The consequences of living longer—Effects of an experimentally extended velvet antler phase on the histomorphology of antler bone in fallow deer (Dama dama)

Antlers are periodically regenerated paired cranial appendages of male deer (both sexes in reindeer) that constitute the fastest-growing bones in the animal kingdom. The annual antler cycle of male deer is linked to testicular activity and largely controlled by seasonal fluctuations of testosterone concentrations in their blood. We studied the effects of an experimental doubling (to eight months) of the velvet antler phase, during which the antlers are covered by skin (velvet), on the histomorphology of antler bone in three adult fallow bucks. Extension of the velvet antler phase in the experimental animals had been caused by administration of the antiandrogen cyproterone acetate (CPA). The distal portions of the antlers from two of the CPA-treated bucks exhibited partial sequestration of the antler cortex, with the separation plane typically located along the border between cortex and spongiosa. It is hypothesized that this was caused by cortical necrosis due to severe ischemia during later stages of the extended velvet antler phase. In places, new cancellous bone had been deposited on the resorption surface of the spongiosa, indicating a regeneration process. Normal fallow deer antlers (“controls”) from this and a previous study, that is, antlers with a timespan of about four months between onset of new antler growth and velvet shedding, exhibited no or only minor bone remodeling and still contained remnants of calcified cartilage in their distal portions. In contrast, the antlers of the three CPA-treated bucks showed evidence (secondary osteons and resorption cavities) of marked bone remodeling along their entire length and lacked remnants of calcified cartilage. Our results underscore that the typical histological features of antler bone reflect its short-lived nature. Antlers are not mechanically loaded during the velvet stage, and it is presently unclear what triggered remodeling activity in the antlers whose lifespan had been experimentally extended.     

The consequences of living longer—Effects of an experimentally extended velvet antler phase on the histomorphology of antler bone in fallow deer (Dama dama)

Antlers are periodically regenerated paired cranial appendages of male deer (both sexes in reindeer) that constitute the fastest‐growing bones in the animal kingdom. The annual antler cycle of male deer is linked to testicular activity and largely controlled by seasonal fluctuations of testosterone concentrations in their blood. We studied the effects of an experimental doubling (to eight months) of the velvet antler phase, during which the antlers are covered by skin (velvet), on the histomorphology of antler bone in three adult fallow bucks. Extension of the velvet antler phase in the experimental animals had been caused by administration of the antiandrogen cyproterone acetate (CPA). The distal portions of the antlers from two of the CPA‐treated bucks exhibited partial sequestration of the antler cortex, with the separation plane typically located along the border between cortex and spongiosa. It is hypothesized that this was caused by cortical necrosis due to severe ischemia during later stages of the extended velvet antler phase. In places, new cancellous bone had been deposited on the resorption surface of the spongiosa, indicating a regeneration process. Normal fallow deer antlers (“controls”) from this and a previous study, that is, antlers with a timespan of about four months between onset of new antler growth and velvet shedding, exhibited no or only minor bone remodeling and still contained remnants of calcified cartilage in their distal portions. In contrast, the antlers of the three CPA‐treated bucks showed evidence (secondary osteons and resorption cavities) of marked bone remodeling along their entire length and lacked remnants of calcified cartilage. Our results underscore that the typical histological features of antler bone reflect its short‐lived nature. Antlers are not mechanically loaded during the velvet stage, and it is presently unclear what triggered remodeling activity in the antlers whose lifespan had been experimentally extended.     

Structure and mineralisation density of antler and pedicle bone in red deer (Cervus elaphus L.) exposed to different levels of environmental fluoride: a quantitative backscattered electron imaging study

The structure and relative degree of mineralisation of antler and pedicle bone of yearling red deer stags exposed either to low or high levels of environmental fluoride were determined by digital quantitative backscattered electron (BSE) imaging. Bone fluoride content (BFC) in antlers (845±86 mg F^?/kg ash, arithmetic mean±S.E.M. ) and pedicles (1448±154 mg F^?/kg ash) of deer from a highly fluoride polluted area in North Bohemia (Czech Republic) were significantly higher (P < 0.001) than those of controls from uncontaminated regions in West Germany (antlers: 206±41, pedicles: 322±52 mg F^?/kg ash). Mean (56.5±4.5%) and maximum (84.9±2.1%) mineralised bone area of the control antlers significantly (P < 0.05 and P < 0.001, respectively) exceeded the corresponding values for the N. Bohemian deer (43.3±1.3 and 73.3±1.9%, respectively), while the pedicles from the 2 groups did not differ significantly. In the pooled antler samples (n = 18), negative correlations existed between BFC and mean (r_s = ?0.62, P < 0.01) as well as maximum (r_s = ?0.69, P < 0.01) mineralised bone area. Morphological imaging revealed a decreased width and an increased porosity of the antler cortex in the N. Bohemian specimens. Mean (148.5±1.7) and maximum (154.2±1.7) BSE‐signal intensities (= grey levels; range between a monobrominated (grey level 0) and a monoiodinated (grey level 255) dimethacrylate resin standard) of the antlers from the controls were significantly higher than those of the N. Bohemian deer (140.7±2.1 and 145.7±2.2, respectively; P < 0.05 for both comparisons). In the pooled antler samples, negative correlations between BFC and mean (r_s = ?0.51, P < 0.05) as well as maximum (r_s = ?0.52, P < 0.05) BSE‐signal intensities were observed. No significant differences in mineralisation density parameters were found for the 2 pedicle samples, and BFC and mineralisation density of the pooled pedicles were uncorrelated. Morphological imaging revealed bone mottling (denoting increased remodelling activity) and frequent occurrence of apparently increased osteocyte lacunae in some of the pedicles from the N. Bohemian deer. It is concluded that the reduced amount of mineralised bone in, and the lower mineralisation density of, the N. Bohemian antlers resulted from a fluoride induced disturbance of bone mineralisation. The rapid growth of antlers leads both to a high mineral demand and a high rate of fluoride uptake during antlerogenesis. This, and the limited lifespan of antlers, which does not allow for a compensation of a delay in the onset or progression of the mineralisation process, renders antler bone particularly susceptible to fluoride. Antlers are therefore considered a useful model for studying fluoride effects on bone formation. Furthermore, analysis of cast antlers enables a noninvasive monitoring of environmental pollution by fluorides.     

Hard fallow deer antler: A living bone till antler casting?

Deer antlers are the only mammalian bone structures which regenerate completely every year. Once developed, antlers are cleaned of the velvet‐like skin. Presently it is believed that due to velvet shedding the blood supply is interrupted in the solidifying antler bone. Histological examinations were made on different parts of fallow deer antlers investigated from the time of velvet shedding till the antler casting. The present study on hard (polished) antlers revealed living bone with regions presenting living osteocytes, active osteoblasts, osteoid seams and even early stages of trabecular microcallus formation, thus indicating to a continuous bone remodeling. A well developed vascular system was found despite the presence of hard antler bone. The pedicle bone exhibits a rich supply of capillaries and vessels connected to the spongy core of the main branch and the compact bone as well. There is evidence that hard fallow deer antlers possess a functioning vascular system that “keeps the antler moist” resulting in a high impact resistance when fights are most frequent. As late as 3 weeks prior to antler casting a large number of living cells were discovered within the antler core. As we have no doubt that parts of the polished fallow deer antler represent a living bone, we have concluded that a sufficient blood supply of the antler core is maintained almost till the time of antler casting by vessels passing through the antler base. Anat Rec 255:69–77, 1999. © 1999 Wiley‐Liss, Inc.     

Hard fallow deer antler: a living bone till antler casting?

Deer antlers are the only mammalian bone structures which regenerate completely every year. Once developed, antlers are cleaned of the velvet-like skin. Presently it is believed that due to velvet shedding the blood supply is interrupted in the solidifying antler bone. Histological examinations were made on different parts of fallow deer antlers investigated from the time of velvet shedding till the antler casting. The present study on hard (polished) antlers revealed living bone with regions presenting living osteocytes, active osteoblasts, osteoid seams and even early stages of trabecular microcallus formation, thus indicating to a continuous bone remodeling. A well developed vascular system was found despite the presence of hard antler bone. The pedicle bone exhibits a rich supply of capillaries and vessels connected to the spongy core of the main branch and the compact bone as well. There is evidence that hard fallow deer antlers possess a functioning vascular system that "keeps the antler moist" resulting in a high impact resistance when fights are most frequent. As late as 3 weeks prior to antler casting a large number of living cells were discovered within the antler core. As we have no doubt that parts of the polished fallow deer antler represent a living bone, we have concluded that a sufficient blood supply of the antler core is maintained almost till the time of antler casting by vessels passing through the antler base.     

Histological studies of bone formation during pedicle restoration and early antler regeneration in roe deer and fallow deer

The purpose of the present study was to examine the process of bone formation in the regenerating cranial appendages of roe deer (Capreolus capreolus) and fallow deer (Dama dama) during the early postcasting period. After the antlers are cast, osteoclastic and osteoblastic activities lead to a smoothing of the pedicle's separation surface, a strengthening of the pedicle bone, and a partial restoration of the distal pedicle portion that was lost along with the cast antler. Initially, bone formation occurs by intramembranous ossification, but early during the regeneration process cartilage is formed at the tips of the cranial appendages, and is subsequently replaced by bone in a process of endochodral ossification. Shortly after the antlers are cast, the cambium layer of the periosteum in the distal pedicle is markedly enlarged, which suggests that the periosteum serves as a cell source for the bone-forming tissue covering the exposed pedicle bone. The histological findings of our study are consistent with the view that the bony component of the regenerating cranial appendages of deer is largely derived from the pedicle periosteum. Based on findings in other bone systems, we speculate that stem cells that can undergo both osteogenic and chondrogenic differentiation are present in the pedicle periosteum. The early onset of chondrogenesis in the regeneration process is regarded as an adaptation to the necessity of producing a huge volume of bone within a short period. This parallels the situation in other cases of chondrogenesis in membrane bones.     

Deer antlerogenic periosteum: a piece of postnatally retained embryonic tissue?

This article reviews the research findings on the piece of periosteum overlying the lateral crest of prepubertal deer frontal bone, known as antlerogenic periosteum (AP). AP was initially discovered by Hartwig and Schrudde in 1974 when searching for the tissue that gives rise to antlers. In their experiment, when AP was transplanted elsewhere on the deer body it formed ectopic antlers. This clearly shows that AP possesses full self-differentiating ability, an attribute that can only be paralleled by embryonic tissue in mammals, like lateral plate mesoderm (LPM). Studies along this line by Goss in the 1980s further demonstrated that AP also holds the patterning information for antler formation. In the 1990s, our group carried out a series of studies on this unique tissue. The results showed that some of the critical features of AP resemble those of embryonic tissues, such as the astonishing growth potential in vivo and in vitro, and rich glycogen content. Histological observations and cell lineage tracing using a genetic marker convincingly demonstrate that pedicles and antlers are the derivatives of AP. Based on these findings, we advanced a hypothesis that AP is a piece of postnatally retained embryonic tissue. Morphological and histological examinations on the presumptive antler growth regions in deer prenatal life showed that the growth of primordial pedicles is initiated in the early pregnant stage (about 55 days) but then ceases (about 100 days) and is subsequently repressed at the late stage of pregnancy. The epidermis overlying the primordial pedicles resembles the apical ectoderm ridge (multicellular layer). These results strongly support our hypothesis. The results from the specific comparison between deer antler formation (from AP in postnatal) and mammalian limb development (from LPM in prenatal) showed that the ontogeny of antlers and limbs are comparable, and that deer antler has the same level of regulative properties as mammalian limbs. We believe that revealing the mechanism underlying the retention of embryonic tissue properties by AP until deer postnatal life will have important implications in biomedical research. Antler formation from AP offers an ideal model to work with in investigating how a self-differentiating system functions.     

Histological structure of antlers in castrated male fallow deer (Dama dama)

Antlers are periodically replaced cranial appendages that, except for the reindeer, are grown only by male deer. The annual antler cycle is controlled by seasonal fluctuations of sex steroid concentrations in the blood, and accordingly castration of male deer causes deviations from normal antler growth. The present study investigated antler histology of castrated fallow bucks (Dama dama). Castration in early spring was followed by casting of the hard antlers carried by the bucks and the growth of a new set of antlers, which remained in velvet permanently. In the following year, numerous bony protuberances developed from the original antler surface. Further growth of these protuberances, which were formed by subperiosteal intramembranous ossification, led to a marked increase in antler diameter in the affected areas. Compared to antlers of intact bucks, the antlers of the castrates showed histological signs of immaturity, suggestive of a reduced bone remodeling and an impairment of the mineralization process. These changes point to the dependence of the above processes on a stimulation by higher levels of sex steroids. Two years after castration, the antlers also developed integumental thickening and showed an initial formation of skin outgrowths. Cystic structures were present in the skin, which were often filled with a presumably sebaceous and/or keratinous material. Formation of intradermal bone or cartilage was not observed in the antlers of the castrated fallow bucks. The histological structure of the skin outgrowths suggested that they were caused by a hypertrophy of the dermal component of the velvet. Due to the localized bone overgrowth, resulting from the periosteal bone apposition onto the original antler surface, skin-lined infoldings originated, which reached deep into the newly formed bone. Our study revealed no indication of invasive/destructive bone growth in the antlers, i.e., of a penetration of the newly formed bone tissue into the pre-existing bone. The hypertrophic bone growth in the antlers of the castrates is compared with other forms of periosteally derived hypertrophic bone formation, including osteomas, in the mammalian skeleton. It is discussed whether the skin and bone outgrowths of the antlers of castrated fallow bucks may be classified as benign tumors.     

鹿茸再生及其蛋白质组学研究进展

背景：鹿茸是目前已知的惟一可以周期性再生的哺乳动物器官。这是一种来源于角柄骨膜并基于干细胞的再生过程。鹿茸又以角柄骨膜致敏区的干细胞作为再生基础,鹿茸及角柄骨膜中的蛋白质组,对于揭开鹿茸所具有的独特生物学活性以及再生机制具有重要的意义。 目的：综述鹿茸再生和目前鹿茸蛋白质组学研究的两种主要途径与研究现状等。 方法：应用计算机在PubMed数据库(http://www.ncbi.nlm.nih.gov/pubmed/)与CNKI数据库(http://www.cnki.net/)中进行检索。在PubMed数据库中的检索词为“deer antler,antler regeneration,antler proteome”;在CNKI数据库中的检索词为“鹿茸再生,鹿茸蛋白质组”。将涉及到鹿茸再生组织学与形态学,鹿茸干细胞以及鹿茸蛋白质组学研究相关的文章找出来,内容无关与重复的文章排除掉,最后纳入43条文献进行综述。 结果与结论：在PubMed与CNKI数据库中通过初检与筛选共找到了43篇文献。鹿茸是能够周期性再生的,并且通过组织学等相关实验表明这种再生是来源于角柄骨膜干细胞的,而角柄骨膜分为致敏区与休眠区,两者是通过与皮肤接触的紧密程度来划分的。正是致敏区骨膜与其所覆盖皮肤的相互作用才最终促使角柄骨膜干细胞发育成完整的鹿茸组织。同时,鹿茸及角柄骨膜中的蛋白质组在这些过程中起着重要作用。通过还原蛋白质谱的完整情况并进一步通过基因工程等后续手段来研究未知蛋白的功能对于了解鹿茸具有的独特生物学活性与再生作用奠定重要基础,同时对于相关蛋白质在哺乳动物器官再生中所具有的调节作用提供参考。 中国组织工程研究杂志出版内容重点：组织构建;骨细胞;软骨细胞;细胞培养;成纤维细胞;血管内皮细胞;骨质疏松;组织工程全文链接：     

Bone turnover associated with antler growth in red deer (Cervus elaphus)

Although it is known that skeletal bone depletion occurs during antler growth in deer, it is not clear whether repletion of the skeleton takes place before or after completion of antler development. This study attempted to correlate repeated scanning electron microscopic measures of ilium and rib bone porosity from six approximately 2‐monthly biopsy samples (using back‐scattered imaging) and biochemical markers of bone turnover (serum hydroxyproline and osteocalcin concentrations) taken for 11 months with antler growth in six red deer stags. No changes were detected in ilium samples but changes in porosity of rib bones and an elevation of the biochemical markers indicated that skeletal depletion occurred during the antler growth period. However, the decrease in rib bone porosity and decline in markers of bone turnover took place before completion of antler growth, indicating that a considerable amount of skeletal repletion could have occurred whilst antlers were also undergoing bone accretion. This latter finding extends the current view of antler growth being accompanied by a form of reversible osteoporosis in the skeleton by showing that there is a period when the antlers and skeleton are both undergoing net bone formation. Anat Rec 256:14–19, 1999. © 1999 Wiley‐Liss, Inc.     

Tissue differentiation and correlated changes in enzymatic activities during primary antler development in fallow deer (Dama dama)

Background: Deer antlers are useful models for studying bone growth and biomineralization in mammals. To achieve a better understanding of the mechanisms underlying the formation of primary cranial appendages in deer, the present study relates the histogenesis of primary antlers to changes in enzymatic (phosphatase) activities in the different tissue zones of this organ. Methods: The growing tips of the primary antlers (4.3 to 5 cm in length) were removed from five fallow bucks, aged about 10 months. Part of the material was processed for light microscopy. The other part was cryofixed, and the different histologically defined regions were analyzed for the activities of alkaline phosphatase (ALP) and tartrate-resistant acid phosphatase (TRAP) as well as for the concentrations of inorganic and organic phosphate. Results and Conclusions: Histologically, the primary antler could in distoproximal direction be divided into eight different zones (dermis; perichondrium; zones of cartilage formation, hypertrophy, mineralization, and degeneration; primary spongiosa; secondary spongiosa). The histological results demonstrate that the elongation of the primary antler proceeded through a modified form of endochondral ossification, resembling that seen during formation of pedicles and secondary antlers. The concentrations of the extractable activities of ALP and TRAP progressively increased from the perichondrium to the zone of cartilage mineralization. Thus, highest activity of TRAP during primary antler formation occurred at an earlier stage of tissue differentiation than in somatic endochondral ossification, where the enzyme is a biochemical marker of osteoclastic activity during bone remodeling. The present results might reflect the presence of osteoclastic precursor cells in the zone of cartilage mineralization as an adaptation to the rapidity of antler growth. Our findings of the contents of extractable ALP, inorganic and organic phosphate in the different tissue zones of the developing primary antler are in good agreement with previous studies analyzing epiphyseal growth plates and point to the fact that ALP causes a rise in inorganic phosphate and the removal of inhibitors for mineralization, like pyrophosphate. © 1995 Wiley-Liss, Inc.     

基于干细胞的器官再生研究模型-鹿茸

背景：鹿茸是惟一能够周期性再生的复杂哺乳动物器官,其再生过程是基于干细胞的存在。研究鹿茸再生机制,探索干细胞在哺乳动物器官再生中的作用对于再生生物学和再生医学研究具有重要的意义。 目的：综述鹿茸再生研究,干细胞及相关因子在鹿茸再生中的作用。 方法：应用计算机检索1994年1月至2012年10月PubMed 数据库(http://www.ncbi.nlm.nih.gov/ PubMed)。检索词为：deer antler;antler regeneration; stem cell,并限定文章语言种类为English。此外还手动查阅相关专著数部。纳入文章所述内容涉及鹿茸再生的组织学、形态学、鹿茸干细胞与微环境研究、相关细胞因子。排除重复研究和纳入标准无关的文章。 结果与结论：共检索文献87篇,最终纳入文献36篇。决定鹿茸发生及再生的关键组织分别为生茸区骨膜和角柄骨膜,这两种组织中的细胞被定义为鹿茸干细胞。鹿茸干细胞上覆盖的皮肤组织构成了这些干细胞活动所需的特定微环境。多种细胞因子如胰岛素样生长因子、性激素、人表皮生长因子、血管内皮生长因子等参与了鹿茸再生及快速生长调控。探索鹿茸干细胞微环境内各组分间相互作用所需的信号因子、阐明其调控机制,对于揭示解鹿茸再生之谜,对了解干细胞在哺乳动物器官再生的作用具有十分重要的意义。     

Future directions in antler research

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? © 1995 Wiley-Liss, Inc.

     

Deer antlers: a zoological curiosity or the key to understanding organ regeneration in mammals?

Many organisms are able to regenerate lost or damaged body parts that are structural and functional replicates of the original. Eventually these become fully integrated into pre-existing tissues. However, with the exception of deer, mammals have lost this ability. Each spring deer shed antlers that were used for fighting and display during the previous mating season. Their loss is triggered by a fall in circulating testosterone levels, a hormonal change that is linked to an increase in day length. A complex 'blastema-like' structure or 'antler-bud' then forms; however, unlike the regenerative process in the newt, most evidence (albeit indirect) suggests that this does not involve reversal of the differentiated state but is stem cell based. The subsequent re-growth of antlers during the spring and summer months is spectacular and represents one of the fastest rates of organogenesis in the animal kingdom. Longitudinal growth involves endochondral ossification in the tip of each antler branch and bone growth around the antler shaft is by intramembranous ossification. As androgen concentrations rise in late summer, longitudinal growth stops, the skin (velvet) covering the antler is lost and antlers are 'polished' in preparation for the mating season. Although the timing of the antler growth cycle is clearly closely linked to circulating testosterone, oestrogen may be a key cellular regulator, as it is in the skeleton of other male mammals. We still know very little about the molecular machinery required for antler regeneration, although there is evidence that developmental signalling pathways with pleiotropic functions are important and that novel 'antler-specific' molecules may not exist. Identifying these pathways and factors, deciphering their interactions and how they are regulated by environmental cues could have an important impact on human health if this knowledge is applied to the engineering of new human tissues and organs.     

Delayed ectopic antler growth and formation of a double-head antler in the metacarpal region of a fallow buck (Dama dama L.) following transplantation of antlerogenic periosteum.

Autologous transplantation of antlerogenic periosteum to the metacarpal region of a fallow buck resulted in the formation of an ectopic pedicle within a few months after transplantation. In the following years the pedicle increased in size, but no ectopic antler development occurred. However, in the ninth year after transplantation a first antler of about 3 cm length grew from the ectopic pedicle. The hard ectopic first antler was not cast, and renewed antler growth, starting in late spring of the following year, led to the formation of a larger subsequent antler of 7.3 cm length. The latter exhibited a distal flattening (palmation) and a rudimentary basal coronet, thereby displaying morphological features typical of the subsequent antlers of fallow bucks. The results of the experiment indicate (1) that transformation from pedicle to antler growth can occur several years after pedicle initiation, when the pedicle has reached a certain threshold size, (2) that species and age specific features of antler morphology tend to be realized even in ectopic locations, and (3) that primary and subsequent antler growth was apparently triggered by cells of the pedicle periosteum, being derived from the transplanted antlerogenic periosteum. The findings thereby emphasize the importance of local factors in the control of pedicle/first antler transformation and antler morphogenesis.     

Bone turnover associated with antler growth in red deer (Cervus elaphus).

Although it is known that skeletal bone depletion occurs during antler growth in deer, it is not clear whether repletion of the skeleton takes place before or after completion of antler development. This study attempted to correlate repeated scanning electron microscopic measures of ilium and rib bone porosity from six approximately 2-monthly biopsy samples (using back-scattered imaging) and biochemical markers of bone turnover (serum hydroxyproline and osteocalcin concentrations) taken for 11 months with antler growth in six red deer stags. No changes were detected in ilium samples but changes in porosity of rib bones and an elevation of the biochemical markers indicated that skeletal depletion occurred during the antler growth period. However, the decrease in rib bone porosity and decline in markers of bone turnover took place before completion of antler growth, indicating that a considerable amount of skeletal repletion could have occurred whilst antlers were also undergoing bone accretion. This latter finding extends the current view of antler growth being accompanied by a form of reversible osteoporosis in the skeleton by showing that there is a period when the antlers and skeleton are both undergoing net bone formation.     

Antler Transformation is Advanced by Inversion of Antlerogenic Periosteum Implants in Sika Deer (Cervus nippon)

Deer antlers offer a unique model for the study of tissue‐specific stem cells and organogenesis, as antler stem cells are confined to the antlerogenic periosteum (AP), a tissue that can be readily located (overlying a frontal crest) and experimentally manipulated. AP consists of an upper fibrous layer and a lower cellular layer. Tissue transplantation and membrane insertion experiments demonstrated that antler formation is triggered by the interactions between AP and the overlying skin. Interestingly, fairly normal antlers can be induced to grow by an inverted AP implant (the AP cellular layer facing the skin) at an ectopic site, raising the question whether the initial inductive signal is derived from the fibrous layer or cellular layer or both. To answer this question, in this study we used eight sika deer stag calves and selected one side of future antler growth region for implanting inverted AP and the contralateral side for noninverted AP as the control. The results showed that implantation of the AP discs in an inverted orientation generated pedicles with final height (17 ± 5.1 mm), less than half the height of those formed from the noninverted AP implants (45 ± 11.7 mm). Critically, antler transformation was initiated from a shorter pedicle, which was formed from the region where the AP cellular layer was brought in close proximity to the overlying skin. Therefore, the AP cellular layer, as opposed to the AP fibrous layer, is likely to be the main source of the initial inductive molecules for antlerogenesis. Anat Rec 293:1787–1796, 2010. © 2010 Wiley‐Liss, Inc.     

Morphometric analysis of osteonal architecture in bones from healthy young human male subjects using scanning electron microscopy

The shape and structure of bones is a topic that has been studied for a long time by morphologists and biologists with the goal of explaining the laws governing their development, aging and pathology. The osteonal architecture of tibial and femoral mid‐diaphyses was examined morphometrically with scanning electron microscopy in four healthy young male subjects. In transverse sections of the mid‐diaphysis, the total area of the anterior, posterior, lateral and medial cortex sectors was measured and analysed for osteonal parameters including osteon number and density, osteon total and bone area and vascular space area. Osteons were grouped into four classes including cutting heads (A), transversely cut osteons (B), longitudinally cut osteons (C) and sealed osteons (D). The morphometric parameters were compared between the inner (endosteal) and outer (periosteal) half of the cortex. Of 5927 examined osteons, 24.4% cutting heads, 71.1% transversely cut osteons, 2.3% longitudinally cut osteons and 2.2% sealed osteons were found. The interosteonic bone (measured as the area in a lamellar system that has lost contact with its own central canal) corresponded to 51.2% of the endosteal and 52.4% of the periosteal half‐cortex. The mean number of class A cutting heads and class B osteons was significantly higher in the periosteal than in the endosteal half‐cortex (P < 0.001 and P < 0.05, respectively), whereas there was no significant difference in density. The mean osteon total area, osteon bone area and vascular space area of both classes A and B were significantly higher (P < 0.001 for all three parameters) in the endosteal than in the periosteal half‐cortex. The significant differences between the two layers of the cortex suggest that the osteoclast activity is distributed throughout the whole cortical thickness, with more numerous excavations in the external layer, but larger resorption lacunae closer to the marrow canal. A randomly selected population of 109 intact class B osteons was examined at higher magnification (350×) to count osteocyte lacuna and to analyse their relationship with osteon size parameters. The distribution frequency of the mean number of osteocyte lacunae increased with the increment in the sub‐classes of osteon bone area, whereas the density did not show significant differences. The number of osteocyte lacunae had a direct correlation with the osteon bone area and the mean osteon wall thickness, as well as the mean number of lamellae. The osteocyte lacunae density showed an inverse relationship. These data suggest a biological regulation of osteoblast activity with a limit to the volume of matrix produced by each cell and proportionality with the number of available cells in the space of the cutting cone (total osteon area). The collected data can be useful as a set of control parameters in healthy human bone for studies on bone aging and metabolic bone diseases.