Collagen Development in Granulation Tissue as Compared With Collagen of Skin and Aorta From Injured and Non‐injured Rats

1981

Granulation tissue was produced by subcutaneous implantation of viscose‐cellulose sponges on rats. The effect of D‐penicillamine 500 mg/kg/day for 10 days on granulation tissue and the connective tissue of skin, bone and aorta was studied by comparison of treated animals with operated and unoperated controls. In addition, the effect of sponge implantation on intact tissues was studied by comparison between the control groups. The amount of granulation tissue was not affected by D‐penicillamine, and the granuloma‐DNA content even increased. D‐penicillamine increased the amount of salt soluble collagen in all tissues consistent with an inhibition of collagen crosslinking as reflected by increased aldehyde content and α/β chain ratio in soluble skin collagen. Skin appeared to be most sensitive. The content of free hydroxyproline and RNA and the RNA/DNA ratio in skin decreased suggesting a decreased collagen biosynthesis. The hydroxylation of proline and lysine was not affected by the treatment. The water percentage of aorta increased during D‐penicillamine treatment, and the 35S‐sulphate uptake in the sulphated glycosaminoglycans was stimulated in all tissues, in granulation tissue mainly in the chondroitin‐4/6‐sulphate fraction. No quantitative glycosaminoglycan changes occurred in granulation tissue. Sponge implantation caused an increase in the amount of salt soluble skin collagen without any change in α/β chain ratio and aldehyde content of purified, soluble collagen. D‐penicillamine plus operation reduced the collagen content of aorta. The effects of D‐penicillamine on connective tissue compounds may be of importance for its antirheumatic efficacy, but the accompanying effect on normal tissues may imply side effects.

Section: Effect Of Operationcontrasting

confidence: 72%

Effect of D‐Penicillamine on Collagen, Glycosaminoglycans, DNA and RNA of Granulation Tissue and Connective Tissue of Skin, Bone and Aorta in Rats

Junker

Helin

1981

“…The salt soluble collagen was extracted 4 times overnight with 0.5 M-NaCI (HANSEN 1975), and in the aortic samples the insoluble collagen was separated from elastin as gelatine after 3 autoclavings at 128" (NEUMAN & LOGAN 1950). The samples of skin and granulation tissue were also extracted with sodium chloride as above after 17 hours incubation at 37' in order to detect any cross-linking in vitro (HANSEN 1975). In all the collagen fractions the 14C-OH-proline activities were determined as described by JUVA & PROCKOP (1966).…”

Section: Collageti Fractionationmentioning

confidence: 99%

Cyclophosphamide Effect on Collagen Metabolism in Granulation Tissue, Skin, and Aorta of Rats

Hansen

1977

Self Cite

Granulation tissue was produced in rats by subcutaneous implantation of viscose cellulose sponges. Cyclophosphamide 10 mg/kg was given daily intraperitoneally to 15 rats from the day of sponge implantation. Fifteen rats served as pair‐fed controls, and 15 rats served as non‐starved controls. After 14 days of treatment the animals were decapitated. Groups of 3 rats were given 50 μci 14C‐proline intraperitoneally 1, 3, 6, 12, or 24 hours before death. The aorta, skin, and granulation tissue were examined. Cyclophosphamide caused no effect on collagen of the aorta, while in skin, the only detectable effect was a decrease in 14C‐OH‐proline biosynthesis. In granulation tissue, cyclophosphamide caused a fall in the dry weight, a decrease in 14C‐proline uptake and 14C‐OH‐proline synthesis, as well as an increase in the alpha‐amino nitrogen to OH‐proline ratio in purified collagen. These results indicate that cyclophosphamide inhibits the synthesis of proteins, including collagen, and inhibits the hydroxylation of proline in collagen. No effect of cyclophosphamide could be detected on the amount of salt soluble OH‐proline or on collagen cross‐linking. The registered effects of cyclophosphamide on granulation tissue is probably of importance with regard to the anti‐inflammatory action of cyclophosphamide. This action as well as the tissue differences in the sensitivity of cyclophosphamide may be of relevance to the clinical application of cytostatics in rheumatological diseases.

“…First, Hyp from additional sources like elastin (Neuman & Logan 1950) and the Clq complement component (Reid 1974) cannot be distinguished by this procedure. However, elastin derived Hyp in normal skin and granulation tissue constitutes only 1-5% of the total Hyp content (Hansen 1975). About 20% of aortic Hyp in rats of almost the same age originates from elastin (Hansen 1975), which therefore was determined separately.…”

Section: Discussionmentioning

confidence: 99%

“…However, elastin derived Hyp in normal skin and granulation tissue constitutes only 1-5% of the total Hyp content (Hansen 1975). About 20% of aortic Hyp in rats of almost the same age originates from elastin (Hansen 1975), which therefore was determined separately. Due to the low plasma concentration of Clq, the error from simultaneous determination of Clq in the tissues is probably small.…”

Section: Discussionmentioning

confidence: 99%

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Effect of Different Doses of D‐Penicillamine and Combined Administration of D‐Penicillamine and Methylprednisolone on Collagen, Glycosaminoglycans, DNA and RNA of Granulation Tissue, Skin, Bone and Aorta in Rats

Junker

Helin

1981

D‐penicillamine (D‐pen) in doses of 20, 100 and 500 mg/kg/day or D‐pen 100 mg/kg/day plus methylprednisolone (MP) 2.0 mg/kg/day was administered daily for 42 days to rats implanted with viscose‐cellulose sponges. Operated, pairfed rats served as controls. D‐pen increased the DNA content of granulation tissue, but had no effect on the amount of tissue produced. In contrast, high dose D‐pen reduced the content of DNA and collagen in skin. A dose related inhibition of collagen crosslink formation occurred in all tissues, particularly in skin, as indicated by increased proportions of extractable collagen with increased α/β chain ratio and aldehyde content. Moreover, low doses of D‐pen increased the hydroxyproline/proline ratio of acid soluble skin collagen, presumably due to solubilization of type III collagen as demonstrated by SDS‐polyacrylamide gel electrophoresis in the presence of 3.6 M urea. These changes were associated with increased skin fragility and edema plus excess elastin deposition in the aorta after high dose D‐pen treatment. Low dose D‐pen stimulated the 35S‐sulphate uptake into the sulphated glycosaminoglycans (GAGs) of granulation tissue without altering their relative amounts, whereas high dose D‐pen reduced the concentration of chondroitin‐4/6‐sulphate in skin. MP antagonized the solubilizing effect of D‐pen on collagen, probably by inhibition of the collagen synthesis. In addition, MP inhibited the cell proliferation and GAG metabolism. Food restriction reduced the DNA content of granulation tissue. The inhibitory effect of D‐pen on the formation of granuloma collagen crosslinks in the presence of unaltered rate of collagen biosynthesis may diminish the amount of fibrotic tissue due to increased degradability of crosslink deficient collagen. Simultaneous administration of MP may facilitate this effect by inhibiting the biosynthesis of collagen. However, long‐term D‐pen treatment seems to increase the susceptibility of normal tissues to mechanical injury.