Evidence-Based Complementary and Alternative Medicine 5 Table 3: Eﬀect of maternal dietary supplementation of elk velvet antler on neuromotor coordination (standing) of rat pups. Dietary group Control EVA Control EVA Time spent standing† Supported (PND7–11) Unsupported (PND16–18) PND 7 0.0 (0.00) 3.4 (0.11)∗∗ 0.0 (0.00) 3.2 (0.12)∗∗ PND 8 4.4 (0.21) 4.2 (0.25) 4.3 (0.23) 4.5 (0.24) PND 9 7.4 (0.31) 8.1 (0.33)∗ 6.8 (0.27) 8.4 (0.40)∗ PND 10 Male 7.2 (0.26) 10 (0.45)∗ Female 7.3 (0.30) 9.8 (0.40)∗ PND 11 PND 16 PND 17 PND18 10.2 (0.65) 11.1 (0.67) 10.5 (0.53) 11.3 (0.56) 6.5 (0.33) 9.8 (0.44)∗ 6.8 (0.28) 10.2 (0.57)∗ 9.6 (0.46) 13.7 (0.61)∗ 8.9 (0.36) 13.3 (0.58)∗ 12.3 (0.55) 18.7 (0.77)∗ 13.6 (0.54) 19.1 (0.70)∗ † Time in seconds during a six-minute observation period. Each value represents the mean of 56 pups ± standard error of mean (SEM). The level of signiﬁcance was set at ∗P ≤ 0.05 or ∗∗P < 0.001. [23, 24]. Furthermore, we assessed potential gender diﬀer- ences as previous studies have noted diﬀerences according to gender in the in utero and/or postnatal development of progeny from dams undergoing experimental dietary or drug interventions [25–30]. Assessment of EVA eﬀects at this lifestage is relevant as EVA contains a number of bioactive substances including polyamines, chondroitin sulfate, minerals (calcium, phos- phorous, magnesium, iron, and potassium), glucosamine, glycosaminoglycans, and hyaluronic acid as well as all 24 amino acids [31, 32]. Furthermore, growing velvet antler contains neutrophin-3 [33] and a number of other growth factors and proto-oncogene mRNA [34]. Cultured antler cells respond to IGF-1 and IGF-2 [35] and display receptors for estradiol and IGF-1 [36]. Deer antler has been shown to contain biologically active molecules including antinar- cotic factor(s) [37], growth promoting polypeptides [38], diaphorase activity [39], and epidermal growth factor [40]. We observed no discernable adverse eﬀects of long-term maternal EVA supplementation on developmental indices of rat oﬀspring. Our results are consistent with Hemmings and Song [2] who reported that EVA supplemented chow had no long-term eﬀect on the oﬀspring of Fischer rats fed EVA chow from the 18th day of gestation to 88 days after birth. This particular study concentrated on the eﬀect of maternal EVA supplementation from the telophase of gestation throughout the lactation period [2]. Our study sug- gests that maternal supplementation prior to and throughout pregnancy causes no teratogenic or adverse outcomes on postnatal physical and neurological developmental parame- ters of oﬀspring. In our study, we used typical physical and neurological development parameters [41–44] to assess the eﬀects of long- term maternal EVA supplementation on postnatal develop- ment. The physical maturation parameters, pinna unfolding, eye opening, and anogenital hair are developmentally and hormonally controlled [45], but were not signiﬁcantly aﬀected in either gender by the dam’s consumption of EVA. On the other hand, the onset and completion of incisor eruption occurred one day earlier in oﬀspring from EVA-fed dams. Incisor eruption is inﬂuenced by epidermal growth factor and transforming growth factor, both of which are found in EVA [46, 47]. Our data suggests that maternal EVA supplementation may modulate the factors leading to incisor eruption in oﬀspring. In the rat, the ﬁrst 2 weeks after birth constitute a period of rapid brain growth, a critical phase in neurobehavioral maturation, which corresponds to the last months of human fetal brain growth [48]. Among the battery of tests used in the examination of neurological reﬂex development, right- ing reﬂex, negative geotaxis, cliﬀ avoidance, and acoustic startle were positively inﬂuenced by EVA supplementation. Furthermore, the number of pups acquiring these skills in the EVA group showed signiﬁcant improvement through the observation period compared to those in the control group. Although the earlier developing exploratory activities such as crawling, head waving [49], and grooming were not aﬀected by EVA, pups of EVA-supplemented dams were signiﬁcantly younger in completing more advanced skills and exhibiting more purposeful movements. For example, the righting reﬂex is thought to measure the development of dynamic postural adjustments that require the integrity of muscular and motor function and adequate acquisition of symmetrical coordination between left and right sides of the body [50]. Both male and female pups suckling dam receiving EVA diet showed earlier onset and completion of this reﬂex as compared with control pups. This result indicates that EVA maternal supplementation beneﬁts the development of dynamic postural adjustments and acquisition of sym- metrical coordination of the body and has a positive eﬀect on the development of these skills. It has been reported that there is a gender diﬀerence in neuronal maturation during brain development [51], but we did not observe a gender diﬀerence in any of the tests of neurological reﬂex development. Furthermore, our data revealed no diﬀerence between control and EVA-supplemented pups in the onset and completion of a battery of reﬂexes that assess maturation of sensory motor cortex and cerebellar and vestibular systems (namely, bar holding ability, vibrissa placing, and visual placing) [52, 53]. 5. Conclusions In summary, our study suggests that long-term maternal dietary consumption of EVA has beneﬁcial eﬀects on physical and neurological development of oﬀspring rats. The text on