Crosslinking

The most utilized reagents are aldehydes such as formaldehyde and glutaraldehyde. Formaldehyde reacts
with the -amino groups of lysine and hydroxylysine residues to an intermediate imine which forms a
crosslink either with tyrosine or with an amid group of asparagine or glutamine [M. Chvapil !.". #ronentahl $.
van $inkle %r. Medical and surgical applications of collagen in& '.(. )all '.*. %ackson +,ds.- .nternational !eview of Connective
Tissue !esearch (cademic /ress 0ew 1ork 2345 pp. 2672. ,. #hor Methods for the treatment of collagenous tissues for
8ioprostheses 9iomaterials 2: +2334- 3;62<;.=. ( ma>or advantage of formaldehyde is the possi8ility of
crosslinking dry collagen material with reagent in the vapor phase instead of treatment in li?uid in
particular a?ueous environments [[33= $. Friess !.%. 9iron *. Foskett The effect of formaldehyde and ethylene oxide
treatment on porous collagen systems /harm. !es. 2@ +2334- *-2;@.
[2<<= !.%. !uderman C.$.!. $ade $.'. *hepard F. "eonard /rolonged resorption of collagen sponges& vapor-phase treatment
with formaldehyde %. 9iomed. Mater. !es. 4 +2345- A756A7;.=.
'ue to the 8rittleness of formaldehyde crosslinked products potential toxicity and adverse reactions arising
from residuals and reversi8le fixation formaldehyde is no longer favored al8eit approved for medical
devices [*.'. Borham Collagen in& '. 9yrom +,d.- 9iomaterials *tockton /ress 0ew 1ork 2332 pp. ;;62AA.=.
Treatment with GTA is a commercially via8le process its efficiency has 8een optimized it is inexpensive
and completed in a relatively short period and provides more sustained long-term dura8ility than
crosslinking with formaldehyde [!.F. Cliver !.(. Brant !.$. Cox (. Cooke ,ffect of aldehyde crosslinking on human
dermal collagen implants in the rat 9r. %. ,xp. /ath. 72 +23:<- ;@@6;@3. ,. #hor Methods for the treatment of collagenous tissues
for 8ioprostheses 9iomaterials 2: +2334- 3;62<;.=.
(s an alternative to aldehyde treatment strong and resistant collagen material can 8e o8tained using
hexamethylendiisocyanate +)'C- as a crosslinker [,. #hor Methods for the treatment of collagenous tissues for
8ioprostheses 9iomaterials 2: +2334- 3;62<;.=.
The epoxy functionality from ethylene glycol diglycidyl ether glycerol polyglycidyl ether methylglycidyl
ether predominantly reacts with the amino groups on lysine much like BT(.
Crosslinking with carbodiimides especially 2-ethyl-5-+5- dimethylaminopropyl-car8odiimide +,'C-
offers the main advantage over aldehydes )'C or epoxy compounds in that these car8odiimides only
facilitate the formation of amide 8onds 8etween car8oxylic and amino groups on the collagen molecules
without 8ecoming part of the actual linkage.
Car8oxylic groups are methylated and converted into hydrazides followed 8y reaction with sodium nitride
to form the acyl azide or can 8e modified with diphenylphosphoryl azide. These azide functionalities
couple with amino groups to give amide crosslinks.
( ma>or handicap of chemical crosslinking agents is the potential toxic effect of residual molecules andDor
compounds formed during in vivo degradation. Therefore alternative physical methods are pursued
including dry heat exposure to ultraviolet or -irradiation [,. #hor Methods for the treatment of collagenous tissues
for 8ioprostheses 9iomaterials 2: +2334- 3;62<;.=.
The effectiveness of UV and chemical crosslinking of collagen membranes as dermal dressings has been
evaluated by measuring their mechanical properties and resistance to collagenase as good indicators of the
fabrication process.[ Lee JE, ark J!, "#ang $%, &im J&, &im J', %ub ". ())*. !haracteri+ation of UV,irradiated
dense-porous collagen membranes. /orphology, en+ymatic degradation and mechanical properties. $onsei /ed J
0(.*1(2*13.4
Tannic acid cross-links collagen 8oth in acid +p) 5.:- and weakly 8asic +p) 4.@- medium. .ntermolecular 8ridging
produces changes in the amide ... .! 8and of the collagen-tannic acid matrices. The higher the amount of tannic acid in
the matrix the slower the enzymatic degradation process. The release of tannic acid from the collagen-tannic acid
matrices is slower for samples o8tained at p) 5.: than for those resulted at p) 4.@ due to the higher amount of
intermolecular 8onds formed.
Type . fi8rillar collagen hydrogel extracted from calf hide 8y 8asic and acid treatments at
AAoC using hydrochloric acid was crosslinked with formic or glutaric aldehyde [M B (l8u
E M "eca !heological 9ehaviour of some Cross-linked Collagen )ydrogels for 'rug 'elivery Fse
,uropean Cells and Materials Gol. 2<. *uppl. 2 A<<; +page :- .**0 2@45-AA7A= The differences in
viscosity and homogeneity of the hydrogel crosslinked with the two aldehydes can 8e explained
8y the complex crosslinking mechanism when glutaraldehyde is used.