Cleft Site Research Articles

Evaluation of the Regenerative Capacity of Demineralized Bone Matrix vs Fat Graft in Alveolar Cleft Model in Albino Rats.

This study was performed to evaluate the regenerative capacity of demineralized bone matrix vs fat graft, both guided by pericardium membrane in alveolar cleft model in albino rats. A total of 72 rats were required in this study. A surgical bone defect with a 7 mm length × 4 mm width × 3 mm depth was created as a model of an alveolar cleft, then the rats were divided randomly into four equal groups each group contained 18 rats: control group (defect only), the membrane group (the defect was covered by the pericardium membrane), the demineralized bone matrix (DBM) group (the defect was filled with DBM guided by pericardium membrane) and fat group (the defect was filled with a fat graft guided by the pericardium membrane). Around 6 rats from each group were euthanized after 2, 4, and 8 weeks. Skulls were scanned with cone beam computed tomography (CBCT) and harvested for histological evaluation with routine H&E immunohistochemical stains (Anti-osteocalcin and Anti-Wnt5a). The data was recorded and statistically analyzed by a two-way ANOVA. The study showed a notable formation of new bone, and expression of OCN and Wnt5a were notably increased by time in the fat group. However, the density of bone grafts and OCN and Wnt5a expression decreased with time in the DBM group. Control and membrane groups showed negative OCN and Wnt5a immune-reactivity in the cleft site. Fat graft results were superior to DBM results with regard to mucosal closure and accelerated bone regeneration, and may represent an effective treatment for alveolar cleft reconstruction. Finding an inexpensive, accessible, biocompatible and easily manipulated treatment for craniofacial reconstruction and fat graft fulfilled the desired aims. Further investigations with prolonged evaluation periods are needed. How to cite this article: Abdel Raouf E, Elsherbini AM, Abdel Salam Yousef Y, et al. Evaluation of the Regenerative Capacity of Demineralized Bone Matrix vs Fat Graft in Alveolar Cleft Model in Albino Rats. J Contemp Dent Pract 2024;25(6):554-562.

Clinical outcomes of gingivoperiosteoplasty for unilateral cleft lip and palate performed in early childhood

Conventional gingivoperiosteoplasty (GPP) performed during infancy adversely affects maxillary development. However, the outcomes of this procedure in early childhood have rarely been reported. Therefore, we examined the postoperative outcomes of GPP conducted in patients aged 1.5 years with unilateral cleft lip and palate (UCLP).This study included 87 non-syndromic patients with complete UCLP who had undergone early two-stage palatoplasty during the 1999–2004 period. The protocol comprised soft palate plasty at 1 year of age and hard palate closure at 1.5 years of age. In the GPP group (n=34), we introduced the GPP procedure during hard palate closure; in the non-GPP group (n=53), the labial side of the alveolar cleft remained intact. We examined computed tomography images taken at 8 years of age to observe bone formation at the alveolar cleft site. We also conducted cephalometric analysis to examine maxillary development at 12 years of age.Bone bridges at the alveolar cleft site were observed in 92% and 5.6% of the GPP and non-GPP groups, respectively. Moreover, 59% of the GPP group did not require secondary alveolar bone grafting (sABG), whereas all the patients in the non-GPP group underwent sABG. No statistically significant differences were noted in the maxillary anteroposterior length (GPP: 45.5±3.7mm, non-GPP: 45.9±3.5mm, p=0.67) and sella–nasion–point A angle (GPP: 75.6±4.5°, non-GPP: 73.8±12.6°, p=0.49).This study’s findings suggest that GPP performed at 1.5 years of age minimises the necessity of sABG and does not exert a negative influence on maxillofacial development.

Cortical Block Grafting Successfully Augments Alveolar Cleft Sites for Dental Implant Placement

BackgroundSupplemental bone grafting is regularly required before dental implant placement in patients with cleft lip and palate (CLP). PurposeThe study purpose was to measure and compare implant osseointegration and changes in graft dimensions following lateral incisor onlay cortical bone grafting in CLP and non-CLP patients. Study design, setting, sampleRetrospective cohort study composed of patients who presented to Boston Children’s Hospital and underwent autogenous onlay cortical block bone grafting at lateral incisor sites from 2015 through 2023. Patients were excluded if the cone beam computed tomography (CBCT) quality was insufficient for accurate measurements. Predictor variableThe predictor variable was CLP status coded as CLP or non-CLP. Main outcome variableThe primary outcome variable was successful implant osseointegration confirmed by a torque of 35 N/cm or more after 3 months of implant healing. Secondary outcomes were change in bone width between pre- and post-operative CBCT scans at lateral incisor sites and the need for additional bone augmentation prior to or during implant placement. CovariatesCovariates were age, sex, cleft location, and time from bone graft to postoperative CBCT and implant placement. AnalysesData analyses were performed using T-tests, Fisher’s Exact Tests, Mann-Whitney U Tests, and Pearson’s correlation. P<0.05 was considered statistically significant. ResultsA total of 22 subjects (16 with CLP) were evaluated. The mean age at the time of graft was 19.3 ± 2.4 years with 52.6% males. Implants osseointegrated at 20 of 22 lateral incisor sites (one CLP failure, one non-CLP failure). There was significant change in bone width after grafting for patients with CLP (p<0.001). Patients with CLP experienced a 3.32 (± 1.80) mm and 2.99 (± 1.61) mm increase in bone width at two different levels. Patients with CLP achieved greater boney changes near the alveolar crest than non-cleft patients (p=0.008) but the change was not significantly different more apically (p=0.86). One subject with CLP required additional grafting during implant placement. Conclusions and RelevanceCortical block onlay bone grafting is a predictable technique to augment lateral incisor sites in patients with cleft lip and palate for placement of a dental implant.

Three-dimensional Analysis of Maxillary Morphology in Infants with Unilateral Cleft Lip and Palate.

To three-dimensionally (3D) analyze the maxillary morphology of infants with unilateral cleft lip and palate (UCLP) and preliminarily classify the alveolar arch to assist in personalization of sequence therapy. Retrospective study. Patients with UCLP referred to outpatients' clinic. 84 nonsyndromic infants with complete UCLP were recruited (58 boys, 26 girls, mean age 29.48 days). Morphometric analysis was conducted on 3D maxillary models. Principal component analysis (PCA) and cluster analysis were combined to classify maxillary phenotypes preliminarily. The Wilcoxon Signed Rank test and the Kruskal-Wallis test were used to compare differences between variables. A P value less than .05 was considered statistically significant. The maxilla was divided into three types: narrow, homogenous and broad, accounting for 9.52%, 23.81% and 66.67% respectively. The alveolar cleft site (median value) was located in 61% of the total length of the alveolar arch. In the comparison of anterior and total alveolar lengths, the non-cleft side had longer alveolar bone than the affected side, a difference of approximately 2 mm. Pairwise comparisons of variables describing alveolar symmetry revealed significant differences in all subjects; whereas type C had poorer arch symmetry than types A and B, mainly in terms of anterior and overall symmetry. In infants with UCLP, the maxillary alveolar arch was inherently asymmetrical with partially bone missing (about 2 mm). Significant differences in alveolar bone morphology and symmetry exist between different types of infants, with individuals with broad clefts (type C, the largest proportion) having the worst maxillary development.

Mandibular canine development as an indicator of maxillary cleft site canine development

Objectives: The objective of this study was to determine if mandibular canine development is an indicator of maxillary cleft site canine development. Material and Methods: A cross-sectional chart review with analysis of cone-beam computed tomography (CBCT) imaging of patients aged 7–12 with a unilateral complete cleft lip and palate who received alveolar bone graft surgery from January 2009 to January 2019. Three-dimensional (3D) measurements of the cleft site canine and ipsilateral mandibular canine were taken. A two-dimensional image was created from the 3D CBCT to simulate a traditional panoramic radiograph. Descriptive statistics and multivariable linear regression analyses were conducted, adjusting for age and gender. Pearson and Spearman correlations were calculated. The main outcome measure was the mandibular canine crown-to-root ratio (C:RR) compared to that of the maxillary cleft site canine C:RR. Results: Five hundred and thirteen patients were reviewed, of which 78 met the inclusion criteria. Adjusting for age and gender, every unit change in mandibular canine C:RR corresponds to 0.69 (95% confidence interval [CI] = 0.42–0.95) change in maxillary canine C:RR, up to a mandibular canine C:RR of 3. Beyond a mandibular C:RR of 3, no change in the maxillary canine C:RR was observed (−0.02; 95%CI = −0.27–0.24). Pearson correlation coefficient was r = 0.65, and the Spearman correlation was P = 0.75. Conclusion: Mandibular canine C:RR is correlated to maxillary cleft site canine C:RR when the mandibular canine C:RR is ≤3.

Radiographic Assessment of Cleft Alveolar Bone Defects: A Preliminary Comparison of Periapical and Cone Beam Computed Tomography Images

Background &amp; Purpose Accurate radiographic assessment of secondary alveolar bone grafting (SABG) is critical in evaluating SABG outcomes in patients with cleft lip and palate. This study compared Periapical (PA) and Cone beam computed tomography (CBCT) grading of SABG outcomes and conducted an analysis of agreement rate between the two imaging modalities. Methods A retrospective chart review was performed of patients who underwent secondary alveolar bone grafting at a single institution. Bone quality of the alveolar cleft site was assessed at three root levels of the adjacent teeth on PA and CBCT images. Results 48 patients had pre- or post-operative images with both CBCT and PA that were appropriate for comparison. A total of 174 alveolar bone sections (three root levels of adjacent teeth in 58 pairs) were graded on CBCT and PA images. Agreement of CBCT grading with PA grading was 41.3% (72/174) overall. CBCT assessments rated bone quality lower than PA grading in 51.2% (89/174) of sections. CBCT had higher grading than PA image grading in 7.5% (13/174) of sections. The weighted Cohen Kappa value for comparison of CBCT and PA readings was 0.17, indicating a low rate of agreement. Wilcoxon signed rank test demonstrated that the difference between grading on CBCT and PA assessments was statistically significant ( P &lt; .001). Conclusions CBCT and PA readings demonstrated an overall low rate of agreement in the assessment of cleft alveolar bone. This investigation suggests that PA radiographs have lower sensitivity in detecting poor bone grafting outcomes.

What is the Success Rate of Repeat Alveolar Bone Graft?

Success rates for alveolar bone grafting range from 30 to 96%. There is limited information regarding the success of repeat grafts. The purpose of this study was to determine the radiographic success rate of repeat alveolar bone grafts. The study designs was a retrospective cohort study of patients who underwent repeat grafting by 1 surgeon over 15years. To be included, subjects had to have: cleft lip and alveolus and a cone-beam computed tomography (CBCT) scan obtained >6months after repeat graft. Patients were excluded if CBCT was inadequate. Predictor variables were sex, age at repeat graft, cleft type, presence of an erupted canine, premaxillary osteotomy at time of repeat graft, presence of a visible oronasal fistula, size of bony defect, presence of a bony palatal bridge, and whether the surgeon who performed the repeat graft also performed the initial graft. The outcome variable was graft success determined using CBCT assessment and defined as a score of >3 out of 4 in each domain: vertical bone level, labiopalatal thickness, and piriform symmetry. The covariates were time from bone graft to CBCT (months) and age at time of CBCT (years). Frequency distributions, relative risk with 95% confidence intervals, medians, and interquartile ranges were calculated. Pearson c2 and Fisher exact tests were performed to determine predictors of outcome. A P<.05 was considered statistically significant. Fifty subjects (54% male) who had repeat bone grafting to 59 cleft sites were included. Median follow-up time from repeat graft to CBCT was 7.0months (interquartile range: 5.9months). The radiographic success rate was 81.4%:91.7% if the same surgeon performed both initial and repeat grafts, but 78.7% if initial graft was completed by another surgeon (P=.43). Despite being a multifactorial issue, extensive and bilateral clefts, the presence of an erupted tooth in the cleft area, a visible oronasal fistula, and concomitant osteotomy of the premaxilla are warning signs of the possibility of failure. Performing repeat alveolar bone grafting by an experienced surgeon appears to increase the chance of success.

Evaluating SWAG and Its Validity When Compared to 3D Imagery of Secondarily Grafted Cleft Sites.

To test validity of 2D Standardized Way to Assess Grafts (SWAG) ratings to assess 3D outcomes of bone grafting (ABG). 43 patients (34 UCLP, 9 BCLP) with non-syndromic complete clefts, bone-grafted at mean age 9yrs/3mos, with available post-graft occlusal radiographs and cone beam computed tomography (CBCT) (taken mean 4yrs/9mos post-ABG). 2D occlusal radiographs rated twice using SWAG by 6 calibrated raters. 12 scores were averaged and converted to a percentage reflecting bone-fill. Weighted Kappas were assessed for SWAG reliability. 3D cleft-site bone volume was calculated by 1 rater using ITK-SNAP. 13 cleft sites were re-measured by the 'one rater' for 3D reliability using Intraclass Correlation Coefficient (ICC). 2D versus 3D ratings were compared using paired t-test, independent samples t-test, Bland-Altman and Linear Regression. Significance level was P = .5. 2D reliability was 0.724 (intra-rater) and 0.546 (inter-rater). 3D reliability was 0.986. Bland-Altman plot comparing 2D vs 3D showed for 45 of 47 graft-sites were within 2 SD's. Mean % bone-fill was 64.11% with 2D and 69.06% with 3D (mean difference = 4.95%) that was a non-significant difference in both t-tests. Regression showed a statistically significant relation between the two methods (r2 = 0.46; P = .0001). 2D SWAG systematically and non-significantly underestimated bone-fill. There was a significant correlation between 2D/3D methods. Bland-Altman analysis illustrated the similarity of the two methods. For comparisons of group (cleft treatment Centers') bone grafting outcomes, the 2D method may suffice as a proxy for the 3D method. However, with individual variation up to 40% in 2D estimates of actual 3D volume, 2D SWAG method cannot be used in place of 3D images.

Structural profile of ligand-based inhibition of bacterial FtsZ

Aim. The idea of the study was to compare and generalize RCSB Protein Data Bank and ChEMBL data in order to establish the structural and biological relationship of experimentaly proved effectors of FtsZ with binding sites. Methods. Literature and database search. Comparison of protein and ligand structures. Protein structure modeling, MD, structural superimposition, etc. Results. The experimental protein-ligand complexes structures of bacterial FtsZ were revised. The structural superimposition of experinental PDB and full-atomic AlphaFold2 models of bacterial FtsZs confirmed their significant structural similarity. Three protein-ligand binding sites were identified by structural alignment. The rating based on database (RCSB Protein Data Bank, ChEMBL, DrugBank, BindingDB, PubChem), patente and literature information on FtsZ-ligand interactions identify perspective sites and main reference compounds. Сonclusions. It was identifyd 3 main protein-ligand binding regions in FtsZ: I. Nucleotide Binding Domain (Ia. Site of GTP/GDP and Ib. MB3 site); II. Site of inter-domain cleft (IDC) and III. Site of coumarin bindig (4HC = 4-hydroxycoumarin). It was indicated that benzamide-binding site, located in the region of inter-domain cleft of FtsZ, demonstrate highest site- and target-specificity.

Predictors of Outcomes in 900 Alveolar Bone Grafts.

Significant discrepancies exist in the reported variables influencing alveolar bone graft outcomes. The purpose of this study was to evaluate graft success and identify outcome predictors in a large patient cohort using an objective cone beam computed tomography assessment tool. Consecutive patients with cleft lip/palate who underwent alveolar bone grafting by 1 surgeon were included. Predictor variables were age at graft, oronasal fistula, canine position, concurrent premaxillary osteotomy, size of cleft, presence of bony palatal bridge, history of failed graft, location of primary repair, and surgeon experience. The outcome variable was graft success, determined using a cone beam computed tomography assessment tool and defined as a score of 3 or 4 (out of 4) in the following domains: vertical bone level, labiopalatal thickness, and nasal piriform symmetry. The sample included 900 alveolar cleft sites (median graft age, 9.9 years). The success rate was 94.6%. Presence of an erupted canine, large cleft defect, or premaxillary osteotomy were independent predictors of graft failure; presence of a bony palatal bridge was associated with graft success ( P < 0.05). Presence of an erupted canine, large bony defect, or premaxillary osteotomy increase the risk of failure, and a bony palatal bridge portends success. Age 12 years or older, visible oronasal fistula, history of failed graft, primary cleft repaired at outside institution, and surgeon experience were associated with higher graft failure, but were not independent predictors when controlling for covariates. Surgeons should be aware that these factors in combination increase the odds of graft failure. Risk, III.

Outcomes of alveolar cleft repair in complete cleft lip and palate after tertiary gingivoperiosteoplasty and segmental Le Fort I osteotomy.

Gingivoperiosteoplasty is often used for reconstruction of alveolar defects in infants with cleft lip and palate. This study aimed to examine outcomes of tertiary gingivoperiosteoplasty, which has not previously been investigated. This prospective study included 11 adults with complete cleft lip and palate (n = 12 sites) who consecutively underwent segmental Le Fort I osteotomy and concomitant gingivoperiosteoplasty for correction of skeletal class III deformity, nasoalveolar fistula and alveolar cleft. Outcomes included clinical and radiographic evaluations of gingivoperiosteoplasty at the time of osteotomy (presence of nasoalveolar fistula, residual cleft defect and unsupported root ratio of cleft-adjacent teeth), and determination of influencing factors for the clinical success of alveolar cleft repair. Study variables included age, gender, pre-surgical orthodontic treatment and alveolar cleft width of cleft-adjacent canine and angulation between cleft-adjacent teeth before surgery and 1-week postsurgery. Posttreatment, no nasoalveolar fistula remained. The residual cleft defect decreased significantly (p < 0.01). The unsupported root ratio of cleft-adjacent teeth did not differ (p > 0.05); eight cleft sites reached Bergland I or II (67% success). One-week postsurgery, the minimal alveolar cleft width of cleft-adjacent canine was significantly less in the success group compared with the failed group (p = 0.01). Tertiary gingivoperiosteoplasty and segmental Le Fort I osteotomy decreased nasoalveolar fistulas and induced alveolar bone formation. The minimal alveolar cleft width immediately after surgery was the major influencing factor of clinical success. Segmental Le Fort I osteotomy with simultaneous gingivoperiosteoplasty efficaciously repairs adult alveolar clefts.

Autologous Tooth Transplantation in Craniofacial Malformations.

To evaluate the applicability of transplanted teeth in young patients with craniofacial anomalies. Observational study. Comprehensive Centre for Cleft Palate and Craniofacial Malformations. Patients with craniofacial anomalies who underwent tooth transplantation. Only children with complete clinical and radiological documentation and a follow-up period of at least 1.5 years were included. Tooth transplantation. Retrospective evaluation of clinical records, pre- and postoperative radiographs, and operative charts. Clinical characteristics of patients, preoperative parameters and postoperative outcome parameters were collected. A total of 17 patients with 23 tooth transplantations were included. The median follow-up period was 6.7 years. The pooled survival and success rates were 91%. Notably, one out of two teeth that were transplanted into the bone grafted alveolar cleft site had to be extracted, which might indicating a higher risk for this procedure. In total, two transplanted teeth had to be extracted during the follow-up period, one due to external resorption and the other one due to perio-endo lesion. One patient needed endodontic treatment due to pulp necrosis. We consider tooth transplantation to be a reliable and suitable procedure in the dental rehabilitation of young patients with craniofacial anomalies and fitting concomitant circumstances. We encourage craniofacial teams to reconsider this option more frequently in appropriate cases.

Characterizing the microbiota of cleft lip and palate patients: a comprehensive review.

Orofacial cleft disorders, including cleft lip and/or palate (CL/P), are one of the most frequently-occurring congenital disorders worldwide. The health issues of patients with CL/P encompass far more than just their anatomic anomaly, as patients with CL/P are prone to having a high incidence of infectious diseases. While it has been previously established that the oral microbiome of patients with CL/P differs from that of unaffected patients, the exact nature of this variance, including the relevant bacterial species, has not been fully elucidated; likewise, examination of anatomic locations besides the cleft site has been neglected. Here, we intended to provide a comprehensive review to highlight the significant microbiota differences between CL/P patients and healthy subjects in various anatomic locations, including the teeth inside and adjacent to the cleft, oral cavity, nasal cavity, pharynx, and ear, as well as bodily fluids, secretions, and excretions. A number of bacterial and fungal species that have been proven to be pathogenic were found to be prevalently and/or specifically detected in CL/P patients, which can benefit the development of CL/P-specific microbiota management strategies.

Comparison of Alveolar Bone Grafting Outcomes using CBCT in Individuals with UCLP Based on the Presurgical Orthodontic Treatment Methods.

The purpose is to evaluate outcomes of alveolar bone grafting based on the pre-grafting orthodontic preparation methods. Retrospective analysis of individuals with unilateral cleft lip and palate. 28 individuals with non-syndromic UCLP from two craniofacial centers, 14 individuals each from XXXX and XXXX. The alignment group underwent maxillary expansion with incisors alignment while the non-alignment group underwent only maxillary expansion for presurgical orthodontic preparation. Initial and post-surgical CBCT scans were compared to observe changes in angulation of the incisor adjacent to the cleft site, alveolar bony root coverage, and bone graft outcomes. In the alignment group, the buccolingual rotation decreased by 32.35 degrees (p = .0002), the anteroposterior inclination increased by 14.01 degrees (p = .0004), and the mesiodistal angulation decreased by 17.88 degrees (p = .0001). Alveolar bony coverage did not change after bone graft in both groups, and no difference was observed between the groups. Chelsea scale showed satisfactory bone graft outcome (category A, C) in 12 cases (85.71%) in the alignment group and 11 cases (78.51%) in the non-alignment group. The volumetric measurement showed the alignment group had better bone fill of 69.85% versus 51.45% in the non-alignment group (p = .0495). Alveolar bony coverage on the tooth adjacent to cleft sites did not change with alveolar bone grafting surgery in either of the alignment and non-alignment group. Presurgical orthodontic alignment does not induce root exposure nor poorer bone grafting outcome.

Prevalence of Dental Anomalies in Taiwanese Children with Cleft Lip and Cleft Palate

The aim of this study was to investigate the prevalence of tooth agenesis, tooth malformation, and eruption patterns of upper canines/first premolars in Taiwanese children. A total of 132 cleft lip and cleft palate (CLCP) patients (82 boys and 50 girls) underwent alveolar bone grafting (ABG) between 2012 and 2022. The patients’ dental records and X-ray images were inspected. We examined dental anomalies, including congenital missing teeth, microdontia, and transposition from the upper canines to the upper first premolars in these CLCP patients. Additionally, we investigated the mean ABG operation age (9.27 ± 0.76 years) of our patient; 40.9% of them received pre-ABG orthodontic treatment at 8.72 ± 0.70 years. Among the 132 cleft subjects, the prevalence of tooth agenesis is 73.5% (97/132). The most frequently missing teeth are the maxillary lateral incisors (right side: 46.2%; left side: 47.0%). In this study, microdontia are found in all the upper incisors, of which the highest percentage (18.9%) is observed in the upper left lateral incisors. The prevalence of upper canine and first premolar transposition is 10.6%. The pattern of tooth agenesis and microdontia of the upper lateral incisors shows a strong correlation with the cleft sites of these CLCP patients in our study. These results may support the idea that the patterns of dental anomalies in CLCP patients are region-specific.

Advanced simulations and screening to repurposing a 3C protease inhibitor against the rupintrivir-resistant human norovirus-induced gastroenteritis

Human norovirus (HuNoV) causes acute viral gastroenteritis in all age groups, and dehydration and severe diarrhea in the elderly. The World Health Organization reports ∼1.45 million deaths from acute gastroenteritis annually in the world. Rupintrivir, an inhibitory medicine against the human rhinovirus C3 protease, has been reported to inhibit HuNoV 3C protease. However, several HuNoV 3C protease mutations have been revealed to reduce the susceptibility of HuNoV to rupintrivir. The structural details behind rupintrivir-resistance of these single-point mutations (A105V and I109V) are not still clear. Hence, in this study, a combination of computational techniques were used to determine the rupintrivir-resistance mechanism and to propose an inhibitor against wild-type and mutant HuNoV 3C protease through structure-based virtual screening. Dynamic structural results indicated the unstable binding of rupintrivir at the cleft binding site of the wild-type and mutant 3C proteases, leading to its detachment. Our findings presented that the domain II of the HuNoV 3C protease had a critical role in binding of inhibitory molecules. Binding energy computations, steered molecular dynamics and umbrella sampling simulations confirmed that amentoflavone, the novel suggested inhibitor, strongly binds to the cleft site of all protease models and has a good structural stability in the complex system along the molecular dynamic simulations. Our in silico study proposed the selected compound as a potential inhibitor against the HuNoV 3C protease. However, additional experimental and clinical studies are required to corroborate the therapeutic efficacy of the compound.

Psychological Distress and Related Factors among Parents having Children with Cleft Lip and Palate Disorder

Background: Raising a child with a cleft lip and or palate disorder (CLPD) brings many challenges to the parents. Having children with CLPD can be a source of parental psychological distress (PPD). Stress experienced by parents may affect the development of a child. This study was designed to assess the prevalence and factors associated with PPD among parents having children with cleft lip and palate attending a major dental hospital in Sri Lanka. Materials and Methods: Cross-sectional study was conducted in cleft lip and palate clinics of Dental Hospital, Peradeniya, Sri Lanka. PPD was assessed using the General Health Questionnaire-30. Site of cleft and associated disabilities were recorded on data recording form using clinical records of the children. Chi-square and multivariate logistic regression analysis were used to analyze the data. Results: Out of 384 parents who accompanied their children to the clinic, 62.5% were mothers. The most common cleft site of the patients was cleft involving lip and palate (32.3%) and the least common site was isolated cleft of the soft palate (8.3%). The most common disability reported was speech problems (41.4%). The prevalence of PPD was 34%. Chi-square statistics showed that PPD was significantly associated with age of the child (P &lt; 0.000), family income (P = 0.011), family structure (P &lt; 0.000), previous knowledge on CLPD (P = 0.045), site of the cleft (P &lt; 0.000), and disabilities associated with particular cleft (P &lt; 0.000). Multivariate logistic regression analysis demonstrated that age of the child, family structure, cleft of the hard and soft palate, feeding difficulties, and speech problems were significant predictors for PPD. Conclusion: PPD is an important factor to consider in managing children having CLPD. This finding provides valuable information for the provision of multidisciplinary approach to manage PPD.

Conformational stability of ageritin, a metal binding ribotoxin-like protein of fungal origin

Ageritin is a ribotoxin-like protein of biotechnological interest, belonging to a family of ribonucleases from edible mushrooms. Its enzymatic activity is explicated through the hydrolysis of a single phosphodiester bond, located in the sarcin/ricin loop of ribosomes. Unlike other ribotoxins, ageritin activity requires divalent cations (Zn2+). Here we investigated the conformational stability of ageritin in the pH range 4.0–7.4, using calorimetric and spectroscopic techniques. We observed a high protein thermal stability at all pHs with a denaturation temperature of 78 °C. At pH 5.0 we calculated a value of 36 kJ mol−1 for the unfolding Gibbs energy at 25 °C. We also analysed the thermodynamic and catalytic behaviour of S-pyridylethylated form, obtained by alkylating the single Cys18 residue, which is predicted to bind Zn2+. We show that this form possesses the same activity and structure of ageritin, but lower stability. In fact, the corresponding values of 52 °C and 14 kJ mol−1 were found. Conservation of activity is consistent with the location of alkylation site on the opposite site of the catalytic site cleft. Inasmuch as Cys18 is part of a structurally stabilizing zinc-binding site, disrupted by cysteine alkylation, our results point to an important role of metal ions in ageritin stability.

Neprosin belongs to a new family of glutamic peptidase based on in silico evidence

Neprosin was first discovered in the insectivorous tropical pitcher plants of Nepenthes species as a novel protease with prolyl endopeptidase (PEP) activity. Neprosin has two uncharacterized domains of neprosin activation peptide and neprosin. A previous study has shown neprosin activity in hydrolyzing proline-rich gliadin, a gluten component that triggers celiac disease. In this study, we performed in silico structure-function analysis to investigate the catalytic mechanism of neprosin. Neprosin sequences lack the catalytic triad and motifs of PEP family S9. Protein structures of neprosins from Nepenthes × ventrata (NvNpr) and N. rafflesiana (NrNpr1) were generated by ab initio methods and comparatively assessed to obtain high-quality models. Structural alignment of models to experimental structures in the Protein Data Bank (PDB) found a high structural similarity to glutamic peptidases. Further investigations reveal other resemblances to the glutamic peptidases with low optimum pH that activates the enzyme via autoproteolysis for maturation. Two highly conserved glutamic acid residues, which are stable according to the molecular dynamics simulation, can be found at the active site of the substrate cleft. Protein docking demonstrated that mature neprosins bind well with potent antigen αI-gliadin at the putative active site. Taken together, neprosins represent a new glutamic peptidase family, with a putative catalytic dyad of two glutamic acids. This study illustrates a hypothetical enzymatic mechanism of the neprosin family and demonstrates the useful application of an accurate ab initio protein structure prediction in the structure-function study of a novel protein family.

Construction of a Pig Alveolar Cleft Model in Imitation of Cleft Lip and Palate Congenital Deformity.

Alveolar cleft repair is a key step in multiple disciplinary treatment for patients with cleft lip/and palate. Although autologous bone grafting has been used worldwide over the past half century, alternative advanced techniques, such as the use of bone substitutes and guided tissue regeneration, have shown their great potentials and have been recommended by a growing number of physicians and surgeons. The employment of new therapeutic approaches and devices in clinical routine requires tremendous experimental efforts and appropriate animal models with similar sizes and sites of deformity to that of human both anatomically and physiologically. The aim of this study is to develop a juvenile porcine model with surgically created alveolar clefts imitating congenital alveolar cleft in the cleft lip and palate. Alveolar defects between second incisor and canine were surgically created in two miniature pigs (unilateral cleft in P1 and P2); bilateral alveolar defects were surgically created between first and third incisor in one miniature pig (P3) using piezo surgery. Pigs were sacrificed (P1 at 1 month after the surgery and P2 at 3 months postoperatively) and the evaluation of defects were performed by assessing result from the computed tomography (CT) scan and histopathological examination. Postoperative CT scan results showed that the size of the defect remained the same, whereas the edge of the defect became irregular 3 months after the surgery. In all pig subjects, histopathological examination found no sign of osteogenesis in the area of defect, indicating that our surgical procedure was successful in establishing porcine models for alveolar cleft in congenital cleft lip and palate. In conclusion, we developed alveolar cleft in porcine models to mimic the size, site, and environment of congenital alveolar cleft in cleft lip and palate. The novel animal model can be employed in pilot studies for the purpose of optimizing the current surgical treatment techniques as well as developing new treatment procedures and test the bone substitute materials. The bilateral model can be applied in further control studies.Impact statementCancellous iliac bone graft was the most popular surgical technique as well as the gold standard to reconstruct alveolar cleft. Nevertheless, several disadvantages exist regarding the additional surgical field of donor side and delayed age of alveolar bone grafting. Bone tissue-engineered strategy offers a promising alternative to address the gap in the current limitation of autologous bone to treat the growing craniofacial skeleton. Among different species of laboratory animals, porcine is suitable for oral and maxillofacial bone and implant-related research, where alveolar defect can be surgically developed simulating the size and site of alveolar cleft occurring together with cleft lip and palate. In this proposal, a reproducible porcine model of alveolar bone defect imitating congenital alveolar cleft during craniofacial growing stage is successfully constructed that will show great potential application in the field of tissue engineering and regenerative medicine. The model for bilateral alveolar cleft can be potentially applied in a controlled study in future.