LINE-1 methylation in cleft lip tissues:influence of infant MTHFR c.677C>T genotype

Objective: To investigate the influence of MTHFR c.677C>T genotype on LINE-1 methylation in lateral and medial tissues from cleft lip (CL). Methods: Forty-five consecutive non-syndromic cleft lip with or without cleft palate (nsCL/P) cases were included in the study. Genomic DNA was extracted from tissues at both sides of cleft lip, and LINE-1 methylation was detected by bisulfite conversion and pyrosequencing. MTHFR c.677C>T genotyping was carried out using the TaqMan genotyping assay. Results: LINE-1 methylation level was significantly higher on medial side of cleft lip compared with lateral side (p = 0.001). This difference was not significantly influenced by the case's sex or cleft type. However, MTHFR c.677C>T genotyping revealed that the difference in LINE-1 methylation across cleft lip was restricted to carriers of C allele of MTHFR c.677C>T and was not apparent in TT homozygous cases (p = 0.027). Conclusion: This integrated analysis supports the previous finding of differences in DNA methylation across the two sides of cleft lip and further suggests a possible role of MTHFR c.677C>T genotype in establishing this difference

Ultrastructural analysis of collagen fibril diameter distribution in cleft lip

Objective: A preliminary study to determine collagen fibril diameter (CF-ED) distribution on medial and lateral sides of cleft lip (CL). Material and Methods: Tissue samples from medial and lateral sides of CL were fixed in 2.5% glutaraldehyde and 1% osmium tetroxide and embedded in Araldite CY212 resin for transmission electron microscopy. The analysis of CF-ED was performed using the ImageJ program. To characterize the packaging of collagen fibrils (CFs) in the two tissues, we estimated the collagen number density (CF-ND) and fibril-area-fraction (FAF). Differences in measurements across the two sides were calculated using Wilcoxon signed-rank test. Results: The CF-ED was statistically significantly (p < 0.001) smaller on the medial side (45.69 ± 7.89 nm) than on the lateral side (54.18 ± 7.62 nm). The medial side had a higher CF-ND and a higher percentage of FAF than the lateral side. Conclusion: Our finding of a smaller CF-ED and higher CF-ND and FAF for the medial side suggests possible differences in size and distribution of CFs between medial and lateral sides of CL. This finding provides knowledge toward underlying tissue biomechanics that may help reconstruction of perioral tissue scaffolds, ultimately resulting in better treatment of patients with oral clefts

Early Secondary Gingivo-Alveolo-Plasty in the treatment of unilateral cleft lip and palate patients: 20 years experience

Since 1988, the Milano surgical protocol has included lip, nose and soft palate repair at 6-9 months of age and closure of the hard palate at 18-36 months together with an Early Secondary Gingivo-Alveolo-Plasty (ESGAP). Prior to 1988 the alveolar cleft was repaired in a third step by bone grafting at 9-11 years of age. The goal of this study was to evaluate the long-term maxillary growth in unilateral cleft lip and palate (UCLP) patients who have undergone ESGAP. Alveolar ossification after ESGAP was also studied in the permanent dentition. Growth assessment was carried out comparing lateral X-ray cephalograms of a UCLP ESGAP sample (15 consecutive patients with a mean age of 18.2 +/-1.2 years) and of the UCLP bone graft sample (10 consecutive patients with a mean age of 18.7 +/-1.1 years) and a sample from the Oslo cleft lip and palate (CLP) centre sample (15 patients with a mean age of 18.1 +/-0.8 years). Alveolar ossification in the sample which had undergone ESGAP was evaluated through a sample of panoramic X-rays of UCLP in the permanent dentition. Alveolar bridging was assessed using a modified Bergland's scoring system. From the results it seems that ESGAP allows for an excellent alveolar ossification, but patients show an inhibition of maxillary growth compared with the secondary bone graft group and with the Oslo group. Although ESGAP allows for early repair of the alveolus together with palate repair, thus eliminating in all patients the need for secondary bone grafting, it seems to have an inhibiting influence on maxillary growth which increases the need for Le Fort I osteotomies. It is however fair to recall, that, even with an adjunctive Le Fort I osteotomy, ESGAP enables the total number of operations to be reduced to three, instead of most European protocols (4-5 procedures

Introduction to Surgical Navigation in Oral Surgery: A Case-Series

The application of surgical navigation in oral and maxillo-facial surgery has been increasing over time. In fact, computer-assisted surgery provides real-time, precise, and accurate position and guidance during surgery. The purpose of our work is to introduce the evolution of surgical navigation in recent decades, describe some technical aspects of this technology, explore new possibilities of application of surgical navigation in oral surgery, and validate the accuracy of computer-assisted surgery. We included four patients in our sample who underwent virtual planning on the cone beam CT data set and surgical navigation using non-invasive fiducial markers. The first patient presented a dislocated orthodontic arch in the soft tissues of the cheek, while the other patients presented supernumerary and impacted dental elements. Among them, two patients were affected by craniofacial synostosis. We evaluated the accuracy of computer-assisted surgery, calculating the discrepancy between the real and virtual target. In all cases, the target registration error was less than or equal to 1 mm. We can affirm that surgical navigation is a valid tool to enhance oral surgery, guaranteeing an undoubted advantage in terms of the reliability and predictability of the results, especially in complex cases

A cephalometric intercenter comparison of patients with unilateral cleft lip and palate: Analysis at 5 and 10 years of age and long term

Objective: To compare the short- and long-term craniofacial growth of patients operated with the Milan protocol to those operated with the Oslo protocol. Design: The Milan sample included 88 patients with unilateral cleft lip and palate (UCLP) at 5 years, 26 at 10 years, and 23 at the end of growth. The Oslo samples included 48 UCLP patients at 5 years, 29 at 10, and 23 at growth completion. Lateral cephalograms were used for comparison. An unpaired t test was run for the 5- and 10-year-old samples. The samples long term were matched for age and sex, and a paired t test was run. Results: There was no significant cephalometric difference in the maxillary prominence at 5 years, a mild but significant difference at 10 years, and again no difference at the end of growth. Nevertheless, at an older age, the need for orthognathic surgery was larger in the Milan sample (26%) than in the Oslo sample (13%). Conclusion: Although no statistically significant differences in the cephalometric measurements were found long term, the need for orthognathic surgery was clinically judged to be larger in the Milan sample. On the other hand, although the Milan protocol seemed to require more final jaw surgery, only the cases that needed an additional orthognathic procedure in the Milan group will undertake a third surgical step, while the Oslo protocol included three surgical steps for all the patients