Foreskin-isolated keratinocytes provide successful extemporaneous autologous paediatric skin grafts.

Severe burns in children are conventionally treated with split-thickness skin autografts or epidermal sheets. However, neither early complete healing nor quality of epithelialization is satisfactory. An alternative approach is to graft isolated keratinocytes. We evaluated paediatric foreskin and auricular skin as donor sources, autologous keratinocyte transplantation, and compared the graft efficiency to the in vitro capacities of isolated keratinocytes to divide and reconstitute epidermal tissue. Keratinocytes were isolated from surgical samples by enzymatic digestion. Living cell recovery, in vitro proliferation and epidermal reconstruction capacities were evaluated. Differentiation status was analysed, using qRT-PCR and immunolabelling. Eleven children were grafted with foreskin-derived (boys) or auricular (girls) keratinocyte suspensions dripped onto deep severe burns. The aesthetic and functional quality of epithelialization was monitored in a standardized way. Foreskin keratinocyte graft in male children provides for the re-epithelialization of partial deep severe burns and accelerates wound healing, thus allowing successful wound closure, and improves the quality of scars. In accordance, in vitro studies have revealed a high yield of living keratinocyte recovery from foreskin and their potential in terms of regeneration and differentiation. We report a successful method for grafting paediatric males presenting large severe burns through direct spreading of autologous foreskin keratinocytes. This alternative method is easy to implement, improves the quality of skin and minimizes associated donor site morbidity. In vitro studies have highlighted the potential of foreskin tissue for graft applications and could help in tissue selection with the prospect of grafting burns for girls.

Study of proliferation and 3D epidermal reconstruction from foreskin, auricular and trunk keratinocytes in children.

OBJECTIVE: Severe burns in children are conventionally treated with split-thickness skin autografts or epidermal sheets. An alternative approach is to graft isolated keratinocytes. We evaluated foreskin and other anatomic sites as donor sources for autologous keratinocyte graft in children. We studied in vitro capacities of isolated keratinocytes to divide and reconstitute epidermal tissue. METHODS: Keratinocytes were isolated from foreskin, auricular skin, chest and abdominal skin by enzymatic digestion. Living cell recovery, in vitro proliferation, epidermal reconstruction capacities and differentiation status were analyzed. RESULTS: In vitro studies revealed the higher yield of living keratinocyte recovery from foreskin and higher potential in terms of proliferative capacity, regeneration and differentiation. Cultured keratinocytes from foreskin express lower amounts of differentiation markers than those isolated from trunk and ear. Histological analysis of reconstituted human epidermis derived from foreskin and inguinal keratinocytes showed a structured multilayered epithelium, whereas those obtained from ear pinna-derived keratinocytes were unstructured. CONCLUSION: Our studies highlight the potential of foreskin tissue for autograft applications in boys. A suitable alternative donor site for autologous cell transplantation in female paediatric burn patients remains an open question in our department. We tested the hypothesis that in vitro studies and RHE reconstructive capacities of cells from different body sites can be helpful to select an optimal site for keratinocyte isolation before considering graft protocols for girls.

Epidermal healing in burns: autologous keratinocyte transplantation as a standard procedure: update and perspective.

BACKGROUND: Treatment of burned patients is a tricky clinical problem not only because of the extent of the physiologic abnormalities but also because of the limited area of normal skin available. METHODS: Literature indexed in the National Center (PubMed) has been reviewed using combinations of key words (burns, children, skin graft, tissue engineering, and keratinocyte grafts). Articles investigating the association between burns and graft therapeutic modalities have been considered. Further literature has been obtained by analysis of references listed in reviewed articles. RESULTS: Severe burns are conventionally treated with split-thickness skin autografts. However, there are usually not enough skin donor sites. For years, the question of how covering the wound surface became one of the major challenges in clinical research area and several procedures were proposed. The microskin graft is one of the oldest methods to cover extensive burns. This technique of skin expansion is efficient, but results remain inconsistent. An alternative is to graft cultured human epidermal keratinocytes. However, because of several complications and labor-intensive process of preparing grafts, the initial optimism for cultured epithelial autograft has gradually declined. In an effort to solve these drawbacks, isolated epithelial cells from selecting donor site were introduced in skin transplantation. CONCLUSIONS: Cell suspensions transplanted directly to the wound is an attractive process, removing the need for attachment to a membrane before transfer and avoiding one potential source of inefficiency. Choosing an optimal donor site containing cells with high proliferative capacity is essential for graft success in burns.

Mutations in the human UBR1 gene and the associated phenotypic spectrum.

Johanson-Blizzard syndrome (JBS) is a rare, autosomal recessive disorder characterized by exocrine pancreatic insufficiency, typical facial features, dental anomalies, hypothyroidism, sensorineural hearing loss, scalp defects, urogenital and anorectal anomalies, short stature, and cognitive impairment of variable degree. This syndrome is caused by a defect of the E3 ubiquitin ligase UBR1, which is part of the proteolytic N-end rule pathway. Herein, we review previously reported (n = 29) and a total of 31 novel UBR1 mutations in relation to the associated phenotype in patients from 50 unrelated families. Mutation types include nonsense, frameshift, splice site, missense, and small in-frame deletions consistent with the hypothesis that loss of UBR1 protein function is the molecular basis of JBS. There is an association of missense mutations and small in-frame deletions with milder physical abnormalities and a normal intellectual capacity, thus suggesting that at least some of these may represent hypomorphic UBR1 alleles. The review of clinical data of a large number of molecularly confirmed JBS cases allows us to define minimal clinical criteria for the diagnosis of JBS. For all previously reported and novel UBR1 mutations together with their clinical data, a mutation database has been established at LOVD.

Infantile hypertrophic pyloric stenosis: are viruses involved?

Infantile hypertrophic pyloric stenosis (IHPS) is characterized by abnormal thickening of the internal circular muscle layer. IHPS is known to be due to a combination of genetic and environmental factors, but its precise causes and pathophysiology are poorly understood. The objective of the study is to determine the prevalence of the principal viruses targeting the respiratory and digestive tracts in children with IHPS. Nasopharyngeal fluids, stools, vomit, and surgical pyloric muscle fragments and swabs were tested by cell culture, viral antigen assay and PCR. IHPS was diagnosed in 23 boys and 8 girls with a mean (± SD) age of 42 ± 15 days (range 20-88 days). There was no seasonal pattern of diagnosis. Twenty-two children (71%) lost weight (mean 246 ± 164 g, range 30-600 g) after the onset of vomiting, and five (16.1%) were dehydrated. Seven (22.6%) infants had been exposed to an infectious contact within 15 days before admission, and one on the day of admission (3.2%). Ear, nose and throat samples and pyloric muscle specimens were negative for all the viruses tested. An adenovirus type 3 was recovered from one stool sample, and RT-PCR was positive for an enterovirus on one vomit sample. This study suggests that the principal viruses targeting the respiratory and digestive tracts are not responsible for IHPS.

Growth in infants in the first two years of life after neonatal repair for unilateral cleft lip and palate.

OBJECTIVE: To evaluate the growth during the first two years of life in infants after unilateral cleft lip and palate neonatal repair. METHOD: All mature infants with nonsyndromic unilateral cleft lip and palate (NSUCLP) born between 2004 and 2007 were included. Information concerning growth was collected. Weight and length at birth, 6, 12, 18 and 24 months of age measurements and data regarding feeding were obtained. RESULTS: Weight and length at birth, 6, 12, 18 and 24 months of age were identical with reference curve values. Children with NSUCLP showed a normal growth at two years. The weight curves lie between 5th and the 50th percentile for girls and between 10th and higher than the 97th percentile for boys. The height curves lie between -1 Standard Deviation and +1 Standard Deviation for girls and 0 and +2 Standard Deviation for boys. CONCLUSION: Feeding difficulties are reported in infants with cleft lip and/or palate CLP/CP. However, the growth in children with NSUCLP and after neonatal cleft lip repair is identical with reference curve values.

[Neonatal computed tomographic maxillary explorations in children with unilateral cleft lip and palate]. / Etudes tomodensitométriques néonatales de l'os maxillaire dans les fentes labiopalatines unilatérales.

The aim was to examine the bony maxillary structures by computed tomographic measurements in newborns with unilateral cleft lip and palate before cheiloplasty. Analysis of maxillary bone was performed and size parameters were measured by computed-tomographic analysis in 12 infants with unilateral cleft lip and palate. We compared the bony maxillary length and the bony maxillary width between the cleft side and the healthy side. For eight patients, the bony maxillary length was different between the cleft side and the healthy side. For three patients, the bony maxillary width was different between the incisor alveolar structure in the cleft side and the healthy side. For six patients, the bony maxillary width was different between the canine alveolar structure in the cleft side and the healthy side. We noted an asymmetry without hypoplasy in bony maxillary structure in newborns before cheiloplasty. The data can serve as the starting point for a control and later evaluation on the efficiency of different therapeutic approaches of alveolar and maxillary development in children with cleft lip and palate.

Early repair for infants with cleft lip and nose.

OBJECTIVE: To study the results 10 years after early surgical cleft lip and nose repair. PATIENTS AND METHODS: We present the outcome of 123 early cleft lip repairs whose condition was managed in a multidisciplinary team according to a strict protocol. We give the observation results of operations of a single surgeon's neonatal surgery over a 10-year period in term of aesthetic assessments and dental arch relationships. One hundred and twenty-three patients were operated on during the first 4 weeks of life; a subgroup of 40 child had been operated on at a week or less of age. RESULTS: The results show good aesthetic assessments and dental relationships, with facial growth appropriate for the age. CONCLUSIONS: We are currently encouraging early cleft lip and nose repair in the full-term baby as the good method of management of newborns with cleft.

Surgical treatment of haemangioma in infants.

Haemangiomas usually can be identified by their clinical course. They are characterised by presentation at birth or shortly thereafter, and a rapid proliferative phase over the first 12 months. The haemangioma then usually stabilises and slowly involutes over a period of 5-7 years. For a long time, surgery has been limited to complicated cases, and correcting after-effects following involution. Nevertheless, aesthetic, psychological or functional prejudices may justify early surgery. We conducted a retrospective study of patients treated between 1995 and 2001. A total of 31 patients with facial and cervical haemangiomas were studied. For each, the type of lesion and its topography, age and operative indications, surgery, postoperative complications and aesthetic and functional results have been considered. Thirty-one haemangiomas were operated. The average age was 30 months (1-60 months). After an average follow-up of 3 years, the results were very good in 20%, good in 66%, and fair in 14% of cases. Early curative surgery of haemangioma before spontaneous involution, and before school-age is justified because of social and psychological considerations in infants and their family.

[Vesicoureteral reflux: diagnosis and management in children]. / Reflux vésico-urétéral: diagnostic et prise en charge chez l'enfant.

OBJECTIVE: Based on a retrospective study of children followed for vesicoureteric reflux, the authors evaluated the role of antenatal diagnosis in the detection and global management of the most frequent uropathy observed in children. MATERIALS AND METHODS: The case files of 180 children followed for reflux over a 4-year period were reviewed. The diagnosis of reflux was based on retrograde cystography. The authors tried to define the main indications for the renal and urinary tract assessment and defined the grade and type of reflux, as well as the therapeutic indications and results. 180 children (105 girls and 75 boys) with vesicoureteric reflux were observed during the study period. The mean age at the time of diagnosis was 26.75 months. RESULTS: Vesicoureteric reflux was usually diagnosed in a context of acute pyelonephritis in 139 children (77%);, at the time of the first episode in 84% of cases. Antenatal diagnosis of reflux during screening for dilated renal pelvis was the second most frequent modality (29 children, 16% of cases). The reflux involved a single urinary tract in 163 children and a duplicated tract in 17 children. Reflux was bilateral in 43% of cases. Reflux was less than grade II in 66% of cases, but an inverse proportion was observed in the case of antenatal diagnosis (62%). 55% of cases were treated surgically and 45% were treated medically. Follow-up of the children showed a low recurrent pyelonephritis rate that was similar in the two groups. CONCLUSION: Acute pyelonephritis remains the leading mode of discovery of vesicoureteric reflux and renal ultrasonography combined with retrograde cystography after a first episode of renal infection is therefore recommended. Antenatal screening may also reveal vesicoureteric reflux. It is difficult to summarize the therapeutic indications for reflux, as the treatment of vesicoureteric reflux is related to the child's age and sex, the grade of reflux, the clinical repercussions of the reflux and certain environmental factors, such as the family and social status.

[Laparoscopic treatment of the nonpalpable testis. Results]. / Traitement laparoscopique des testicules impalpables. Résultats.

OBJECTIVE: The purpose of laparoscopy in the management of the non palpable testis is to provide information regarding testicular presence and location to facilitate overall surgical management. MATERIALS AND METHODS: We report our experience with laparoscopic orchiopexy to treat 39 nonpalpable testes in 32 patients, patient age ranged from 2.3 years to 14 years (average 4.18 years). We retrospectively reviewed the medical records of all patients for a 5-year period. RESULTS: At laparoscopy 11 testes were at the internal inguinal ring, these patients underwent one-stage laparoscopic orchiopexy without division of the spermatic vessels. 18 high intra-abdominal testes underwent two-stage Fowler-Stephens orchiopexy. One Patient underwent laparoscopically assisted orchiectomy for atrophy, 9 testes were absent. At follow-up 6, 12 and 24 months after 26 of 29 (89%) testes are without atrophy, and in acceptable scrotal position. CONCLUSIONS: The low incidence of complications and 89% success rate underscore the feasibility of laparoscopic orchiopexy. It is our procedure of choice for the management and treatment of nonpalpable testis.