112 N. 8/umenkranrz and G. Asboe-Hansen Table I. Molar ratio Hyp/Hyl as calculated from ana/yses oj collagen in a variety oj tissues from ,,arious ,·ertebrates
No. of lagen Col
samples type
i\1a11
Skin 138
Tendon I
Bone 4
Aorta thoracica 2 Annulus fibrosus (5) 35
Outer 17
Inner 18
Nucleus pulposus (5) 16
Ca11ilage 4
Seal Skin Rat Skin. ventral
Skin. dorsal Tcndon, tail Cartilage Mouse Skin, ventral Skin. dorsal Skin. tail Tendon Birds:
Chicke11 Skin Tendon Calvaria Cartilage
Pe11g11i11
Skin Fishes:
Trigla gumard11s Skin
Swim bladder Rays Dorsal fin Gad11s morrlwa
Skin Bone
2
4 4 2 2
6 6 2 2
4 2 2 2
r +111 I I 1+11 Il Il
l+Ill
I+ III I Il
l+lll
1+111 1+111 I
1+111 1 I Il
Molar ratio Hyp/Hyl 16.5 ±0.5 15.7 22.1 ±0.5 14.7 ±2.7 9.45± 1.27 10.3 ±1.0
8.64±0.8 7.77±0.8 7.4 ±0.2 18.9 ±0.3 15.5 ±0.6 16.5 ±0.4 11.0 ±0.7 6.9 ±0.2 19.0 ±0.4 18.0 ±0.6 15.2 ±1.2 15.2 ±1.0
14.8 ±0.3 13.8 ±0.4 16.0 ±0.3 4.8 ±0.2 20.0
14.5 11.6 7.5 5.4 10.9 7.6
evaporated separately (4) and analysed for Hyp and Hyl.
using lhe automated procedures of Blumenkrantz & As
boe-Hansen (2, 3).
From amino acid analyses of purified collagen of dif
ferent tissues reported by various authors. the molar ratio Hyp/Hyl was obtained.
Acta Oen11awr,·11,•r /S1od'10/111) 58
Calcula1io11s
Results were express ed as µg Hyp or Hy! per I O mg drie.:
and defatted tissue (DDT). and the Hyp/Hyl molar rat was calculated. The ratios were compared and diffe
ences evaluated by using Studenfs 1-test.
RESULTS
In Table I. the molar ratios Hyp/Hyl of the sample·
analysed are presented. The ratios of samples o human skin. tendon. bone. aorta, cartilage. annulu•
fibrosus (AF), and nucleus pulposus (NP) are pre
sented. together with a similar analysis of tissues o other vertebrates. The differences between th.
molar ratios Hyp/Hyl of skin, bone and tendo,·
were found statistically non-significant. while th
difference between the ratios of the mentioned ti, sues and cartilage was significant (P<0.01).
The difference in Hyp/Hyl molar ratio betwee·
AF and NP was statistically significant (P<0.01 On comparison of human cartilage with NP ratio, the difference was statistically non-significant (P 0.05). The ratios of the aorta thoracica ar
also significantly different from that of cartilag.
(P<0.05).
The ratios calculated on the basis of the amin acid analyses of various tissues pe1formed by othe authors are given in Table Il. The molar ratil' Hyp/Hyl of types I and III collagen were signif cantly higher than those of types Il and IV. Th finding is attributable to their lower content of H, in relation to Hyp. It was found in our own ana yses. performed on dried and defatted tissue. - well as by calculation from the amino acid analyst:-
of purified collagen.
DISCUSSION
Normal cartilage produces only type Il collagen culture. whereas slices of osteoarthritic cartilag::
also synthesize type 1 (25). The fact that the pol peptides isolated from urine of patients with Paget disease show the same Hyp/Hyl ratio as norrn.11 human bone (Table Il) confirms the nation that t polypeptides mentioned originate from the remode ling of bone which takes place in this disease ( 1�
The molar ratio Hyp/Hyl in urine may thus be useful indicator of collagen resorption, breakdo"
or synthesis of incompletely cross-linked collage Although the collagen of skin. tendon. bone, a
-.. ble Il. Molar ratio Hyp, rifled collagen and subun
=Rachitic, NR=non-rachitic
·sue/organ endon (tail)
�ndon
�ndon ..-n .n 'l4IJl
.:(111 _;bn
er er ::.e
-e (diaphysis) -ne (diaphysis)
"e (diaphysis) -,e (diaphysis)
-;iJage �e --tilage
�,physis) _-jJage
?{physis) --.iJage
""llwth plate) -.ilage -owth plate)
-..ilage (nasal) --:ilage
:}nx: hyaline) -.ilage ..r: elastic)
-ulage
'Yiliscus; fibrous)
�ge -.Jage (larynx) :::.tlus fibrosus eus pulposus .:ment membrane
-meruli) -'"11ent membrane
"lleruli) .-nent membrane
=ent membrane -..Je)
::ient membrane .,,eruli) .-::ient membrane
::neruli)
""lent membrane
capsule)
Vertebr Rat Man Dog Man Rat Calf (3 mo Calf
(3 mo.
Calf (9 mo.
Calf (9 mo Chick Lung-fö Man Man Chick(R Man Chickr.-;
Chick(R Chick(�
Chick Chick Chick Chick(R Chick('\
Chick(R Chick(:-0 Ox Pig Pig Pig Chick Pig Pig Ox Pig Ox Man Man Man Man
Hydroxyproline to hydroxylysine molar rario 113 Table Il. Molar ratio Hyp/Hyl as calcu/ated from amino acid analysis performed by various authors on ed as µ.g Hyp or Hyl per JO mg drie '.'urified collagen and subunirs
DDTI. and ratios were compared and the Hyp/Hyl molar ran difTe· R=Rachitic, NR=non-rachitic
<i,r,; Student's 1-test. Iso-
lated Col-
col- lagen Ra-
Tissue/organ Vertebrate lagen a, /311 /312
"•
/322 type tio AuthorRESULTS
..
Tendon (tail) Rat 13.6 19.3 22.0 11.7 8.2 Piez et al. (20)
lf ratios Hyp/Hyl of the sample, Tendon Man 14.0 Piez & Linkins (21)
ri1ed. The ratios of samples or Tendon Dog 17.0 Kefalides ( 14)
. bone. ao11a, cartilage. annulu, Skin Man 16.0 20.0 20.0 13.4 10.8 10.3 Bomstein & Piez (6)
Skin Rat 16.1 22.3 24.8 14.1 10.7 Piez et al. (20)
nucleus pulposus (NP) are pre- Skin Calf I 11. I Bailey & Sims ( I)
JJ a similar analysis of tissues ot (3 mo. old)
The differences between the Skin Calf III 17.0 Bailey & Sims (I)
(3 mo. old)
'fyl of skin, bone and tendor Skin Calf I 13.3 Bailey & Sims (I)
.:all} non-significant. while tht (9 mo. old)
Skin Calf III 19.7 Bailey & Sims (I)
the ratios of the mentioned tis- (9 mo. old)
a!> significant (P<0.01). Skin Chick I I 5. 36 Trelstad et al. (25)
, Hyp/Hyl molar ratio between Skin Lung-fish 13.7 Eastoe (7)
Liver Man I 18.5 Rojkind & Palomo (22)
�t.istically significant (P<0.01) Liver Man III 22.6 Rojkind & Palomo (22)
1uman cartilage with NP ratios. Bone Man 19.6 Krane et al. ( 15)
Bone (diaphysis) Chick(R) 10.6 Toole et al. (24)
-tatistically non-significant (P> Bone (diaphysis) Chick(NR) 15.7 Toole et al. (24)
of the aorta thoracica are Bone (diaphysis) Chick(R) 9.2 Toole et al. (24)
ifferent from that of cartilage Bone (diaphysis) Chick(NR) 12.4 Toole et al. (24)
Bone Chick 18.5 Miller et al. (18, 19)
Bone Chick 12.5 Miller et al. ( 18, 19)
1ted on the basis of the amino Cartilage Chick 4.5 Trelstad et al. (25)
ious tissues performed by other Cartilage Chkk(R) 4.8 Toole et al. (24)
(epiphysis)
.n Table Il. The molar ratios Cartilage Chick(NR) 5.4 Toole et al. (24)
and III collagen were signifi- (epiphysis)
Cartilage Chick(R) 4.4 Toole et al. (24)
hose of types Il and IV. This (growth plate)
e 10 their lower content of Hy) Cartilage Chick(NR) 4.5 Toole et al. (24)
h was found in our own anal- (growth plate)
Cartilage (nasal) Ox 4.5 Lee-Own & Anderson
dried and defatted tissue, as ( 17)
n from the amino acid analyses Cartilage Pig Il 4.3 Eyre & Muir (9)
(larynx: hyaline)
Cartilage Pig
(ear; elastic) Il 5.3 Eyre & Muir (9)
Cartilage Pig I 10.2 Eyre & Muir (9)
Il (meniscus: ftbrous)
iSCUSSCON Cartilage Chick Il 4.5 Lane & Weiss {16)
Carti lage (lary nx) Pig 4.7 Eyre & Muir (8)
duces only type Il collagen in Annulus fibrosus Pig 6.6 Eyre & Muir (8)
.:ö of osteoarthritic cartilage Nucleus pulposus Pig 4.3 Eyre & Muir (8)
Basement membrane Ox IV 2.7 Sato et al. (23)
l eS). The fäet that the poly- (glomeruli)
n urine of patients with Paget's Basement membrane Ox IV 2.8 Sato et al. ( 23)
.me Hyp/Hyl ratio as normal (glomeruli)
Basement membrane Man IV 3.0 Sato et al. (23)
f. confirms the nation that the Basement membrane Man IV 2.8 Sato et al. (23)
1ed originate from the remodel- (tubule)
J.es place in this disease ( 15). Basement membrane Man IV 2.65 Kefalides ( 12)
(glomeruli)
� Hyl in urine may thus be a Basement membrane Man IV 3.1 Kefalides ( I 3)
'lägen resorption. breakdown. (glomeruli)
Basement membrane IV 3.0 Kefalides & Denduchis
1ple1ely cross-linked collagen. (!ens capsule) (Il)
;en of skin, tendon. bone. and
I Aua DcmrnWl'C/Jl'r !Stu('k/ro/mJ 58
