Falk Birkenfeld

Local treatment of meniscal lesions with vascular endothelial growth factor.

BACKGROUND The healing potential in the avascular regions of the meniscus is very limited, and improving the vascularity might be a reasonable way to improve healing. Vascular endothelial growth factor (VEGF) is one of the most potent proangiogenetic factors. We hypothesized that the local application of VEGF(165) would (1) improve the healing of a lesion in the avascular region of the meniscus, (2) induce angiogenesis in both the avascular and vascular regions, and (3) increase the amounts of VEGF mRNA and VEGF. METHODS In eighteen sheep, the medial menisci were cut longitudinally in the avascular region and were sutured. Three groups were established depending on the suture material: (1) uncoated Ethibond, (2) Ethibond coated with VEGF(165) and its carrier Poly(D,L-Lactide) (PDLLA), and (3) Ethibond coated with PDLLA. The contralateral medial menisci served as a control group. Each of the three suture type groups included six animals. After eight weeks, the sheep were killed, and the menisci were examined macroscopically. Immunohistochemistry of Factor VIII and VEGF and real-time reverse-transcription polymerase chain reaction (RT-PCR) of VEGF mRNA were performed. Additionally, the VEGF release kinetics from the VEGF/PDLLA-coated suture were evaluated in vitro. RESULTS In this model, VEGF did not improve meniscal healing. It did not increase angiogenesis in the avascular or vascular region, the VEGF concentration, or the amount of VEGF mRNA. VEGF release from the coated suture peaked on Day 3 and was nearly zero on Day 9. CONCLUSIONS The local application of VEGF(165) as eluted from suture did not increase meniscal angiogenesis or improve meniscal healing. In addition, there was no effect on the amount of VEGF mRNA and VEGF. The VEGF carrier (PDLLA) may have been inadequate because of the short duration of VEGF supply.

Detection of vascular endothelial growth factor (VEGF) in moderate osteoarthritis in a rabbit model

INTRODUCTION Vascular endothelial growth factor (VEGF) is detectable in later stages of human osteoarthritis (OA), but not in the healthy articular cartilage. Due to its capacity to increase matrix metalloproteinases and to decrease their inhibitors (tissue inhibitors of metalloproteinases or TIMPs) VEGF seems to play an important role in the development of osteoarthrosis. In late stages of osteoarthritis, invasion of blood vessels from the subchondral growth plate, synovitis with angiogenesis and osteophyte growth is observable. Several studies have revealed a central role for VEGF in all these phenomena. In order to investigate whether VEGF participates in early changes of OA or may even possess characteristics of a marker of OA, we developed an experimental posttraumatic OA New Zealand White rabbit animal model. MATERIALS AND METHODS In four skeletally mature New Zealand White rabbits, OA was induced by joint instability after transsection of the anterior cruciate ligament in both knees. After eight weeks the animals were killed. OA was verified histologically using the Mankin scale. Expression of VEGF was detected by immunohistochemistry and RT-PCR. Proteoglycans were evaluated by using HE and safranin-O staining. Four non-surgically treated animals acted as a control. RESULTS The mean Mankin score was 5.11 (±2.14), corresponding to a moderate OA. VEGF and VEGF transcripts were detectable in the cartilage of early experimental posttraumatic OA rabbits. Control samples remained negative for VEGF mRNA and protein. DISCUSSION The results of this study are promising concerning the role of VEGF as a diagnostic marker. VEGF could further be participated in early changes of OA. A therapeutic approach by modulation of VEGF production could be a possibility for the future.

Forces affecting orbital floor reconstruction materials – A cadaver study

INTRODUCTION The objectives of this study were: (i) to evaluate the applied force and the displacement of the orbital contents after orbital floor reconstruction using artificially aged reconstruction materials in fresh frozen human heads and (ii) to analyze the puncture strength of the materials. MATERIAL AND METHODS Six fresh frozen human heads were used, and orbital floor defects in the right and left orbit were created by 3.0 J direct impacts on the globe and infraorbital rim. The orbital floor defect sizes and displacements were evaluated after a Le-Fort-I osteotomy. RESULTS The orbital floor defect sizes were 208.3(SD, 33.4) mm(2) for the globe impacts and 221.8(SD, 53.1) mm(2) for the infraorbital impacts. The forces on the incorporated materials were approximately 0.003 N and 0.03 N for the PDS-foil and collagen membrane, respectively. The displacements of the materials were +0.9 mm and +0.7 mm for the PDS-foil and collagen membrane, respectively. The puncture strengths of the PDS-foil and collagen membrane decreased from approximately 70 N and 12 N at week 1 to approximately 5 N and 1.5 N at week 8 of artificial aging. CONCLUSION The force applied to the orbital content is minimal, and the puncture strengths of the artificially aged materials are more than sufficient for the measured forces.

Mechanical properties of collagen membranes: Are they sufficient for orbital floor reconstructions?

INTRODUCTION The most common reconstruction materials for orbital floor fractures are PDS (polydioxanone) foil and titanium meshes. These materials have advantages and disadvantages. Therefore, new materials are needed to improve surgical outcomes. MATERIALS AND METHODS Three resorbable collagen membranes (Smartbrane(®), BioGide(®), Creos(®)) were tested for their mechanical properties (puncture strength) in mint and artificially aged (3, 6, 8 weeks) conditions and were compared to PDS foil, titanium meshes (0.25 mm, 0.5 mm) and human orbital floors (n = 7). RESULTS The following puncture strengths were evaluated: human orbital floor, 0.81 ± 0.49 N/mm(2); 0.25 mm titanium mesh, 5.36 ± 0.25 N/mm(2); 0.5 mm titanium mesh, 16.08 ± 5.17 N/mm(2); Smartbrane, 0.74 ± 0.31 N/mm(2); BioGide, 1.65 ± 0.45 N/mm(2); and Creos, 2.81 ± 0.27 N/mm(2). After artificial aging, the puncture strengths were significantly reduced (p ≤ 0.05) at 3, 6 and 8 weeks as follows: Smartbrane, 0.05 ± 0.03 N/mm(2), 0.03 ± 0.02 N/mm(2), and 0.01 ± 0.01 N/mm(2), respectively; BioGide, 0.42 ± 0.06 N/mm(2), 0.41 ± 0.12 N/mm(2), and 0.32 ± 0.08 N/mm(2), respectively; and Creos, 2.02 ± 0.37 N/mm(2), 1.49 ± 0.42 N/mm(2), and 1.36 ± 0.42 N/mm(2), respectively. CONCLUSION The tested materials showed sufficient puncture strength for orbital floor reconstruction in mint condition. Moreover, after artificial aging, the Creos and BioGide membranes showed sufficient resistance, while Smartbrane showed equivocal data after eight weeks. Therefore, collagen membranes have adequate properties for further in vivo investigations for orbital floor reconstructions.

Maximum forces applied to the orbital floor after fractures.

Abstract The objective of this study was to measure the force on and displacement of completely detached intraorbital tissue from thebony orbit, as a worst-case scenario after orbital trauma, to preserve the maximum load and predict the necessary strength of reconstruction materials. Six fresh-frozen human heads were used, and orbital floor defects in the right and left orbits were created by the direct impact of 3.0 J onto the globe and infraorbital rim. The orbital floor defect sizes and displacements were evaluated after performing a Le Fort I osteotomy. In addition, after the repositioning of the completely detached intraorbital tissue, the forces and displacements were measured. The mean orbital floor defect sizes were 208.3 (SD, 33.4) mm2 for globe impacts and 221.8 (SD, 53.1) mm2 for infraorbital impacts. The mean intraorbital tissue displacement after the impact and before repositioning was 5.6 (SD, 1.0) mm for globe impacts and 2.8 (SD, 0.7) mm for infraorbital impacts. After repositioning, the displacements were 0.8 (SD, 0.5) mm and 1.1 (SD, 0.7) mm, respectively. The measured forces were 0.10519 (SD, 0.00958) N without the incorporation and approximately 0.11128 (SD, 0.003599) N with the incorporation of reconstruction materials. The maximum forces on the completely detached orbital tissue were minimal (∼0.11 N) and suggest the use of collagen membranes as reconstruction materials for orbital floor defects, at least in medium-sized fractures.

Deficiency of the DSPP-cleaving enzymes meprin α and meprin β does not result in dentin malformation in mice

Formation of dentin requires the maturation of procollagen I and the proteolytic processing of the dentin sialophosphoprotein (DSPP). These cleavage events can be facilitated by the metalloproteinases meprin α and meprin β as well as by bone morphogenetic protein 1 (BMP-1). All three enzymes have been shown to play important roles during collagen I maturation in vivo and their potential in cleaving DSPP was demonstrated in vitro. Hence, it has been discussed whether meprin α, meprin β, BMP-1 or all three are crucial factors in the onset and progression of dentin-related diseases and this issue is addressed here. In this study, we compare the incisors and molars of meprin α (Mep1a-/-)- and meprin β (Mep1b-/-)-deficient mice with wild-type (WT) controls on the macroscopic and microscopic level. The dentin was evaluated towards the bone mineral density, dentin volume, calcification and collagen matrix integrity. Using immunohistochemistry, we could identify meprin β, BMP-1 and DSPP/DSP in the pre-dentin of WT mice. Nevertheless, no significant dentin malformation was observed in Mep1b-/- or Mep1a-/- deficient mice.

Forces charging the orbital floor after orbital trauma.

Abstract The objectives of this study were (i) to evaluate different fracture mechanisms for orbital floor fractures and (ii) to measure forces and displacement of intraorbital tissue after orbital traumata to predict the necessity of strength for reconstruction materials. Six fresh frozen human heads were used, and orbital floor defects in the right and left orbit were created by a direct impact of 3.0 J onto the globe and infraorbital rim, respectively. Orbital floor defect sizes and displacement were evaluated after a Le Fort I osteotomy. In addition, after reposition of the intraorbital tissue, forces and displacement were measured. The orbital floor defect sizes were 208.3 (SD, 33.4) mm2 for globe impact and 221.8 (SD, 53.1) mm2 for infraorbital impact. The intraorbital tissue displacement after the impact and before reposition was 5.6 (SD, 1.0) mm for globe impact and 2.8 (SD, 0.7) mm for infraorbital impact. After reposition, the displacement was 0.8 (SD, 0.5) mm and 1.1 (SD, 0.7) mm, respectively. The measured applied forces were 0.061 (SD, 0.014) N for globe impact and 0.066 (SD, 0.022) N for infraorbital impact. Different fracture-inductive mechanisms are not reflected by the pattern of the fracture. The forces needed after reposition are minimal (∼0.07 N), which may explain the success of PDS foils [poly-(p-dioxanone)] and collagen membranes as reconstruction materials.

Forces charging the orbital floor after fractures.

The objective of this study was first to establish a method to measure forces and displacement of the orbital content in defects of the orbital floor in truncated fresh and unfixed heads and second to characterize reconstruction materials with regard to punctuation strength and compression. Orbital floor defects (10 × 20 mm and 15 × 20 mm; 3 mm behind the orbital rim) were prepared after Le Fort I osteotomy. The values of force and displacement were recorded in 6 freshly frozen human heads. In addition, the punctuation strength of 2 reconstruction materials (polydioxanone [PDS] foil and collagen membrane) was evaluated using a Zwick Z010 TN1 universal testing machine. The forces of the orbital content (28.41 [SD, 1.6] g) applied to the defects of 10 × 20 mm and 15 × 20 mm with an intact periorbita were 0.04 (SD, 0.003) N (0.0002 MPa) and 0.07 (SD, 0.02) N (0.0002 MPa), respectively, and with a split periorbita were 0.06 (SD, 0.03) N (0.0003 MPa) and 0.08 (SD, 0.06) N (0.00026 MPa), respectively. The displacement values without reconstruction materials of the 10 × 20-mm and 15 × 20-mm defects were 0.94 (SD, 0.7) mm and 1.2 (SD, 0.5) mm, respectively. The PDS foil could withstand forces of 118.9 (SD, 14.1) N (0.375 MPa), and the collagen membrane could withstand forces of 44.5 (SD, 5.3) N (0.14 MPa). This is the first study to report forces charging the orbital floor. The presented results support the use of PDS foils and collagen membranes as reconstruction materials for orbital floor defects, at least in smaller and medium-sized fractures.

Changes in human mandibular bone morphology after heat application

PURPOSE Intraosseous heat development is always a problem during bone surgery performed using rotary burs and ultrasound devices. However, only few data exist regarding the morphological effects of applied heat on bone surfaces. METHODS We used 24 human mandibular bone specimens of the mental region from six body donators. Three body donators were fixed in ethanol and the others were stored frozen. Heat application to the bone surfaces at temperatures of 40 degrees C, 50 degrees C, 60 degrees C and 100 degrees C for 1 min respectively, was performed under controlled conditions using an iron heater, and followed by examination using (i) scanning electron microscopy (SEM), (ii) demineralized paraffin sections, and (iii) cryostat sections (both HE staining). RESULTS There was no difference in the morphology or histology between fixed or unfixed bone specimens. The bone surface was smooth in both groups at 40 degrees C and 50 degrees C of heat application. Applications of 60 degrees C and 100 degrees C induced a rough-textured surface with small cavities visible with SEM and demineralized HE staining. The bone appeared to be unaffected at lower planes. The frozen HE histology could not be evaluated. Although useful in other studies, here the sections were broken and displaced on the glass slide. Therefore, this technique is not recommended by the authors. CONCLUSION Our findings suggest the applicability of SEM for bone surface morphology and demineralized paraffin sections (HE staining) for frontal plane evaluation. Fixed and non-fixed bone specimens seem to be equal in their morphology and can both be used in these kinds of studies.

Highly porous hydroxyapatite with and without local harvested bone in sinus floor augmentation: a histometric study in pigs

OBJECTIVE Sinus floor augmentation with autologous bone is an accepted treatment option in dental implantology. In this study, an entirely synthetic, nano-structured, hydroxyapatite-based bone substitute material (SBSM, NanoBone(®); Artoss, Rostock, Germany) was supplemented with a mixture of locally harvested bone to enhance osteogenesis. METHODS Bilateral sinus augmentation procedures were performed in eight domestic pigs using the lateral window technique. On the right side (control), 2.6 ml of SBSM was used, and on the left side (test), 2.6 ml of SBSM with additional 15% (390 μl) autologous bone was used. At the time of augmentation, a titanium implant (ITI(®)) was inserted from a laterocaudal direction. After 3 months, the sites of augmentation were removed and examined in non-decalcified sections by microradiography and fluorescence microscopy of sequentially labelled specimens and histometry. RESULTS On both sides, a significant amount of newly formed bone was observed. However, a statistically significant difference in the bone-implant contact was observed in the control group (median, 28.9%) compared with the test side with the additional autologous bone (median, 40.6%) (P = 0.01). Different bone density was achieved from the coronal to apical surfaces (medians, 54.6%, 9.6%, and 27.5%) compared with the test side (medians, 55.2%, 40.6%, and 44.2%). The median of augmentation height was 8.6 mm on the control side and 11.5 mm on the test side (P = 0.01). Bone apposition was observed in both groups after 15 days. CONCLUSION The SBSM shows acceptable results in sinus floor augmentation. The additional use of locally harvested autologous bone enhances bone density and osseointegration of the implants.