September, 1967

390 T h e Journal o[ P E D I A T R I C S

Cellular growth of human placenta

IIL Intrauterine growth failure

Placentas from in[ants with intrauterine growth [ailure were examined [or total organ
weight, desoxyribose nucleic acid ( D N A ) , ribonucleic acid ( R N A ) , and protein. In
eight in[ants with severe maI[ormations, the placentas were entirely normal
when compared with those [rom physically normal infants of comparable gestationaI
age. In seventeen in[ants with low birth weight but with no maI[ormations there
was a proportional reduction in weight o[ the placenta and in its DNA and protein
contents; there was, however, a marked elevation o[ R N A content. Thus
protein/DNA was normal whereas the R N A / D N A ratio was elevated. It is suggested
that this increased ratio may be a mani[estation o[ placental insufficiency.

Myron Winick, M.D.

NEW YORK~ N. Y,

T ~ I E T O T A L organ content of DNA is
a measure of the number of cells at any
given time? The protein/DNA ratio is a
measure of individual cell size, and the
quantity of RNA per cell may be expressed
as an R N A / D N A ratio. During growth any
or all of these quantities or ratios m a y
change in response to various environmental
stresses. For example, malnutrition in neo-
natal rats results in fewer cells of normal
size in all organs studied. 2 Congenital heart
disease in young children is associated with
a reduction in both number of cells and cell
size in skeletal muscle2 Increased caloric
intake in neonatal rats is accompanied by an
increase in number of cells in various organs,
From the Cornell University Medical College,
I300 York Ave.
Supported by grants [rom The National
Foundation No. 1270, the Ross Laboratories
No. 15-0532082, and the Nutrition
Foundation No. 357.
Vol. 71, No. 3, pp. 390-395
with cell size increasing only in brain. 4 The
number of cells is also increased in the pla-
centas of diabetic mothers. 5 R N A / D N A ra-
tios have been increased under a variety of
conditions.
Although intrauterine growth failure has
been ascribed to "placental insufficiency" due
to a "faulty intrauterine environment," the
placentas from these infants are histologi-
cally normal2 Moreover, no biochemical ab-
normality has ever been demonstrated.
This study investigates the possibility of
a biochemical abnormality by comparing to-
tal weight, protein, DNA, and R N A content
in placentas from infants too smalt for ges-
tational age with the same parameters in
placentas of normal infants.
MATERIALS AND METHODS
The placentas from 25 infants who were
more than two standard deviations below
Volume 71
Number 3
the mean weight for gestational age were
examined. 7 Expected date of confinement
was established from the date of the last
menstrual period. The infants were divided
into two groups: one containing those with
major congenital malformations; the other,
those with no obvious physical defect other
than size. Fig. 1 identifies the infants in the
two groups. There were eight patients in
the first group: two with anencephaly; three
with microcephaly and multiple malforma-
tions, one of whom had proved rubella; and
three with multiple severe skeletal deformi-
ties. Seventeen patients comprised the second
group, in which low birth weight and small
stature were the only physical deviations.
Twelve of the mothers of these infants had
uneventful pregnancies; four had mild to
moderate pre-eclampsia, and one had severe
pre-eclampsia.
Placentas were obtained immediately upon
delivery and processed as previously de-
scribed. 7 DNA, RNA, and protein were de-
termined by methods identical to those pre-
viously described. 8 Lines on all graphs' in-
3,500 - -
3,000 -
2,500 -
E
2,000 -
..~ 1,500
1,000 -
50o
0 [
10
Fig. 1.* S y m b o l s : triangles, n o p h y s i c a l a b n o r m a l i t i e s ; circles, severe c o n g e n i t a l m a l f o r m a t i o n s .
Lines i n d i c a t e r a n g e of n o r m a l f o r 50 h u m a n p l a c e n t a s J
* S y m b o l s a r e t h e s a m e for Figs. 1 to 5.
Intrauterine growth [ailure 39 1
dicate the upper and lower ranges of normal
values as previously determined on 50 nor-
mal human plaeentass
RESULTS
Placental weight. Fig. 2 shows that the
placentas from the eight infants with con-
genital defects were of no~xnal weight for
gestational age. The placentas from the 17
infants without defects were small for gesta-
tional age. However, when these data are
compared to the infant weight, the placentas
from both groups were actually large. Thus,
in infants whose growth failure is accom-
panied by severe malformations, there is no
decrease in placental weight, whereas pla-
centas from infants with only intrauterine
growth failure do show retarded growth.
This retardation, however, is not as great as
in the fetus.
Placental DNA. Total organ DNA and,
therefore, total cell number were normal in
all 25 placentas when compared to DNA
content of normal placentas of comparable
weight (Fig. 3). Since the weight of the pta-
, [ f [ p 1 J
20 30 40
Weeks of gestation
392 Winick The Journal o[ Pediatrics
September 1967

600 -

500

4OO
E

.~,
3oo
9 A AA
A
9 AA

~- 200 -

/
i00 -

o I , I 1 I I
o 10 20 30 40
Weeks of gestation

Fig. 2.

1,400 - -

~ o

./
1,200 -

1,000

E
<
N 8oo -

600 -
o_

400 -

2OO

, I , I , I I
100 200 300 400 5O0
Placental weight/gms)

Fig. 3.

centas in the malformed group was normal,
DNA content was, therefo.re, normal for
gestational age. However, since placental
weight is decreased in fetal intrauterine
growth failure unassociated with abnormali-
ties, the DNA content of the placenta is also
decreased proportionately. Thus, the smaller
placenta contains fewer cells than the one
of normal size does.
Placental Protein. The total placental con-
tent of protein was comparable to that of
normal placentas of similar weight (Fig. 4).
The protein/DNA ratio is, therefore, nor-
mal, indicating that dry-cell size is essen-
Volume 71
Number 3
32-
28
24
20 -
~- 16
~_ 12 -
&OA/~////
8 -
4 -
o f I I
0 100 200
Placental weight.(gms)
Fig. 4.
dally the same in both groups. Therefore, it
can be assumed that the smaller placentas
from the infants with intrauterine growth
failure and no malformations contained cells
of normal size but had fewer of them than
did the larger placentas of the infants with
malformations.
Placental RNA. Total organ RNA con-
tent was normal in the eight placentas from
infants with congenital malformations,
whereas it was markedly elevated in all 17
infants without malformations (Fig. 5). Both
RNA concentration and R N A / D N A ratios
were normal in placentas from the eight in-
fants with intrauterine growth failure and
associated malformations. RNA concentra-
tion and R N A / D N A ratios in the 17 infants
with intrauterine growth failure but no mal-
formations were markedly elevated.
DISCUSSION
Infants with intrauterine growth failure
can be divided into two groups depending
on the presence or absence of major malfor-
mations. These data suggest that these clin-
ical types may be etiologically different. In
infants with major malformations and in-
Intrauterine growth [ailure 3 93
9 9
, I , I , F
300 400 500
trauterine growth failure, placentas tend to
be of normal weight and have normal nu-
cleic acid and protein content. By contrast,
placentas from infants born with only intra-
uterine growth failure are small for gesta-
tional age. These placentas contain decreased
quantities of DNA. Cell number is reduced.
Protein content is also reduced proportion-
ally to the reduction in weight and DNA
content. The protein/DNA ratio is, there:-
fore, normal, indicating that dry-cell size
is normal. The reduction in placental weight
for gestational age is, therefore, accompanied
by a concomitant reduction in cell number,
with the cell size remaining unaffected.
DNA synthesis and cell division have
been shown to cease before growth ceases
in both rat 9 and human placentas. 7 In hu-
man placenta there is no further increase in
cell number after approximately the thirty-
sixth week of gestation. Since cell number
is reduced in these placentas, it would ap-
pear that, whatever the stimulus to retarded
growth is, it must already be operating
prior to the thirty-sixth week of gestation.
One can conclude, then, that the causes of
faulty intrauterine growth with no other
3 94 Winick
900 ---
800 -
70O
60O
500
400
3OO
2OO
I00
IOO
Fig. 5.
abnormalities affect both placenta and fetus.
They are operating before the thirty-sixth
week of gestation and are accompanied by
a reduction in the expected number of pla-
cental cells.
In addition, there is a selective increase
in RNA content in these placentas. Thus
the R N A / D N A ratio is markedly elevated.
At present, the significance of an elevated
R N A / D N A ratio is unknown. During exper-
imental cardiac hypertrophy due to mechani-
cal factors, 1~ and uterine hypertrophy due
to hormonal factors, 11 R N A / D N A ratios in-
crease. Experimental nerve stimulation is
accompanied by an increase in the ratio in
the excited muscle, 12 and human bone mar-
row has shown a similar rise in association
with various malignant neoplasms2 a It would
appear that a rise in the R N A / D N A ratio
is a nonspecific change occurring in response
to a variety of stimuli. These data, then, in
no way suggest the nature of the basic pla-
cental abnormality. However, they do dem-
onstrate that an abnormality does exist with-
in the placenta. The increased RNA/DNA
ratio is not dependent on a lack of fetal
growth alone, since it does not occur when
The Journal o[ Pediatrics
September 1967
z~
A
A
A
A
AA
h AAA 2
9 9
~ 9 9
I , I L I , I
200 300 400 500
Placental weight (gins)
growth is impaired in conjunction with
congenital malformations. It is possible that
this is actually a biochemical manifestation
of "placental insufficiency" in the placentas
of infants with intrauterine growth failure
and no other abnormalities.
At times it is difficult, because of faulty
dates, to know if a newborn infant is truly
premature or a full-term infant manifesting
intrauterine growth failure. Apart from clini-
cal differences in these infants, differentia-
tion may be possible on the basis o~ an ele-
vated R N A / D N A ratio in the placentas.
SUMMARY
Placentas from 25 infants with intra-
uterine growth failure were examined for
total weight and nucleic acid and protein
content. In eight infants with concomitant
severe malformations, placental weight and
the protein, DNA, and RNA contents were
all within the normal range. In 17 infants
of abnormally small size but with no. malfor-
mations, there was a proportional reduction
of placental weight and of the protein and
DNA contents, but a paradoxical increase
in i:he RNA content. Protein/DNA ratios
Volume 71
Number 3
were normal in the infants of the latter
group, whereas R N A / D N A ratios were
markedly elevated.
These data indicate that intrauterine
growth failure in infants without gross mal-
formations is associated with reduced pla-
cental weight accompanied by fewer cells
which are normal in size:. The amount of
RNA per cell ( R N A / D N A ratio.) is in-
creased. It is suggested that this increased
ratio may be a manifestation of placental
insufficiency.
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1. Enesco, M., and Leblond, C. P.: Increase
in cell number as a factor in the growth
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2. Winick, M., and Noble, A.: Cellular response
during malnutrition at various ages, J. Nutri-
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3. Cheek, D. B., Graystone, J., and Mehrizi,
A.: The importance of muscle cell number in
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Johns Hopkins Hosp. 118: 140, 1966.
4. Winick, M., and Noble, A.: Cellular response
with increased feeding of neonatal rats, J.
Nutrition 91: 179, 1967.
Intrauterine growth [ailure 3 95
5. Winick, M., and Noble, A.: Cellular growth
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