VELVETANTLERREVIEW

Velvet Antler
Its historical medical use, performance enhancing effects and pharmacology
By Dr. John S. Church

Dr. John Church, Game Farm Manager for Canadian Rocky Mountain Resorts, is
responsible for the construction of game farm facilities and the purchase, care and
management of wapiti, bison, white–tailed deer and reindeer. Dr. Church received both
his B.Sc. in Wildlife Management and his Ph.D. in Rangeland and Wildlife Resources
from the University of Alberta. In between these studies, he received his M.Sc. in
Biology: Applied Animal Behavior at Dalhousie University in Halifax. In addition to his
current position, Dr. Church has worked as a researcher at the Centre for Agricultural
Diversification at Dawson Creek, BC working with bison, and as an instructor in the
Diversified Livestock program at Lakeland College in Vermilion.

1. Historical Use of Velvet Antler

The importance in traditional Chinese medicine of the advancement of health and the
prevention of ill health is in direct disagreement with western medical practice, which is
more impressed with the treatment of ill health (Fulder 1980a). In fact the entire culture
of traditional Eastern medicine is one of the quest for health rather than the treatment
of ill health (Brekhman 1980; Kaptchuk and Creacher (1987)). Historical literature in
both Chinese and Korean describes antler as soft growing tissue with velvet, and highly
regarded the efficacy of antler as preventative medicine. Currently, there is an
expanding stake in medicinal products which are alternative in nature and have tonic
effects or effects on well–being. Holistic medicine is one area where velvet antler has
traditionally found a niche in oriental medicine. It has also been used historically in the
specific treatment of a number of conditions including anemia, arthritis, impotence,
mynoxenia, dysfunctional uterine bleeding, dizziness and vertigo, insomnia, amnesia,
wounds and pain (Kong and Ko 1987; Yoon 1989).

The use of velvet antler as a medicine and the momentousness of sexual well-being in
Chinese tradition, have consummated in velvet antler being regarded by western
commentators as an aphrodisiac. This characterization is unfortunate since in western
countries, it has resulted in velvet antler being ignored as a serious candidate for
pharmacological activity or application. In this regard, it is fairly ironic that a Korean
doctor (Yoon 1989) observes that about 10% of velvet antler users are children. In
Korea, antler is regarded as a fundamental component in herbal medicine, used for its
preventative and restorative functions.

It is theorized that deer antler amplifies the body’s metabolism in general, preserves
and renews injured organs and tissues (accelerating healing and recovery from injury),
assists immune and phagocytic functions (anti-inflammation, anti-arthritis, anti-stress),
moderates the aging process, has hypotensive-vascular effects, and ameliorates both
gonadotrophic and thyroid function. This report will attempt to address the scientific
validity of these traditionally held beliefs.

2. What is Velvet Antler?

The name "antler" comes from the Latin word "anteoculae" meaning "in front of the
eyes". Antlers are appendages of the skull, created from a thick bony core and upheld
on permanent skin covered pedicles (protuberance of the frontal bone). The creation
of antlers by males is observed after pedicle development from the periosteum of the
frontal bones of almost all members of the deer family Cervidae (Goss 1983).

Unlike horns in cattle, antlers are cast off every year. Deer or cervids such as caribou,
wapiti and moose grow antlers while cattle or bovids including mountain goats, bighorn
sheep, bison and pronghorn antelope possess horns. With the exception of reindeer
and caribou (Rangifer tarandus), only the males grow antlers.

3. Velvet Antler Composition

The developing antler is composed of an aggregate of distinct cell types including
fibroblasts, chondroblasts, chondrocytes, and osteocytes (Banks and Newberry 1982).
Growing antler tips are composed of minute millimeters of undifferentiated
mesenchymal cells that begin to differentiate very abruptly as cartilaginous tissue.
Afterward the cartilage is replaced by bone, under the influence of testosterone and its
metabolites, and the velvet is shed leaving mature hard antler (Fenessy and Suttie
1985). Consequently when velvet antler is harvested at suitable stages for use as high
quality oriental medicines, it is actively growing cartilage-type tissue which is not of
uniform composition, that is sought. Chemical identification of antler is currently being
explored by Canadian scientists to identify the active components, and to locate the
quality and criterion of antler and antler by-products by utilizing chemical markers
(Sunwoo et al. 1995).

The rate of mineralization or calcification of velvet antler is commonly referred to as a
gauge of the probable pharmacological quality, with heavily calcified velvet antler being
downgraded. The constituents of dry matter analysis of velvet antler demonstrate that
collagen, calcium, phosphorus, and magnesium increase upward, while protein and
lipids decrease downward from the tip to the base of the main beam in growing antler.

However this largely depends on the stage of growth as is indicated by the relative
mineral content in the lower section of velvet antlers cut at different stages of growth
after casting. The effect of stage of development on lipid content is also significant.
Amino acid contents stated as a percentage of total protein and lipid is considerably
higher in the tip section, from which the antler grows. The concentrations of uronic acid,
sulfated glycosaminoglycan, and sialic acid decrease from the tip portion downward
towards the base of the growing antler. The tip segment has the best proportions of
tyrosine and isoleucine and the smallest proportions of glycine and alanine. Linolenic
acid was discovered in the tip segment only.

Recent studies at the University of Alberta, Canada has shown that velvet antlers
contain chondroitin sulfate as a major glycosaminoglycan with small amounts of keratin
sulfate, hyaluronic acid, and dermatan sulfate (Sunwoo et al. 1997). Even more
recently, the same researchers in Canada have extracted and characterized
proteoglycans from the cartilaginous portion of velvet antler from wapiti, and found two
types of proteoglycans including large chodroitin sulfate proteoglycan and small
proteoglycan, decorin (Sunwoo 1998).

Research back in 1988 established that chondroitin sulfate A is an extremely potent
anti-inflammatory agent. There are convincing opinions that there is substantial
difference in mineralization between species of deer at the same stage of growth, but
this has not been quantified. The compositional changes from the tip to the base are
reflected in both Chinese (Wang and Zhou 1991) and Korean (Yoon 1989) medical
systems which broadly classify the various parts of velvet antler. The tip is referred to
as the wax piece, the next section is the blood piece, and finally the bottom is known as
the base or bone piece (Fennessy 1992). Once the velvet antler is harvested, blood
quickly seeps away from the tip region, although inverting the antler will help alleviate
this problem. Depending on the drying methods, the dried product can have
considerable blood in this section. However the traditional Chinese drying methods
result in the tip remaining empty of blood; therefore its often categorized as the wax
piece.

Dr. Peter Fennesy, general manager of the Invermay Research Centre in Otago, New
Zealand has stated that initial research data indicates that elevated levels of a natural
growth hormone called insuline–like growth factor (IGF-1) exists in the blood of deer
during the antler growth cycle as well as receptors to IGF-1. As human beings age,
growth hormone levels decline along with IGF-1, which results in muscular atrophy.
Velvet antler is most likely an unrefined source of IGF-1 that can improve muscular
development. Cell culture studies have discovered that the administration of IGF–1 and
2 can have a significant effect on the cells in velvet antler. These growth factors
augment cell division in undifferentiated cells in the fibroblast zone, the growing tip and
the cartilage zone. These finding indicate that IGF- 1 and 2 are likely important
facilitators for antler growth. The significance of these factors to the cell regeneration
processes in humans has recently been a source of much speculation.

Subsequent studies at Oxford University in England has resulted in the discovery that
IGF-1 increases the release of alkaine phosphatase and cell growth in the distal antler
tips of male red deer (Cervus elaphus). This growth factor increases the rate of cell
division in the inner layer of the perichondrium, the reserve mesenchyme and the
cartilage zone. The biochemistry that contributes to the rapid growth of velvet antlers
probably has undiscovered medical potential for humans with regards to increasing cell
growth and repair.

4. Traditional Medicinal Uses of Antler

In Oriental medicine, the different sections of velvet antler have assorted uses. The
upper two sections are often used as preventative tonics in children while the middle
portion is often used to treat arthritis and osteomyelitis. The lowest part of velvet antler
is often administered to older people to help prevent calcium. Velvet antler has also
been used in childbirth to assist delivery, anemia, menopausal disorders, impotence
and spermatorrhea.

As a medical product, velvet antler is dried, processed and used in a variety of
treatments. Traditional methods of processing antler were designed to avoid spoilage
during the slow drying process.

Processing generally involves repeated immersions in boiling water followed by drying
using either heat or open air drying. Although no details are given, studies cited by
Russian scientists Yudin and Dubryakov (1974) state that boiling of the antlers, one of
the traditional steps commonly used is contraindicated in terms of its effects on
pharmacological activity. There are many different forms in which dried antler is
processed into for use, including slices, powders and extracts. Velvet antler in Asia is
often included as a single component of prescription medicine while other over-the-
counter preparations that also include velvet antler are combined with other traditional
medicines, especially herbs.

5. Performance Enhancing Effects of Velvet Antler

Velvet antler has often been regarded as having performance enhancing effects on the
human body. There is scientific evidence from a number of studies that have revealed
such effects in both animals and humans. For example, Brekhman et al. (1969) showed
that pantocrin increased the working capacity of mice. Russian scientists Yudin and
Dubryakov (1974) have reported that control athletes on an exercise cycle performed
15 kg/m of dynamic work whereas those given pantocrin increased this considerably to
74 kg/m and those given rantarin (a preparation of reindeer antler) increased to 103
kg/m. In a like manner, the athletic performance in a 3000m run was enhanced
following patocrin administration (Brekhman et al. 1969). According to Russian scientist
Korobkov (1974, cited by Fulder 1980b) with regards to the use of velvet antler in
athletes, the action is primarily aimed at accelerating the restorative processes after
intensive activity and at increasing the body’s resistance to unfavorable external
influences. In essence, pantocrin and other naturally occurring substances in velvet
antler have served to accelerate the body’s natural restorative processes.

For well over a decade, Dr. Arkady Koltun, MD, Ph.D., Chairman of the Medical
Committee for the Russian BodyBuilding Federation, has conducted research into
anabolic agents that are known to improve performance, strength, and musculature in
atheletes. In studies with Russian kayakers, weightlifters, bodybuilders and powerlifters,
Dr. Koltun found that velvet antler has both myotropic (increases muscular strength)
and neurotropic (nerve strengthening) properties. He also found properties in antler
that are beneficial in treating infectious disease, fatigue and hypertension.

The performance enhancing effects of velvet antler are likely the results of increasing
the circulating levels of androgens in the blood of these atheletes. There is now
considerable evidence for the gonadotrophic effects of velvet antler. Androgens
(testosterone and its metabolites) are known to stimulate the development of seminal
vesicles and the prostate gland of sexually immature neonate rats, or retard the
degeneration of these organs in newly castrated animals. Velvet antler preparations
pantocrine and rantarin have all been shown to have androgenic effects.

Haematopoietic effects of velvet antler have been demonstrated in numerous
experiments. Preparations of velvet antler have been shown to stimulate red blood cell
synthesis and increase erythropoietic activity in cases of drug induced anemia in
rabbits and rats. It seems likely that such erythropoietic activity may well be responsible
for at least part of the stamina-improving effects of velvet antler preparations in
distance runners. In this sense, the responses would be similar to those ascribed to
blood-doping where an athlete in re-transfused with his own blood prior to competition.

6. Pharmacological Effects of Velvet Antler

The documented effects of velvet antler in studies with laboratory animals are
numerous, generated mainly from the former Soviet Union, as well as Korea, China,
Japan, Hong Kong, New Zealand and recently Canada. Much of the Russian work is
concerned with the extracts of pantocrin or rantarin.

The reported pharmacological effects and evidence for bioactivity include the following:

·         stimulating and tonic effects

·         androgenic/gonadotrophic effects

·         haemotopoietic effects

·         hypotensive and cardiovascular effects

·         anti-stress effects

·         growth-stimulating effects

·         retardation of aging

·         accelerated recovery from injury

·         anti-tumor effects

·         anti-cholesterol effects

The biological activity is highly correlated with several of the extract components
including pentose sugars, free amino acids, free fatty acids and phospholipids. The
Russian extract pantocrin has demonstrated hypotensive effect in animals under
anesthesia. The effect is transient, causing a drop in arterial pressure of up to 50%.
The hypotesive effects of the alcohol extract pantocrine are likely due to the presence
of lysophophtidyl cholines.

Some further evidence of potent pharmacological activity of velvet antler or antler
preparations include evidence that treatment with velvet antler can protect against
shock or stress. For example, Kang (1970) reported that antler pre-treatment has
reduced cell degradation in rats subjected to heat stress, cold stress or electric shock.
Yudin and Dubryakov (1974) reported that rantarine alleviated the adverse effects of
stress in normal stress-related responses such as hypertrophy of the adrenals,
involution of the thymus gland and reductions in the weight of the liver and kidneys
when laboratory animals were administered the extract.

Wang et al. 1988 claims that it is the polysaccharide content that is responsible for the
anti-ulcer effects of velvet antler preparations. Kim and Lim (1977) cited Russian
studies showing that treatment of patients with rantarin prior to surgery for
gastrointestinal tumors resulted in reduced stress responses in rantarin-treated
patients. Velvet antler treated rats have also been shown to better tolerate carbon
tetrachloride-induced liver damage with some evidence of different responses with
velvet of different sources, presumably due to the preparations being from velvet
harvested at different stages of growth. Beubenick (1986) mentioned that an extract
from the growing antler tip section facilitates healing of epidermal wounds in rats. Thus
from a variety of sources, there would appear to be good evidence for the efficacy of
velvet antler preparations in the treatment and alleviation of stress related conditions.

In Korea, a study was conducted to evaluate the nutritive value of velvet antler on
blood cholesterol levels in rats. The blood cholesterol level was significantly reduced in
the rat when the diet was supplemented with velvet antler. In addition, body weight gain,
feed intake and feed efficiency remained unchanged, proving lowered cholesterol
levels were not due to these factors. Korean researchers have determined that feeding
velvet antler to broiler chickens resulted in a small but significant increase in growth
rate and food conversion efficiency over an 8 week period. Interestingly, the weight of
the testes was significantly increased while the thyroid weight was decreased.

Studies in Japan (Wang et al. 1988) have shown marked effects of velvet antler
preparations on biochemical parameters related to aging in senescence-accelerated
mice (SAM), a model for senility. The hot water extract of velvet antler was administered
for 8 days. Treated mice showed significant improvements in parameters normally
associated with senility, including an increase in plasma testosterone. The effects were
generally observed only in the SAM strain and not in the control strain of mice,
suggesting that velvet preparation may exert an anti-aging effect in senile animals.

Wang et al. (1988) demonstrated that mice subjected to chloroform damage to the liver
that is caused by an increase in free radicals could be alleviated by treatment with
velvet antler. Further studies (Wang et al. 1988) revealed a direct effect on the rate of
protein synthesis in the liver and kidney apparently mediated by an increase in RNA
polymerase activity (RNA polymerase regulates RNA transcription from nuclear DNA).
The studies carried out by Wang and his associates (1988 a,b,c) appear to be careful,
well thought out and very credible. These studies provide a good starting point for
further work in this area. Effects such as those reported by Wang et al. (1988b) in the
kidney (and in the liver) are also produced by androgens, again suggesting that some
more intensive research directed towards the steroid-like activities of velvet antler
preparations would be beneficial.

Prostaglandins discovered in velvet antler have been recognized as anti-inflammatory
components that reduce the body’s reaction to injury, swelling, infection, pain and
arthritis. Also, the collagen in velvet has been demonstrated as a healing agent when
ingested and when applied as a topical skin treatment. Studies conducted in China on
an extract of antler have shown anti-inflammatory properties by reducing acute and
chronic inflammation in rats. The extract reduced ascorbic acid and cholesterol
contents in the adrenal glands and decreased the serum hydrocortisone level in rats.
The results of these studies indicate that the antler contains anti-inflammatory and
other agents that are beneficial for reducing the body’s response to arthritis and injury
and cardiovascular health.

In biochemical studies conducted at the Oriental Medicine Research Center of the
Kitasato Institute in Tokyo, Japan, polysaccharides have been identified in velvet antler
that tend to reduce the blood’s tendency to clot and to thin the blood. This effect
indicates that antler would contribute to improved circulation, decreased risk of stroke
and improved general cardiovascular health.

Japanese researchers have also investigated the effects of pantocrine on the recovery
of rats and rabbits from an induced whiplash-type injury. Pantocrine treatment
enhanced glycolysis in nervous tissue, an effect actually specific to neural tissue
(Takikawa et al. 1972 a,b). There is also support for such effects from double-blind
study in humans suffering from cervical injuries, where pantocrine treatment aided
recovery (Uelki et al. 1973).

Li and Wang (1990) cited Chinese studies showing that treatment of rats with a velvet
antler extract resulted in marked increases in the numbers of monocytes, suggesting
the presence of components that might affect the immune system. In New Zealand,
researchers have found that extracts from velvet antler have reduced tumor cell growth
(Suttie et al. 1994) and may in the future be utilized in the fight against cancer. Anti-
tumor activity of antler and antler fermented in Bacillus P-92 were demonstrated in
mice. Fermentation increases the amount of free amino acids, polypeptides and other
compounds that produce healthful effects. The survival rate of mice with tumors
increased from 25 to 40 percent. The neutrophil levels in the mice were increased 2 to
3 fold for antler and 3 to 4 fold for fermented antler. The higher levels of neutrophils
increased the body’s ability to resist injury and disease. Results suggest that
fermentation increases some of the health benefits of velvet antler.

Recently, Canadian researchers at the University of Alberta, have demonstrated that
the glycosaminoglycans in the water soluble fractions of velvet antlers have growth
promoting effects on cells (Sunwoo and Sim 1996). The researchers at the University of
Alberta observed a number of interim results from the consumption of velvet antler
extracts in addition to enhanced cell and whole animal growth including; anti–stress and
anti-inflammatory properties, increases in HDL (desirable) cholesterol and increases in
red blood cell counts (Sim et al. 1995a, Sim et al. 1995b, Sunwoo, 1988, Sunwoo et al.
1995, Sunwoo et al. 1997, Sunwoo and Sim 1996).

7. Scientific Explanation for Velvet Antler

Clearly the case for the pharmacological or bioactivity of velvet antler is very strong.
However, there is not yet a unifying hypothesis to explain the many and varied effects
of velvet antler in different animal species. The hypotensive effects have been
explained as at least partly due to the actions of choline compounds. Choline
compounds are not unique to velvet antlers. Other facets of biological activity ascribed
to velvet antler are not so easy to explain, although Wang et al. (1985 cited by Wang et
al. 1988) states that the anti–ulcer effects of velvet antler preparations is due to the
presence of various polysaccharides. Velvet antler likely contains peptide growth
factors (e.g. epidermal growth factor EGF), but concentrations would be low and would
the concentrations retain their biological activity through processing? In respect to
growth factors, however, EGF has been shown to replace estrogen in the stimulation of
female genital tract development, a phenomenon that raises fascinating questions
about the interrelationships between steroids and peptide growth factors. Steroids and
growth factors may survive processing but to date there has been no systematic
evaluations of the steroid composition of velvet antler published in the scientific
literature. However, it seems most unlikely that steroids present in the velvet antler
would be solely responsible for the observed androgenic effects. Rather compounds
present in the antler are inducing steroid synthesis in the treated animals, presumably
via effects on the hypothalamus or pituitary gland and then on the adrenal or testis.

Fulder (1980) proposed a general theory to explain the effects of these "antifatigue
substances" which include pantocrin, in that the biologically active components are
generally glycosides, where the active chemical groups are linked to sugar molecules.
Fulder proposes that the primary site of action of the glycosides is the hypothalamus
and the pituitary gland. The most commonly used glycoside in western medicine is
digitoxin, originally isolated from the plant commonly referred to as foxglove, which is
well known and has medically accepted and potent effects of the cardiac system. This
area of the glucoside/glycoside link is potentially very important and one where future
studies might provide more insight into the nature and efficacy of some of the
compounds present in many of the traditional medicines of the East.

8. Future Directions for Velvet Antler Research

A more scientific understanding of the bioactive components of velvet antler is
necessary to define that nature of the compounds and their effects in animal systems.
This is necessary to define the effects of drying and processing methods on bioactivity
and to maintain and improve product quality. It is also necessary in the search for new
bioactive compounds which may be unique to velvet antler and which could provide
new insights into the control of differentiation, growth and metabolism. One of the prime
objectives must be to develop in vitro systems to assay the bioactivity of velvet antler
preparations. This may be difficult in the sense that some of the reported effects of
velvet antlerwould appear to be dependent on an integrated whole animal system.
There is also the possibility that some of the effects are due to the synergistic effects of
two or more components present in the velvet antler. The whole area, though clearly
one of considerable complexity, is likely to be very rewarding.



9. Scientific References of Velvet Antler

Adams, J. L. 1979. Innervation and blood supply of the antler pedicle of the Red deer.
N Z Vet J. 27: 200-201.

Bae, D. S. 1977. Study on the effect of antler on growth of animals. III. Effect of antler
on the ability of spermatogenesis of cocks fertilization. Korean J Anim Sci 19: 407-412.

Banks, W. J. and J. W. Newberry. 1981 Light microscope studies of the ossification
proccess in developing antlers. In Antler Development in Cervidae. ed. R. D. Boone.
Caesar Kleberg Wildlife Research Institute. Kingsville Texas. pp 231-260.

Bubenik, G. A., Bubenik, A.B. 1986. Phylogeny and ontogeny of antlers and neuro-
endocrine regulation of the antler cycle - a review. Saeugetierk. Mitt. 33(2/3): 97-123.

Bubenik GA, Schams D, White RJ, Rowell J, Blake J, Bartos L Comp Biochem Physiol B
Biochem Mol Biol 1997 Feb;116(2):269-277 Seasonal levels of reproductive hormones
and their relationship to the antler cycle of male and female reindeer (Rangifer
tarandus). Department of Zoology, University of Guelph, Ontario, Canada.

Seasonal levels of LH, FSH, testosterone (T), estradiol, progesterone (P), and prolactin
(PRL) were determined in the plasma of five adult bulls, and five barren and four
pregnant cows of Alaskan reindeer (Rangifer tarandus), which were sampled every 3
weeks for 54 weeks. The male reproductive axis was sequentially activated; LH peaked
in May-June (2 ng/ml), FSH in June (51 ng/ml), and T in September (11.8 ng/ml). LH
levels in females reached a maximum in both groups at the end of August (the
beginning of the rut). Seasonal variation in FSH was minimal in pregnant cows, but
exhibited one elevation (41 ng/ml) in barren ones in November. T levels in cows
remained at barely detectable levels. The decrease of T values observed in both
groups in December and March was not significant. PRL peaked in May in cows (135
ng/ml pregnant, 140 ng/ml non-pregnant) and in June in bulls (92 ng/ml). Estradiol was
highest in bulls in the rut (August), in non-pregnant cows in January and in pregnant
cows in April, shortly before parturition. P levels in the pregnant cows rose from
September and peaked (9 ng/ml) shortly before parturition in April. In the non-pregnant
females P values increased and decreased several times before peaking (5 ng/ml) in
March. In the males, the variation of T and estradiol levels correlated relatively well with
the antler cycle but in the females the variation of neither estradiol, progesterone nor T
appeared to be related to mineralization or casting of antlers.

Breckhman, J. T., Y. L Dubryakov and A. L. Taneyeva. 1969. The biological activity of
the antlers of deer and other deer species. Ivestio Sibirskogo Ordelemia Akalemi Nank
SISR. Biological Series No. 10 (2):112-115

Breckhman J. T. 1980. Man and biologically active substances: The effects of drugs,
diet and pollution on health. Translated by J. H. Appleby. Pargamon Press, Oxford.

Chen X, Jia Y, Wang B Chung Kuo Chung Yao Tsa Chih 1992 Feb;17(2):107-110
Inhibitory effects of the extract of pilose antler on monoamine oxidase in aged mice.
[Article in Chinese] Academy of Traditional Chinese Medicine and Materia Medica, Jilin
Province, Changchun.

It was demonstrated that the water extract of Pilose Antler (WEPA) showed a higher
inhibitory effect on MAO-B activities in the liver and brain tissues of aged mice, but
nearly no effect on NAO-A. WEPA could significantly increase the contents of 5-HT, NE
and DA in the brain tissues of aged mice. In vitro experiments revealed that the
inhibition of WEPA on MAO-B was competitive, but on MAO-A was of mixed-type.

Elliott JL, Oldham JM, Ambler GR, Bass JJ, Spencer GS, Hodgkinson SC, Breier BH,
Gluckman PD, Suttie JM Endocrinology 1992 May;130(5):2513-2520 Presence of
insulin-like growth factor-I receptors and absence of growth hormone receptors in the
antler tip. Ruakura Agricultural Centre, Ministry of Agriculture and Fisheries, Hamilton,
New Zealand.

Red deer antler tips in the growing phase were removed 60 days after the
recommencement of growth for autoradiographical studies and RRAs. Sections were
incubated with radiolabeled GH or insulin-like growth factor-I (IGF-I), with or without
excess competing unlabeled hormones, and were analyzed autoradiographically. There
was negligible binding of [125I]GH in any histological zone of antler sections. [125I]IGF-I
showed highest specific binding in the chondroblast zone to a receptor demonstrating
binding characteristics of the type 1 IGF receptor. The lowest specific binding of [125I]
IGF-I was to prechondroblasts. RRAs on antler microsomal membrane preparations
RRAs on antler microsomal membrane preparations confirmed the absence of GH
receptors and the presence of type 1 IGF receptors found by autoradiography. These
findings suggest that IGF-I may act in an endocrine manner in antler growth through a
receptor resembling the type 1 IGF receptor. The presence of type 1 receptors in the
chondroblast zone implicates IGF-I involvement in cartilage formation through
matrixogenesis. There is no support for IGF-I having a major role in mitosis in the antler.

Elliott JL, Oldham JM, Ambler GR, Molan PC, Spencer GS, Hodgkinson SC, Breier BH,
Gluckman PD, Suttie JM, Bass JJ J Endocrinol 1993 Aug;138(2):233-242 Receptors for
insulin-like growth factor-II in the growing tip of the deer antler. Department of Biological
Sciences, University of Waikato, Hamilton, New Zealand.

Insulin-like growth factor-II (IGF-II) binding in the growing tip of the deer antler was
examined using autoradiographical studies, radioreceptor assays and affinity cross-
linking studies. Antler tips from red deer stags were removed 60 days after the
commencement of growth, and cryogenically cut into sections. Sections were incubated
with radiolabelled IGF-II, with or without an excess of competing unlabelled IGF-II and
analysed autoradiographically. Radiolabelled IGF-II showed high specific binding in the
reserve mesenchyme and perichondrium zones, which are tissues undergoing rapid
differentiation and cell division in the antler. Binding to all other structural zones was
low and significantly (P < 0.001) less than binding to the reserve
mesenchyme/perichondrium zones. Radioreceptor assays on antler microsomal
membrane preparations revealed that the IGF-II binding was to a relatively
homogeneous receptor population (Kd = 1.3 x 10(-10) mol/l) with characteristics that
were not entirely consistent with those normally attributed to the type 2 IGF receptor.
Tracer binding was partly displaceable by IGF-I and insulin at concentrations above 10
nmol/l. However, affinity cross-linking studies revealed a single band migrating at 220
kDa under non-reducing conditions, indicative of the type 2 IGF receptor. These results
indicate that, in antler tip tissues, IGF-II binds to sites which have different binding
patterns and properties from receptors binding IGF-I. This may have functional
significance as it appears that, whilst IGF-I has a role in matrix development of
cartilage, IGF-II may have a role in the most rapidly differentiating and proliferating
tissues of the antler.

Fennessy, P. F. and J. M. Suttie. 1985. Antler growth: Nutritional and endocrine factors.
In: Biology of Deer Production. Wellington, Royal Soc. NZ.

Fennessy, P F 1991 Velvet antler: the product and pharmacology. Proc. Deer Course
for Veterinarians (Deer Branch of the NZ Vet Assoc). 8 169-180

Feng JQ, Chen D, Esparza J, Harris MA, Mundy GR, Harris SE Biochim Biophys Acta
1995 Aug 22;1263(2):163-168 Deer antler tissue contains two types of bone
morphogenetic protein 4 mRNA transcripts. University of Texas Health Science Center
at San Antonio 78284-7877, USA.

Previously we isolated a bone morphogenetic protein 4 (BMP-4) cDNA from human
prostate cancer cells and found that the 5' noncoding exon 1 of this BMP-4 cDNA was
different from that of human bone cell BMP-4 cDNA. Recently we identified two
alternate exon 1s, 1A and 1B, for BMP-4 gene by reverse transcription-polymerase
chain reaction (RT-PCR) assays from fetal rat calvarial osteoblasts. In order to further
examine alternate exon 1 usage in the BMP-4 gene, we screened deer antler tissue
cDNA library. We isolated two types of cDNA clones encoding BMP-4 from this deer
antler cDNA library. Sequencing of these clones have revealed a single open reading
frame encoding a 408 amino acid protein. Comparison of 5' noncoding exon 1 portion
of these cDNA sequences with those of human bone and prostate BMP-4 cDNA
sequences and mouse BMP-4 genomic DNA sequence demonstrated that deer antler
tissue expresses both exon 1A and 1B containing BMP-4 mRNA transcripts. This
suggests that BMP-4 gene may contain alternate promoters or alternate splicing sites
in deer antler tissue.

Feng JQ, Chen D, Ghosh-Choudhury N, Esparza J, Mundy GR, Harris SE Biochim
Biophys Acta 1997 Jan 3;1350(1):47-52 Bone morphogenetic protein 2 transcripts in
rapidly developing deer antler tissue contain an extended 5' non-coding region arising
from a distal promoter. Department of Medicine, University of Texas Health Science
Center at San Antonio 78284, USA.

To understand the regulation of the BMP-2 gene expression, we recently isolated the
BMP-2 gene from a mouse genomic library and characterized the exon-intron structure
and promoter. RNase protection assay using poly (A)+ RNA of mouseosteoblasts
demonstrates that two regions in BMP-2 gene are protected by antisense mouse BMP-
2 RNA probes. These results demonstrate that BMP-2 gene utilizes two alternative
promoters, a distal and a proximal promoter. In the present study we demonstrate that
BMP-2 mRNA from rapidly growing deer antler tissue has an extended 5' non-coding
region compared with the human and rat BMP-2 mRNA. The extended 5' non-coding
region in the deer mRNA represents transcripts from the upstream distal promoter. This
is the first evidence of a natural BMP-2 mRNA from a bone-forming tissue that most
likely initiated from the distal transcription start site.

Fulder, S. 1980a. The hammer and the pesstle. New Scientist. 87 (1209): 120-123

Fulder, S. 1980b. The drug that builds Russians. New Scientist 87 (1215): 516-519.

Garcia RL, Sadighi M, Francis SM, Suttie JM, Fleming JS J Mol Endocrinol 1997 Oct;19
(2):173-182 Expression of neurotrophin-3 in the growing velvet antler of the red deer
Cervus elaphus. Department of Physiology and Centre for Gene Research, Otago
School of Medical Sciences, Dunedin, New Zealand.

Antlers are organs of bone which regenerate each year from the heads of male deer. In
addition to bone, support tissues such as nerves also regenerate. Nerves must grow at
up to 1 cm/day. The control of this rapid growth of nerves is unknown. We examined
the relative expression of neurotrophin-3 (NT-3) mRNA in the different tissues of the
growing antler tip and along the epidermal/dermal layer of the antler shaft of the red
deer Cervus elaphus, using semi-quantitative reverse transcription-polymerase chain
reaction. Expression in the tip was found to be highest in the epidermal/dermal layer
and lowest in the cartilaginous layer in all developmental stages examined. These data
correlate well with the density and pattern of innervation of these tissues. Along the
epidermal/dermal layer of the antler shaft, expression was highest in the segments
subjacent to the tip and lowest near the base, arguing for differences in the temporal
expression of NT-3 in these segments. The expression of NT-3 in cells isolated from the
different layers of 60-day antlers did not mirror that observed when whole tissues were
used and may suggest regional specificity of NT-3 expression within antler tissues.

Goss, R. J. 1983. Deer antlers. Regeneration, Function, and evolution. Academic Press
Inc., Orlando FL (ISBN 0-12-293080-0), 336p.

Goss RJ Anat Rec 1995 Mar;241(3):291-302 Future directions in antler research.
Division of Biology and Medicine, Brown University, Providence, Rhode Island 02912,
USA.

Through a series of interrogatories, unsolved problems of antler evolution, anatomy,
development, physiology, and pathology are probed, with commentaries, on the
following prospects for future research: 1. How could these improbable appendages
have evolved mechanisms to commit suicide, jettison the corpse, and regenerate new
ones every year? 2. By what developmental processes are antlers able to prescribe
their own morphogenesis with mirror image accuracy year after year and in some cases
produce deliberate asymmetries? 3. What causes the scalp to transform into velvet skin
as a deer's first antlers develop? 4. Why do healing pedicle stumps give rise to antler
buds instead of scar tissue? 5. How is the unprecedented rate of antler elongation
related to the diameter and length of the structure to be grown? 6. How come wound
healing by pedicle skin is held in abeyance for several months until new growth
resumes? 7. How is it that tropical deer regenerate antlers at any time of year, while in
temperate zones deer do so in seasonal unison? 8. How do deer find enough calcium
to make such massive antlers in only a few months? 9. What is the nature of the bizarre
tumors that some antlers grow following castration?

Gray, C. M., Taylor, M.L., Horton, M.A., Loudon, A.S.I., and Arnett, T.R. 1989. Studies
with cells derived from growing deer antler. J. Endocrinol. 123: 91.

Gray C, Hukkanen M, Konttinen YT, Terenghi G, Arnett TR, Jones SJ, Burnstock G,
Polak JM Neuroscience 1992 Oct;50(4):953-963 Rapid neural growth: calcitonin gene-
related peptide and substance P-containing nerves attain exceptional growth rates in
regenerating deer antler. Department of Anatomy and Developmental Biology,
University College, London, U.K.

Deer antler is a unique mineralized tissue which can produce very high growth rates of
> 1 cm/day in large species. On completion of antler growth, the dermal tissues which
cover the antler are shed and the underlying calcified tissue dies. After several months
the old antler is discarded and growth of a new one begins. It is known that deer antlers
are sensitive to touch and are innervated. The major aims of this study were to identify
and localize by immunohistochemical techniques the type of innervation present, and to
find out whether nerve fibres could exhibit growth rates comparable to those of antler.
We have taken tissue sections from the tip and shaft of growing Red deer (Cervus
elaphus) antlers at three stages of development; shortly after the initiation of regrowth,
the rapid growth phase, and near the end of growth. Incubation of tissue sections with
antisera to protein gene product 9.5 (a neural cytoplasmic protein), neurofilament
triplet proteins (a neural cytoskeletal protein), substance P and calcitonin gene-related
peptide (both of which are present in and synthesized by sensory neurons) showed the
presence of immunoreactive nerve fibres in dermal, deep connective and
perichondrial/periosteal tissues at all stages of antler growth. The sparse distribution of
vasoactive intestinal polypeptide-like immunoreactivity was found in dermal tissue only
at the earliest stage of antler development. Nerve fibres immunoreactive to
neuropeptide Y, C-flanking peptide of neuropeptide Y and tyrosine hydroxylase, all
present in postganglionic sympathetic nerves, were not observed at any stage of antler
growth. Nerves expressing immunoreactivity for any of the neural markers or peptides
employed could not be found in cartilage, osteoid or bone. These results show that
antlers are innervated mainly by sensory nerves and that nerves can attain the
exceptionally high growth rates found in regenerating antler.

Ha, H., S. H. Yoon, et al. 1990. Study for new hapatotropic agent from natural
resources. I. Effect of antler and old antler on liver injury induced by benzopyrene in
rats. Proc. Japanese Soc. Food & Nutrition 23: 9.

Han, S. H. 1970. Influence of antler (deer horn) on the enterochromaffin cells in the
gastrointestinal mucosa of rats exposed to starvation, heat, cold and electric shock. J.
Catholic Medical College 19: 157-164.

Hattori, M., X-W Yang, S. Kaneko, Y. Nomura & T. Namba. 1989. Constituents of the
pilose antler of Cervus nippon. Shoyakugaku Zasshi 43: 173-176.

Huang SL, Kakiuchi N, Hattori M, Namba T Chem Pharm Bull (Tokyo) 1991 Feb;39(2):
384-387 A new monitoring system of cultured myocardial cell motion: effect of pilose
antler extract and cardioactive agents on spontaneous beating of myocardial cell
sheets. Research Institute for Wakan-yaku (Traditional Sino-Japanese Medicines),
Toyama Medical and Pharmaceutical University.

Effects of various cardioactive agents and a water extract of the pilose antler of Cervus
nippon var. mantchuricus on periodic beating of cultured myocardial cell sheets were
examined by using an image analyzing system. Norepinephrine increased the beating
rate and the beating amplitude, whereas digoxin and forskolin enlarged only the
beating amplitude. Verapamil and propranolol decreased both the beating rate and the
beating amplitude. The water extract of the pilose antler showed no remarkable effects
in a standard medium (2.1 mM Ca2+). However, it significantly increased the beating
amplitude when the beating was suppressed by replacement with a low calcium medium
(0.5 mM Ca2+). A similar effect was found for 70% ethanol-soluble and -insoluble
fractions of the extract.

Ivankina NF, Isay SV, Busarova NG, Mischenko Tya Comp Biochem Physiol [B] 1993
Sep;106(1):159-162 Prostaglandin-like activity, fatty acid and phospholipid composition
of sika deer (Cervus nippon) antlers at different growth stages. State Medical Institute,
Blagoveschensk, Russia.

1. The alteration of lipid composition has been shown to take place at different stages
of antler growth. 2. The greatest amounts of phospholipids and polyunsaturated fatty
acids have been found during the most intense soft antler growth period. 3. The
bioregulators of lipid origin which are prostaglandins of A, B, E and F groups have
been found at the same stage.

Kang, W. S. 1970. Influence of antler (deer horn) on the mesenteric mast cells of rates
exposed to heat, cold or electric shock. J. Cathol. Med. College 19: 1-9.

Kaptchuk, T. and M. Croucher. 1987. The Healing Arts: Exploring the Medical Ways of
the World. New York, Summit Books.

Kim, Y. E., D. K. Lim, et al. 1977. Biochemical studies on antler (Cervus nippon
taiouanus) V: A study of glycolipids and phosholipids of antler velvet layer and
pantocrin. Korean Biochem. J. 10: 153-164.

Kim, K. W. and S. W. Park. 1982. A study of the hemopoietic action of deer horn
extract. Korean Biochem. J. 15: 151-157.

Kim, Y. E. and K. J. Kim. 1983. Biochemical studies on antler (Cervus nippon
taiouanus). VI. Comparative study on the effect of lipid soluble fractions of antler
spponge and velvet layers and pantocrin on the aldolase activity in the rat spinal
nerves. Yakhak Hoeji 27: 235-243.

Kim, K. B. and S. I. Lee. 1985. Effects of several kinds of antler upon endocrine
functions in rats. Kyung Hee Univ Med. J. ?8: 91-110.

Ko KM, Yip TT, Tsao SW, Kong YC, Fennessy P, Belew MC, Porath J Gen Comp
Endocrinol 1986 Sep;63(3):431-440 Epidermal growth factor from deer (Cervus
elaphus) submaxillary gland and velvet antler.

Epidermal growth factor (EGF)-like activity was isolated for the first time from the
submaxillary gland (SMG) and the velvet antler of red deer (Cervus elaphus) by a
combination of Sephadex gel or DEAE-Sephacel and IMAC columns in succession. The
semipurified cervine EGF-like activity (cEGF), with specific activity of 4.7 ng/micrograms
protein from the velvet tissues, can generate a completely parallel competitive binding
curve against mouse EGF in both radioreceptor assay (RRA) and radioimmunoassay
(RIA). Mitogenic activity of EGF from both tissues was demonstrated by stimulating the
incorporation of [3H]thymidine in two different cell lines of fibroblast culture in a dose-
dependent manner. The velvet layer may be the site of EGF synthesis outside the SMG.

Kong, Y., K. Ko, et al. 1987. Epidermal growth factor of the cervine velvet antler. Acta.
Zool. Sin., 33: 301-308:

Kaptchuck, T. and M. Creacher 1987. The healing arts: Exploring the medical ways of
the world. Summit Books, New York, 176 pages.

Lewis LK, Barrell GK Steroids 1994 Aug;59(8):490-492 Regional distribution of
estradiol receptors in growing antlers. Animal and Veterinary Sciences Group, Lincoln
University, Canterbury, New Zealand.

This study of estrogen receptors (ER) was carried out to confirm their presence and to
determine their localisation in antler bones. Partially grown antlers were amputated
from red deer (Cervus elaphus) stags, the skin removed, and samples taken of
periosteum, cartilaginous tissue including perichondrium, and bone. Capacity and
binding of free ER in the samples were calculated by Scatchard analysis of data
obtained from a radioreceptor assay which utilised [3H]estradiol as tracer. High affinity
ER (ka 1.3-3.4 x 10(10)/M) were detected in all tissues sampled with the exception of
bone. Receptor capacity ranged from 12-74 fmol/mg protein, ranking the tissues for
capacity in the following descending order: periosteum, cartilage, calcified cartilage.
These results demonstrate the presence of ER in growing antlers and indicate regional
localization of the receptors within these structures. The absence of ER in bone tissue
within the antler suggests that the effect of estradiol on stimulation of mineralization in
this tissue is indirect and must occur via its binding to the non-calcified tissues of
antlers, e.g., periosteum, perichondrium, and cartilage.

Li C, Waldrup KA, Corson ID, Littlejohn RP, Suttie JM J Exp Zool 1995 Aug 1;272(5):
345-355 Histogenesis of antlerogenic tissues cultivated in diffusion chambers in vivo in
red deer (Cervus elaphus). AgResearch, Invermay Agricultural Centre, Mosgiel, New
Zealand.

In a previous study we showed that formation of deer pedicle and first antler proceeded
through four ossification pattern change stages: intramembranous, transition, pedicle
endochondral, and antler endochondral. In the present study antlerogenic tissues
(antlerogenic periosteum, apical periosteum/perichondrium, and apical perichondrial of
pedicle and antler) taken from four developmental stages were cultivated in diffusion
chambers in vivo as autografts for 42-68 days. The results showed that all the
cultivated tissues without exception formed trabecular bone de novo, irrespective of
whether they were forming osseous, osseocartilaginous, or cartilaginous tissue at the
time of initial implant surgery; in two cases in the apical perichondria from antler group,
avascularized cartilage also formed. Therefore, the antlerogenic cells, like the
progenitor cells of somatic secondary type cartilage, have a tendency to differentiate
into osteoblasts and then form trabecular bone. Consequently, the differentiation
pathway whereby antlerogenic cells change from forming osteoblasts to forming
chondroblasts during pedicle formation is caused by extrinsic factors. Both oxygen
tension and mechanical pressure are postulated to be the factors that cause this
alteration of the differentiation pathway.

Marchenko LI, Kats MA Vrach Delo 1975 Aug;8:135-136 Anaphylactic shock as a
response to subcutaneous administration of pantocrine. Article in Russian

Miller SC, Bowman BM, Jee WS Bone 1995 Oct;17(4 Suppl):117S-123S Available
animal models of osteopenia--small and large. Division of Radiobiology, School of
Medicine, University of Utah, Salt Lake City 84112, USA.

Animal models of osteopenia are reviewed. Endocrine excess or deficiency conditions
include ovariectomy, orchidectomy, glucocorticoid excess and other endocrine states.
Seasonal and reproductive cycles are usually transient and include pregnancy and
lactation, egg-laying, antler formation and hibernation. Dietary conditions include
calcium deficiencies, phosphate excess and vitamin C and D deficiencies. Mechanical
usage effects include skeletal underloading models. Aging is also associated with
osteopenia in many species.

Muir, P. D., Sykes, A.R., Barrell, G.K. 1988. Changes in blood content and histology
during growth of antlers in red deer, Cervus elaphus, and their relationship to plasma
testosterone levels. J. Anat. 158: 31-42.:

Narimanov AA, Kuznetsova SM, Miakisheva SN Radiobiologiia 1990 Mar;30(2):170-174
The modifying action of the Japanese pagoda tree (Sophora japonica) and pantocrine
in radiation lesions. [Article in Russian]

A study was made of the effect of Sophora japonica and pantocrine on irradiated (2.5
Gy) human lymphoblastoid cells. The radioprotective effect was manifested with the
preparations injected separately after irradiation. The highest radioprotective effect
was produced by the mixture of the preparations, the injection 15 min after irradiation
being more effective than preinjection. The protective effect of the agents was studied
on mongrel mice after the administration thereof for the purposes of protection
protection-and-treatment and treatment. Sophora japonica and pantocrine were shown
to increase the survival rate of lethally exposed mice (LD90/30) when administered in a
combination 5-15 min before irradiation and when used for the purposes of protection--
and--treatment: 53.3% and 50% of animals, respectively, survived by day 30 following
irradiation. DMF was 1.25.

Price JS, Oyajobi BO, Nalin AM, Frazer A, Russell RG, Sandell LJ Dev Dyn 1996 Mar;
205(3):332-347 Chondrogenesis in the regenerating antler tip in red deer: expression
of collagen types I, IIA, IIB, and X demonstrated by in situ nucleic acid hybridization and
immunocytochemistry. Department of Human Metabolism and Clinical Biochemistry,
University of Sheffield Medical School, U.K.

The annual regrowth of antlers in male deer is a unique example of complete bone
regeneration occurring in an adult animal. Growth is initiated at the distal antler tip,
which is similar to the epiphyseal growth plate in some respects. However, there is
some debate as to whether this process represents "true" endochondral ossification.
As part of the characterization of the developmental process in pre-osseus antler
tissue, we have studied, by in situ hybridization, the spatial expression of mRNAs for
types I, II, and X collagen. Viewed in a coronal plane, type I procollagen mRNA was
observed in skin, the fibrous perichondrium, and the densely cellular area immediately
adjacent to the perichondrium. Below this area, as cells began to assume a columnar
arrangement and coincident with the appearance of a vasculature and synthesis of a
cartilaginous matrix, transcripts for types I, IIA, IIB procollagen and X collagen were
detected. Further down in the cartilage zone, the pattern of type I procollagen mRNA
expression was altered. Here, the signal was detected only in a morphologically distinct
subpopulation of small, flattened cells within the intercellular matrix at the periphery of
the columns of chondrocytes. The alternative splice form of type II procollagen mRNA
(IIA), characteristic of chondroprogenitor cells (Sandell et al. [1991] J. Cell Biol. 114:
1307-1319), was expressed by a subset of cells in the upper region of the columns,
indicating that this zone contains a population of prechondrocytic cells. Positive
hybridization to type IIA was most abundant in these cells. In contrast, transcripts for the
other procollagen splice form (IIB) and type X collagen were expressed by
chondrocytes throughout the whole of the cartilage region studied. The translation and
export of type II collagen and type X collagen were confirmed by detecting specific
immunoreactivity for each. The spatial distribution of immunoreactivity for collagen
types II and X was consistent with that of corresponding mRNAs. These data
demonstrate for the first time the distinct pattern of expression of genes for major
cartilage matrix macromolecules, the expression of the differentially spliced form of type
II procollagen mRNA (IIA), and specifically the co-localization of types II and X collagen
in the developing antler tip. Taken together, they strongly indicate that antler growth
involves an endochondral process.

Ramirez V, Brown RD Comp Biochem Physiol A 1988;89(2):279-281 A technique for
the in vitro incubation of deer antler tissue. Caesar Kleberg Wildlife Research Institute,
Texas A&I University, Kingsville 78363.

1. A procedure for the in vitro incubation of velvet deer antler tissue was developed.
Biopsy samples were collected in June with a trephine from 2 adult white-tailed deer
and incubated in modified BGJb medium up to 48 hr. Calcium (Ca) and hydroxyproline
(OH-proline) concentrations in the tissue were determined.

2. A significant increase (P less than 0.05) in Ca was exhibited at 4 and 8 hr of
incubation, and, after replenishment of media, at 48 hr.

3. Hydroxyproline concentrations continued to rise throughout the duration of the
incubation period and were significantly higher than controls (P less than 0.05) at 16,
24, and 48 hr. 4. Results suggest antler tissue can be incubated in vitro with the
protocol described, although length of incubation may vary with parameter measured.

Rucklidge GJ, Milne G, Bos KJ, Farquharson C, Robins SP Comp Biochem Physiol B
Biochem Mol Biol 1997 Oct;118(2):303-308 Deer antler does not represent a typical
endochondral growth system: immunoidentification of collagen type X but little collagen
type II in growing antler tissue. Rowett Research Institute, Bucksburn, Aberdeen, U.K.
gjr@rri.sari.ac.uk

The collagen isotypes present at early (6 week) and late (5 month) stages of growing
deer antler were isolated and identified. Pepsin-digested collagens were separated by
differential salt fractionation, SDS-PAGE and Western blotting and subsequently
identified by immunostaining. Cyanogen bromide digestion of antler tissue was used to
establish a collagen type-specific pattern of peptides, and these were also identified by
immunoblotting. Collagen type I was found to be the major collagen in both early- and
late-stage antler. Collagen type II was present in the young antler in small amounts but
was not confined to the soft "cartilaginous" tip of the antler. Collagen type XI was found
in the pepsin digest of the young antler, but collagen type IX was not present at either
stage of antler growth. Collagen type X was found in the young antler in all fractions
studied. Microscopic study showed that the deer antler did not possess a discrete
growth plate as found in endochondral bone growth. Unequivocal immunolocalization of
the different collagen types in the antler were unsuccessful. These results show that,
despite the presence in the antler of many cartilage collagens, growth does not occur
through a simple endochondral process.

Sadighi M, Haines SR, Skottner A, Harris AJ, Suttie JM J Endocrinol 1994 Dec;143(3):
461-469 AgResearch, Invermay Agricultural Centre, Mosgiel, New Zealand Effects of
insulin-like growth factor-I (IGF-I) and IGF-II on the growth of antler cells in vitro.

The effects of insulin-like growth factors -I and -II (IGF-I and -II) on the growth of
undifferentiated (fibroblast zone) cells from the growing tip of red deer velvet antlers
and from cells 1.5 cm distal to the growing tip (cartilage zone) were investigated in
primary cell culture. The addition of IGF-I or IGF-II to the medium of cultures
preincubated in serum-free medium for 24 h increased the rate of [3H]thymidine uptake
in a dose-dependent manner in both cell types, with maximal stimulation occurring when
1 nM-30 nM was added. The addition of IGF-II to the incubation medium containing IGF-
I did not cause a further increase in [3H]thymidine uptake in either cell type over and
above each growth factor alone, indicating that there were unlikely to be synergistic
effects of IGF-II on the mitogenicity of IGF-I. Binding studies were carried out using 3 x
10(5) fibroblast zone cells and cartilage zone cells after they had been incubated in
serum-free medium for 24 h. 125I-Labelled IGF-I (10(-9) M) in a final volume of 200
microliters was added to each culture and incubation carried out at 4 degrees C for a
further hour. 125I-Labelled IGF-I bound specifically to both fibroblasts and cartilage
zone cells; binding was displaced by both unlabelled IGF-I and by IGF-I antibody.

Sempere AJ, Grimberg R, Silve C, Tau C, Garabedian M Endocrinology 1989 Nov;125
(5):2312-2319 Evidence for extrarenal production of 1,25-dihydroxyvitamin during
physiological bone growth: in vivo and in vitro production by deer antler cells. Centre
d'Etudes Biologiques des Animaux Sauvages (CNRS), Beauvoir-sur-Niort, France.

The development of deer antler follows a pattern similar to that described for
mammalian endochondral ossification and has been proposed as a suitable model for
studies of bone growth. We investigated seasonal changes in the plasma
concentrations of 1,25-dihydroxyvitamin D [1,25-(OH)2D] and calcium and the activity
of alkaline phosphatase in relation to the antler cycle during 1 yr in 4 captive roe deer
and measured these biological parameters in 27 wild roe deer during their antler cycle.
A significant elevation of 1,25-(OH)2D in peripheral plasma, with no parallel increase in
the concentration of its precursor 25-hydroxyvitamin D, was observed to accompany
the rapid growth phase of the antler cycle in captive (P less than 0.001) and wild (P
less than 0.025) deer. During the same phase there was a gradient in levels of 1,25-
(OH)2D in antler vs. jugular blood (P less than 0.01). In addition, velvet cells in culture
proved to have the ability to convert 25-hydroxyvitamin D3 into a more polar derivative,
which was indistinguishable from true 1,25-(OH)2D3 with regard to its chromatographic
properties, its UV absorbance at 254 nm, and its ability to bind to the 1,25-(OH)2D3
receptors present in chick intestinal cytosol. These in vivo and in vitro results strongly
suggest that local production of 1,25-(OH)2D by the antler cells does occur in vivo and
may contribute to the increase in plasma 1,25-(OH)2D during bone growth.

Suttie, J. M., P. D. GLuckman, et al. 1985. Insulin like growth factor 1: antler stimulating
hormone? Endocrinol. 116: 846-848:

Suttie, J. M., P. F. Fennessy, et al. 1989. Pulsatile growth hormone, insulin-like growth
factors and antler development in red deer (Cervus elaphus scoticus) stags. J.
Endocrinol. 121: 351-360.

Suttie, J. M., P. F. Fennessy, et al. 1991. Antler growth in deer. Proc. Deer Course for
Veterinarians (Deer Branch, NZ Vet Assoc) 8: 155-168.

Suttie, J. M., I. D. Corson, et al. 1991. Insulin-like growth factor 1, growth and body
composition in red deer stags. Anim. Prod. 53: 237-242.

Sutti, J. M., Fennessy, P. F., Haines, S. R., Sadighi, M., Kerr, D.R. and Issacs, C. 1994.
The New Zealand velvet antler industry: Background and research findings.
International symposium on Cervi Parvum Cornu. KSP Proceedings. Oct. &, 1994.
Seoul, Korea, pp 86-135.

Sim, J. S., Sunwoo, H. H. and Hudson, R. J. 1995a. Cell growth promoting factors in
water-soluble fraction of Canadian elk (Cervus elaphus) antler. page 111, 1st
International Conference on East-West Perspectives on Functional Foods, Singapore,
September, 26-29, 1995.

Sim, J. S., Sunwoo, H. H., Hudson R. J. and Kurylo, S. L. 1995b. Chemical and
pharmacological characterization of Canadian elk (Cervus elaphus) antler extracts.
page 68, 3rd World congress of medicinal acupuncture and natural medicine,
Edmonton, Alberta, Canada, August 10-12-1995.

Sunwoo, H. H. Nakano, T. Hudson, R. J. and Sim, J. S. 1995. Chemical composition of
antlers from wapiti (Cervus elaphus). J. Agric. Food Chem. 43: 2846-2849.

Sunwoo, H. H. 1998. Isolation and characterization of proteoglycans in growing antlers
of wapiti (Cervus elaphus). Chapter 8 In Chemical characterization of growing antlers of
Wapiti (Cervus elaphus). Ph. D. thesis, University of Alberta.

Sunwoo, H. H., Nakano, T. and Sim, J. S. 1997. Effect of water soluble extract from
antlers of wapiti (Cervus elaphus) on the growth of fibroblasts. Can. J. Anim. Sci. 77:
343-345.

Sunwoo, H. H. and Sim, J. S. 1996. Chemical and pharmacological characterization of
Canadian elk (Cervus eoaphus) antler extracts. 96–World Federation Symposium of
Korean Scientists and Engineers Association, June 28 – July 4, 1996, Seoul Korea,
WFKSEA Prodeedings 96: 706-713.

Takikawa, K., N. Kokubu, et al. 1972. Studies on experimental whiplash injury. II.
Evaluation of Pantui extracts, Pantocrin as a remedy. Folia Pharmacol. Japon. 68: 473-
488. [Article in Japanese]

Takikawa, K., N. Kokubu, et al. 1972. Studies on experimental whiplash injury. III.
Changes in enzyme activiation of cervicxal cords and effect of Pantui extracts,
Pantocrin as a remedy. Folia Pharmacol Japon. 68: 489-493.

Wang, B. X., X. H. Zhao, et al. 1988. Effects of repeated administration of deer antler
extract on biochemical changes related to aging in senescence-accelerated mice.
Chem. Pharm. Bull. 36: 2593-2598.

Wang, B. X., X. H. Zhao, et al. 1988. Stimulating effect of deer antler extract on protein
synthesis in senescence-accelerated mice in vivo. Chem. Pharm. Bull. 36: 2593-2598.

Wang, B. X., X. H. Zhao, et al. 1988. Inhibition of liquid peroxidation bu deer antler
(Rokujo) extract in vivo and in vitro. J. Med. Pharm. Soc. for WAKAN-Yaku 5: 123-128.

Wang BX, Zhao XH, Qi SB, Yang XW, Kaneko S, Hattori M, Namba T, Nomura Y Chem
Pharm Bull (Tokyo) 1988 Jul;36(7):2593-2598 Stimulating effect of deer antler extract
on protein synthesis in senescence-accelerated mice in vivo.

Wang BX, Zhou QL Yao Hsueh Hsueh Pao 1991;26(9):714-720 Advances in the
chemical, pharmacological and clinical studies on pilose antler. [Article in Chinese]

Wang BX, Liu AJ, Cheng XJ, Wang QG, Wei GR, Cui JC Yao Hsueh Hsueh Pao 1985
May;20(5):321-325 Anti-ulcer action of the polysaccharides isolated from pilose antler.
[Article in Chinese]

Wang BX, Chen XG, Xu HB, Zhang W, Zhang J Yao Hsueh Hsueh Pao 1990;25(9):652-
657 Effect of polyamines isolated from pilose antler (PASPA) on RNA polymerase
activities in mouse liver. [Article in Chinese] Department of Pharmacology, Academy of
Traditional Chinese Medicine, Changchun.

The incorporations of [3H] leucine into protein and [3H] uridine into RNA in mouse liver
were increased when PASPA was given to mice at a dose of 30 mg/kg for 4 successive
days. The RNA polymerase activity, especially the RNA polymerase II activity in the
solubilized liver nuclear fraction of PASPA-treated mice was also increased. In vitro
experiment demonstrated that PASPA increased the RNA polymerase activity
significantly in mouse liver nuclei at a concentration of 1 microgram/ml. These results
suggest that the enhancement of RNA polymerase activities, particularly RNA
polymerase II activity, induced by PASPA treatment is responsible for the increase in
synthesis of protein and RNA in mouse liver tissue.

Wang BX, Chen XG, Zhang W Yao Hsueh Hsueh Pao 1990;25(5):321-325 Influence of
the active compounds isolated from pilose antler on syntheses of protein and RNA in
mouse liver. [Article in Chinese] Department of Pharmacology, Academy of Traditional
Chinese Medicine and Materia Medica of Jilin Province, Changchun.

The polyamines of pilose antler (PASPA) consist of putrescine (PU, 70.9%), spermidine
(SPD, 26.3%) and spermine (SP, 2.8%). The incorporations of [3H] leucine into protein
and [3H] uridine into RNA in mouse liver tissue were increased when PASPA was given
orally to mice at the dose of 30 mg/kg for 4 successive days. The incorporations of [3H]
leucine into liver protein and [3H] uridine into the cytosolic and nuclear RNA were also
increased by treatment with PU (21 mg/kg). In addition, the RNA polymerase activity in
the solubilized liver nuclear fraction of PU (21 mg/kg)-treated mice was increased. SPD
only promoted the synthesis of protein in mouse liver tissue at the dose of 8 mg/kg.
However, SP showed no effect on the synthesis of protein and RNA polymerase activity
under the used dose (1 mg/kg). The results suggest that PASPA is the main active
substance responsible for the promotion of the synthesis of protein and RNA in mouse
liver.

Yoon, P. 1989. The effect of deer horn on the experimental anemia of rabbits. Journal
Pharmaochemical Society Korea. 8: 6-11.

Yudin, A. M. and Y. L. Dubryakov 1974. A guide for the preparation and storage of
uncalcified male antlers as a medicinal raw material. In Reindeer antlers, Academy of
Sciences of the USSR. Far East Science Center. Vladivostock.

Zhao QC, Kiyohara H, Nagai T, Yamada H Carbohydr Res 1992 Jun 16;230(2):361-372
Structure of the complement-activating proteoglycan from the pilose antler of Cervus
nippon Temminck. Oriental Medicine Research Center, Kitasato Institute, Tokyo, Japan.

An anti-complementary polysaccharide, DWA-2, isolated from an unossified pilose
antler of C. nippon Temminck by digestion with pronase, gel filtration, and affinity
chromatography, consisted mainly of GalNAc, GlcA, IdoA, and sulfate in the molar
ratios 1.0:0.6:0.3:0.8, and small proportions of Man, Gal, GlcNAc, and protein (4.5%).
Methylation analysis, NMR spectroscopy, and degradation with enzymes indicated that
DWA-2 contained chondroitin sulfate A-, B-, and C-like moieties. DWA-2 showed potent
anti-complementary activity, and crossed immunoelectrophoresis indicated that it
cleaved complement C3 in the absence of Ca2+ ion. Digestion of DWA-2 with
chondroitinase ABC or ACI reduced the anti-complementary activity to a low level, but
digestion with chondroitinase B reduced the activity by approximately 40% and the
enzyme-resistant fraction still showed a significant activity.

Zhao D, Zhang X, Zhou F, Wei Z, Tian H Chung Kuo Chung Yao Tsa Chih 1990 Jan;15
(1):37-39 Relation of Fourier transform infrared spectroscopic characteristics of pilose
antler and its traditional quality grade. [Article in Chinese] Beijing Institute for Drug
Control.

The relationship between FTIR characteristics of Pilose Antler and its traditional quality
grade was studied and a rule governing its quality value "Z" was found. We have thus
advanced a new objective target for preparing Pilose Antler tablets and powder.

Zhang ZQ, Zhang Y, Wang BX, Zhou HO, Wang Y, Zhang H Yao Hsueh Hsueh Pao
1992;27(5):321-324 Purification and partial characterization of anti-inflammatory
peptide from pilose antler of Cervus nippon Temminck. Department of Pharmacology,
Academy of Traditional Chinese Medicine and Materia Medica of Jilin Province,
Changchun.

An anti-inflammatory compound was purified and isolated from pilose antler of Cervus
nippon Temminck by dialysis, gel filtration and ion-exchange chromatography
techniques. HPLC and N-terminal amion acid analysis identified the compound as a
homogeneous peptide. The peptide is composed of 68 amino acids and its molecular
weight as determined by amino analysis, is about 7200.

Zhiliaev EV, Dobriakov IuI Klin Med (Mosk) 1995;73(5):77-78 Experience in the use of
rantarine in the treatment of internal diseases. [Article in Russian]

Zioupos P, Wang XT, Currey JD J Biomech 1996 Aug;29(8):989-1002 Experimental and
theoretical quantification of the development of damage infatigue tests of bone and
antler. Department of Biology, University of York, U.K.

This study concerns the development of damage (as measured by a reduction in
elastic modulus) in two kinds of bones differing considerably in their degrees of
mineralisation: laminar bone from bovine femur and osteonal bone from red deer
antler. Antler bone is much tougher than 'ordinary' bone and its failure properties have
been investigated in: (i) monotonic tensile tests and (ii) creep rupture experiments.
Tensile fatigue is another way of examining how damage develops in bone. The
development of damage in the present fatigue tests was non-linear with the cycle
number, the degree of non-linearity was dependent on the level of stress and followed
a clearly different course for bone and antler. Antler was a more damage-tolerant
material, being able to achieve a reduction in the final modulus of elasticity, just prior to
failure, three times greater than ‘ordinary’ bone. The evolution of damage is quantified
by an empirical and a graphical method and by the use of Continuum Damage
Mechanics (CDM) expressions. The CDM method shows important conditions, found in
antler, but not in bone, that seen necessary for achieving stable fractures and
consequently producing very tough materials.