Chapter One

1. Introduction:
This study explored changes that occurred to palatal rugae area during orthodontic treatments. They are considered to be irregular, asymmetric ridges of the mucous membrane of hard palate. Extending laterally from the incisive papilla and the anterior part of the median palatal raphe (Sumathi et al, 2012;Anukool et al,2011; Manashvini et al, 2008;Standring, 2008 and Baily et al,1996). According to the Glossary of Prosthodontic Terms, rugae are anatomical folds or wrinkles (usually used in the plural sense); the irregular fibrous connective tissue located on the anterior third of the palate. They are also called plica palatinae or rugae palatine.(Saraf et al, 2011). It is assumed that the rugae facilitate food transport through the oral cavity, prevent loss of food from the mouth, and participate in food crushing. Because of the presence of tactile and gustatory receptors, rugae contribute to perception of taste, mechanical food qualities, and tongue position(Bhullar et al, 2011). Researchers have always shown interest in the evolutionary development of the folds of tissue found in the roof of the human mouththe palatine rugae (Anukool et al, 2011). Winslow (1955) seems to have been the first to describe them, and the earliest illustration of them is probably by Santorini in 1775. After that a lot of researchers considered palatal rugae or plica palatine in their studies and researches to get deeper understanding of their importance of their presence in mammals and most specifically in humans.

Palatal rugae patterns have been studied extensively in multivariate populations. Many of the previous studies investigated gender-related and ethnicity-related differences in palatal rugae patterns (Kapali et al, 1997). Most importantly, some of the previous studies explored the individuality of palatal rugae patterns ( Shukla et al,2011;Saraf et al,2011). The findings of such studies suggested that palatal rugae are highly individual and can be used as a tool in individual identification. However, some events can contribute to changes in rugae pattern, including trauma, extreme finger sucking in infancy and persistent pressure with orthodontic treatment and dentures. (Eboh, 2012; Sassouni, 1957). The current study is the first to shed the light on morphometric changes occurred due to orthodontic treatment including different criteria such as segmentation, unification as expression for the change in number, change in orientation, change in shape, change in length, change in volume, displacement of lateral end (in anteroposterior and mediolateral direction) and displacement in medial end (in anterioposterior and mediolateral direction). Furthermore, the percentages of casts that had every one of these changes will be counted. In addition, the other goal of this study is to further test the individuality of palatal rugae and their value as a tool in individual identification. This goal is achieved by analyzing the morphometric changes that take place in palatal rugae patterns as a result of orthodontic treatment. Moreover, the researcher examines whether such changes will compromise our ability to match the same palatal rugae pattern before and after orthodontic treatment. The outcome of the study provides more knowledge about the stability of palatal rugae against orthodontic treatment. It will tell us whether orthodontic treatment will affect the use of palatal

rugae in human identification. It will test the use of palatine rugae patterns as an alternative fingerprint in the Jordanian population. This theses is divided in to six chapters, where chapter one is an introduction for the subject, talking about development of palate and palatal rugae anatomically and histologically, the second chapter will talk about previous studies were done on this field of study. The third chapter will contain information about our methodology, materials and sample collection. The fourth chapter will present the results of the study. Chapter five contains the discussion of the results. The last chapter, which is chapter six, includes recommendations and suggestions, and conclusion.

1.1 Development of the Palate and Palatal Rugae.

The palate is the structure that separates the oral from nasal structures. It has two key stages of development during embryonic (primary), and an early fetal (secondary) involving the fusion of structures and a key epithelial to mesenchymal transition in a process known as Palatogenesis. The process begins in the sixth week but is not completed until the 12th week. The critical period of development of the palate is from the end of the sixth week until the beginning of the ninth week.(Bailey and Miller, 1921).

It is formed by the fusion of both right and left maxillary processes and medial nasal process and composed of two main parts: -The Primary Palate It is formed by two parts formed by the merging of two nasal processes. Early in the sixth week, it begins to develop from the deep part of the intermaxillary segment of the maxilla. Initially, this segment is a wedge-shaped mass of mesenchyme between the internal surfaces of the maxillary prominences of the developing maxillae. It represents only a small part of the adult hard palate (the part anterior to the incisive fossa) where Bone gradually develops by intramembranous ossification in the primary palate, forming the premaxillary part of the maxilla, which lodges the incisor teeth). - The Secondary Palate It is divided in two anatomical parts: a-anterior hard palate - ossified (contributions from the maxilla and palatine bones), b-posterior soft palate - muscular. It is formed by the 7th to 8th weeks

of development, its form from two mesenchymal projections that extend from the internal aspects of the maxillary prominences. The growth of maxillary processes from inside gives rise to lateral palatine processes. It will grow medially and fuse with each other with the inferior border of the nasal septum, and then it can be classified as hard palate. The medial epithelial seam at the edges of the palatine shelves breaks down, allowing for the fusion of the palatine shelves. The remaining part is invaded by muscle and is called the soft palate. The fusion between the nasal septum and the palatine processes begins anteriorly during the ninth week and is completed posteriorly by the 12th week, superior to the primordium of the hard palate. Concurrently, bone extends from the maxillae and palatine bones into the lateral palatine processes to form the hard palate. The median palatine raphe indicates the line of fusion of the lateral palatine processes and terminates anteriorly at an oval prominence (Paul et al, 1997), the incisive papilla, which is located 2 cm from the central incisors (Mustafa et al, 2012; Hoggan and Sadowsky, 2001; Morlang,1982). In human embryos, rugae are relatively prominent and occupy most of the length of the palatal shelves at the time of their elevation (Bhullar et al, 2011). The palatal rugae appear toward the 3rd month of intrauterine life (Gandicota et al, 2012; Amaski et al, 2008), distinguished in human embryos of 32-mm Crown-rump length (CRL) next to the incisive papilla (Gandikota et al,2012). At the 550 mm stage of embryonic development, there are five to seven rather symmetrically disposed ridges, with the anterior ones beginning at the raphe, the others more laterally. Towards the end of intra-uterine life, the pattern of rugae becomes less regular, posterior ones disappearing and those anterior become considerably more pronounced and compressed (Bhullar et al, 2011).

1.2 Anatomy and Histology of Palate and Palatal Rugae Area

In the hard palate, anteroposteriorly a thin central groove is bordered on each side by a crest, the palatal raphae. From this crest, laterally three to seven smaller crests emerge on each side. These crests are called palatal rugae (Indira et al,2012).These rugae are just behind the incisive papilla and considered to be rigid and tangentially radiating out(Hemanth et al, 2010). Palate is known as an extremely sensitive region in the oral mucosa and is involved, along with the tongue, in food ingestion and mastication (Luke ,1988; Sakamoto et al. 1989). In the palatal mucosa, a variety of mechanoreceptors are distributed site-specifically, such as free nerve endings, simple corpuscles, Ruffini corpuscles, and Merkel cellneurite complexes. In particular, Merkel cellneurite complexes are known to be densely distributed in the epithelial ridges of the palatal mucosa (palatine rugae).Merkel cells were found by F. S. Merkel in 1875 as chromophobic cells which occur together with Langerhans cells and melanocytes in human epithelium; they are characterized morphologically by polymorphic profiles, cytoplasmic processes, and

intracytoplasmic special granules(Ishizaki et al, 2006). The anatomical position of the rugae in the mouth remains unchanged in its position throughout life. Although there is no consensus on this issue.it is relatively stable and resists decomposition for a few days after death (Indira et al, 2012). The anatomical position of the palatal rugae inside the oral cavity is surrounded by cheek, lips, tongue, teeth, and a buccal pad of fat. All these afford some protection in case of fire and high impact trauma. Rugae are amongst the best protected position in the mouth that remains unchanged throughout life(Hemanth et al, 2010), morphologically individualizing soft tissue structures in the body, which are preserved after death and also accessible during life(Indira et al, 2012). Coslet et al. reported the clinical removal of palatal rugae is not permanent and that when removed, the rugae returned several months later(Patil and Acharya, 2008; Coslet et al,1980).

Histologically, the rugae are covered with stratified squamous (layered scales), mainly parakeratinized epithelium with small rete ridges and submucosa consisting of fibrous connective tissue. Similar to the adjacent tissue of the palate, CK20 positive cells were frequently observed in the cells of the basal cell layer and were particularly aggregated in the tip of the rugae(,Indira et al, 2012,Bhullar et al, 2011; Patil and Acharya, 2008;Ishizaki et al, 2006). The orientation and forms of palatial rugae are governed by the core fibers running anteroposteriorly in concentric circles below each rugae. Fibroblasts and collagen fibers, then accumulate in the connective tissue beneath the thickened epithelium and assume distinct orientation. The core within the palatal rugae of humans contains elements that are believed to contribute to the maintenance of its shape. The main structural element of a rugae contains glycosaminoglycans, which by its hydrophilic nature causes the tissue to swell, and contributes to the maintenance of the shape of a rugae throughout life. (Rajan et al, 2013; Gandikota et al, 2012; Indira et al, 2012; Ishizaki et al, 2006; Bharath et al, 2011; Bhuller at al, 2011; Amasaki et al, 2008; Motabagani, 2006; Thomas, 1984). The palatal rugae at birth are well trainees with a typical orientation pattern and during adolescence they acquire the final feature shape of each individual. Once formed, they may experience changes in their size due to growth of the palate, but their shape is maintained(Gandikota et al, 2012).

Figure 1 : Histology of palatal rugae shows rugae are covered with stratified squamous), mainly parakeratinized epithelium with small rete ridges.
James K, Pauline F, Steele, Nancy Avery .Oral development and nd histology.2 edition. Thieme medical publishers.1994 page 253.

1.3 Physiological Function of Palatal Rugae

The anatomic position of these transverse ridges aid the physiological function of palatal rugae area in oral swallowing, suction in children. In neonates, the rugae which usually corrugate the hard palate assist gripping of the nipple when suckling. Taste perception by improving the relationship between food and the taste receptors in the dorsal surface of the tongue. They participate in speech speciallys and sh phonemes and in the medico-legal identification process (Eboh, 2012; Amasaki et al, 2008; Almeida et al, 1995).

1.4 Significance of Palatal Rugae. Rugae patterns have been studied for various purposes, with reports being published mainly in the fields of anthropology, comparative anatomy,genetics, forensic odontology, prosthodontics, and orthodontics (Hemanth et al,2010).

1.4.1 Significance of Palatal Rugae in Post Mortem Forensic Identification. There are four legally admissible methodologies used to identify human remains (Thomas and Kotz, 1984) visual identification, fingerprints or footprint identification, dental identification, and

DNA evidence (Simmons et al;1987). These are the most commonly used techniques in forensic identification allowing fast and reliable identification.

The palatine rugae possess unique characteristics that can be used in circumstances when it is difficult to identify a dead person through fingerprints or dental records (Bansode and Kulkarni, 2009). One of the main focuses of the forensic odontologist is identification of an individual. Dental identification can be used as the sole method of identifying a deceased person. Dental identification is based on the comparison of antemortem and postmortem records. The records collected to identify a decedent should be accurate and totally inclusive of objective findings (Filhoet al, 2009). It was concluded that palatine rugae dont undergo pronounced changes under such massive thermal insults (Winslow,1732). It is suggested that rugae pattern is as unique to a human as are his or her fingerprints and it retains its shape throughout life (Bhullar,2011). The anatomical position of the rugae inside the mouth - surrounded by cheeks, lips, tongue, buccal pad of fat, teeth and bone - keeps them well-protected from trauma and high temperatures. Palatine rugae can resist postmortem decomposition changes for up to seven days after death (Patil and Acharya, 2008). Moreover, the stability of palatine rugae have been examined in cases of third degree burns. Thus, they can be used reliably as a reference landmark during forensic

identification (Buchner, 1985). Palatal rugae have been considered relevant for human identification due to its stability, which is equivalent to the fingerprint. In that it is unique for each ruga pattern, palatal rugae appear to

possess the features of an ideal forensic identification parameter, that is, uniqueness, postmortem resistance, and stability (Hoggans and Sadowsky, 2001).

Muthusubramanian et al (2005) did a study to examine the extent of palatine rugae preservation for use as an identification tool in burn victims and cadavers. The study results showed that among the subjects with third-degree panfacial burns, 93 percent of the palatine rugae were normal. They concluded that the palatine rugae could be used as a reference landmark during forensic identification of individual. In forensic identification, , use of dental records, visual identification and fingerprints and DNA comparisons probably are the most common techniques used allowing fast and secure reliable identification. When identification cannot be established by fingerprinting or by analysis of dental arches (dental records data), the palatal rugae can be considered as a source of comparative material (Limson and Julian,2004). Many victims of natural disasters such as fires and floods, have also been identified by dental means. Furthermore, many criminal investigations have included the use of dental evidence (English et al, 1988).

1.4.2 Palatal Rugae Area Added to Complete Dentures in Prosthodontics .

Restoring patients' speech is an important goal in complete denture fabrication. For those patients who have difficulty with their speech patterns, accommodating to the introduction of prosthesis, texture in the palatal region may prove helpful. Such problem might be related to changed relationship between the tongue and the new topography of the palatine vault. It is suggested to reproduce the original shape of palate as one of the solutions to relief pain encountered by patient. This also contributes to reduce speech problems resulted from wearing these prostheses (Palmer, 1978).

Methods of incorporating palatal rugae in a newly fabricated and existing complete denture are studied with many researchers (Gitto et al, 1999).

1.4.3 Significance of Palatal Rugae in Orthodontics as Internal Dental Cast Reference Points for Quantification of Tooth Migration: Palatal rugae may serve as suitable reference points from which the clinician can derive the reference planes necessary for longitudinal cast analysis, since dental casts are three dimensional (3-D) records of malocclusion that have been used successfully during diagnosis and treatment planning for orthodontic patients (Rajan et al, 2013). Hoggan and Sadowsky (2001) also investigated the use of the palatine rugae as reference points for measuring tooth movement in a manner comparable with cephalometric superimpositions. The results showed no statistical differences between the mean incisor and molar movement measured cephalometrically. The tooth movement measured relative to the medial and lateral end of the third

palatine ruga. So they concluded that these rugae area can be used successfully to assess anteroposterior tooth movements. Christou and Kiliardis (2001) evaluated the vertical changes in the medial aspects of the rugae and concluded that these changes over time are due to the alterations in the vertical positioning of maxillary incisors and increase in lower face height. Positional changes of posterior teeth in the anteroposterior direction are relevant to the diagnosis and correction of sagittal occlusal abnormalities and arch length discrepancies, which is another clinical significance of palatal rugae area (Bhuller et al, 2011). Furthermore, Yang et al (2013) studied rugae area as it is considered a stable reference points for superimposing 3-dimensional models before and after orthodontic treatment, so they measured 3-dimensional changes for children over 9 years old. Results came up with no significant changes occurred and then they are considered stable .So used as reference points for superimposing and evaluating changes during orthodontic treatment. In the literature, Palatal rugae have been used as reference points for many purposes such as evaluating tooth movement pre- and post-orthodontic treatment .There are also indications that various types of orthodontic treatments may have different effects on the rugae, e.g. non extraction treatment, premolar extraction treatment (different combinations of extraction sequences) and orthopedics maxillary expansion (Hoggan and Sadowsky, 2001; Ong and Woods, 2001).

Although the use of study model comparisons or the superimposition of images of study models to evaluate tooth movement has been attempted, results of these studies have been difficult

to interpret because of the lack of available evidence of stable landmarks (Jones, 1991; Rossouw et al,1991;Van der Linden 1978; Van der Linden 1974).

Recently some researchers have focused on the use of palatal rugae as suitable landmarks, but the results of these studies are not consistent (Mavropoulos et al,2006;Mavropoulos et al 2004;Miller et al,2003;Hoggan and Sadowsky 2001; Ong and Woods,2001Bailey et al 1996;Almeida et al 1995;Grove and Christensen 1988;Simmons et al 1987; Van der Linden 1978;Peavy and Kendrick 1967).

Although teeth are more durable than other parts of the body, identification via dental records also may prove to be inaccurate, because dental treatment might have been performed between the creation of a dental record and the persons death. Other methods of identification like DNA profiling are of limited use because of the high costs and the time-consuming nature of the method which make it particularly unsuitable for large populations.(Venegas et al,2009).

Chapter Two:
2. Literature Review
This literature review discusses four dominant fields regarding studies were done concerning palatal rugae area. Starting the first part with studies that inspected literature that discussed individuality of palatal rugae. The second part explored variations of palatal rugae in different genders and ethnicities. The third part tried to investigate literature of morphometric changes induced by orthodontic treatment. The fourth part consisted of studies included matching between pre and post treatment casts, in order to be considered as a method of human identification.

2.1 Individuality of Palatal Rugae. Palatoscopy or palatal rugoscopy, is the name given to the study of palatal rugae. From a forensic perspective, this is used primarily to establish a person's identity. Palatal rugae area is considered to be highly individual and consistent throughout human life (Indira et al, 2012).

The application of palatal rugae patterns for personal identification was first suggested by Winslow in 1700s then Allen in 1889. But as palatal Rugoscopy, it was first proposed in 1932 by a Spanish investigator called (Trobo Hermosa) ( Sharma et al,2009; Shriram and Meena,2009). In the same year, the earliest illustration of palatal rugae was made by Santorins who made a drawing depicting three continuous wavy lines that crossed the midline of palate (Simmons et al, 1987). So, Lysell (1955) was unsure if the rugae could be used for identification aspects but Sassoni (1957) stated that it's possible to device a classification based on symmetry, number and shape of papilla. When he tested his classification, he was able to identify a person without difficulty (Sassouni et al, 1957).

Researchers had found difficulty in the task of classification of the rugae patterns due to the subjective nature of observation and interpretation within and between observers. Since the study of Lysell (1955) specific anatomical investigations on palatal rugae patterns had been reported by many researchers. Numerous classifications had been devised by several authors to record the palatal rugae patterns; among all, the Silva, Carrea and Lysell, classifications are often used in recording the patterns (Barath et al, 2011).

In 1937 Carrea conducted a detailed study and established a method to classify palatal. In1955 Lysell postulates that palatal rugae patterns have genetic background (Lysell, 1955).This man as well as Sassouni (1957) who just has said that no two palates are alike in their configuration. Furthermore, the palatal print does not be changed with time or age, so in general he believes that

these rugae are unchanged throughout human life, but found that this doesnt apply in every aspect (Filho,2009; Sassouni,1957; Lysell,1955).

Several studies were done and still being done in the past and nowadays. They have revealed statistically proved result, in which the rugae patterns are highly individualistic. Although it was described as being still in infancy in the field of forensic odontology, their uniqueness to individuals has been recognized clearly as providing a potentially reliable source of identification (Sharma et al,2009; Kapali et al, 1997; English et al, 1988; Thomas and Kotze, 1983).

The studies done by de Angelis (2011) in Italy, Hermosilla et al (2009), Shukla et al(2011) and several others were all in agreement that the individuality of palatine rugae were sufficient for their use as method of identification. This was supported by the fact that the morphology of palatine rugae is constant throughout life (Bhullar, 2011; Abdel-Aziz and Sabet, 2001; Fahmi et al, 2001; Almeida et al, 1995; Peterkova et al, 1987; Vander Linden, 1978; Lysell,1955).

In a case study, Breault et al (1999) reported on a single case of a 22-year-old white male. A free gingival graft was placed on the edentulous (toothless) buccal gingival corresponding to tooth #23. The graft was harvested from the palate and included the palatal rugae. After two months, the recipient site displayed prominent ridges and a gingivoplasty was performed. After nine years, the subject returned for treatment and the palatal rugae had reformed in the graft area. This study showed that the rugae can be transplanted to different parts of the body. If transferred to other areas of the anterior palate, rugae can easily be confused in identification .Therefore, they are considered as permanent and unique to each person, and can establish identity through

discrimination (via casts, tracings or digitalized rugae patterns) (Anukool, 2011;Hoggan and Sadowsky, 2001; English et al, 1988). Indira et al (2012) studied palatal rugae pattern among five pairs of twins (non identical). They showed different patterns, although some similar (but not identical) forms are observed on specific location in two pairs of twins. Such findings were consistent with the study conducted by Ritter and Chae (cited in English et al, 1988) which strongly suggested that palatal rugae pattern are unique for each person .I t also questioned the role of genetic influence in determining the rugae pattern in twins. The concept about the individuality of palatine rugae was supported by more recent studies using more technological equipments. It seems to be discussed by Saxena et al (2010). They used the three dimensional stereoscopy techniques in studying the anatomy of palatine rugae and mentioned that there were no similarities in the numbers of primary rugae in both sides of each cast and stressed on the concept of palatine rugae individuality.

2.2 Variations of Palatal Rugae in Different Genders and Ethnicities.

Specific anatomical investigations on palatal rugae patterns had been reported by many authors. They reported that the number of rugae remains unchangeable during life, the size and detailed arrangement changes with palatal growth and that there are differences among ethnicities with or without differences between genders (Eboh, 2012; Filho,2009;Kapali et al, 1997; Dohke and Osato, 1994; Kashima, 1990).

Thomas and Kotze (1983) in their literature highlighted the difficulties in observing, classifying and interpreting the limitless and minute variations in palatal rugae and emphasized the necessity for standardizing the procedures in recording. After a thorough review on all classifications from the literature, the method of identification used in this study (Thomas et al, 1983) was the most practical and easy to apply compared with other methods (Bharath et al, 2011).

Thomas and Kotze were able to discern different rugae patterns in southern African populations implying different genetic origins in their studies in 1983, they found that rugae were unique and individualistic to each ethnic group and it can be used successfully for genetic research (Bhullar, 2011; Thomas and Kotze, 1983).

While Kepali et al (1997), through several approaches aimed to investigate changes in rugae patterns with age in Australian aborigines, and compared patterns between aborigines and Caucasians. They find that the number of primary rugae in Australian aborigines was higher than in Caucasians. Furthermore, the mean number of rugae in aborigines was higher than reported for other ethnic groups. However, Caucasians tend to show higher proportions of rugae that are longer than 10 mm compared with aborigines.

In 1989 Houser et al investigated 117 dental casts of individuals from Swaziland; impressions were obtained, and then poured in plaster. Prominent rugae were observed on the dental casts, the palatal rugae area were similar since this population was very homogenous.

Dohke and Osato (1994) indicated that Japanese females had fewer rugae than Japanese males. In contrast, Saraf et al (2011) compared palatal rugae patterns between Indian males and females. They reported no significant difference in palatal rugae between both sexes in terms of total number or various length measurement. But according to shape, the converging type was statistically greater in number in females whilst the circular type was statistically greater in number in males which is opposite to earlier studies.

Kepali et al (1997) hadnt any significant difference in the number of primary rugae among Australian males and Australian females. On the other hand, Dohke and Osato in 1994 give opposite results, it might be due to the fact that secondary and fragmentary rugae were not included in the study of (Saraf et al, 2011; kepali et al, 1997), and it is the secondary rugae that Dohke and Osato consider in their study, which lead to sex differentiation.

In the study of Preethi et al (2007) that was done on western and south India population, straight pattern was the more found and circular group was not. Kashima (1990) did a study involved Indian and Japanese children. It reported that Japanese children had more primary rugae than Indians.

Sumathi et al (2011) did a study comparing palatal rugae between individuals, where they were found through the whole work, no identically similar palatal rugae patterns appeared in two individuals. According to the most predominant pattern, it was the wavy pattern followed by the straight one, curved, branched, and the least was the circular. There was no significant difference among males and females in this population.

It had been noted that there is astatically significant association between rugae forms and ethnicity (Hermosilla et al, 2009; Kepali et al, 1997). Also in 2012, a study was done by Eboh , intended to describe the shape and gender distribution of palatal rugae and their prevalence in the studied population. It was observed that the dominant rugae form among Urhobo ethnic group was line one, then sinuous patterns, it fails to give any sex significance. Both studies of Kashima (1990) and Eboh (2012) were met.

According to difference in palatal rugae related to gender, many studied variations in this area between males and females. Most of them revealed lack of sex variations. However, indicated a slight difference in number of rugae between both sexes (Saraf et al,2011).

Another study was done by Shetty et al (2011) that attempts to determine the number of different patterns of rugae, and to see if there is any gender differences present in Mangelore population. There was no significant difference among sexes; the predominant shape of rugae among females was curved and straight, while wavy among males. These observations were in contrast to Swetha et al (2005) who report that Mysorean males and Tibetal females had more rugae than their respective counterparts (Sharma et al,2009). Furthermore, in this study circular shaped rugae didnt show any significant gender differences, while Fahmi and Al-Shamrani (2001) study among Saudi males and Saudi females shows an increased incidence of circular shaped rugae in females rather than in males.

Madhankumar et al (2013) make a study with a Cross sectional design in 2009 to evaluate the gender differences with regard to the shape of the palatal rugae and identify the most predominant pattern. They collected 135 students aged 17-25yrs to participate in the study. Based on gender they were divided into two groups comprising of 62 male and 73 female students respectively. Maxillary impressions were poured in dental stone .These casts were utilized to analyze and study the variation in rugae pattern based on classification by Thomas and Kotze. The data obtained were tabulated and analyzed using IBM SPSS 19.0 version. The incidence of specific rugae pattern and its association with gender were analyzed using Chi-square test. They resulted in noticing that straight and curve forms were

2.3 Literature of Morphometric Changes Induced by Orthodontic Treatment. The stability of palatal rugae as a landmark for dental cast analysis is reported by (Patil et al, 2008; Almeida et al, 1995 and Bailey et al, 1996) that form, layout, and characteristics are not affected by the eruption of the teeth or their loss, but sometimes palatal rugae adjacent to the alveolar arch slightly change their position after tooth extraction (Hemanth et al, 2012). On the other hand, other studies showed that some events can contribute to changes in their patterns such as trauma, extreme finger sucking in infancy, and persistent pressure with orthodontic treatment (Kapali et al, 1997). Peavy and Kinderick (1967) demonstrated that the form, layout and characteristics were not affected by the eruption of teeth or their loss, but there were times that some changes occur to the position of palatal rugae near to the alveolar ridge after tooth extraction (Eboh,2012; Peavy & Kendrick,1967) However, Limsoon and Julian (2004) and Baily et al( 1996) said that extractions can produce a local effect on the directions of palatal rugae.

Shukla et al (2011) and Bansode and Kulkarni (2009) had found that some changes do occur in the rugae during orthodontic treatment and the morphology of palatal rugae remains stable throughout life(Hamanth et al,2010 ).

In a study of changes occurring in 15 patients who underwent extraction of four premolars, Hausser (1989) observed orthodontically treated patients. He concluded that the lateral edges of the rugae moved forward about one half the distance of the migration of the adjacent teeth, while the medial rugae were not affected.

On the other hand, Simmons and Colleagues (1987) used the longitudinal database of the Child Research Council of Denver to examine the anteroposterior stability of the medial rugal region. Their analysis indicates that the medial rugae region increased significantly in anteroposterior length, but not uniformly between the sexes. The authors concluded that such changes were characteristic of general craniofacial growth and suggest that the rugae region is responding to the differential growth of the underlying bone. Thus, the authors established that the medial rugal landmarks did not appear to be a stable reference point for tooth migration research.

Peavy and Kendrick (1967) reported that the lateral ends of the rugae that terminated close to the teeth followed the movement of the teeth in the sagittal plane, but not in the transverse plane as was said by (Bhuller et al, 2011). Van der Linden (1978) evaluated the changes in the position of posterior teeth in relation to palatine rugae. He selected 65 normally growing children (aged 6 to 16 years), and six orthodontically treated patients. The maximum mean change in distance

between the rugae in the anteroposterior plane was 0.41 mm. The authors noted larger movements at both the medial and lateral rugae points in the orthodontically treated patients.

Anukool et al (2011) in his study didnt observe change in shape of palatal rugae between pretreatment and post treatment cases.

Bailey et al (1996) presented the results of a double-blind study in 57 adult patients. The maxillary casts of pre- and post-orthodontic treatment are examined. Two groups of patients, an orthodontic extraction group (where two maxillary premolars were extracted) and a non-extraction group, were assessed. Statistically significant changes in rugae were noted only in the extraction group. Though statistically significant changes occurred, the medial and lateral points of the third rugae were not considered clinically significant; it can be used for anatomic reference points in dental cast analysis.

Almeida et al (1995) purported the first rugae as the most stable, whereas Bailey et al (1996) describes the third rugae as the most stable. These findings presented further contradiction to the use of rugae in identification due to possible changes over time and after events. Taken together, these two studies highlighted the discrepancy in the stability of the palatal rugae after orthodontic treatment and extractions.

2.4 Studies Concerned Matching Pre and Post Treatment Casts as a Method of Human Identification.

In 1983 Thomas et al gave a classification (which is most applicable in most studies) for the rugae identification included number, length, shape and unification. So, shapes are classified in to curved, wavy, straight and circular. Unification is divided in to further converge and diverge (Anukool et al, 2011; Thomas and Kotze, 1983).

In 2009 Filho and his colleagues, made a study aims to consider palatal rugae as source of human identification beside other parts related to, in addition to the presence or absence of teeth as complementary method. Their sample consisted of 100 models and their respective negative. Three examiners participated of the research. Digital models are examined by means of the Photoshop 7.0.1 program. The results, led to a percentage of 100% certainty on the identification, so it made the identification possible foe the whole sample. And this led to conclude that it is possible to achieve the human identification by means of palatal rugae provided that it has a previous database. This provides significant advantages in criminal cases (Filho et al, 2009).

In the same year, 2009, Shriram and his colleagues made a study with an objective to compare rugae patterns in pre and post-orthodontic treatment cases as a way of determining stability of palatal rugae through orthodontic treatment. So, 30 preoperative and postoperative dental casts were selected. Thirty casts were randomly selected for the present study. The 30 postoperative casts were mixed with the 30 randomly selected casts. Thirteen examiners were selected as evaluators and instructed to match the 30 preoperative dental casts with the 60 dental casts (30 postoperative and 30 randomly selected casts). The case numbers of those that were correctly matched were noted. The result indicates that during fixed orthodontic treatment, some changes occur, but not in the rugae pattern. The 13 examiners achieved 90% correct matches. So, it was

concluded that Palatal rugae patterns are unique to an individual. Therefore, used for individual identification in forensic odontology.

Palihar et al (2010) conducted a study to compare the palatal rugae in pre and post-orthodontic treated cases in term of their relation to each other, and to median plane for their uniqueness and stability. The results showed that there is no significant difference between pre and postorthodontic cases, so they concluded that palatal rugae area remain stable throughout life. Consequently, changes that occur in adolescent stage affect length only.

In a double blind study, English et al. (1988) selected 25 orthodontic cases with pretreatment and post treatment dental casts. The casts came from subjects over 14 years of age. Then the 25 post orthodontic casts were placed within the 100 casts. Seven dentists and 2 dental assistants were chosen as evaluators. Five of the dentists with varying forensic experience in identification were assigned as individual evaluators. Two teams were also formed; both teams had no forensic experience. Each team was asked to compare them to the 125 casts for matches. The time required for the comparison, coupled with the correct percentage, was recorded. Eight investigators correctly matched 100% of the casts and one investigator correctly matched 88%. From this study, it was suggested that palatal rugae can be used for identification purposes. It also demonstrated that changes that occur from orthodontic movement, extractions, aging, and palatal expansion do not modify the rugae enough to prevent identification.

Then Anukool et al (2011) conducted a study in India aimed to judge the use of palatal rugae in pre and post- orthodontic cases and determine their reliability. The classification given by

(Thomas et al, 1983) was used for rugae identification. No statistically significant difference was found. This proved that rugoscopy identify an individual even after orthodontic treatment also. Thus, palatal rugae hold potential as a supplementary tool, along with the dentition, to establish the identity of an individual.

In the same year, Shukla and his colleagues (2011) wrote a research that aimed to determine the stability of palatal rugae before and after orthodontic treatment. They selected 50 orthodontic cases with preand post-treatment casts, and 50 casts were randomly selected as variables. In the pre and post-treatment group, changes in transverse measurements were significantly different for lateral points of first rugae, in addition, anteroposterior changes were significant for the distances between first and second rugae. All inter-point measurements of third rugae were stable in posttreatment casts. Thirty blinded examiners compared 50 trimmed preorthodontic casts to similarly prepared one hundred casts for possible matches based on pattern of rugae. The percentages of correct matches for examiners had a median of 90%. The matching ascertained that the morphology of palatal rugae remains stable throughout life although some changes occur. Subsequently, points associated with the third palatal ruage were the most immutable over a person's life and could be used as a reference to evaluate changes in teeth positions happened during orthodontic treatment (Shukla et al, 2011).

Stavrianos et al (2012) examined 50 orthodontic cases where their treatment lasted from 18 months to four years. Then those 50 were mixed with other 100 casts that were randomly selected. The whole of 150 casts were given to five researchers to identify similarities between them, four of the researchers had given 100% match for all 50 casts, and the last one matched 47 of them.

Therefore, they resulted that palatal rugae area is a unique personalized forensic evidence for recognizing persons.

Shetty et al ( 2013)also conducted a study that aims to investigate palatal rugae patterns in females and males, and to evaluate the stability of these patterns in pre and postoperative orthodontic cases, Fifty patients were selected for this study (25Males and 25 females). From the above sample, 10 males and 10 females had undergone orthodontic treatment, and their casts were retrieved for sex determination analysis and stability of rugae patterns pre and posttreatment. They resulted that some Changes can occur in bony structures during fixed orthodontic treatment, but rugae patterns remain stable. Kappa statistics and Chi square test were used to analyze agreement between the two evaluators, and 95% correct matches were achieved. So they concluded that Palatal rugae are unique to every individual and can be used as an indicator in forensic odontology. (Madhankumar et al, 2013) Babu et al (2013) aimed in their study that was done to study, analyze and report the significant palatal rugae patterns among West Godavari District population. The results then were compared with those patterns reported in literature to give an extra identification in cases of crime or mass disasters. They select 100 pre-orthodontic casts. Thomas et al (1983) classification which included the number, shape and unification patterns of rugae was used. The results represented that wavy pattern is the significantly predominant pattern of PR. They concluded that Palatal rugae patterns are definitely associated with regional variation and can aid as an additional tool in forensic identification procedures. Several studies have subsequently used palatal rugae as a stable reference points for assessment of orthodontic tooth movement (Mavropoulos et al, 2005; Ashmore et al, 2002).

During literature review in Middle East countries three studies were done which concerned the morphometric analysis of palatal rugae in United Kingdom of Saudi Arabia, Egypt, and the third one was in Jordan, where all of them revealed palatal rugae individuality. For using orthodontic treated cases to asses and evaluate palatal rugae area as a land mark for individuality, a study was done in Egypt by Abd el Aziz and Sabet (2001) .They studied 50 pre and post orthodontic casts to measure the stability of palatal rugae area, there were some changes after orthodontic treatment. Results were insignificant by comparing pre and post results. It seemed that the lateral third rugae is the most stable points and can be used for identification (Mustafa et al, 2012; Abdel-Aziz and Sabet, 2001; Fahmi and Al-Shamrani, 2001; El-Fotoh and El-Sharkawy, 1998).

Chapter Three: Methodology

3.1 Sample Collection The study was done in the department of anatomy at Jordan University of Science and Technology, Irbid, Jordan. The study tried to evaluate post orthodontic treatment changes occur in palatal rugae area of palate. So, the researcher collected 50 pre orthodontics and 50 post orthodontics dental casts from a private clinic in Irbid, in the north of Jordan. Dental casts of the upper jaw are used as records of malocclusion. They are of great asset to orthodontists during diagnosis, treatment planning and monitoring improvements for orthodontic patients. Hence, serve

as replicate for palatal rugae area. Abdel-Aziz and Sabet (2001) mentioned that dental casts used for analysis of orthodontic treatment changes is an important goal for orthodontic and forensic researchers. 3.2 Material and Equipment The study used maxillary dental Casts, they were sent to the lab to be duplicated. Then, using Hb pencil, rugae area was delineated under adequate light. Every pair of casts (represent pre and post for every patient) was given a serial number but not shown to the evaluators. A vernier caliper was used to measure the length of each rugae, (this instrument measures of accuracy .05mm and easy to be used), it's both edges were put on both end of the rugae. The distance between the two points was read with its scale to give us the length to measure whether any change occurred.

3.3. Classification Criteria of Palatal Rugae Palatal rugae were classified by many classifications during literature. Here in this study, the researcher applied the classification given by Thomas et al, which composed of three parts. The first part classifies palatal rugae according to their length. Those rugae, which have a length of more than 5 mm were referred to as primary rugae. Secondary rugae were those that have a length of 3 to 5mm, whilst fragmentary rugae were those to have less than 3mm. The second part classifies palatal rugae according to their orientation. The orientation of each rugae is determined by the relationship between the origin (medial end) and the termination (lateral end) points of each rugae.

Rugae are considered anterior rugae when the termination point is anterior to the origin point. Posterior rugae when the termination point is posterior to it. When both points are on the same level it's considered horizontal. If rugae changes direction for more than one time, it is considered irregular rugae.The third part classifies palatal rugae according to their shape in to one of six major types: straight, curved, wavy, circular, diverging and converging. Straight rugae run directly from their origin to termination. Curved rugae have a simple crescent shape that is curved gently. When curved rugae have any bend at origin or termination, they are considered wavy rugae. Circular rugae display a definite continuous ring formation. Rugae that begin from the same origin medially and split laterally are classified as diverging, whereas, these rugae that begin from more than one origin, but they unite at their lateral portions, are classified as converging rugae. This classification was used to know how to give names to the palatal rugae, and to record changes if occurred in them after orthodontic treatment in the right manner. Furthermore, to make our next classification that is based on another ten criteria easier to deal with. So, these ten criteria were considered to be studied and evaluated, followed by comparing between pre orthodontics and post orthodontics casts as follows: a. Segmentation; represents one or more rugae in the pre treatment cast that split in to two or more than that in the post treatment cast. b. Unification; represents two or more rugae area in the pre treatment cast that become one in the post treatment cast.So the number of rugae here is decreased. c. Change in orientation; represents any change in one or more rugae in their direction. d. Change in shape; it meant that one or more of the rugae changed its way that it looks like form circular to crescent shape for example. e. Change in length; is one or more of the rugae could be elongated or shortened.

f. Change in prominence; is one or more of the rugae could be changed to more bulbous or be thinned or it can be more or less obviously recognized. g. Displacement of lateral end in mediolateral direction; a movement of lateral end of one or more rugae in either sides to be nearer or far away of the alveolar ridge. h. Displacement in medial end in mediolateral direction; a movement of medial end of one or more rugae in either sides to be nearer or far away of the center line. i. Displacement in medial end in anterioposterior direction; a movement of medial end of one or more rugae in either sides to be nearer or far away of the central incisors. j. Displacement of lateral end in anteroposterior direction; a movement of lateral end of one or more rugae in either sides to be nearer or far away of the alveolar ridge that they were in the pre orthodonticscasts .

3.4. Data Analysis 3. 4.1. Palatal Rugae Changes: The data were then manually transferred to an Excel file on the computer by using every pair of cast's serial number. Changes occurred were recorded in every post orthodontic cast regarding all criteria we asked for. Then the researcher gave the number 0 for the criterion that didnt change in post treatment cast, and number 1 for the criterion that had changed. The frequencies of how many casts that did change or got number 1for every criterion were counted. At the end of treatment, the percentages of casts that had changes were calculated. Then this excel file will be transformed in to SPSS program to give us frequencies of changes happened in palatal rugae after orthodontic treatment.

3.4.2. Test of Individuality Regarding individuality of palatal rugae area, the researcher compared every one of the pretreatment casts of the fifty casts with the other forty nine casts. Three main categories were overlooked including both their right, left halves and Medline of the entire palate. This was done to observe any pairs had the same rugae pattern .So the researcher did 1225 comparison between the 50 pre treatment casts. Some of them might had the same number of rugae but not the same shapes and orientations. Similarly, every post treatment cast was also compared with the other post treatment casts .1225 comparisons were done again in order to assess the individuality of palatal rugae even after orthodontic treatment.

3.4.3 Matching Test The researcher blocked out teeth in the pre orthodontics casts, duplicated post treatment casts and the original post treatment casts (as shown in figure 2) in a standardized manner according to English et al (1983) technique to be recognized without the help of the occlusion but only based on the shape of palatal rugae themselves. This was done in a way that involved the teeth and vestibule and edentulous area to ensure their influence did not exist anymore. The researcher asked ten evaluators to match between the pre orthodontics and post orthodontics cast related for the same person to ascertain the uniqueness of this area. So, the evaluators selected the closest match based on the pattern of rugae. A sample of ten was chosen randomly from the table of random digit numbers. Every evaluator was asked to match this sample of ten of the pre orthodontics cast with their similar of the fifty original post orthodontics ones. Then the same evaluator matched ten of

the pre orthodontics casts with their right matches of the duplicated post orthodontics casts. Finally he/she will match ten of the original post orthodontics casts with their counterparts from all duplicated post orthodontics casts. All samples were chosen randomly, and every evaluator did his trials without knowing the right matches, just by looking for palatal rugae area. Furthermore, every evaluator did his trials without being seen by other evaluators.

Figure (2): Duplicated post orthodontic casts blocked out 3.5 Statistical Analysis The right matches for all evaluators were recorded for the three trials. The means and standard deviations for trials were calculated. After that, data were transformed in to SPSS program and

paired t-test was used to detect any significant difference between pre and post treatment data for all groups we measured.

Chapter Four: Results

Since our study is a descriptive one, we will try to achieve the first aim of our study which is to calculate frequencies and percentages of morphometric changes occurred in post treatment casts regarding the criteria we choosed to study. Then regarding the individuality test in the second section, which is our second aim, was done to verify an accuracy of identification based on rugae pattern. Finally the matching test which is the second part of the second aim was done to ascertain the stability of palatal rugae despite of changes that occur throughout person life. In addition, to assess weather changes occurred in palatal rugae due to orthodontic forces hamper identification. 4.1 Morphologic Changes Induced by Orthodontic Treatment. 4.1.1 Frequencies and Percentage of Palatal Rugae Changes Related to Segmentation

Here in this section we got 11 casts out of 50 that were had segmentation, and this means that 11 casts that had one or more rugae that were divided in to two parts, giving us 22% ,while those hadnt changes were 78%, as shown in table1. 4.1.2 Frequencies and Percentage of Palatal Rugae Changes Related to Union 10 casts of the fifty were had this union criteria, that means two or more rugae were unified and became one. This gives us a percent of 20%, and those without changes were 80%, as shown in table 1. 4.1.3 Frequencies and Percentage of Palatal Rugae Changes Related to Orientation Regarding this criteria 3 casts only showed changes in orientation, and this gives us a percent of 6%,while those who didnt change were 94%.This is very low percent for change which is considered to be clinically insignificant, as shown in table 1. 4.1.4 Frequencies and Percentage of Palatal Rugae Changes Related to Shape Regarding this criteria 3 casts only showed changes in orientation, and this gives us a percent of 6%, while those who didnt change were 94%.also this criterion resembles low percent of change therefore, high percent of stability as shown in table 1. 4.1.5 Frequencies and Percentage of Palatal Rugae Changes Related to Length. I n this criterion 14 casts were changed in length either in increased or decreased pattern.This is

with percent of 28%, while those didnt change give a percent of 82% as shown in table 1.

The next four criteria are generalized somehow looking for all rugae as one component without making specifications for every rugae just to give an idea of stability and individuality of palatal rugae area for clinical significance only not for statistical significance. 4.1.6 Frequencies and Percentage of Palatal Rugae Changes Related to Displacement of Lateral End in Anteroposterior Direction 30 casts showed anteroposterior changes due to displacements of lateral ends of palatal rugae and this had a percent of 6o%, while those cast didnt displaced showed 40% as shown in table 1.

4.1.7 Frequencies and Percentage of Palatal Rugae Changes Related to Displacement of Lateral End in Mediolateral Direction For this criterion 28 casts showed displacement, and this gave a percent of 56%, leaving 44% for those casts didnt have any displaced rugae as shown in table 1. 4.1.8 Frequencies and Percentage of Palatal Rugae Changes Related to Displacement in Medial End in Anterioposterior Direction 27 casts out of fifty had displaced medial ends of palatal rugae anteroposteriorly giving a percent of 54%. Other group with no displacement got 46%as shown in table 1. 4.1.9 Frequencies and Percentage of Palatal Rugae Changes Related to Displacement in Medial End in Mediolateral Direction Only 10 casts regarding this criterion had displacement in this direction to give a percent of 20%, casts those with no displaced rugea gave a percent of 80% as shown in table 1.

4.1.10 Frequencies and Percentage of Palatal Rugae Changes Related to Change in Prominence Here in this criterion 27 casts showed change in prominence either decreased or increased after orthodontic treatment. This gave a percent of 54% while other casts which were stable in volume had a percent of 46% as shown in table 1.

Table 1: Frequencies and Percentage of morphometric Changes of Palatal Rugae after treatment.

Morph.change Segmentation Unification Orientation Shape

frequency 11 10 03 03

percentage% 22% 20% 6% 6%

Length Disp.lat.A-P Disp.lat M-L Disp.med.A-P Diap.med.M-L Prominence

14 30 28 27 10 27

28% 60% 56% 54% 20% 54%

In our sample of study 16 cases out of 50 were had extraction for first premolars. This gave a more extra space needed to be closed which give a more possible changes in palatal rugae area at the end of treatment .At least three changes occurred in any one of them, although they all shared the change in palatal rugae area related to displacement of lateral end of palatal rugae both in anteroposterior and mediolateral directions. Regarding the non-extraction group four of these 34 post treatment casts had no change in any of these criteria at all .In both extraction and non-extraction cases no cast involved changes in all criteria. The casts that contained the highest percent of changes had changed 7 of these criteria but according to matching test still could be correctly matched, representing those changes as clinically insignificant. 4.2 Individuality Test Regarding this test the researcher tried to conduct a qualitative comparison between pairs of casts. By taking one and compare it with the remaining 49 casts. When pre orthodontics casts were

compared with each other, no two were alike or looks the same of others; even when comparing the left with the right side for the same cast they were different from each other and can be easily differentiated. Numbers of trials or comparisons were 1225, and was validated by another examiner to make this step more accurate .This comparison aimed at detecting any identical palatine rugae patterns. Furthermore, post orthodontic casts were compared in the same manner. Another 1225 comparisons were done. No two similar casts existed. The entire palates of each pair of casts were compared first then each half of the palate was compared to its counterpart in the second cast. The results showed that we did not have any identical palatine rugae pattern either completely or partially in our sample. This indicates that palatine rugae patterns among Jordanians are very individualized. Post treatment casts comparisons revealed that individuality of palatal rugae area ascertained despite of morphometric changes occurred.

Table (2): Individuality test Area of comparison Number of comparisons Pre Entire palate Left side rugae Right side rugae 1225 1225 1225 Post 1225 1225 1225 Number of similarities Pre 0 0 0 post 0 0 0

4.3 Matching Test Talking about the first trial which was between the pre and post treatment casts, the mean for it for all evaluators was 90% with a standard deviation 1.054, the second trial mean was 94%, and the standard deviation was 0 .966, and 99% was the mean of the third trials that were matched correctly by the same evaluators with a standard deviation of 0.316 as shown in table 3. Regarding this matching test when comparing trials of pre orthodontics casts with the post orthodontics ones and the trial of post orthodontics with the duplicated post treatment casts (when p value less than .05) there was a statistical significant difference between them, by using paired t test ,it was calculated to be 2.46. While by comparing the first trial with that of pretreatment casts with the duplicated post treatment casts, it was obviously seen that they were not statistically significant.

Table (3): T-test for the First and the Third Trials.

Type of trial Pre and post Post and copy

Mean 90 % 99%

St.D 1.054 0.316

df 18

t 2.46

P value 0.05

Chapter Five: Discussion

The researcher in this chapter discusses the results of the study in three parts. The first part discusses the results of the morphometric changes which occurred to palatal rugae after orthodontics treatment. The second part discusses the results of individuality test, and the third part discusses the results of matching test, and analyses the results of evaluators matching of pre and post treatment casts.

This study is the first to be conducted in Jordan for detection of morphometric changes resulted after orthodontics treatment. Estimation of orthodontics treatment changes is a step forward for better treatment results, thus stability of palatal rugae is evaluated, and this emphasized the importance of palatal rugae in accordance to previous literature. Our aims in this study were to test the uniqueness and individuality of palatal rugae among Jordanian population. Furthermore, evaluate the reliability palatal rugae as forensic tool for identification through certain morphological changes resulted post treatment. 5.1 Criteria of Palatal Rugae that Have Changed and its Effect on Palate Morphology Regarding the first morphologic change which is segmentation, (the splitting of the palatal rugae in to two parts) just occurred in 22% of casts. This wasnt studied in the literature, but it can be seen that although this type of change occurred, still didnt affect the individuality of palatal rugae. Orthodontics treatment strategies involves certain procedures regarding increasing or decreasing the width of palate by expansion or retraction(ong and wood,2001). These ways can act on soft tissue by stretching or compressing, this can lead to fragments in these rugae and splitting of its continuity (Damstra et al,2009). So, its suggested that fragmentation appeared as if this rugae became two parts very near to each other (Ashmore et al, 2002). Also regarding the second change, just 20% of casts were unified and united to become one. This could have the same explanation of the first. Movement of teeth and soft tissues in certain way that lead to compression of tissues and being unified .They could be very near to each other after teeth moved and appeared as one rugae .

The previous result didnt agree with the study of Parihar et al(2010) who concluded that changes occur in the position of rugae in orthodontics cases are statistically insignificant, and doesnt affect their unification. In our study these changes were significantly affected matching efficiency. Stavriano et al (2012) stated in their study that number of palatal rugae remains stable throughout life .Changes that happened as a result of orthodontics therapy may be seen only to their length. This may not agree with our results regarding the first two morphological changes. In the literature, consensus of opinion is that the rugae remain fairly stable in number and morphology except when there is trauma, such as orthodontic tooth movement, which may modify their alignment. (Paliwal et al, 2010; Houser et al, 1989; Jacob and shalla, 1987). Regarding changes in orientation and shape, just 6% of casts for each of them showed this change. It was very limited involving only one terminal part of one rugae. These results agreed with a study were done by Hopkinz and Harris (2003) suggested that there are Breadths of the individual rugae changed in different directions after orthodontic treatment. Consequently, they considered them as movable points. On the contrary to this, Anukool et al (2012) concluded that no change in shape of palatal rugae was observed when comparing pretreatment with post treatment cases. Incidence of change in rugae shape was low when compare pre with post treatment casts as reported by other studies (kapali ,1997; Robison ,1998)

Stuart et al (2005) concluded that Palatal rugae have been shown to be highly individualistic and consistent in shape throughout the life. It is highly suggested that palatal rugae pattern is

unique to human being, as his fingerprints. Also changes that resulted from orthodontic movement, extraction, aging, and palatal expansion did not modify the rugae enough to hamper identification, which didnt agree with our study. On the other hand, in another study, post treatment casts werent taken in consideration because of changes in orientation in margins of lateral rugae has been found. These results might support our findings (Goyal and Goyal, 2013).

Length, the fifth morphological changes were observed in 28% of casts, not necessarily to be increased after orthodontics treatment, but certain casts their rugae were decreased in length , and that was depending on the type of treatment carried on, whether it was rapid maxillary expansion which contribute to increase in length due to stretching, or maxillary constriction and retraction of anterior segment by either using functional or removable appliances(Peavey and Kendrick,1976).

This agreed with the study of Bansode and Kolkarni (2009), who used in their study arch expansion technique and gave a result of definite increase in the length of rugae, but these teeth movements did not significantly alter their pattern.

On contrary, Shetty et al (2011) aimed to evaluate reliability of palatal rugae as forensic tool. They tried to assess stability of palatal rugae in particular ethnic group. Results showed that palatal rugae remain unchanged after orthodontic intervention.

Parihar et al (2010) mentioned that median rugal region increased significantly in anteroposterior length with growth. These measurements were observed as being greater in

orthodontic treated cases. These results evaluated palatal rugae stability and individuality independent of sex and age.

Percentage of palatal rugae changes related to displacement of lateral end in anteroposterior and mediolateral and medial end in anterioposterior directions were 60%, 56% and 54% respectively. It means that more than half of our sample had these changes; in addition, all the extraction group casts shared the first and the second of them in our sample. That gave an idea about more spaces to be closed, so, more teeth movement that could be miss identified as rugae displacement. This agreed with Almeida et al (1995), where he considered extraction of two premolars creates more space for movement, which changes the lateral and medial rugae nearest to extracted premolars. In non extraction cases ,stable medial ends of rugae in m-l direction . lateral points appeared less stable. In addition, they pointed that no significant difference between type of treatment modality and its effects on changes resulted after orthodontics therapy. Furthermore, Vanderlanden (1978) said that lateral rugae move with teeth, unlikely, he was unable to support this due to his small sample. Peavey and Kinderik (1967) said that the closer the rugae are to teeth, the more prone they are to stretch in the same direction of associated teeth movement. The influence of orthodontic treatment was observed on positions of lateral rugae either in anteroposterior or mediolateral directions. Moyers et al (1988) reported that the distance between the functional occlusal plane and rugae increased overtime, which agreed with us, but this change is not easy to be attributed to occlusal plane or rugae itself.

In the study of Shukla et al (2011), the rugae points were unstable in the transverse direction with respect to the lateral points of the first rugae. These results concurred with Bailey et al (1996) who believed that extraction of the first premolars creates a large space for distal retraction of the maxillary anterior teeth, which affect the positions of the lateral points of the first rugae, thus, changing the transverse distance between them. In contrary to this, he observed stability of medial rugae points in transverse direction, which didnt agree with our results.

In our study 20% of the whole sample showed changes in medial rugae in transverse direction . But her in Shukla et al study they resulted that None of the medial points of the first rugae were affected for the transverse values. This finding is in congruence with previous studies by Housser (1989) who concluded that lateral edges of the rugae move forward with the migration of teeth in extraction cases but felt that the medial ruga points were unaffected. These results didnt agree with our study which showed changes in medial rugae points. Damstra et al (2009), corresponded with our results ,he suggested adding rapid maxillary expansion to fixed appliance therapy. This will cause changes in transverse measurements of medial ends of rugae.

Some investigators have evaluated the stability of the palatal rugae during orthodontic treatment. In patients treated with a functional appliance or headgear, the medial rugae points appeared more stable than the lateral rugae points. In the comparison of tooth movement measured by cephalometric superimposition relative to the rugae points on study models. T he medial points of the third palatal rugae are suggested to be suitable reference points for the assessment of anteroposterior tooth movement. Conversely, another study reported that the lateral points of the

third rugae are the most stable. Thus, identification of the most stable area is still controversial (Abdel-Aziz and Sabet, 2001;Hogan and Sadowsky,2001; Almeida et al, 1995).

Simmons et al. (1987) found that the medial rugae were unstable reference points. (Christov and Kiliaridis, 2008) investigated medial rugae in both transverse and anterioposterior directions and observed their change over time. In contrast, Lebret (1962) reported that the distances between the rugae landmarks were almost constant, especially for rugae points near the median raphe. Peavy and Kendrick (1967) and Almeida et al. (1995) found that the lateral ends of the rugae were greatly affected by movement of the teeth, but the medial rugae appeared to be a suitable anatomic landmark for the construction of stable reference points for longitudinal cast analysis (Devreese et al, 2007).

Several studies in the literature observed displacements occurred in rugae. Many of them ascertained their stability. They considered changes due to teeth movement and migration ,or changing in arch circumference increasing or decreasing. Other studies tried to postpone taking post treatment casts enough time after completing their treatments. It was based on their theory that palatal soft tissues are stretched during treatment and changed in dimensions. Relapse will take place after a while and return back to their normal position as if before starting treatment (Damstra et al,2009;Cotton,1987).

Changes related to prominence of rugae after treatment, 54% were changed in their size, whether increased or decreased and this doesnt agree with Anukool et al (2012) results, where

they observed no change in rugae size after treatment. On the other hand, it agreed with the results of Hopkinz and Harris (2011) that anterior rugae were narrowed but the distal ones were stretched. Taking whole criteria in general, Souza lima (1964) assessed pre and post treatment casts, and concluded that no change in morphology or arrangement of palatal rugae occurred.

5.2 Individuality Test

The results of this test proved individuality of palatal rugae among Jordanian population. It showed no identical pairs when compared every with each other. Pre treatment and post treatment casts compared, which gave 1225 comparisons for each. These results were agreed with most studies that were concerned with the individuality. They ascertained the uniqueness of palatal rugae area despite of being similar in the same number of rugae. Their shapes and orientations differed from one person to another. Most of the literature agreed that the individuality of palatine rugae is sufficient for their use as a method of identification. This was supported by the fact that the morphology of palatine rugae is constant throughout life (Hemanth et al, 2010; Almedia et al, 1995). Patil et al (2011) considered palatal rugae as permanent and stable land marks that could be used for person discrimination in forensic identification. Having the same results, Shetty et al (2013), found the palatal rugae are characteristic enough to be used in discrimination.

Furthermore, (Bansode and Kulkarni, 2009; Ernence and Rener 1999; Elfotoh et al, 1998; English et al, 1988) cohered their results with others where they ascertained the use of palatal rugae pattern for use in human identification in forensic odontology.

Regarding Jordanian population, one study was done to prove individuality of palatal rugae and ascertained their use in forensic identification was done by (Mostafa et al, 2012).the results of their study revealed individuality of palatal rugae area in people without considering any changes might inflect on them. On the other hand our results proved individuality not only regarding pre treatment casts. It ascertained the individuality of post treatment casts despite changes induced by orthodontic treatment.

5.3. Matching Test

In the present study when matching between pretreatment and post treatment casts, the results showed significant difference between the trials of pre orthodontics with post orthodontics casts. While matching post treatment casts with those duplicated post treatment casts yielded statistically insignificant results. These results showed that changes occurred after treatment decreased matching efficiency and affected percentage of right matches. On the other hand, the results with duplicated casts showed higher percent of correct matches attributed to high accuracy level they kept. which increased levels of correct matching ability. Our results in this parts declared that morphological changes of palatal rugae during orthodontic treatment decreased matching efficiency .Therefore, hampered human identification.

In literature, most studies included matching trials between pre and post treatment casts, were done correctly without difficulties with almost 100%. Parihar et al (2010) noticed that all casts were matched in ease, thus, he considered them as unique and don't change. Stavirianos et al (2012) conducted his matching test with four evaluators got 100% of right matches and the fifth one 94% which is statistically in significant. Bansode and Kulkarni (2009) showed in their study results close from our study ranged from 76.6% to 96.6% with 13 evaluators participated.they concluded that despite of changes in rugae area during orthodontic treatment,the morphology of palatal rugae remains stable throughout life. Anukool et al (2011). In his study, argued that pretreatment and post treatment casts can be easily matched with each other quite easily, thus palatal rugae pattern are unique for individual and do not change. These results suggest the usefulness and easy reproducibility of comparing palatal rugae patterns for personal identification of cases. Intra-observer discrepancies might affect the easily of right matching, where the observer didnt make an effort to focus on rugae area after slight orthodontic changes, or didnt look for the whole pattern, instead of that he just took one part of rugae and looked for it, which can lead to mismatch and variations in their knowledge regarding oral anatomy and in their experience with forensic identification, However, some examiners had problems during matching, which may be due to complex rugae patterns, overlapping of rugae, and poorly demarcated rugae (Bansodeand Kulkarni, 2009). Apart from problems of intra-observer discrepancies in reading rugae patterns, there is no doubt that even greater discrepancies could exist between observers e.g. differences in forensic experience. The existence of this unreliability brings into question the present usefulness of descriptive rugoscopy in fields such as forensic science. Although researchers have confirmed the

potential value of rugae in personal identification. It is important that exact reproductions of patterns (for example, casts or photographs) are available and being updated moreover to depend on them accurately. Furthermore, classification systems are refined so they are reliable but relatively simple to apply.

Chapter Six : Conclusion

This chapter aimed at summarizing the important points in this thesis. It is considered an important study because it's one of the empirical studies, which studied changes occurred due to orthodontic treatment including different criteria, e.g., segmentation, union as expression for the change in number, change in orientation, change in shape, change in length, change in volume,

displacement of lateral end (in anteroposterior and mediolateral direction)and displacement in medial end (in anterioposterior and mediolateral direction)also. Furthermore, it is the first study concerned all these morphometric changes together.

In addition, the other goal of this study was to further test the individuality of palatal rugae and their value as a tool in individual identification. This goal was achieved by analyzing the morphometric changes that take place in palatal rugae patterns as a result of orthodontic treatment. Moreover, the researcher examined whether such changes will compromise our ability to match the same palatal rugae pattern before and after orthodontic treatment.

This study dealt with changes that occurred to palatal rugae area through orthodontic treatments. Orthodontic treatment and tooth movement have a significant effect on the stability of Palatal rugae as investigated by previous researchers. Many studies in literature have found certain landmarks on the rugae to be stable reference points for measuring tooth movements during orthodontic treatment, but still there are controversial about this in literature.

The rugae morphological pattern may be useful in forensic science in case of mutilation when compared with other parts of the body. Palatal rugae form an intrinsic and integral pattern for every single individual and can also help in sex determination. The ease of reproducibility andlower level of variation makes palatal rugae a potential toolin forensic odontology.

Morphometric changes which could occur as a result of orthodontic changes were studied. These changes affected matching reliability. This justified the need for more updated antimortem

records when identification is needed. However, other methods of recording the rugae pattern are possible for identification purpose, which might include photographs, palatal prints or impressions, or computerized tomography of the rugae pattern. In addition, individuality of palatal rugae in Jordanian population was assessed by comparing all pre orthodontics casts with each other which gave us 1225 comparisons, which agreed with all studies for different population in literature. The study reports that orthodontic treatment inflicts changes on palatal rugae. Such changes can potentially compromise human identification in cases where palatal rugae records are not updated. The study continues to suggest that palatal rugae patterns are highly individual. Even after orthodontic treatment induced changes, palatal rugae patterns remain individualized. Thus, they hold potential as a supplementary tool, along with the dentition, to establish the identity of an individual, so they considered as a reliable guide in forensic identification when updated records are available.

Although this study is very important in literature, it is limited by using a small sample which is 50 pre and 50 post casts, in addition, it is limited by the contribution of evaluators with varying forensic experience. Nevertheless, in the light of previous results the researcher recommends that larger samples should be examined in detail to further validate the findings of this type of study and come to definitive conclusions. Likewise, further studies need to be researched on varying population in different areas.

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Appendix A
Changes occurred post orthodontic treatment: Segmentation

Pre treatment cast without segmentation.

Post treatment cast with segmentation in one of the rugae .

Appendix B
Changes in post orthodontics treatment: Union

Pre treatment cast with rugae that is separated from each other.

Post treatment casts with these rugae are united together and become one rugae.

Appendix C
Changes in post orthodontic treatment: Shape

Pre treatment cast with straight shape rugae.

Post treatment cast with the same rugae that is being curved.

Appendix D
Changes in post orthodontic treatment: Orientation

Pretreatment cast posteriorly oriented rugae.

Post treatment cast with anteriorly oriented rugae.

Appendix E
Changes in post orthodontic treatment: Length

Pre treatment cast with this length.

Post treatment cast with decreased length of rugae.

Appendix F
Change in post orthodontic treatment: Displacement of lateral rugae A-P

Pretreatment cast with lateral rugae placed.

Post treatment cast with more anteriorly placed lateral rugae

Appendix G
Changes in post orthodontic treatment: Displacement of lateral rugae M-L

Pretreatment cast with lateral rugae placed .

Post treatment cast with lateral rugae are more medially displaced.

Appendix H
Change in post orthodontic treatment: Displacement of medial rugae A-P

Pre treatment cast with these dimensions of palatal rugae

Post treatment cast with medial rugae are more anteriorly displaced

Appendix I
Change in post orthodontic treatment: Displacement of medial rugae M-L

Pretreatment cast of palatal rugae.

Post treatment cast with medial rugae are more laterally displaced.

Appendix J
Changes in post orthodontic treatment: Prominence

Pretreatment cast with a good prominent rugae .

Post treatment cast with decreased prominence of rugae.