Morphol. Neel - Scand. 8 (1970) 143 ~ 160 A New View on the Control of the Morphogenesis of the Skull J. VAN LIMBORGH Department of Anatomy and Embryology, University of Amsterdam Received for publication June 15, 1970 INTRODUCTION During the past few decades, there has been a veritable revival of the study of the skull. Whereas the older studies in this field usually concerned the solution of -volutionary, hereditary, racial and typological problems, the more recent studies fly related to the development and growth of the skull. These studies have considerably extended our knowledge of the fundamentals of skull biology. In sscordance with the results obtained, the ideas about the way in which the morphogenesis of the skull is controlled, are profoundly changing. It is this change of ideas which, here, will be discussed. This discussion will centre around three important problems facing us in respect of the morphogenesis of the skull. Since these problems derive directly from the morphogenetic phenomena we may observe, we will commence witha general survey of the normal development and growth of the skull. SKULL DEVELOPMENT The embryonic development of the skull starts comparatively late, when the primordia of all other structures of the head, such as the brain, the cerebral nerves, the eyes, the otic structures. the blood vessels and the muscle primordia, have already developed. But only then, between and around these structures, and also in the pharyngeal arches, condensations of mesenchyme appear in which we immedi- ately recognize the main contours of tie skull. The condensed mesenchyme in the area of the cranial base and in the pharyngeal arches, too, differentiates into cartilage. Thus, the cartilaginous skull primordium, the chondrocranium (Fig. 1) develops, which grows rapidly. During this process, the condensed mesenchyme is reduced to a thin layer, the perichondrium, that covers the surface of thé ciitilage. From the cartilages of the pharyngeal arches, which will be left out of discussion, develop mainly the’ ossicles of the ear, the hyoid bone and the laryngeal skeleton (Fig, 1), The cartilaginous base of the skull consists of a middle part, which joins in front with the nasal capsule and laterally with the otic capsules.144 J. VAN LIMBORGH Soon, in the basal cartilage the first centres of endochondral ossification appear the cartilage is destroyed and replaced by bone, These centres increase both in number and in size. Gradually, most of them merge, making the contours of the various bones and their parts visible. The cartilaginous areas left between the ossified parts, are the synchondroses. Almost simultaneously with the formation of the chondrocranium, begins the differentiation of the desmocranium: in the mesenchymal condensations of the calvaria and of the facial areas, centres of intramembranous ossification develop These centres, rather small in number, increase rapidly and soon take the shape o: the bones to be formed in these areas (Fig. 1). As the centres grow, both between and around them the quantity of condensed mesenchyme decreases. The narvow strips remaining between the bones, are the sutures; the membranous layer of mesenchyme covering the bones elsewhere forms the periosteum. froeta bane parietal bone [Px xs goae ‘oo maj of the sphonotd bone ner ofthe aphancid bone preryooid proces oral capisle erolle ps oe noxillo styloid proces male aygorotic Bone Machel comiage onaible -sorau mejus of the sphere mondibulor Hoarent ees ‘coma minus ofthe hyatd bone protd cantloge 1. Schematic drawing of the skull of a 12 weeks’ embryo, Dotted white and shade chondrocranium; white and dotted grey: desmocranium. SKULL GROWTH ‘The growth of the skull tissues in the foetal end postnatal periods of life is, in fat! aditect continuation of the embryonic developmental processes. In the chor cranium, in particular the cranial base, growth occurs as a result of growth of ! synchondroses, followed by endochondral ossification. Most of the synchondresANEW VIEW ON THE CONTROL OF THE MORPHC SIS OF THE SKULL 145 «ill present at the time of birth, close between the second and fourth year of life. The lest to ossify is the sphenooccipital synchondrosis which closes around the seventeenth yeur. By then. the growth of the cranial base has been completed. In 2 desmocranium, the calvaria and the facial skeleton, growth is due to continued ccuramembranous ossification: the growing periosteal and sutural tissues deposit zw bone on the pre-existing bone, so that each of the bones increases and the sutures decrease in size. These growth processes continue till about the twentieth year. Then, the periosteum stops growing and the sutures begin to ossify From these facts, itis clear that there are two ~ but no more than two — growth mechanisms, namely the endochondral and the intramembranous bone growth. During skull growth, however, there is not only considerable activity on the part of the boneforming cells, the osteoblasts, but also of the bonedestroying cells, the osteoclasts, as was recently demonstrated convincingly (Enlow, 1968). Thanks to the latter cells, pre-existing spaces in the skull, like the brain cavity and the orbits, grow with the other structures, and new cavities such as the paranasal sinuses n be formed. In addition. both the osteoblasts and the osteoclasts contribute to the modelling and remodelling processes, due to which the growing skull undergoes continual changes in shape and structure (Krogman, 1961). ‘There is in these changes no unifomnity, nor do they proceed simultaneously. The degree of bone deposition and bone resorption differs from place to place, and locally from time to time. too. A clear picture of the changes occurring, can be obtained by just considering the growth of the skull as a whole (Fig. 2). During foetal life, the neuroeranium (formed by the calvarja and the cranial base) grows much more rapidly than does the splanchnocranium, i.e. the facial skeleton, In the wsbom, the jaws especially’ are still relatively small. After birth, the neurocranium continues to grow rapidly till the fifth year; but then, growing becognes a very slow process indeed. The growth of the facial boties Proceeds postnatally more gradually, ut it continues till adulthood has been reached. As a result, from the Fifth year awards the size-ratio shifts in favour of the splanchnocranium, whilst — more- ser — this pare of the skull gradually occupies a position more directly underneath che neurocranium. Thus, chenges are taking place not only in size and shape, but slso in the topographié relationships. Ih spite of these changes, however, other observations indicate that the various skull parts grow in a,cq-ordinatéd manner. The original pattern of the skeleton is ‘ways maintained, and in broad outline the skull grows concentrically, the station- biological centre lying in the body of the sphenoid bone. Also, several angular ‘ons between various skull parts remain very constant. These observations show that the processes of endochondral and intramembran- ous bone growth are properly co-ordinated, but that at the same time many changes in the proportional relationships between the various skull parts occur. These changes, however, show a close correlation with the growth of the other structures of the head. For example, the phase af rapid growth of the neurocranium coincides with the period in which the brain develops rapidly, and the rater uniform growth the facial skeleton 2s a whole, correlates to a high degree with the gradual growth end the increasing use of the muscles of mastication. Apparantly, the growth146 J. VAN LIMBORGH a; @ 4FOETAL 5 FOETAL 6 FOETAL 8 FOETAL MONTHS MONTHS. MONTHS MONTHS. NEWBORN 2 YEARSA NEW VIEW ON THE CONTROL OF THE MORPHOGENESIS OF THE SKULL 147 ADULT 2.2. A scties of pho occurring during gr aphs illustrating the changes in size and shape of th th skull148 J.VAN LIMBORGH processes of the sku! are therefore not only co-ordinated with each other but also with the growth of the other stiflctures of the héad. PROBLEMS The three main problems concerning the control of the morphogenesis of the skull can now be formulated as follows: 1. Are there, in the embry onie phase, any causal relationships between the develop: ment of the skull on the one hand, and the presence of the primordia of the other head structures on the other; 2. How is, within.the skull once it is formed, co-ordination between the endochon- dral and the intramembranous bone growth brought about; and 3. In which way is co-ordination between the growth of the skull and that of the other structures realized? . Before trying to diswer these questions, the various categories of controlling factors which should be considered, will be defined. CONTROLLING FACTORS Among the categories of controlling factors to be distinguished are, firstly, the intrinsic genetic factors; ie. the genetic factors which are inherent to the: skull tissues themselves. Secondly, there is the category of epigenetic factors. These are genetically determined factors which manifest themselves by an indirect way, ¢ is by intervention or intermediary of influences effected by other structures. Characteristic of these influences is, that their effects will only be normal, if the structures from which they emanate are normal, too: every morphological or functional modification of these structures, whether spontaneous or experiment will result in a modification of their effects. The epigenetic factors may originate from adjacent structures and have a local influence ~ for exemple brain, eyes, and so on — or may be produced by distant structures and have®general influence - es, for example, the sex hormons. A final category is formed by the environments! factors in the true sense of the word: factors originating from the external environment. Also here local influences, such as local external pressure and muscle forces, and general influences (food and oxygen supply, etc.) may be distinguishe On the way the various controlling factors operate, at this moment only restricted data are available. With regard to the intrinsic genetic factors, we may suppose that they operate through intracellular regulatory mechanisms, in whic the RNA transcription is one of the most important steps. In respect of the gen factors, both epigenetic and environmental, we may assume that they affect metabolic processes of the cells, which they can either stimulate or inhibit. The nature of the local factors is still unknown, but recent studies seem to indicate # these factors, again both epigenetic and environmental, have — at least in part — th? character of electric fields that, according to their charge, will stimulate theA NEW VIEW ON THE CONTROL OF THE MORPHOGENE! SOF THE SKULL 149) activities of the osteoblasts or of the osteoclasts (Yasuda, 1954; Bassett and Becker, 1962; Becker, Bassett and Bachmann, 1964; Bassett, Pawluk and Becker, 1964; Bassett, 1968). CONTROL OF SKULL DEVELOPMENT Until a few years ago, there was a general conviction that the embryonic develop- ment of the skull is entirely controlled by intrinsic genetic factors (Fig. 3). PROCESS: CONTROL intrinsic genetic factors local epigenetic factors CRANIAL DIFFERENTIATION locel environmental factors general environmental factors Fig. 3, Diagram showing the hitherto generally accepted view on the control of embryonic skull differentiation, [At first sight, there seem to be few grounds for arguing this viewpoint, but closer study of the data now available does raise well-founded doubts. In fact, the most important consequence which genetic fixation of the characteristics of form and structure in the prospective skull tissues themselves should have, namely a high degree of individuality of the skull undef circumstances, is not materialized. Both the findings in spontaneous malformations as well as the results of many experi- ments carried out in normal embryos, prove beyond any doubt that there exist close relationships between the development of the skull and the presence and the condition of the primordia of the other head structures. Good examples of these relationships are those existing between eye and orbit. If there is no eye primor- dium, there will be no orbit. If there is only a single eye primordium, but a single orbit will develop. If two eye primordia lie close together, two contiguous orbits vill develop. Orbits only develop in their normal position, if the eye primordia, too, are normally located. If there is abnormal width between these primordia, the orbits, too, will develop with abnormal space in between. If there are three eye primordia, three orbits will develop. Around abnoanally enlarged eye primordia, enlarged orbits will develop, whilst around too small an eye primordium, too small an orbit will develop (Weiss and Amprino, 1940; Bellaires, 1955; Coulombre and Crelin, 1958;a.0.). These observations show that the development of orbits as such, as well as their number, position and size, entirely depend on the presence, the150 J. VAN LIMBORGH number, the position and the size of the eye primordia, There is nothing to indicate that orbits tend to differentiate independently. A similar absence of individual tendencies to differentiation has also been established for many other parts of the mesenchyme! embryonic skull primordiim (Schowing, 1961; a.o.). It is the adjacent other structures of the head which determine the presence, position and form of these skull parts. Accordingly, we may conclude that these adjacent structures exert powerful morphogenetic in- fluences; and since the development of the skull, normally, proceeds to a heredi tary, species-specific pattern, we may class these influences in the category of local epigenetic factors (van Limborgh, 1967, 1968, 1969), Other experiments (Benoit, 1956; a.0.) have shown, however, that in the condensed skull mesenchyme intrinsic genetic factors are not entirely lacking. But they are small in number and determine general characteristics only, such as the potency to differentiate into cartilage or bone and, probably, also the relationships between this potency of differentiation and time or age. In conclusion, the data now available force us to believe that the processes of skull differentiation are mainly controlled by local epigenetic factors, originating from adjacent other structures of the head, and furthermore by a few instrinsic genetic factors. An additional role of minor importance may, perhaps, be attributed to general epigenetic and environmental factors (Fig. 4). PROCESS CONTROL wv fectors” intrinsie genetic factors ZA locel epigen “_- = General epigenetic factors / CRANIAL DIFFERENTIATION J =e lecal environmental foctors ~~ general environmental factors.’ Fig. 4. Diagrarn showing the new view on the control of embryonic skull differentiation. ” CONTROL OF SKULL GROWTH With regard to the control of skull growth, modern literature advances three theories that are particularly relevant, viz. an oldge one —which,as fasas U know, for the lst time was formulated by Sicher in 1952 and the more recent theories of Scott and Moss respectively. According to the older theory (Fig. 5), the growth of the skull is largely controlled by intrinsic genetic factors. Only the modelling of the surface-con'i- guration of the bones and the internal structure of the bone tissue, too, would beANEW VIEW ON THE CONTROL. OF THE MORPHOGENESIS OF THE SKULL 151 PROCESS CONTROL - < Z “intrinsic genetic factors CHONDROCRANIAL . ‘GROWTH local epigenetic factors le ean |e DESMOCRANIAL local environmental factors GROWTH general epigenetic factors general environmental factors Fig. 5. Diagram showing the classic view on the control of skull growth. sabjeet to additional influences from local environmental factors, including muscu- iat forees. Aside from the last influences, the growth of the skull would therefore be independent, and the apparently existing correlation between the growth of the skull on the one hand and the growth of the other structures of the head on the other, would not be founded on any relationship of dependence but would be aconsequence of the fact that the growth of all the structures of the head is etically attuned to each other. Another characteristic feature of this theory is, iat 28 regards their significance for the control of the growth of the skull, equal is ascribed to all the osteogenic tissues, i.e. cartilage, sutures and periosteum. Scott (1953, 1956, 1958, 1962), however, assumes that intrinsic growth-con- srolling factors are only present in the cartilage and in the periosteum, claiming that owth of the sutures is secondary, ie. entirely dependent on extra-sutural in- PROCESS CONTROL < = ic genetic foctors | CHONDROCRANIAL é GROWTH 4 dj local epigenetic fectors J general epigenetic Factors suturel | DesMo- | SEN) | <—~—wsmmmemameaad loco! environmental foctors | CRANIAL | GROWTH a ia pesiutel general environmental factors grow Fig. 6. Diagram showing Scott"s view on the control of skull growth.152 J.VAN LIMBORGH fluences. In addition, the cartilaginous parts of the skull should be seen as centres of growth. According to this theory, the growth of the sutures is regulated on the one hand by the growth of the synchondroses and on the other hand by the growth of the head structures adjacent to the skull. In this way, the growth of the skull in itself would be properly co-ordinated and also keep in step with the growth of the other structures. Additionally, sutural growth may be modified by local environ- mental influences (Fig. 6), Moss and his co-workers (1959, 1960, 1961, 1962, 1964, 1966) have lately come up in strong defence of the theory that the growth of the skull is entirely secondary. This view is based on the theory of the functional cranial components developed by van der Klaauw (1941, 1946, 1948, 1951, 1952), according to which the skull is essentially composed of units, the size, shape and position of which are determined by their functions. Moss introduced the conception of “functions! matrix”, meaning those adjacent structures, to the presence and functions of which the cranial components as defined by van der Klaauw, are related. Moss claims that the growth of the various functional cranial components — whether they are formed by endochondral or intramembranous ossification is immaterial — is largely dependent on the growth and function of the functional matrices; that is, in ovr terminology, dependent on local epigenetic factors. A considerable additional in- fluence may be exerted by local environmental factors. On the other hand, Moss denies the existence of any intrinsic regulatory mechanism in the growing sk tissues (Fig. 7). ‘These three relevant theories prove to be at variance with each other on severs! points, and it seems therefore worthwhile to test them again on their value. We then find that the theory as formulated by Sicher (1952), according to which the growth of the skull would be highly independent, should be seriously question ed. Though it may be true that skull growth is to alarge extent genetically determined, this does by no means have to imply that the guiding genetic factors are inherent to the skull tissues themselves; these factors could operate indirectly. through local epigenetic factors, as is the case in embryonic development. Nor is the fact that comparative anatomical studies have shown that in vertebrates there aie PROCESS CONTROL Intrinste genetic factors CHONDROCRANIAL GROWTH general epigenetic factors DESMOCRANIAL ae local environmental Factors GROWTH general environmental factors locel epigenetic Factors Fig. 7. Diagram showing Moss’ view on the control of skull growth.