Effects of the methylene chain length of chemically introduced crosslinks on the properties of collagen

Watanabe, Kazuo; Nakagawa, Junko; Ebihara, Tetsuya; Okamoto, Yasushi

doi:10.1016/s0032-3861(97)00046-3

Cited by 11 publications

(8 citation statements)

References 16 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…We have ascertained that the length that can span over one collagen helix pitch ( > 9.5 A 17 ) is necessary for the cross-linker to efficiently introduce the crosslink into the collagen molecule. 18 The cross-linking reagents employed in this study react with the amino group of the protein (Scheme 1), and the maximum lengths between two imino nitrogen atoms interposing each chemical cross-link are presumed to be within the range of 11 to 12 A. Therefore, we selected DMS and APTS in order to examine the structural effects on the properties of the cross-linked SCLs.…”

Section: Resultsmentioning

confidence: 99%

Comparison of Properties of the Collagen Cross-Linked with Aliphatic or Aromatic Reagent

Watanabe

1997

Polym J

Self Cite

View full text Add to dashboard Cite

KEY WORDSCollagen I Cross-Link I Cross-Linkage Structure I Denaturation Temperature I Helix Regeneration ICollagen is a very useful protein for constructing artificial organs. 1 -6 When collagen is employed as a biomaterial, chemical cross-links are usually introduced into the collagen molecule to stabilize the collagen triple-helical structure, because a remarkable change in the properties of collagen is caused by the disintegration of the triple-helical structure. 7 -10 In order to prepare a fine cross-linked collagen, the relationship between cross-linkage structure and the properties of the cross-linked collagen was previously examined using six kinds of cross-linking reagents. 11 Heat-denatured collagens cross-linked with aliphatic reagents regenerated more triple helix than those cross-linked with aromatic reagents by cooling. In the present study, cross-linked collagens were prepared using two kinds of cross-linking reagents that have quite different molecular structures, and several properties of these collagens were extensively investigated. EXPERIMENTAL MaterialsCollagen (solubilized collagen with lime, SCL) was isolated from steer hide using an alkaline solution. 12 Dimethyl suberimidate dihydrochloride (DMS) was obtained from Wako Pure Chemical Industries, Ltd., and 1-acetoxypyrene-3,6,8-trisulfonylchloride (APTS) was from Lambda. Sample PreparationThe preparation of the collagen cross-linked with DMS (DMS-SCL) was carried out according to the procedure of our previous investigation, 11 but various amounts of DMS were added to each SCL solution. The amount of the DMS bound to SCL was estimated by quantifying the number of free amino groups in the cross-linked SCL using trinitrobenzenesulfonic acid. 13 The degree of cross-linking was expressed as the percentage of the closed amino group of SCL by binding the cross-linker. A DMS-SCL solution for spectroscopic measurements was prepared as follows. The lyophilized DMS-SCL was added to 20 mM acetic acid and was continuously stirred at 4oC overnight. The DMS-SCL solution was then filtered, and the concentration was adjusted to 0.01% based on the microbiuret method. 14 The preparation of the collagen cross-linked with pyrenyl reagent (Py-SCL) was performed as follows. 286SCL solution, the concentration of which was about 0.075%, was prepared using 0.1 M phosphate buffer, pH 10. The pyrenyl cross-linking reagent, APTS, was dissolved in acetone, and then various quantities of the acetone solution were continuously and uniformly added to each SCL solution with gentle stirring. The mixture was continuously stirred in the dark at 4°C for 17 hours. After the cross-linking reaction had been completed, excess glycine was added to the reaction mixture in order to obstruct the free active groups of APTS by binding glycine. The mixture containing glycine was stirred for additional 3 hours. The reaction mixture was then dialyzed against 10 liters of I M acetic acid and five changes of 15 liters of 5 mM acetic acid. The dialysate was lyophilized and stored below -20°C...

show abstract

Section: Resultsmentioning

confidence: 99%

Comparison of Properties of the Collagen Cross-Linked with Aliphatic or Aromatic Reagent

Watanabe

1997

Polym J

Self Cite

View full text Add to dashboard Cite

show abstract

“…Several sugar alcohols (polyols) have been promoted as potential sugar substitutes in caries limitation [9,10]. In collagen, chemical cross-links have been introduced to stabilize the structure of collagen [11][12][13][14][15][16]. The stability of calf skin collagen (CSC) type I during thermal and chemical denaturation in the presence of glycerol was investigated [17].…”

Section: Introductionmentioning

confidence: 99%

Stability of collagen with polyols against guanidine denaturation

Usha

Ramasami

2008

Colloids and Surfaces B: Biointerfaces

View full text Add to dashboard Cite

“…Recently, dimethylsuberimidate (DMS) has been used for cross-linking collagen instead of GTA [ 1 4] because the cytotoxic effects of DMS are lower than those of GTA [ 1 5] . We have investigated the properties of the collagens cross-linked with various chemical reagents including DMS and found that DMS is an effective reagent for cross-linking collagen [16,17].…”

Section: Introductionmentioning

confidence: 99%

Adhesion of macrophages on collagen irradiated with ultraviolet light

Watanabe

Koyama

1999

Journal of Biomaterials Science, Polymer Edition

Self Cite

View full text Add to dashboard Cite

Properties of collagen irradiated with ultraviolet (UV) light were examined using the techniques of sodium dodecyl sulfate-polyacrylamide gel electrophoresis, spectroscopic measurements, and cell adhesion assay. Both photopolymerization and photodegradation of the collagen appeared to occur with UV irradiation because the aggregation of collagen and disintegration of the triple-helical structure were observed. The formation of the cross-links between the tyrosine residues in collagen by photoreaction was presumed. The adhesion of macrophages (Mphis) on a polystyrene plate was suppressed by coating the plate surface with collagen. Anti-adhesive activity of collagen on Mphi cells decreased with UV irradiation. However, appreciable anti-adhesive activity remained in the modified collagen even when collagen was irradiated with UV light for 24 h.

show abstract

Effects of the methylene chain length of chemically introduced crosslinks on the properties of collagen

Cited by 11 publications

References 16 publications

Comparison of Properties of the Collagen Cross-Linked with Aliphatic or Aromatic Reagent

Comparison of Properties of the Collagen Cross-Linked with Aliphatic or Aromatic Reagent

Stability of collagen with polyols against guanidine denaturation

Adhesion of macrophages on collagen irradiated with ultraviolet light

Contact Info

Product

Resources

About