BMP4、TGF-β3基因多态性与环境暴露在非综合征性唇腭裂发生中的交互作用

目的 探讨环境暴露因素、骨形态生成蛋白4(BMP4)基因、转化生长因子β3(transforming growth factor beta-3,TGF-β3)基因之间的交互作用在非综合征性唇腭裂(nonsyndromic cleft lip and cleft palate,NSCLP)发生中的可能作用.方法 通过问卷调查获取环境暴露资料.用聚合酶链反应(PCR)-限制性片段长度多态性(restriction fragment length polymorphism,RFLP)技术对对照组(200例)和NSCLP组(200例)各基因位点的多态性进行检测.采用多因子降维法(multifactor djmensionality reduction,MDR)分析基因之间、基因与环境之间的交互作用关系,并对筛选的交互作用关系用Logistic回归进行验证.结果 BMP4 T538C、TGF-β3 C641A和TGF-β3G15572-三个单核苷酸多态性(single nucleotide polymorphism,SNP)位点间的交互作用与NSCLP的发生无关联.基因与环境交互作用分析发现,BMP4 T538C与母亲妊娠早期被动吸烟、母亲妊娠早期感染史对NSCLP的发生具有交互作用;TGF-β3G15572-与母亲妊娠早期被动吸烟、母亲妊娠早期感染史、父亲知晓妊娠前吸烟、父亲知晓妊娠前饮酒、母亲妊娠早期补充维生素对NSCLP的发生具有交互作用.经Logistic回归验证,结果一致.结论 NSCLP是基因与环境因素共同作用的结果,易感基因多态性影响着个体对环境因素的反应,研究它们之间的相互关系对阐明NSCLP的病因及发病机制具有重要意义.     

BMP2 and BMP4 variations and risk of non-syndromic cleft lip and palate

ObjectiveNon-syndromic cleft lip with or without cleft palate (CL/P) is one of the most common congenital anomalies and arises from the interaction of environmental and genetic factors. The objective of this study was to investigate the association between the BMP2 (bone morphogenetic protein 2) and BMP4 (bone morphogenetic protein 4) polymorphisms with non-syndromic CL/P to clarify the potential role of these genes in the etiology of CL/P in Iranian population.DesignThe allelic and genotypic frequencies of BMP2 rs235768 A > T and BMP4 rs17563 T > C polymorphisms were determined in 107 unrelated Iranian subjects with non-syndromic CL/P and 186 control subjects using PCR and RFLP methods, and the results were compared with healthy controls. A p-value of <0.05 was considered statistically significant.ResultsThe BMP2 rs235768 AT genotype was significantly higher (P = 0.009, OR = 3, 95% CI = 1.3–7.0) in the CL/P (59.8%) than the control group (33.3%). Similarly, the BMP4 rs17563 TC genotype were significantly higher (P = 0.008, OR = 3.7, 95% CI = 1.4–9.9) in the CL/P (70.0%) than the control group (44.6%).ConclusionThe BMP2 rs235768 A > T and BMP4 rs17563 T > C polymorphisms could be considered as the risk factor for non-syndromic CL/P in Iranian population.     

Non-syndromic cleft lip with or without cleft palate in Asian populations: Association analysis on three gene polymorphisms of the folate pathway

ObjectiveOrofacial clefts (OFCs) are one of the most common birth defects in humans. They are the subject of a number of investigations aimed at elucidating the bases of their complex mode of inheritance involving both genetic and environmental factors. Genes belonging to the folate pathway have been among the most studied. The aim of the investigation was to replicate previous studies reporting evidence of association between polymorphisms of folate related genes and the occurrence of non-syndromic cleft lip with or without cleft palate (NSCL/P), using three independent samples of different ancestry: from Tibet, Bangladesh and Iran, respectively.DesignSpecifically, the polymorphisms rs1801133 of MTHFR, rs1801198 of TCN2, and rs4920037 of CBS, were tested.ResultsA decreased risk of NSCL/P was observed in patients presenting the C677T variant at MTHFR gene (relative risk for heterozygotes = 0.53; 95% confidence interval [C.I.] = 0.32–0.87). The investigated polymorphisms mapping at TCN2 and CBS genes did not provide any evidence of association.ConclusionOverall, these results indicate that NSCL/P risk factors differ among populations and confirm the importance of testing putative susceptibility variants in different genetic backgrounds.     

Revisiting straight-line repair in unilateral complete cleft lip: a comparison with rotation-advancement repair

Rotation-advancement repair (RAR) has been the most widely used technique for unilateral cleft lip repair. We recently used a straight-line repair with medial orbicularis muscle lengthening (SLR-ml) technique, based on the hypothesis that it could minimize the postoperative scar appearance without causing s short-lip deformity when muscle reorientation is performed correctly. A retrospective cohort study was conducted on unilateral complete cleft lip patients who underwent cheiloplasty between 2009 and 2017. Two cheiloplasty techniques were compared: RAR and SLR-ml. Outcomes were evaluated by assessing follow-up photographs using three methods: (1) glance impression on a five-point scale, (2) Manchester Scar Scale, and (3) indirect anthropometry. Seventy-one patients were analysed: 41 in the RAR group (28 male, 13 female) and 30 in the SLR-ml group (15 male, 15 female). The glance impression (P = 0.506) and Manchester Scar Scale (P = 0.347) scores did not differ between the groups. According to the symmetry ratio (cleft side value/non-cleft side value), vertical lip height (P = 0.804), horizontal lip length (P = 0.881), and Cupid’s bow width (P = 0.122) did not differ significantly between the groups. The preoperative lip height discrepancy was not correlated with the postoperative vertical lip height. The SLR-ml method can be regarded as a successful tool for symmetric repair of unilateral cleft lip.     

Dental implant placement in alveolar cleft patients: a retrospective comparative study on clinical and aesthetic outcomes

The aim of this retrospective study was to assess the clinical and aesthetic outcomes, and patient satisfaction, following dental implant therapy in cleft patients. Implant survival, changes in marginal bone level, pocket probing depths, plaque and bleeding indices, aesthetics, and patient satisfaction were assessed in 17 alveolar cleft patients and 17 matched controls. At follow-up (mean 72.4 ± 46.4 months), one implant had been lost in the cleft group. Mean marginal bone loss at follow-up was −0.4 ± 0.4 mm in cleft patients and −0.2 ± 0.4 mm in controls. Aesthetics of the peri-implant soft tissues (pink aesthetic score) were less favourable (P = 0.025) in cleft patients (5.0 ± 1.9) than in controls (6.5 ± 1.7), while peri-implant parameters were comparable in the two groups. Overall patient satisfaction was 8.6 ± 0.9 in cleft patients and 8.9 ± 1.1 in controls (P = 0.331). In cleft patients, no difference in aesthetics was observed between patients who received additional bone augmentation at 3 months prior to implant placement and those who did not (P = 0.092). Dental implant therapy in cleft patients is associated with high implant survival, minor marginal bone loss, healthy peri-implant soft tissues, and high patient satisfaction. Only the aesthetics of the soft tissues was worse in cleft patients compared to augmented non-cleft patients.     

Association between BMP4 gene polymorphisms and cleft lip with or without cleft palate in a population from South China

ObjectivePrevious studies have suggested an association between several polymorphisms of the BMP4 gene and susceptibility to non-syndromic cleft lip with or without cleft palate (NSCL/P) in various populations. However, this association may vary according to ethnic group and the form of NSCL/P. This study analyzed the association between the BMP4 gene polymorphisms rs762642, rs17563, and rs10130587 with the risk of cleft lip only (CLO), cleft palate only (CPO), and cleft lip with palate (CLP) in a population from South China.MethodsThis case-control study included 165 patients with NSCL/P (53 patients with CPO, 52 with CLO, and 60 with CLP) and 52 healthy volunteers. Peripheral blood samples were collected from all subjects to genotype the rs762642, rs17563, and rs10130587 polymorphisms by direct sequencing. Genotype and allelic frequencies of these polymorphisms were compared between healthy volunteers and patients with various forms of NSCL/P.ResultsThe genotype and allelic frequencies of rs762642 differed significantly between subgroups (CPO and CLP) and normal controls, whereas a significant difference was observed only in the CLO subgroup for the rs17563 polymorphism and in the CLO and CLP groups for the rs10130587 polymorphism. In addition, we identified a novel association of a BMP4 gene polymorphism, which was in linkage disequilibrium with the rs10130587 polymorphism, with CLO and CLP.ConclusionThe BMP4 gene polymorphisms rs762642, rs17563, and rs10130587 exhibit different associations with different forms of NSCL/P, suggesting that different forms of NSCL/P may have different etiologies.     

Mandibular effects of maxillary distraction osteogenesis in cleft lip and palate

Maxillary distraction osteogenesis (DO) is a reliable treatment for severe maxillary deficiency in cleft lip and palate (CLP). The objective was to analyze its long-term effects on the mandible. A retrospective study of 24 CLP treated with maxillary DO using the Polley and Figueroa technique was done; patients were followed for more than 4 years. Preoperative (T0), 6–12 months postoperative (T1), and ≥4 years postoperative (T2) cephalometric radiographs were evaluated. A classical cephalometric analysis was used to assess treatment stability, and a Procrustes superimposition method was used to assess local changes in the shape of the mandible. The mean age of patients at T0 was 15.4 ± 4.1 years. SNA increased at T1 and T2 (P < 0.001), with no significant relapse between T1 and T2, indicating stability at 1 year after treatment (T0 = 72.4 ± 5.3°; T1 = 81.3 ± 6.2°; T2 = 79.9 ± 6.1°). SNB, facial angle, gonial angle, and symphyseal angle remained stable. Long-term analysis of the mandible demonstrated a minimal counter-clockwise rotation of the body (mandibular plane = −0.2 ± 3.2°) and ramus (−0.6 ± 4.3°). Maxillary DO in CLP had no significant effect on the shape or rotation of the mandible. The maxillary advancement remained stable after 1 year.     

Association between MAFB rs17820943 and rs6072081 polymorphism and risk of nonsyndromic cleft lip with or without cleft palate: a meta-analysis

While there have been previous studies examining the relation between the rs17820943 and rs6072081 polymorphisms in the v-maf musculoaponeurotic fibrosarcoma oncogene homolog B (MAFB) gene and rates of nonsyndromic cleft lip with or without cleft palate (NSCL/P), at present the results of these studies have been inconsistent. This meta-analysis therefore aimed to conduct a more robust assessment of the association between the MAFB rs17820943 and rs6072081 polymorphisms and NSCL/P risk. The Embase, Web of Science, PubMed, the China National Knowledge Internet (CNKI), and Wanfang databases were systematically searched to identify relevant studies. In total, five studies incorporating 2769 patients and 2885 controls were identified assessing the rs17820943 polymorphism and three studies incorporating 1242 patients and 1310 controls assessing the rs6072081 polymorphism were identified. This analysis revealed the MAFB rs17820943 and rs6072081 polymorphisms to be linked to a significantly reduced NSCL/P risk (rs17820943: C vs T: OR = 0.76, 95% CI = 0.70-0.82; CC vs CT: OR = 0.75, 95% CI = 0.67-0.85; CC vs TT: OR = 0.58, 95% CI = 0.49-0.67; CC + CT vs TT: OR = 0.67, 95% CI = 0.59-0.77; CT + TT vs CC: OR = 1.43, 95% CI = 1.28-1.60; rs6072081: A vs G: OR = 0.77, 95%CI = 0.68-0.86; AA vs AG: OR = 0.76, 95%CI = 0.64-0.90; AA vs GG: OR = 0.58, 95%CI = 0.45-0.74; AA + AG vs GG: OR = 0.68, 95%CI = 0.54-0.84; AG + GG vs AA: OR = 1.40, 95% CI = 1.19-1.65). The results of the present meta-analysis indicate that in an East Asian population, for both rs17820943 and rs6072081 were associated with NSCL/P.     

Timing of palate repair affecting growth in complete unilateral cleft lip and palate

ObjectiveTo evaluate the facial morphology characteristics of patients with complete unilateral cleft lip and palate (UCCLPs) who had undergone cleft palate repair at different times.DesignThis study included 46 nonsyndromic UCCLPs and 38 age and sex matched non-cleft patients. 35 cephalometric measurements were used to evaluate the facial morphology. Student’s t-test, one-way ANOVA and rank sum tests were used for comparison. Significant difference was defined at 95% level.ResultsThe data showed that UCCLPs who had palatoplasty between 7 and 12 years had greater PMP-A, PMP-ANS, Ba-ANS, Ba-A, Ba-N-ANS than those operated on before 4 years of age, and UCCLPs who had palatoplasty at 4–12 years had smaller Y-axis angle than those operated on before 4 years of age.ConclusionsThe maxillary sagittal length increased gradually as von Langenbeck repair was delayed. UCCLPs who underwent palate repair using von Langenbeck technique at 4–12 years had a more protrusive maxilla and less clockwise rotated mandible than those repaired before 4 years. UCCLPs operated using the von Langenbeck technique at 4–12 years had better head-face morphology than those operated on before 4 years. There was no difference in facial morphology among UCCLPs with palate repair at 4–12 years.     

Otitis media with effusion in Nigerian children with cleft palate: incidence and risk factors

Otitis media with effusion is common in children with cleft palate, and the aim of this study was to find out its incidence and risk factors in Nigerians. We prospectively studied 84 patients (42 with cleft palate and 42 control subjects); 27 were male and 15 female, who were age and sex matched with control subjects. The extent and size of the clefts were measured using a dental cast and Vernier calipers, and the otitis media was diagnosed with otoscopy and tympanometry. The mean (SD) age of the groups was 11 (7) months (range 1–33). Twelve children in the cleft group had otitis media compared with three in the control group. Infants and boys were more likely to be affected. There was a significant association between age (p = 0.02), sex (p = 0.01), and size of cleft (p = 0.00). However, only the size of the cleft was confirmed to be an independent predictor, with children who had extremely wide clefts being more likely to develop otitis media than those with narrow clefts (OR = 8.71, 95%CI = 1.07 to 70.5).We conclude that the incidence of otitis media with effusion was higher among children with cleft palate than among those who did not have a cleft. Infants had a higher incidence than older children, and boys had a higher incidence than girls. Age, sex, and the size of the palatal cleft were significantly associated with otitis media, but not the extent of clefting.     

Current concepts on the effect of environmental factors on cleft lip and palate

The aim of this study was to investigate the effect of environmental factors, such as tobacco, alcohol and folic acid intake, obesity, stressful events, low blood levels of zinc and fever during pregnancy, on the incidence of cleft lip and/or palate (CL ± P). An electronic search was performed in the Cochrane Reviews, the ISI Web of Knowledge, PubMed and Scopus, along with a manual search to identify other relevant case–control and cohort studies. Quality assessments and an evaluation of publication bias were undertaken. Statistical heterogeneity was examined, and odds ratios (ORs) and 95% confidence intervals (CI) estimated using the random effects model. Of 372 articles initially retrieved, 28 studies were selected as eligible for meta-analysis. No evidence of publication bias was found using funnel plot analysis and the Egger linear regression method. Many studies were classified as low quality due to inadequate case–control data. On the basis of this research, maternal factors most associated with CL ± P were: tobacco (OR 1.48), alcohol (OR 1.28), folic acid intake (OR 0.77), obesity (OR 1.26), stressful events (OR 1.41), low blood zinc levels (OR 1.82), and fever during pregnancy (OR 1.30). Folic acid intake by the mother reduced the risk of CL ± P in offspring (OR 0.77).     

Bite force of children with repaired unilateral and bilateral cleft lip and palate

ObjectiveTo assess the bite force (BF) of children with repaired cleft lip and palate (CLP).DesignChildren aged 6–12 years, with and without CLP, were divided into the following 5 groups: (1) control group (CON): 34 children without CLP (17 female, 17 male, mean age 8.2 ± 1.4); (2) cleft lip group (CL): 31 children with cleft lip involving the pre-maxilla (15 female, 16 male, mean age 9.7± 1.3); (3) unilateral CLP group (UCLP): 36 children with complete unilateral CLP (11 female, 25 male, mean age 9.4± 1.6); (4) bilateral CLP group (BCLP): 32 children with complete bilateral CLP (11 female, 21 male, mean age 9.5± 1.7); and 5) cleft palate group (CP): 17 children with complete cleft palate (9 female, 8 male, mean age 9.4± 1.6). Briefly, in this clinical trial, BF was assessed before alveolar bone grafting with a gnathodynamometer (IDDK, Kratos, Cotia, SP, Brazil). For CON, BCLP, CL and CP groups, BF was obtained in the anterior and posterior region of the maxilla. For the UCLP group, BF was assessed in the anterior and posterior regions of both segments. Differences among groups were evaluated by ANOVA test, and Tukey’s test was used to assess any correlations among variables (P < 0.05).ResultsUnexpectedly, no differences of BF were observed among CON and any of the cleft groups. However, a stronger BF was observed in the CL group when compared to the UCLP and BCLP groups. Next, no differences were observed between the cleft side and the noncleft side in the UCLP group. Lastly, in all groups, BFs from the anterior region of the maxilla were less when compared to the posterior regions.ConclusionThe BF of children with CLP is no different from children without CLP.     

Rs12941170 at SOX9 gene associated with orofacial clefts in Chinese

ObjectiveNon-syndromic orofacial cleftings (NSOCs) are considered as complex trait, which results from genetic and/or environmental modifiers. Current findings could only explain small portion of the NSOCs. SOX9 gene plays an important role during craniofacial development in animal models and the Pierre Robin sequence (PRS). However, its role in non-syndromic clefts remains unknown.DesignIn this study, we selected eight SNPs in and around SOX9 gene to make maximum coverage, and genotyped them by using RFLP-PCR and ligase detection reaction (LDR) methods to test its associations among 151 NSOCs (53 NSCLP, 52 NSCLO and 46 NSCPO) from Western Han Chinese population.ResultsAllelic TDT results showed that G allele at rs12941170 of SOX9 was under-transmitted among NSOCs (p = 0.00014, OR = 0.55 and 95%CI: 0.40–0.75), which could indicate that the G allele is protective against NSOCs; parent-of-origin effect analysis showed that G allele at rs12941170 was maternally under-transmitted (p = 0.002), while there was no statistically difference between the maternal and paternal transmission of it. To test if the adjacent SNPs travel together from parents to the affected individual, we carried out the sliding window haplotype analysis, it is interesting to find that the haplotypes carrying the G allele at rs12941170 also was under-transmitted for NSOCs, NSCL/P, NSCLP and NSCPO (lowest p = 0.00033).ConclusionsThis study suggested that G allele at rs12941170 was protective, which could decrease the risk for NSOCs from Western Han Chinese population, and it will provide new reference for future research and genetic counseling in NSOCs.     

Bone regeneration with BMP-2 and hydroxyapatite in critical-size calvarial defects in rats

BackgroundThis study describes the effect of bone formation by BMP-2 (bone morphogenetic protein-2), a bone formation inducer, with or without hydroxyapatite (HAP) application to critical-size defects in rat calvarial bone.Material and methodsTwenty male Wistar rats were divided into four groups of 5 animals each: control, HAP, BMP, and mixed BMP/HAP. A Critical-size defect of 4 mm was made using a trephine in the calvarial bone and, after that, BMP and/or HAP was applied to the defect according to the grouping. Defects were evaluated radiographically and histologically using ImageJ color analyzer software at 4 weeks postoperatively.ResultsThe histological data were more precise than the radiologic data due to the white color of the porous-type HAP material. The highest radiopacity was noted in the mixed BMP/HAP group (162.07 ± 9.06), followed by the HAP group (133.15 ± 21.8), then the BMP group (100.79 ± 8.27), and, lastly, the control group (54.45 ± 8.39). After subtracting the white background and using ImageJ for histological analysis, the highest rate of osteochondrogenesis was in the mixed BMP/HAP group (85.29% ± 8.21), and then the BMP group (77.34% ± 7.39), followed by the HAP group (59.82% ± 11.23), and, lastly, the control group (40.27% ± 7.44). Differences in the values between groups were then analyzed using confidence intervals (CI) of 95 and 99%.ConclusionWithin 4 weeks, the mixed BMP/HAP group showed the highest level of bone induction, especially compared to the BMP group, but this was non-significant; even with a 95% CI, the result was negative. This reveals that BMP alone can be applied, with a final result the same as that seen in the mixed BMP/HAP group. BMP and HAP, both being osteoinducting agents, even though they differ from a material classification point of view, have a positive effect on osteogenesis.     

Evaluation of airflow characteristics before and after septoplasty in unilateral cleft patients with a deviated nasal septum: a computational fluid dynamics study

The aim of this study was to evaluate changes in airflow characteristics before and after septoplasty in unilateral cleft lip and palate (UCLP) patients using computational fluid dynamics (CFD) models. The study was designed as a prospective cohort study involving pre- and postoperative computed tomography data from 12 UCLP patients with a deviated nasal septum who underwent septoplasty. CFD analysis of nasal airflow was conducted to study changes in velocity, pressure, volume, nasal resistance, and wall shear stress of the nasal domain before and after surgery. The study results demonstrated a statistically significant difference in pressure drop after septoplasty: median 116.10 Pa (interquartile range (IQR) 749.02 Pa) preoperative compared with 43.39 Pa (IQR 349.01 Pa) postoperative (P =  0.004). Maximum wall shear stress was found to be approximately three times lower after septoplasty: median 6.15 Pa (IQR 1908.62 Pa) preoperative versus median 2.51 Pa (IQR 540.06 Pa) postoperative (P = 0.002). Changes in nasal resistance were also found to be statistically significant: median 460.59 Pa·s/l (IQR 1946.99 Pa·s/l) preoperative versus median 166.61 Pa·s/l (IQR 694.08 Pa·s/l) postoperative (P = 0.04). These values demonstrate significant changes in flow dynamics after surgery indicative of a more uniform airflow pattern and stabilization of the nasal mucosa.     

Nasolabial shape and aesthetics in unilateral cleft lip and palate: an analysis of nasolabial shape using a mean 3D facial template

The aim of this study was to determine the amount of deviation in nasolabial shape in patients with a cleft compared with an average non-cleft face, and to assess whether this difference is related to nasolabial aesthetics. Three-dimensional stereophotogrammetric images of 60 patients with a unilateral cleft were used. To quantify shape differences, four average non-cleft faces were constructed from stereophotogrammetric images of 141 girls and 60 boys. Three-dimensional shape differences were calculated between superimposed cleft faces and the average non-cleft face for the same sex and age group. Nasolabial aesthetics were rated with the modified Asher-McDade Aesthetic Index using a visual analogue scale (VAS). Mean VAS scores ranged from 51.44 to 60.21 for clefts, with lower aesthetic ratings associated with increasing cleft severity. Shape differences were found between cleft faces and the average non-cleft face. No relationship was found for the VAS, age, and sex, except that a lower VAS was related to a higher nose and lip distance between the superimposed cleft and average non-cleft faces for nasal profile (P =  0.02), but the explained variance was low (R² = 0.066). In conclusion, except for nasal profile, nasolabial aesthetics were not influenced by the extent of shape differences from the average non-cleft face.     

Postoperative lip and nose forms following primary bilateral cleft lip repair selecting one/two stage surgery in comparison to those of healthy children

PurposeTo analyze the short-term outcomes for the lip and nasal forms after primary treatment following our surgical strategy for bilateral cleft lip with/without palate (BCL ± P) repair selecting one- or two-stage surgery at Kagoshima University Hospital.Patients and methodsTwenty-one patients with BCL ± P were treated and followed up over 1 year (1–6 years). Patients underwent primary lip repair by one- or two-stage surgery depending on the prolabium height, medial-upward advancement of nasolabial components, and vestibular expansion using two cleft margin flaps. The postoperative lip and nasal forms were longitudinally measured using serial color photographs and were compared to those of 18 age- and sex-matched healthy Japanese children.ResultsPostoperative lip form showed the upper lip height and vermilion mucosal height were significantly increased compared with the preoperative values. The vermilion/cutaneous lip height ratio was improved to the same level as that of controls at 3 years of age. Symmetry of lip and nasal forms was successfully achieved postoperatively, but the nasal height was still smaller than that of controls.ConclusionsOur surgical management strategy for BCL ± P will provide well-balanced and symmetrical lip and nasal forms, except for the nasal height, without damaging an infant's prolabium.