Mark P. Mooney

The osteogenic potential of adipose-derived stem cells for the repair of rabbit calvarial defects

Introduction: Bone replacement is often necessary during reconstruction of craniofacial anomalies or trauma. Adipose-derived stem cells (ASCs) possess osteogenic potential and are a promising cell source for bone tissue engineering. The present study was designed to assess the osteogenic potential and utility of using ASCs to regenerate bone in a rabbit calvarial defect model. Methods: Rabbit ASCs were seeded on gelatin foam (GF) scaffolds and induced in osteogenic medium containing bone morphogenetic protein (BMP)-2. Thirty-four 8-mm calvarial defects were randomly treated with autograft, no treatment, GF scaffold, GF + ASCs, or GF + osteoinduced ASCs. After 6 weeks, calvaria were harvested and underwent histologic and radiologic analyses to compare healing between the treatment groups. Results: Defects treated with autograft underwent complete healing. Radiologically, there were no significant (P > 0.05) differences in healing among empty defects, and those treated with GF alone or GF plus osteoinduced ASCs. Osteoinduced ASCs exhibited significantly (P < 0.05) greater healing than noninduced ASCs. Conclusion: Preimplantation osteoinduction of ASCs enhances their osteogenic capacity. Lack of a significant osteogenic effect of ASCs on calvarial healing at 6 weeks may be secondary to use of noncritical-sized defects. Larger defects would likely demonstrate the osteogenic potential of ASCs more definitively.

Testing the critical size in calvarial bone defects: revisiting the concept of a critical-size defect.

Background: There is a clinical need for bone replacement strategies because of the shortfalls endemic to autologous bone grafting, especially in the pediatric patient population. For the past 25 years, the animal model that has been used to test bone replacement strategies has been the calvarial critical-size defect, based on the initial size of the bone defect. This study was undertaken to test the concept of the critical size in several different models. A review of the theoretical and scientific bases for the critical-size defect was also undertaken. Methods: Two different rodent species (including 28 adult mice and six adult rats) were used to assess bone healing by means of two-dimensional radiographic analysis after creating small bone defects using different surgical techniques. Results: Defects in mice that were smaller than critical-size defects (1.8-mm diameter) were shown to heal a maximum of 50 percent 1 year postoperatively. Small defects (2.3-mm diameter) in the rat skull showed approximately 35 percent healing after 6 weeks. Neither the choice of rodent species nor the maintenance of the dura mater significantly affected calvarial bone healing. Conclusions: These results suggest that calvarial bone healing is not well described and much more data need to be collected. Also, after a review of the existing literature and a critique of the clinical applicability of the model, it is suggested that the use of the term “critical-size defect” be discontinued.

Bone morphogenetic protein 2 therapy for craniofacial surgery.

Between 2 and 10 years of age, the developing craniofacial skeleton poses a significant reconstructive challenge. Local autogenous bone is largely unavailable, distant bone grafts are fraught with significant morbidity and limited yield, and alloplastic materials are incompatible with the growing calvarium and facial skeleton. Bone morphogenetic protein (BMP) 2, a member of a class of proteins first noticed in the 1960s to promote bone deposition in soft tissues, offers a potential solution to the bone shortage historically faced by the pediatric craniofacial surgeon. A review of English language literature was conducted from the 1960s to the present. Attention was focused on BMP-2s osteoinductive mechanism, basic science and translational laboratory findings, and multidisciplinary clinical experiences. Bone morphogenetic protein 2 has been embraced by spine surgeons, is gaining in popularity for long-bone repair, and is making its way into the plastic surgery literature. Bone morphogenetic protein 2 may provide a basis for an off-the-shelf tissue-engineered bone construct that is compatible with the growing craniofacial skeleton while free from the morbidities of distant graft harvest. Questions remain, however, regarding the safety and efficacy of this compound in the context of pediatric craniofacial surgery. In an effort to facilitate the clinicians risk-benefit analysis of this emerging technology, we present a primer on the basic science of BMP-2, a discussion of possible morbidities associated with its use, a review of laboratory and clinical trials with this substance to date, and an analysis of strategies to maximize its efficacy in craniofacial surgery.

Development of a strain of rabbits with congenital simple nonsyndromic coronal suture synostosis. Part II: Somatic and craniofacial growth patterns.

In the March 1993 issue of The Cleft Palate-Craniofacial Journal we reported a female rabbit born in our laboratory with complete bilateral coronal suture (CS) synostosis. This follow-up study presents our attempts to breed the animal and establish a strain of craniosynostotic rabbits. The second part of this study presents longitudinal somatic and craniofacial growth data in offspring with coronal suture synostosis. Serial growth data from 72 animals were collected for the present study. The sample consisted of 11 animals (10 offspring and the original female) with complete nonsyndromic unilateral (plagiocephalic) or bilateral (brachycephalic) CS synostosis, 19 animals with partial CS synostosis, and 42 unaffected control litter mates. At 10 days of age, all animals had radiopaque amalgam markers placed on either side of the frontonasal, coronal, anterior lambdoidal, and sagittal sutures. Body weights and serial lateral and dorsoventral head radiographs were taken at 1.5 (10 days), 6, 12, and 18 weeks of age. All animals showed similar body weights at 1.5 weeks of age, while completely synostosed animals exhibited a slight (about 12%), but significantly (p < .001) lowered body weight by 18 weeks of age. Results revealed that by 1.5 weeks of age the completely synostosed animals already exhibited brachycephalic cranial vaults, midfacial hypoplasia, and increased flattening of the cranial base compared to unaffected siblings. This pattern continued through 18 weeks of age, with the partially synostosed animals exhibiting intermediate morphologies.(ABSTRACT TRUNCATED AT 250 WORDS)

Orbicularis oris muscle defects as an expanded phenotypic feature in nonsyndromic cleft lip with or without cleft palate

Nonsyndromic cleft lip ± cleft palate is a complex disease with a wide phenotypic spectrum; occult defects of the superior orbicularis oris muscle may represent the mildest subclinical form of the lip portion of the phenotype. This study used high‐resolution ultrasonography to compare the frequency of discontinuities in the OO muscle in 525 unaffected relatives of individuals with nonsyndromic cleft lip  ±  cleft palate versus 257 unaffected controls. OO muscle discontinuities were observed in 54 (10.3%) of the non‐cleft relatives, compared to 15 (5.8%) of the controls—a statistically significant increase (P = 0.04). Male relatives had a significantly higher rate of discontinuities than male controls (12.0% vs. 3.2%; P = 0.01); female relatives also had a higher rate of discontinuities than female controls, but the increase was not statistically significant (8.9% vs. 7.4%; P = 0.56). These data confirm the hypothesis that subepithelial OO muscle defects are a mild manifestation of the cleft lip phenotype. Identification of subepithelial OO muscle defects may be important in a clinical setting, as a means of providing more accurate recurrence risk estimates to relatives in cleft families. Furthermore, the expansion of the cleft lip  ±  cleft palate phenotypic spectrum should improve the power of genetic studies.

Histological definition of the vomeronasal organ in humans and chimpanzees, with a comparison to other primates

The vomeronasal organ (VNO) is a chemosensory structure that has morphological indications of functionality in strepsirhine and New World primates examined to date. In these species, it is thought to mediate certain socio‐sexual behaviors. The functionality and even existence of the VNO in Old World primates has been debated. Most modern texts state that the VNO is absent in Old World monkeys, apes, and humans. A recent study on the VNO in the chimpanzee (Smith et al., 2001b ) challenged this notion, demonstrating the need for further comparative studies of primates. In particular, there is a need to establish how the human/chimpanzee VNO differs from that of other primates and even nonhomologous mucosal ducts. Histochemical and microscopic morphological characteristics of the VNO and nasopalatine duct (NPD) were examined in 51 peri‐ and postnatal primates, including humans, chimpanzees, five species of New World monkeys, and seven strepsirhine species. The nasal septum was removed from each primate and histologically processed for coronal sectioning. Selected anteroposterior intervals of the VNO were variously stained with alcian blue (AB)‐periodic acid‐Schiff (PAS), PAS only, Gomori trichrome, or hematoxylin‐eosin procedures. All strepsirhine species had well developed VNOs, with a prominent neuroepithelium and vomeronasal cartilages that nearly surrounded the VNO. New World monkeys had variable amounts of neuroepithelia, whereas Pan troglodytes and Homo sapiens had no recognizable neuroepithelium or vomeronasal nerves (VNNs). Certain unidentified cell types of the human/chimpanzee VNO require further examination (immunohistochemical and electron microscopic). The VNOs of P. troglodytes, H. sapiens, and New World monkeys exhibited different histochemistry of mucins compared to strepsirhine species. The nasopalatine region showed great variation among species. It is a blind‐ended pit in P. troglodytes, a glandular recess in H. sapiens, a mucous‐producing duct in Otolemur crassicaudatus, and a stratified squamous passageway in all other species. This study also revealed remarkable morphological/histochemical variability in the VNO and nasopalatine regions among the primate species examined. The VNOs of humans and chimpanzees had some structural similarities to nonhomologous ciliated gland ducts seen in other primates. However, certain distinctions from the VNOs of other primates or nonhomologous epithelial structures characterize the human/chimpanzee VNO: 1) bilateral epithelial tubes; 2) a superiorly displaced position in the same plane as the paraseptal cartilages; 3) a homogeneous, pseudostratified columnar morphology with ciliated regions; and 4) mucous‐producing structures in the epithelium itself. Anat Rec 267:166–176, 2002.

Osseous guided tissue regeneration using a collagen barrier membrane.

The formation of mature, fibrous tissue in surgical osteotomy sites during the healing process can produce clinically undesirable results such as nonunion or encapsulation of alloplastic implants. The techniques of guided tissue regeneration have been used to ameliorate this problem by presenting a barrier to the invasion of fibrous tissue elements into the wound-site clot. The most frequently used barrier material, polytetrafluoroethylene is effective, but suffers the disadvantage of requiring surgical removal after clot organization is completed. A biocompatible, resorbable membrane that will effectively control the type of tissue that can invade and organize a clot would be advantageous, because it would not require surgical removal. In the present study, the efficacy of a collagen membrane to effect guided tissue regeneration in a rabbit zygomatic arch osteotomy model was tested. Complete, bilateral narrow (1 mm) or wide (5 mm) vertical osteotomies were created in eight adult New Zealand white male rabbits. On one side, the wound site was surrounded by a collagen barrier membrane prior to closure, while the other side was left uncovered (control side). Four animals were killed at 2 and 4 weeks postoperatively for gross radiologic and histologic examination of the wound site. The wide osteotomy sites without a barrier membrane showed invasion by fibroblasts resulting in fibrous nonunion, while the contralateral sides with the barrier membrane showed no fibrous tissue ingrowth and bony union by 4 weeks postoperatively. Although narrow wound sites without the barrier membrane showed fibrous tissue formation, the perimeter of the defects showed some new bone deposited at the periosteal surface, bridging the osteotomy site and producing osseous union.(ABSTRACT TRUNCATED AT 250 WORDS)

Nonsynostotic occipital plagiocephaly: factors impacting onset, treatment, and outcomes.

Background: Nonsynostotic occipital plagiocephaly remains a diagnosis of concern in infancy. This study evaluates factors affecting the onset, treatment, and outcomes of nonsynostotic occipital plagiocephaly. Methods: A retrospective chart review and telephone survey were performed. A posterior occipital deformation severity score was used. Factors such as demographics, behavioral and helmet therapy, feeding patterns, torticollis, multiple gestation pregnancies, prematurity, and congenital nonsynostotic occipital plagiocephaly were evaluated. Results: One hundred five infants were identified. Of these, 95 percent were Caucasian, 93 percent were from two-parent households, and 70 percent were from households earning more than

A rabbit model of human familial, nonsyndromic unicoronal suture synostosis. II. Intracranial contents, intracranial volume, and intracranial pressure

50,000. Repositioning was attempted in 95 percent, and 45 percent progressed to helmet therapy. When comparing change in posterior occipital deformation severity score with helmet therapy to repositioning, a difference was found (p < 0.05). Forty-nine percent of patients were breast-fed, and when compared with the general population, a difference was found (p < 0.05). Twenty percent of infants had torticollis, and when compared with population norms, a difference was found (p < 0.05). Twelve percent of patients were twins, and when compared with population norms, more twinning occurred (p < 0.05). Congenital nonsynostotic occipital plagiocephaly was found in 10 percent of patients and did not result in an increased risk of progression to helmet therapy. Conclusions: This study demonstrates trends that may predict additional risks for developing nonsynostotic occipital plagiocephaly, including torticollis, plural births, and increased socioeconomic affluence. In addition, the nonsynostotic occipital plagiocephaly cohort was breast-fed less than the general population, demonstrating that breast-feeding may be preventative, as breast-fed infants are repositioned more frequently and sleep for shorter periods. As in other studies, cranial molding helmet therapy was more effective in correcting nonsynostotic occipital plagiocephaly than repositioning alone.

A Successful Algorithm for Limiting Postoperative Fistulae following Palatal Procedures in the Patient with Orofacial Clefting

This two-part study reviews data from a recently developed colony of New Zealand white rabbits with familial, nonsyndromic unilateral coronal suture synostosis, and this second part presents neuropathological findings and age-related changes in intracranial volume (ICV) and intracranial pressure (ICP) in 106 normal rabbits and 56 craniosynostotic rabbits from this colony. Brain morphology and anteroposterior length were described in 44 rabbit fetuses and perinates (27 normal; 17 synostosed). Middle meningeal artery patterns were qualitatively assessed from 2-D PCC MRI VENC scans and endocranial tracings from 15, 126-day-old rabbits (8 normal, 7 rabbits with unicoronal synostosis). Brain metabolism was evaluated by assessing 18F-FDG uptake with high-resolution PET scanning in 7, 25-day-old rabbits (3 normal, 4 with unicoronal or bicoronal synostosis). Intracranial contents and ICV were assessed using 3-D CT scanning of the skulls of 30 rabbits (20 normal,10 with unicoronal synostosis) at 42 and 126 days of age. Serial ICP data were collected from 66 rabbits (49 normal; 17 with unicoronal synostosis) at 25 and 42 days of age. ICP was assessed in the epidural space using a Codman NeuroMonitor microsensor transducer. Results revealed that cerebral cortex morphology was similar between normal and synostosed fetuses around the time of synostosis. Significantly (P<0.05) decreased A-P cerebral hemisphere growth rates and asymmetrical cortical remodeling were noted with increasing age in synostotic rabbits. In addition, rabbits with unicoronal suture synostosis exhibited asymmetrical middle meningeal artery patterns, decreased and asymmetrical brain metabolism, a “beaten-copper” intracranial appearance, significantly (P<0.05) decreased ICV, and significantly (P<0.01) elevated ICP compared with normal control rabbits. The advantages and disadvantages of these rabbits as a model for human familial, nonsyndromic unicoronal suture synostosis are discussed, especially in light of recent clinical neuropathological, ICV, and ICP findings recorded in human craniosynostotic studies.