Scientific Reference for Velvet Antler Research

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

Archer, R. H., and P J. Palfreyman. 1983. Properties of
New Zealand Deer Velvet, Part I: Search of the Literature
Vol I.Massey University and Wrightson NMA Ltd.

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.

Barnett, M.L.; D. Gombitchi; D.E. Trentham. A pilot trial of
oral type II collagen in the treatment of juvenile rheumatoid
arthritis. Arthritis & Rheumatism, 1996; 39 (4): 623-628.

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.

Conte, A.; M. de Bernardi; L. Palmieri; P Lualdi; G.
Mautone; G. Ronca. Metabolic fate of exogenous chondroitin
sulfate in man. Arzneim-Forsch./Drug Res 1991; 41(11):
76~77 I.

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.

Fisher, B.D.; M. Gilpin; D. Wiles. Strength training
parameters in Edmonton police recruits following
supplementation with elk velvet antler (EVA). University of
Alberta. I 998.

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.

Gerrard, D.F; G.G. Sleivert; A. Goulding; S.R. Haines: J.
M. Suttie. Clinical evaluation of New Zealand deer velvet
antler on muscle strength and endurance in healthy male
university athletes.

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.

Josephson, D. Concern raised about performance enhancing
drugs in the US. BMJ I 998;3 17:702 (12 September).

Kalden, J.R., and J. Sieper. Oral collagen in the treatment of
rheumatoid arthritis. Arthritis and Rheumatism, 1998; 41(2):
191-194.

Kamen, B. Red Deer Antler Velvet: Growth Hormone
Connection, and More! Health Sciences Institute. 1998; 2(8):
1-2.

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:

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.

Morreal, P; R. Manopulo; M. Galati; L. Boccanera; G.
Saponati; L. Bocchi. Comparison of the antiinflammatory
efficacy of chondroitin sulfate and diclofenac sodium in
patients with knee osteoarthritis. J Rheumatol 1996; 23:1
385-I 391.

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.

Palmieri, L.;A. Conte; L. Giovannini; P Lualdi; G. Ronca.
Metabolic fate of exogenous chondroitin sulfate in the
experimental animal. Arzneim-Forsch Drug Res I 990; 40
(l):319-323.

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.

Setnikar, I.; C. Giacchetti; G. Zanolo 1986.
Pharmacokinetics of glucosamine in dog and in
man.Arzneim.-Forsch. Drug Res; 36 (I): 729-735.

Setnikar, I.; R. Palumbo; S. Canali; G. Zanolo 1993.
Pharmacokinetics of glucosamine in man.Arzneim.-Forsch.
Drug Res; 43(l I):1109-1113.

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.

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.

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.

Trentham, D.E.; RA. Dynesius-Trentham; F.J. Orav; et al.
1993, Effects of oral administration of type II collagen on
rheumatoid arthritis. Science 261:1 727-1730.

Wang, B. 1996, Advances in research of chemistry,
pharmacology and clinical application of pilose antler.
Proceedings of the 1996 International Symposium on Deer
Science and Deer Products. I4-31.

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.

Yasui, N., and M.E. Nimni. 1998. Cartilage collagens. In:
Collagen, Volume I. M.E. Nimmi, ed. Boca Raton: CRC
Press. 225-24 I.

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.