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The origin of bone formed by heterotopic periosteal autografts.

PURPOSE: This study tested the hypothesis that a significant amount of the new bone produced by heterotopic periosteal autografts is derived osteoinductively because proliferating periosteal cells express the bone morphogenetic protein (BMP). MATERIALS AND METHODS: Rabbit ulnar and radial periosteum were autografted as free grafts (FGs) to the forelimb musculature, and as millipore diffusion chambers grafts (MDCGs) to the rectus abdominus muscle. The grafts were recovered at 3, 5, 7, 14, and 28 days postoperation, fixed in 4% paraformaldehyde, demineralized in 0.6N HCL, and 4.0 microns paraffin-embedded sections were immunostained with monoclonal antibody against recombinant human (rh) BMP-2. RESULTS: Sections from FGs recovered 5 to 28 days postoperatively exhibited cartilage and bone; fibrous tissue, cartilage, bone, and osteochondroid differentiated within MDCGs. Although BMP-2 was expressed by mesenchymal cells, osteoblasts, osteocytes, and osteoclasts, none of the MDCGs produced the osteoinductive signature of transmembrane bone formation. CONCLUSIONS: These observations indicated that the larger fraction of the new bone produced by heterotopic periosteal autografts is derived from the graft cells.

Animals↗

The spatiotemporal expression of TGF-beta1 and its receptors during periosteal chondrogenesis in vitro.

Transforming growth factor-beta1 (TGF-beta1) has been shown to stimulate chondrogenesis in periosteal explants cultured in agarose suspension. TGF-betas exert their cellular effects through a heteromeric cell membrane receptor complex consisting of TGF-beta type I and type II receptors. In this study, the spatial and temporal expressions of the type I receptor (TbetaR-I), type II receptor (TbetaR-II) and endogenous TGF-beta1 in periosteal explants cultured in vitro were examined using reverse transcription polymerase chain reaction (RT-PCR) and immunohistochemistry. The temporal changes in the expression of the TbetaR-I and TbetaR-II mRNAs correlated with that of TGF-beta1. Exogenous administration of TGF-beta1 upregulated the expression of both receptors and of the TGF-beta1 ligand in a biphasic pattern. The earlier peak of upregulation was observed at 7 days in culture. A later peak of upregulation was seen at 42 days, at which time cartilage formation reached a maximum. Immunohistochemical studies demonstrated co-localization of TbetaR-I and TbetaR-II simultaneously among the same cells expressing TGF-beta1. TGF-beta1 treatment increased the expression of TGF-beta1, TbetaR-I and TbetaR-II in mesenchymal cells in the cambium layer at 7 days in culture. Small round chondrocytes showed widely distributed immunoreactivity of TGF-beta1, TbetaR-I and TbetaR-II in the 42-day explants treated with TGF-beta1. These observations support the hypothesis that TGF-beta1 regulates the initiation and formation of cartilage during periosteal chondrogenesis.

Activin Receptors, Type I↗

Interleukin-1 alpha and epidermal growth factor synergistically enhance the release of collagenase by periosteal connective tissue in vitro.

The effects of recombinant human interleukin-1 alpha (IL-1 alpha) and murine epidermal growth factor (EGF) on the release of collagenase were studied in an in vitro model system using periosteal explants from rabbit calvariae. Following an incubation period of 72 h it was shown that IL-1 alpha in combination with EGF (IL-1 alpha + EGF) induced a synergistic increase in the amount of collagenase released by periosteal explants. This increase appeared to be at least 10-fold. Most of the enzyme was present in a latent form since the increase in enzyme activity was only detectable after activation by APMA and the molecular weight as determined in immunoblots corresponded to the latent form of this enzyme. Incubations carried out with IL-1 alpha alone resulted in a 2- to 4-fold increase of total enzyme activity, whereas the amount of collagenase in media of EGF-treated periosteal did not surpass control values. A neutralizing anti-IL-1 alpha antibody completely blocked the enhanced release of collagenase as induced both by IL-1 alpha and by IL-1 alpha + EGF. Indomethacin partially prevented the IL-1 alpha + EGF-induced increase in enzyme release, suggesting the involvement of prostaglandins. The amount of tissue inhibitor of metalloproteinases (TIMP) as determined by ELISA was slightly elevated in culture media obtained from all cytokine-treated explants. Comparable results were obtained by Western blot analysis as well as by a functional bioassay. It is suggested that the concomitant presence of the cytokines IL-1 alpha and EGF may play an important role in collagenase-mediated degradation of collagen.

Animals↗

Bone engineering on the basis of periosteal cells cultured in polymer fleeces.

Large bone defects caused by severe trauma, infection or tumor resection are still a major challenge for orthopaedic surgery. The key concept for successful bone regeneration consists of combining the osteoinductive effect of osteogenic cells with a suitable carrier structure to promote osteoblastic differentiation and optimal matrix production. Therefore, periosteal cells cultured in polyglycolic-polylactid acid (PGLA) fleeces were investigated for their osteogenic differentiation and used to repair critical size bone defects in a rabbit model. Periosteal cells were isolated from New Zealand White rabbits and expanded in vitro. Osteogenic differentiation was investigated by analysis of alkaline phosphatase and osteocalcin production in vitro depending on culture conditions and passage number. Cells were seeded into PGLA fleeces. After further cultivation, tissue constructs were examined histologically and by immunohistochemistry for cell distribution and osteogenic differentiation. These constructs of defined size were used to repair critical size calvarial defects (group I) in rabbits compared to a defect repair with polymers only (group II) or to untreated defects (group III). Bone healing was evaluated after 4 weeks by radiodensitometry and a special histological scoring system. For early evaluation, radiodensitometry was not sensitive enough to detect differences in calcification. However, on histologic examination the group with cell/fleece constructs revealed intense formation of uncalcified bone. The mean defect closure of the experimental group I was 65%, compared to control groups II and III with 31% and 22%, respectively. The established methods of 3-D-cell culture and ex-vivo transplant assessment proved to be a valuable tool for quality assurance. The results demonstrate that the combination of periosteal cells and polymer fleeces is a tissue engineering approach, which may have clinical applications in various fields of reconstructive surgery.

Journal Article↗

Periosteal infusion of bupivacaine/morphine post sternal fracture: a new analgesic technique.

BACKGROUND AND OBJECTIVES: Sternal fracture pain is severe and is difficult to alleviate due to the forces acting on the chest wall during respiration. We describe a continuous infusion regional analgesic technique for pain due to sternal fracture. CASE REPORT: A 47-year-old woman presented with a spontaneous sternal fracture, precluding effective coughing. Diclofenac and increasing doses of opioids did not give adequate pain relief and led to opioid toxicity. Two brief periods of analgesia were achieved with deep subcutaneous infiltration of bupivacaine. An epidural catheter was positioned periosteally, and an infusion of bupivacaine was commenced at 5 mL/h, achieving long-lasting analgesia. The bupivacaine concentration was reduced in a stepwise fashion from 0.5% to 0.25% and was changed to levobupivacaine after 3 days. Adding morphine (5 mg/60 mL levobupivicaine) permitted a reduction in infusion rate. The catheter was removed after 14 days because a local infection developed that resolved uneventfully with antibiotic therapy. CONCLUSIONS: Continuous infusion of local anesthetic and opioid to a sternal fracture site using a periosteally positioned catheter led to successful analgesia and hence improved respiratory function. Clinicians should consider placing a periosteal catheter when pain associated with sternal fracture cannot be adequately controlled with conventional methods.

Analgesics, Opioid↗

Prefabricated vascularized periosteal grafts using fascial flap transfers.

The purpose of this study was to demonstrate the feasibility of prefabricated vascularized periosteal grafts using fascial flap transfers. Fifty rats were divided into two (Group A, underwent periosteal wrapping, and Group B underwent fascial wrapping). Fascial flap from the medical leg, which were nourished by the saphenous artery and vein, were used for all grafts. In Group A, the fascia was transferred onto the proximal tibial periosteum. Two weeks postoperatively, the elevated fascioperiosteal flap was wrapped around a 4-mm hydroxapatite cube. In Group B, the fascial flap was immediately wrapped around the hydroxyapatite cube. In both groups, the hydroxapatite-wrapped cube was transferred to the inguinal region. Three days postoperatively, five rats from group A were sacrificed. On histologic examination, these five showed no necrosis of the fascioperiosteal flap and no bone formation. The remaining 45 rats were sacrificed 5 weeks postoperatively. Microangiography was performed on five rats in Group A, revealing neovascularization of the hydroxyapatite. On histologic examination, new bone formation was confirmed in 15 of the remaining 20 rats in Group A, and in three of the 20 rats in Group B. These experiments demonstrated that the prefabricated vascularized periosteal graft, which has good osteogenic capacity, could be carried out using a fascial flap transfer.

Angiography↗

Hydraulic elevation of the periosteum: a novel technique for periosteal harvest.

Periosteum has been promoted as a potential substrate for tissue engineering. Its principal virtues are that it has a source of pluripotential mesenchymal cells and chondrogenic growth factors located in the cambium layer, and it can serve as a template for directional evolution of neo-tissue. The clinical use and in vitro study of periosteum-derived neo-tissue has been limited by the level of surgical skill required for harvest. Precise surgical technique, task-specific experience, adequate volume of procedures, and general surgical expertise are required for optimal harvest using the traditional periosteal elevator method. This report describes an easily mastered technique that preserves viability while providing the harvest of relatively large amounts of periosteum. Skeletally mature New Zealand white rabbits (11 males/20 tibias; 4 females/8 tibias; approximate weight 3.5 kg) and one Yucatan miniature pig were used for harvest of periosteum from the tibia using the traditional periosteal elevator and the developed hydraulic elevation approach. Histologic examination of the periosteal explants obtained by the developed method showed preservation of the cambium layer containing the progenitor cells necessary for the generation of neo-cartilage. This technique provides a simple method of harvesting large segments (>5 cm x 1 cm) of periosteum in a single procedure and may facilitate better exploitation of periosteum in tissue engineering.

Animals↗

The effect of periosteal preservation on osteogenesis in a canine rib autograft model: tetracycline fluorescence incident photometry.

In the first portion of this study, we describe a new technique, tetracycline fluorescence incident photometry (TFIP) for quantifying new bone formation. In this portion, we use routine histology, Microfil vascular injection, and tetracycline incidence fluorescence (TFIP) to determine the effect of periosteal preservation on bone formation in canine rib autografts. We found that periosteal preservation significantly enhanced new bone formation in both cortical and trabecular bone (83-70% of control) when compared with autografts without the periosteum (76-60% of control) (p < 0.05). In addition, periosteal preservation favorably influenced graft revascularization. We found that the periosteum of the transferred rib autografts was made up of 3 distinct layers: (1) the inner (cambial) layer of osteogenic cells, (2) the middle (fibrous) layer of osteogenic reserve cells, and (3) the outer vascular network of arterioles and venules, which communicate with the trabecular vessels internally. This outer vascular network has not been previously described in transferred bone grafts and is responsible for early graft revascularization.

Animals↗

Orbital cellulitis with periosteal elevation.

Computerized tomography scan evidence of periosteal elevation in patients with orbital cellulitis is interpreted in the current medical literature as an indication of subperiosteal abscess. We present three such cases in which surgical drainage yielded clear fluid or granulation tissue rather than pus. A fourth case resolved on antibiotic therapy alone. Cases of periosteal elevation that resolve without surgery may represent inflammatory effusion, infections of lesser virulence, or propagation of granulation tissue rather than true abscesses. We suggest that periosteal elevation seen in patients with orbital cellulitis should represent a relative rather than an absolute indication for drainage surgery.

Abscess↗

Treatment of chondral lesions in advanced osteochondritis dissecans: a comparative study of the efficacy of chondrocytes, mesenchymal stem cells, periosteal graft, and mosaicplasty (osteochondral autograft) in animal models.

Management of chondral lesions in osteochondritis dissecans remains a challenge. This study investigated the efficacy of periosteal graft, osteochondroidal autograft, autologous chondrocyte and mesenchymal stem cell transplants in the treatment of chondral lesions in animal models. Full-thickness articular cartilage defects were created in the weight-bearing surface of the medial femoral condyle in 20-week-old NZW rabbits. A total of 56 knees were randomly divided into four groups as follows: group 1, transfer of cultured chondrocytes; group 2, transfer of cultured mesenchymal stem cells; group 3, repair by periosteal graft; and group 4, mosaicplasty. All of the contralateral knees served as control. Gross, histologic, and biomechanical examinations at 36 weeks after the operation showed that the cultured chondrocytes and mesenchymal stem cells had comparable enhancing effects on the repair of chondral defects in advanced osteochondritis dissecans, whereas mosaicplasty did well initially and periosteal graft did less favorably.

Animals↗

Periosteal chondroma: MR characteristics.

PURPOSE: The purpose of this study was to describe the MR characteristics of periosteal chondroma. METHOD: MR images of 12 proven cases of periosteal chondroma were analyzed with reference to tumor morphology and size. MR features were correlated with radiographic and pathologic findings. RESULTS: Tumor size ranged from 1 to 7 cm in maximum diameter with a mean value of 2.6 cm. On MR images, a soft tissue mass at the bone surface with pressure erosion of adjacent cortical bone could be identified in all cases. All lesions were bordered by a hypointense rim (100%) and frequently showed a lobulated configuration (75%). Edema of medullary bone or soft tissues was not observed in any of the cases. Signal intensity of cartilaginous tumor tissue was typically hypo-or isointense relative to muscle on T1-weighted (100%) and hyperintense relative to fat on T2-weighted (92%) and T2*-weighted (100%) MR images. Radiographically significant calcifications of the tumor matrix, present in half of the cases, caused focal signal loss on MR images of all pulse sequences. Contrast enhancement was observed predominantly at the periphery of the lesions (100%), which on pathologic examinations typically contained fibrovascular bundles, surrounding the cartilage lobules. CONCLUSION: Periosteal chondroma appears to have a relatively typical MR appearance, which reflects the histologic composition of the lesion. In addition to radiography, MRI therefore can substantially aid in the preoperative diagnosis of this rare bone lesion.

Adolescent↗

Combined encephaloduroarteriosynangiosis and bifrontal encephalogaleo(periosteal)synangiosis in pediatric moyamoya disease.

OBJECTIVE: We compared the surgical results of simple encephaloduroarteriosynangiosis (EDAS) and EDAS with bifrontal encephalogaleo(periosteal)synangiosis for the treatment of pediatric moyamoya disease. METHODS: Data for 159 children (up to 15 yr of age, 76 boys and 83 girls) who underwent indirect revascularization procedures for the treatment of moyamoya disease between 1987 and 1998 were retrospectively reviewed. Group A patients underwent simple EDAS (n = 67). Group B patients underwent EDAS with bifrontal encephalogaleo(periosteal)synangiosis (n = 92). The surgical results for each group were compared, in terms of clinical outcomes, neuroimaging changes, extent of revascularization evident on angiograms, and hemodynamic changes evident on single-photon emission computed tomographic scans. The average follow-up periods were 45 months for Group A and 22 months for Group B. RESULTS: The overall clinical outcomes and neuroimaging changes tended to be better for Group B. In terms of single-photon emission computed tomographic changes of the whole brain after surgery, Group B patients exhibited more favorable outcomes than did Group A patients (62 versus 36%, P = 0.003). The surgical results for the anterior cerebral artery territory were significantly better for Group B than for Group A, with respect to outcomes of anterior cerebral artery symptoms (81 versus 40%, P = 0.015), revascularization on angiograms (79 versus 16%, P < 0.001), and hemodynamic changes on single-photon emission computed tomographic scans (70 versus 52%, P = 0.002). The incidences of postoperative infarctions were not significantly different between the two groups. CONCLUSION: EDAS with bifrontal encephalogaleo(periosteal)synangiosis is a more effective surgical modality for the treatment of pediatric moyamoya disease, compared with simple EDAS, because it covers both the middle cerebral artery and anterior cerebral artery territories of the brain.

Adolescent↗

Revascularized periosteal grafts--a new method to produce functional new bone without bone grafting.

Rib periosteum was transplanted to the groins of 9 dogs. In half of the periosteal grafts, no microvascular anastomoses were done (free grafts); at 6 weeks after grafting they had become resorbed. The other periosteal grafts were revascularized by microvascular anastomoses of the intercostal vessels to local muscular vessels; at 6 weeks those with confirmed vascular patency had all formed substantial amounts of new bone. Five cm, full-thickness defects were created in the tibias of 10 dogs. The control animals (without grafting) did not heal in two months. However, the experimental dogs, with vascularized periosteal grafts in the defects regenerated their tibias with healthy new bone by 6 weeks--and were walking on them then.

Animals↗

Palatal periosteal response to surgical trauma.

Investigation of the response to surgical procedures on periosteum from different skeletal sites indicates differences in potential for bony repair. The response of palatal periosteum to surgical trauma was investigated following two procedures in 12 young dogs. Results were seen histologically and by fluorescence of tetracycline and CDAF bone labels. Palatal periosteum elevated and replaced showed evidence of a proliferative cellular response of the inner periosteal layer and renewed osteogenic activity on the third day postoperatively. The immediate posttrauma period was characterized by the formation of an expanded medullary zone (reactive medullary zone), after which osteogenesis appeared to return to normal. Repair bone in the resection area was not related primarily to the periosteal flap but always appeared at and extended at and extended from the edges of the resection cavity. In; this model, palatal periosteal elevation interrupted osteogenesis for 2 or 3 days and stimulated an intense cellular proliferation. Free palatal periosteum overlying a resection pocket was not seen to spontaneously for bone within a 3-week period.

Animals↗

Free periosteal graft repair of maxillary clefts in adolescents.

Free periosteal grafting of the maxillary cleft in adolescents is an alternative, simplified method of reconstructing secondary maxillary cleft deformities, particularly the sunken nostril sill. The historical background of bone grafting, periosteal flaps, and free periosteal grafts in infants, along with recent studies of adolescent growth patterns of the maxilla and cleft side dentition, provides the rationale for our protocol. Fourteen patients were operated on ranging in age from 9 to 30. They were studied clinically by Panorex x-rays and some by CT scans. New bone bridging the maxillary cleft with resultant improvement in the cleft deformities was found routinely in patients under 15 years of age. Some patients exhibited greater cosmetic improvement than others.

Adolescent↗

Periosteal flaps and grafts in primary cleft repair: a follow-up study.

A long-term follow-up study of maxillary clefts operated on primarily with the maxillary periosteal flap (67 patients) and the free tibial periosteal graft (23 patients) showed a definite bone bridge in 64 versus 85 percent of the patients. Still, secondary bone grafting was indicated in both in over 70 percent. Lateral crossbite was observed in all and anterior crossbite (mostly dental) was seen in over 80 percent of both groups. There were no statistically significant differences in the cephalometric angular measurements between the groups. Inside the maxillary periosteal flap group the maxillary growth was more retarded in unilateral and especially bilateral complete clefts than in alveolar clefts only. The maxillary growth seemed to be on average better than in primarily bone-grafted materials. The primary use of periosteum had been abandoned in our unit already in 1974, because it did not fulfill the expectations of prevention of maxillary collapse, lesser need for secondary bone grafting, and better midfacial growth.

Adolescent↗

Experimental use of fibrin glue to induce site-directed osteogenesis from cultured periosteal cells.

The purpose of this study was to determine whether a combination of fibrin glue and cultured periosteal cells will result in new bone formation at heterotopic sites in nude mice. Growing cells and developing matrices surrounding periosteal explants from the diaphyses of radii of newborn calves were minced and mixed with fibrin glue in a syringe. The cell/matrix-fibrin glue admixture was then injected into the subcutaneous space on the dorsum of athymic nude mice. After 12 weeks of implantation, gross morphology and histologic investigations showed newly formed bone structures in all cell/matrix-fibrin glue admixtures, but none in fibrin glue injected alone and used as control samples. Osteopontin, a protein important in bone development, was identified by a Western blot assay of the cell/matrix-fibrin glue composite. This study supports the feasibility of initiating site-directed formation of bone structures at heterotopic tissue sites by means of injection of cultured periosteal cells and matrix in a fibrin glue carrier.

Animals↗

Evolving acoustic characteristics of the canal wall down cavities due to neo-osteogenesis by periosteal flap.

OBJECTIVE: To determine whether the acoustic properties of the canal wall down cavity would come closer to those of the normal ear canal as the cavity is lined with periosteal flap. STUDY DESIGN: Prospective study. SETTING: Tertiary referral center. PATIENTS: Twenty one subjects who underwent canal wall down mastoidectomy and total ossicular replacement prosthesis tympanoplasty were selected. INTERVENTIONS: Canal wall down cavities were lined with postauricular periosteal flap. The acoustic properties of 21 canal wall down cavities (mastoidectomy ears) and 16 contralateral normal ears (control ears) were measured with a hearing aid fitting system. MAIN OUTCOME MEASURES: Frequency, amplitude, bandwidth, and Q factor of the resonance curve along with the loss in amplitude below 1 kHz were compared in a paired-wise manner between the first and third months and the third and sixth months postoperatively in mastoidectomy ears, and also between mastoidectomy and control ears. RESULTS: In the mastoidectomy ears, the peak resonance frequency and bandwidth significantly increased at the end of the third and six months as the volume of the cavity decreased ( < 0.05). However, the amplitude and Q factor of the resonance curve remained stable. At the end of the first month, all parameters were different in both groups ( < 0.05). At the end of the sixth month, the frequency and bandwidth of the resonance curve in both groups were not different. The mastoidectomy ears had some decibel-amplitude loss below 1 kHz. Only its frequency changed as the cavity became smaller ( < 0.05). CONCLUSIONS: New bone formation facilitated by a periosteal flap in the posterior part of the CWD cavity behind the facial ridge may provide acoustic properties similar to those of the normal ear canal.

Acoustic Impedance Tests↗