Evaluation of giant arachnoid granulations with high-resolution 3D-volumetric MR sequences at 3T.

PURPOSE: To evaluate the contribution to the diagnosis of the giant arachnoid granulations (AGs) of three-dimensional (3D) high-resolution magnetic resonance (MR) imaging sequences such as T2-weighted sampling perfection with application optimized contrasts using different flip-angle evolution (SPACE) and post-contrast T1-weighted magnetization prepared rapid gradient echo (MPRAGE). MATERIALS AND METHODS: Patients with 45 giant AGs were included in this retrospective study. All the patients were performed 3D T2-weighted SPACE and contrast enhanced MR venography sequences, as well as conventional cerebral MR imaging sequences. Post-contrast T1 weighted MPRAGE sequence were performed on 38/45 patients. All cerebral MR examinations were reviewed by the 2 neuroradiologists. Each GA was evaluated carefully to assess location and mean diameter. RESULTS: The most common location for giant AGs was at both transverse sinuses. Fluid signal feature within the giant AGs was not isointense to CSF on SE T1 and FLAIR MR imaging in 32 of 45 giant AGs. There were cerebral herniation into AG in 10 (22.2 %) of 45 giant AGs. 33 (73.3 %) of 45 giant AGs had central vein finding into AG in contrast enhanced MR venography. Signal void phenomenon into AG in 3D T2-weighted SPACE MR sequence was identified in 28 (62.2 %) of 45 giant AGs. CONCLUSIONS: Fluid within giant AGs had no completely CSF-like signal intensity on conventional and 3D high-resolution MR imaging sequences. Majority of CSF-incongruent fluid within giant AGs on conventional sequences is mostly due to intra-AG CSF flow.

The effect of dextrin-rhEGF on the healing of full-thickness, excisional wounds in the (db/db) diabetic mouse.

Chronic wounds, such as ulceration of the lower limb, represent a significant clinical challenge in today's ageing society. With the aim of identifying improved therapeutics, we have previously described a bioresponsive, dextrin-recombinant human epidermal growth factor conjugate (dextrin-rhEGF), that (i) protects rhEGF against proteolytic degradation by human chronic wound fluid; and (ii) mediates rhEGF release by α-amylase, capable of stimulating increased proliferation/migration in normal dermal and chronic wound fibroblasts; and keratinocytes, in vitro. The aim of this study was to extend these findings, by investigating the effects of dextrin-rhEGF on wound healing in the (db/db) diabetic mouse, a widely used in vivo model of delayed wound healing. Standardised, full-thickness excisional wounds, created in the dorsal flank skin, were treated topically with succinoylated dextrin (50 µg/mL), rhEGF (10 µg/mL) or dextrin-rhEGF (1 or 10 µg/mL). Treatments were applied immediately after injury and subsequently on post-wounding, days 3 and 8. Wound healing was assessed macroscopically, in terms of initiation of neo-dermal tissue deposition and wound closure (including wound contraction and re-epithelialisation), over a 16 day period. Wound healing was assessed histologically, in terms of granulation tissue formation/maturity; cranio-caudal wound contraction and wound angiogenesis (CD31 immuno-staining), using tissues harvested at day 16. Blood samples were also analysed for α-amylase and rhEGF concentrations. In this established impaired wound healing model, the topically-applied dextrin-rhEGF significantly accelerated wound closure and neo-dermal tissue formation at the macroscopic level; and significantly increased granulation tissue deposition and angiogenesis at the histological level (p<0.05), relative to untreated, succinoylated dextrin and rhEGF alone controls. Overall, these findings support the further development of bioresponsive polymer conjugates, for tissue repair.

Use of platelet-rich plasma under autogenous onlay bone grafts.

OBJECTIVES: To analyze the healing of autogenous onlay bone grafts in three different situations, focusing on the interface area. MATERIAL AND METHODS: Sixteen rabbits underwent autogenous bone graft surgeries in the calvaria. The block bone grafts were positioned in three different situations: direct contact between bone graft and receptor bed, graft interposed by particulate bone, and graft interposed by platelet-rich plasma (PRP). After 7, 15, 30, and 60 days, the specimens were retrieved for histological and morphometric evaluation. RESULTS: All groups healed uneventfully and presented incorporation of the grafts after 30 days. A slightly more evident new bone formation could be observed in the PRP group in the first analyzed period, and an earlier maturation of bone in the last period, although no statistically significant differences were achieved. CONCLUSION: The use of additional material between the bone graft and the receptor bed when using the onlay technique must be carefully considered, taking into account the size of the reconstruction and the cost/benefit relation. The addition of PRP in between autogenous bone blocks and the receptor bed did not confer significant benefit for the new bone formation and healing on the calvaria of bone of rabbits.

Experimental studies on hydroxyapatite powder-carboxymethyl chitin composite: injectable material for bone augmentation.

We developed a hydroxyapatite (HA) powder-carboxymethyl chitin composite (HA-CMC composite) that can be injected with a 14G needle by adding distilled water. We prepared Materials I (HA = 57.0 wt%) and II (HA = 40.2 wt%) and examined their biocompatibility and osteoconductivity. With a 2-mm skin stab, the material was injected on the calvarial bone of rats. The periosteum was denuded blindly in half of the cases and preserved in the other half of the cases. Simultaneously, the material was injected subdermally into the abdominal skin to examine diachronic volume alteration of the material. Our results indicated that the new materials had biocompatibility as high as that achieved with previously developed HA materials. The difference in HA concentration did not influence the osteoconductivity, but the periosteum and the soft tissue on the cranium seemed to be an obstacle to bone ingrowth. On the other hand, the volume alteration was significantly smaller in Material I than in Material II. This composite may be especially useful in facial bone augmentation because it can be injected with only a small skin stab. When used for that purpose, the periosteum of the host bone should be denuded to facilitate bone ingrowth, and Material I will be preferable to Material II in terms of the maintenance of the initial volume.

Granulación por fusión en mezcladores granuladores de alta velocidad / Melt granulation in high shear mixer

La granulación por fusión en un solo paso es una técnica alternativa basada en la utilización de un agente aglutinante sólido que funde a temperaturas entre 50-80 ºC, en lugar de solventes acuosos u orgánicos. Esta propiedad permite emplear esta técnica para la formulación de fármacos sensibles a la humedad, evitar el controvertido uso de solventes orgánicos y acortar el proceso de granulación en términos de tiempo y energía al eliminar la fase de secado. Asimismo, una adecuada selección del agente aglutinante y del resto de excipientes permitirá el desarrollo y elaboración de formas farmacéuticas tanto de liberación inmediata como de liberación controlada. Uno de los equipos más empleados en la granulación por fusión es el mezclador granulador de alta velocidad, que permite realizar esta técnica en un solo paso. Además, una vez optimizado el proceso, permite su aplicación a escala industrial. En este artículo se realiza una descripción de la técnica de granulación por fusión y las variables que influyen en dicho proceso en un mezclador granulador de alta velocidad

Histologic examination of pedicled buccal fat pad graft in oral submucous fibrosis.

PURPOSE: This report evaluates the wound healing process of buccal fat pad (BFP) grafted on a defect of the buccal mucosa for oral submucous fibrosis (OSF). PATIENTS AND METHODS: Sixteen patients with limitation of mouth opening under the diagnosis of OSF were surgically treated by cutting the fibrotic bands on the buccal mucosa (10 bilateral and 6 unilateral). The defects created were then covered by a BFP graft. The bulging BFP was trimmed postoperatively on a weekly basis until it was fully epithelialized. The specimens were stained and examined microscopically. RESULTS: Inflammatory cell infiltrate, blood vessel congestion, and fibrinous exudates covering the BFP were obvious by week 2. At week 3, blood vessel congestion and fat cell number decreased markedly. Evidence of stratified squamous epithelium with parakeratosis was seen in the margin of the BFP graft. At week 4, the number of fat cells decreased significantly and the original BFP was almost completely replaced by granulation tissue. The original BFP was fully covered by stratified squamous epithelium by week 5. CONCLUSIONS: The BFP graft has been widely used for covering exposed defects created by fibrotic band excision for the improvement of mouth opening limitation. The healing process was documented microscopically by weekly observation.

Adenovirus-mediated VEGF(165) gene transfer enhances wound healing by promoting angiogenesis in CD1 diabetic mice.

It has been previously shown that vascular endothelial growth factor (VEGF) plays a central role in promoting angiogenesis during wound repair and that healing-impaired diabetic mice show decreased VEGF expression levels. In order to investigate the potential benefits of gene therapy with growth factors on wound repair, a replication-deficient recombinant adenovirus vector carrying the human VEGF(165) gene (AdCMV.VEGF(165)) was topically applied on excisional wounds of streptozotocin-induced diabetic mice. Treatment with AdCMV.VEGF(165) significantly accelerated wound closure when compared with AdCMV.LacZ-treated, as well as saline-treated control mice, by promoting angiogenesis at the site of injury. Our findings suggest that AdCMV.VEGF(165) may be regarded as a therapeutic tool for the treatment of diabetic ulcers.

Impaired wound healing in mice deficient in a matricellular protein SPARC (osteonectin, BM-40).

BACKGROUND: SPARC is a matricellular protein involved in cell-matrix interactions. From expression patterns at the wound site and in vitro studies, SPARC has been implicated in the control of wound healing. Here we examined the function of SPARC in cutaneous wound healing using SPARC-null mice and dermal fibroblasts derived from them. RESULTS: In large (25 mm) wounds, SPARC-null mice showed a significant delay in healing as compared to wild-type mice (31 days versus 24 days). Granulation tissue formation and extracellular matrix protein production were delayed in small 6 mm SPARC-null wounds initially but were resolved by day 6. In in vitro wound-healing assays, while wild-type primary dermal fibroblasts showed essentially complete wound closure at 11 hours, wound closure of SPARC-null cells was incomplete even at 31 hours. Addition of purified SPARC restored the normal time course of wound closure. Treatment of SPARC-null cells with mitomycin C to analyze cell migration without cell proliferation showed that wound repair remained incomplete after 31 hours. Cell proliferation as measured by 3H-thymidine incorporation and collagen gel contraction by SPARC-null cells were not compromised. CONCLUSIONS: A significant delay in healing large excisional wounds and setback in granulation tissue formation and extracellular matrix protein production in small wounds establish that SPARC is required for granulation tissue formation during normal repair of skin wounds in mice. A defect in wound closure in vitro indicates that SPARC regulates cell migration. We conclude that SPARC plays a role in wound repair by promoting fibroblast migration and thus granulation tissue formation.

Vanadate ingestion increases the gain in wound breaking strength and leads to better organized collagen fibers in rats during healing.

Repair of incision wounds closed by suturing is evaluated by the progressive gain in wound breaking strength. Previously the closure of open wounds in rats ingesting vanadate, an inhibitor of tyrosine phosphate phosphatases, was shown to occur with deposition of more uniformly organized collagen fiber bundles. The hypothesis of this study was that deposition of more uniformly organized collagen fibers would enhance the gain in wound breaking strength of incisional wounds. Six adult rats received vanadate-supplemented saline drinking water for 1 week before placement of two 6-cm, parallel, suture-closed wounds on their backs. Six control rats received identical wounds and were given saline drinking water. The drinking water regimen was continued for 1 week after wounding, and then wound strength was tested with a tensiometer and tissue samples were obtained for histologic evaluation. Wound breaking strength doubled in vanadate-treated rats compared with controls. Bright-field and polarized light microscopy showed that the connective tissue matrix of granulation tissue from control rats was oriented perpendicular to the surface of the skin. In contrast, the connective tissue matrix of granulation tissue from vanadate-treated rats was oriented parallel to the skin surface. The gap in granulation tissue between the edges of the wounds in the vanadate-treated rats was greater than that in controls. Electron microscopy showed that wounds in the vanadate-treated contained uniform collagen fibers that were 20 percent greater in diameter and more evenly spaced than they were in controls. It is proposed that these changes in the organization of collagen fibers within incisional wounds were responsible for the increased wound breaking strength observed in rats ingesting vanadate.

Arachnoid granulations on high-resolution MR images and diffusion-weighted MR images: normal appearance and frequency.

Arachnoid granulations (AGs), protrusions into the cerebral venous sinus lumen, have been reported on cerebral venography, contrast enhanced CT, and conventional MR imaging. Although thin-sliced high-resolution MR images and diffusion-weighted images are frequently obtained, there have been no detailed reports concerning AGs on these images. In this study, the frequency and positional distribution of AGs in the transverse sinus was investigated on thin-sliced high-resolution MR images, and their appearance on diffusion-weighted MR images was evaluated. At least one AG was found in 107 of 151 subjects (70.9%). No statistically significant differences were noticed between males and females or between the right and left sides. No significant correlations between age and size or between age and the number of AGs were noted. On diffusion-weighted images, all AGs showed iso-intensity to normal brain tissue, which was higher than the reported signal intensity of arachnoid cyst and lower than that of epidermoids. In conclusion, AGs are normal structures that are frequently found in the cerebral venous sinuses on high-resolution MR images. Knowledge regarding their frequency and normal appearance would be helpful to avoid confusion between pathological processes and AGs. It is also important to know that AGs are frequently found even in the younger population.

Vascularization into a porous sponge by sustained release of basic fibroblast growth factor.

Vascularization into a poly(vinyl alcohol) (PVA) sponge was investigated using basic fibroblast growth factor (bFGF). This growth factor was impregnated into biodegradable gelatin microspheres for its sustained release and then the bFGF-containing microspheres or free bFGF were incorporated into PVA sponges. Following subcutaneous implantation into the back of mice, the bFGF-containing gelatin microspheres induced vascularization in and around the sponge to a significantly greater extent than that of free bFGF from 3 days after implantation. Significant ingrowth of fibrous tissue into the sponge was also observed when bFGF-containing microspheres were added to the sponge in contrast to free bFGF. Tissue ingrowth occurred into the deeper portion of the sponge over time while it accompanied formation of new capillaries. Empty gelatin microspheres had no effect on vascularization and the level of fibrous tissue ingrowth into the sponge was similar to that of the control group. It was concluded that incorporation of gelatin microspheres containing bFGF into the PVA sponge was effective in prevascularization of the sponge pores.

MRI of arachnoid granulations within the dural sinuses using a FLAIR pulse sequence.

The purpose of the study was to assess the signal intensities of arachnoid granulations within the dural sinuses using the FLAIR sequence for differentiation of space-occupying lesions in and adjacent to the dural sinuses. We retrospectively reviewed MR images of the brain of 1118 consecutive subjects, ranging in age from 0 to 93 years (mean 57.2 years). Nodules within the dural sinuses with signal intensities similar to that of cerebrospinal fluid (CSF) on both T1 and T2 weighted images were defined as arachnoid granulations. The location, signal intensity on T1 weighted spin echo (SE), T2 weighted fast SE and FLAIR images, the impression on the inner table of the skull, and the size of the lesion were assessed. 112 subjects (10.0%), age range 4-89 years old (mean 58.9 years), were found to have 134 arachnoid granulations. The commonest location was the transverse sinus, with 115 granulations (85.8%). The prevalence of the granulations showed a peak in the sixth decade of age. All granulations were isointense relative to CSF on T2 weighted images and almost all lesions were isointense relative to CSF on T1 weighted images. On FLAIR images, 90.3% of the granulations were isointense relative to CSF and the other 9.7% granulations were slightly hyperintense compared with the CSF. 21 (15.7%) subjects showed impressions on the inner table; one case involved the outer table. In conclusion, arachnoid granulations were isointense or slightly hyperintense relative to CSF on FLAIR. FLAIR images are helpful in differentiating arachnoid granulations from other dural sinus lesions or skull lesions which have an intensity similar to that of CSF on T1 weighted and T2 weighted images.

Connective tissue formation in subcutaneous cellulose sponge implants in the rat. The effect of the size and cellulose content of the implant.

Granulation tissue formation was studied in viscose cellulose sponges with different cellulose contents and sizes after subcutaneous implantation in rats. Samples were removed and studied histologically and histomorphometrically 1-16 weeks after implantation. The implants with lower cellulose content and smaller size were invaded by more cells and filled with connective tissue more rapidly than those with the higher content and larger size. In larger sponge implants the beneficial effect of the lower cellulose content was more conspicuous.

Dermal microvasculature and tissue selective thinning techniques (ultrasound and water-jet) of short-time expanded skin in dogs.

Certain reconstructive procedures, like auricular reconstructions, require thin and well-vascularized skin. The aims of this study were to analyze if the increased survival of expanded skin flaps was due to morphologic changes of the dermis, if thinning of short-time expanded skin was possible without harm to the microcirculation and if tissue selective cutting methods could be used to resect subcutaneous fat without damaging its vessels. Eighty-two 200-ml expanders were implanted into the trunk regions of 26 beagles and filled immediately with sterile saline. In the first series of experiments, the expansion was terminated after intervals of 0.5-5 weeks and dermal vessels were analyzed morphometrically. In the second series the expanded flaps were raised after 2 weeks and thinned solely surgically or with the additional use of an ultrasonic knife or with cutting by water jet. In contrast to sham flaps, the expanded skin showed only very few areas of necrosis and these were located superficially in most cases. The relative volume of the dermal vessels and their quantity showed a significant increase after the expansion. Additionally, the subcutaneous tissue could be thinned down to 0.4 mm with the water-jet-cutter. Findings demonstrated that the method used could create a well-vascularized skin flap of minimal thickness that could be very helpful for special reconstructive procedures.

Type V collagen in experimental granulation tissue.

To evaluate the spatial and temporal expression of type V collagen in a wound healing model, subcutaneously implanted viscose cellulose sponges in rats were used to induce granulation tissue formation. Analyses on granulation tissue were carried out on days 3, 5, 8, 14, 21, 30, 59 and 84. Acid soluble collagens were extracted and the relative amount of type V collagen was quantified by SDS-PAGE. Specific antibodies to type I, III and V collagens were used in immunohistochemistry and specific RNA probes to proalpha1(I), proalpha1(III) and proalpha1(V) collagen in in situ hybridization. Type V collagen content increased relative to type I and III collagens up to day 8 and remained at the same level for up to the three months. Type V collagen was expressed strongly in blood vessel walls as seen in immunohistochemistry. In situ hybridization showed that all of the three types of collagen were expressed mostly in fibroblast-like cells and also in rounded cells, especially type V collagen. In conclusion, type V collagen was seen in the wound healing model in increasing amounts from day 3 onwards, its localization being highly associated with blood vessels in granulation tissue and it was synthesized by fibroblast-like and rounded cells.

Inhibition of wound contraction with locally injected lathyrogenic drugs.

BACKGROUND: Previous studies using systematically administered lathyrogens to inhibit wound contractures have produced inconsistent results. The purpose of this study was to investigate the effects of lathyrogenic drugs on wound contraction when injected locally. METHODS: Two symmetrical full-thickness wounds were made on the dorsum of either side of hairless (hr/hr) mice; thus, each animal served as its own control. Animals were divided into groups receiving daily local injections of beta-aminopropionitrile or D-penicillamine, or both beta-aminopropionitrile and D-penicillamine and normal saline vehicle (control side) for 5 or 10 days. The rate of contraction was determined by serial measurements of the surface area of each wound during the treatment period. At the end of the treatment period, the wounds were excised en bloc with the chest wall and prepared for blinded histological analysis. Granulation tissue thickness, number of fibroblasts in granulation tissue per unit area, number of inflammatory cells (neutrophils, lymphocytes, macrophages and mast cells) in subjacent muscle per unit area, and collagen deposition in subjacent muscle were determined. RESULTS: Wound contraction, granulation tissue thickness, and collagen deposition in subjacent muscle were decreased only in wounds treated with beta-aminopropionitrile plus D-penicillamine. Collagen deposition in subjacent muscle was also decreased in wounds treated with D-penicillamine alone. Neither drug alone nor the combination affected the number of inflammatory cells in subjacent muscle. Body weight was not affected by the experimental procedures. CONCLUSIONS: The combination of beta-aminopropionitrile and D-penicillamine is potentially useful for inhibiting contracture formation when injected locally.

Effect of basic fibroblast growth factor on the healing of defects in the canine anterior cruciate ligament.

The effect of basic fibroblast growth factor (bFGF) on the healing of partial anterior cruciate ligament (ACL) lacerations was investigated in 17 adult mongrel canines. The defect was created in the midsubstance of both ACLs using a biopsy punch. In the bFGF group, a bFGF-impregnated pellet was sutured to the infrapatellar fat pad close to the defect. In the control group, the same pellet without bFGF was used. The healing process was evaluated macroscopically and histologically at 1, 3, 6, and 24 weeks postoperatively. In the bFGF group, a pannus-like tissue which contained abundant blood vessels extended into the defect from the adjacent synovial tissue in the early postoperative phase. Histological examination of the tissue which filled the defect revealed initial remodeling processes with a decreased number of cells and better orientation of the collagen fibers at 6-24 weeks. On the other hand, in the control group, poor healing processes were observed at each examination period. This study demonstrated that the application of a bFGF-impregnated pellet may enhance the healing potential of a partially injured ACL. Enhanced neovascularization and the formation of granulation tissue induced by bFGF early in the healing process accounted for the increased healing response.

Autologous pericranial graft resurfacing after high condylectomy and discectomy of the temporomandibular joint in rabbits.

PURPOSE: This study tested the effects of an autologous pericranial graft placed over the condyle on healing after high condylectomy and discectomy. MATERIALS AND METHODS: Sixteen young adult New Zealand white rabbits were divided into four test groups and a control group. Three animals from the test group and one control animal were killed at 2, 4, 8, and 12 weeks postoperatively. Each experimental animal received a high condylotomy and discectomy bilaterally. In addition, on one side a pericranial graft was placed on the shaved bone and immobilized with a tissue adhesive. RESULTS: The results showed better healing of the articular defects in the joints where grafts had been placed, including earlier and more complete soft tissue covering, more substantial neochondrogenesis, and better organization of the regenerated articular cartilage. CONCLUSION: Pericranial grafts may be helpful in facilitating healing after high condylectomy. However, technical improvements in the method of tissue fixation may be necessary.

The use of cryopreserved human skin allografts in wound healing following Mohs surgery.

BACKGROUND: Immediate reconstruction has become the preferred approach to management of full-thickness cutaneous defects following microscopically controlled excision (MCE) of tumors. In a minority of patients, however, large reconstructive procedures are contraindicated, and a long-term biological dressing that stimulates healing while minimizing wound care is desirable. OBJECTIVE: To assess the utility of cryopreserved human skin allografts (HSA) in wound care and wound healing following Mohs surgery. METHODS: Sixteen patients were treated with HSA following MCE and followed postoperatively for evidence of infection, involution, or survival of HSA, and granulation tissue production. Follow-up was 2-26 months. RESULTS: The use of HSA resulted in one of three general outcomes: rapid healing and rejection, subsequent full-thickness skin grafting, or persistence of HSA during prolonged healing. CONCLUSIONS: HSA are a safe alternative to immediate reconstruction in a carefully selected population of skin cancer patients. They minimize wound care while providing continuous wound coverage during healing, and are an efficient bridge to full-thickness skin grafting.