cellular elements, and this phenomenon could be used as a peculiar test to prognosticate the process of wound healing.