Synovial mesenchymal stem cells promote healing after meniscal repair in microminipigs.

OBJECTIVE: The induction of synovial tissue to the meniscal lesion is crucial for meniscal healing. Synovial Mesenchymal stem cells (MSCs) are an attractive cell source because of their high proliferative and chondrogenic potentials. We examined whether transplantation of synovial MSCs promoted healing after meniscal repair of extended longitudinal tear of avascular area in a microminipig model. DESIGN: Longitudinal tear lesion was made in medial menisci and sutured in both knees, and then a synovial MSC suspension was administered for 10 min only in unilateral knee. The sutured meniscus was evaluated morphologically and biomechanically at 2, 4, and 12 weeks. The behavior of transplanted MSCs was also examined. RESULTS: The meniscal healing at 12 weeks was significantly better in the MSC group than in the control group; macroscopically, histologically and by T1rho mapping analysis. Transmission electron microscopic analysis demonstrated that the meniscus lesion was occupied by dense collagen fibrils only in the MSC group. Biomechanical analysis revealed that the tensile strength to failure of the meniscus higher in the MSC group than in the control group in each microminipig. Synovial tissue covered better along the superficial layer from the outer zone into the lesion of the meniscus in the MSC group at 2 and 4 weeks in each microminipig. Synovial MSCs labeled with ferucarbotran were detected in the meniscus lesion and adjacent synovium by MRI at 2 weeks. CONCLUSION: Transplantation of synovial MSCs promoted healing after meniscal repair with induction of synovium into the longitudinal tear in the avascular zone of meniscus in pigs.

Preparation and microstructure analysis of chitosan/hydroxyapatite nanocomposites.

Chitosan/hydroxyapatite (HAp) composites with a homogeneous nanostructure have been prepared by a co-precipitation method. According to TEM observations, HAp crystallites in the composites formed elliptic aggregations 230 nm in length and 50 nm in width. The typical length of the aggregations corresponded approximately to that of a chitosan molecule. The size of the constituent HAp crystallites was found to be predominantly 30 nm in length and 10 nm in width, and the c-axes were well aligned in parallel with the chitosan molecules in the respective aggregations. The growth of the HAp crystallites is considered to occur at nucleation sites, most probably forming the complexes with amino groups on chitosan with calcium ions. The compact composites obtained have been found to be mechanically flexible, and this flexibility has been improved further by heating at 120 degrees C for 20 min in an autoclave with saturated steam pressure.

A study of induction of nerve regeneration using bioabsorbable tubes.

A silicone tube (S-tube) was packed with CPLA (copolymer of poly-L-lactide) fibers (S-tube+CPLA) or collagen fibers (S-tube+fiber). Two types of tube were prepared from a collagen sheet (Col-tube) and a bioabsorbable atelocollagen membrane for guided tissue regeneration (GTR-tube). They were packed with collagen fibers or films (Col-tube+fiber, GTR-tube+fiber and GTR-tube+film). Bridge grafting (15 mm in length) was performed with these tubes in a rat sciatic nerve model. Specimens were harvested after 8 weeks. Minifascicles were formed in the open space between the CPLA fibers in the S-tube+CPLA group. Regenerated axons were also formed in the degenerated collagen fibers in the S-tube+Col group. Immunocytochemistry evaluation revealed that Schwann cells invaded the space in the absorbing collagen fibers. Histologic analysis of the regenerated axons in the groups with Col-tubes or GTR-tubes revealed that both the Col-tube and the GTR-tube packed with collagen fibers were effective in providing a scaffold for regenerating nerve tissue.

The biocompatibility and osteoconductive activity of a novel hydroxyapatite/collagen composite biomaterial, and its function as a carrier of rhBMP-2.

A hydroxyapatite/type I collagen (HAp/Col) composite, in which the hydroxyapatite nanocrystals align along the collagen molecules, has been prepared. The biocompatibility, osteoconductive activity, and efficacy as a carrier of recombinant human bone morphogenetic proteins (rhBMPs) of this novel biomaterial were examined. The composite material was implanted in the backs of Wistar rats, and specimens were collected for histological observations until week 24. In a second experiment, other samples of the composite material (5 x 5 x 10 mm3) were drilled and immersed in a solution of rhBMP-2 (0, 200, 400 microg/mL), and subsequently grafted in radii and ulnae in beagle dogs. As a control, three unfilled holes were left in one radius and ulna. X-ray images were prepared, and specimens collected for histological observation at weeks 8 and 12. Histological findings of the composites grafted in rats showed that the surface of the material was eroded as a result of macrophage infiltration. X-ray images and histological findings for the composites implanted in dogs support the idea that HAp/ Col has a high osteoconductive activity and is able to induce bone-remodeling units. In cases where the implants are grafted at weight bearing sites, treatment with rhBMP-2 at a dose of 400 microg/mL may be useful to shorten the time needed until bone union has occurred.

Development of an orthodontic simulator for measurement of orthodontic forces.

In this research we developed an orthodontic simulator for analysis of orthodontic forces distribution in the case of the continuous arch technique. In order to take into account vertical force, besides horizontal force and occlusogingival moment on instrumented artificial tooth, tooth root was designed as a ring load cell. Fixing each artificial tooth onto combined mechanical stages enabled variation of tooth anteroposterior position and inclination related to occlusal plane. Calibration of forces and moment of each instrumented artificial tooth showed linearity of outputs and provided the generalized inverse matrix for evaluation of forces and moments from measured strain data. A simulation to retract the anterior teeth was accomplished, and distribution of forces and moment acting at the bracket of each tooth were determined. The average accuracy was 82% and 97% in the horizontal and vertical direction, respectively, by comparing the applied forces and the evaluated ones.

[Mechanical behavior of the cement layer of a cast crown--effect of the mechanical properties of casting alloy].

Experimental crowns with uniform thickness at their axial surfaces were made of dental casting gold-silver-palladium alloy (12% gold) and dental casting gold alloy (Type IV). They were cemented on model teeth for abutment and their deformations under applied loads were measured with strain gages. The effect of axial metal thickness on the crown's rigidity and the cement fracture strength were examined. Among the structures of cemented crowns and abutment teeth, that of the axial metal thickness of 0.3 mm had cement failure at the smallest loads. As the metal thickness was increased, the crown's deformation decreased and the cement failure load increased. For a thickness greater than 0.7 mm, however, the increase in failure load was not obvious. The difference of the casting alloy had no significant effect on the cement fracture. Accordingly, the rigidity of dental prostheses should be one of the most important design factors for preventing failure under occlusal load, and it is recommended that the thickness of the axial surface should be more than 0.7 mm.

Fatigue behavior of the zinc-phosphate cement layer.

PURPOSE: Fatigue behavior of luting cement film between model fixed partial dentures (FPD) and their dies was investigated to gain a better understanding of the mechanical properties of zinc-phosphate cement. MATERIALS AND METHODS: Fifty-six brass dies were made, half of them with grooves and half without grooves. A cantilever FPD for each die was cast in Au-Pd-Ag alloy. Four linear strain gauges were attached to the marginal portions of each retainer. The FPDs were cemented to the dies using zinc-phosphate cement. A fatigue test machine was used to cyclically load the specimens with 50 or 100 N for 5.5 x 10(5) cycles (1 Hz) in 37 degrees C water. During the fatigue test, strain measurements were made after every 0.5 x 10(5) load cycles to discover if strain response was altered. After completion of the fatigue test, ultimate tensile strength of each specimen was measured and statistically analyzed by 2-way analysis of variance. RESULTS: With or without grooves, strain responses were the same at every periodic strain measurement throughout the 50-N fatigue test. For the 100-N fatigue test, strain responses for the specimens without grooves were different before and after cyclic testing, but there was no difference for the specimens with grooves. There were no significant differences in tensile strength among the specimens after the fatigue tests. CONCLUSION: No fatigue fracture was observed in zinc-phosphate cement film after repetitive loading. Repetitive loading may improve stress transmission within the zinc-phosphate cement film.

Cell culture test of TCP/CPLA composite.

Thermoplastic polymers filled with a ceramic powder (here described as a "composite") of beta-tricalcium phosphate (TCP) and copoly-L-lactide (CPLA) was prepared by a heat-kneading method. The TCP/CPLA composite obtained had a three-point bending strength of 51.26 +/- 634 MPa, a Young's modulus of 5.18 +/- 1.11 GPa, and a fracture strength of 52.64 +/- 2.975 MPa, which is sufficient for usage as an artificial bone implant material. According to cell culture tests using MC3T3-E1 cells, the TCP/CPLA composite showed no cytotoxicity and the cells were in close contact with the surfaces of the composite without any observable morphological change. These results suggest that a composite consisting of osteoconductive TCP and biodegradable CPLA is suitable as a bioactive artificial bone material.

Synthetic collagen fibers coated with a synthetic peptide containing the YIGSR sequence of laminin to promote peripheral nerve regeneration in vivo.

The usefulness of collagen fibers and the YIGSR sequence (Tyr-lle-Gly-Ser-Arg) of laminin for nerve regeneration were examined in vivo. Type I collagen gel (G-group), Type I collagen fibers (F-group), Type I collagen fibers coated with laminin (L-group) or the YIGSR sequence (Y-group) were packed into silicone tubes, 15 mm long, and transplanted to the sciatic nerves of Wistar rats. Empty silicone tubes were used as the control. The animals were sacrificed 8 weeks after transplantation. Bridging of the nerve was confirmed in the F-(7/12), Y-(7/10) and L-group (6/10), but no bridging was observed in any of the animals of the G- and control group. Nerve regeneration among the space of collagen fibers was observed, and it was suggested that fibroblasts infiltrated the gap in the substance of the degenerated collagen fibers were followed by Schwann cells on the basis of immunocytochemistry. The number of myelinated axons per regenerated tissue in the tube (density), and total area of myelinated axons per measured regenerated tissue in the tube (% axon area) in each the L- and Y-group were significantly higher than that in the F-group (P < 0.05). These results suggest the possibility of obtaining adequate nerve regeneration with new artificial materials only.

Deformation of restoration and fracture of luting cement film.

OBJECTIVES: Functional occlusal loading can generate complicated stresses in the luting cement film between a restoration and its abutment. Such stresses may lead to cement fractures, which can promote cement dissolution and induce clinical problems. The purpose of this study is to determine the function of the cement film under loading, and to clarify the relationship between the deformation of the restoration and the cement fractures. METHODS: Eccentric loads were applied onto model extension bridges which were cemented on brass dies with zinc phosphate cement. Strain measurements were recorded using of a strain gauge method, ultimate strength by tensile tests, leakage by dye penetration tests and cement fractures by ultrasonic microscopic analyses. Additionally, the bridges which were just seated on their dies without cementation were loaded, and strain measurements were recorded. RESULTS: The cases for linear strain increment with loading showed high tensile strength and minimal dye penetration. The cases with non-linear strain behavior showed low tensile strength and considerable dye penetration. However, no fracture was observed in the occlusal cement film in both cases. The cement fracture first occurred at the opposite marginal region to the loading side. CONCLUSIONS: One of the significant functions of the cement film is diminishing the deformation of restorations during loading. The deformation of the restoration affects initiation and propagation of the cement fracture considerably.

Surface strain on crown and luting cement fractures.

This study investigated fractures of the cement layer between a crown and a die. Model crowns were luted on dies with zinc phosphate cement and various magnitudes of load were applied. Surface strain on the crown was measured using strain gauges. Seal was evaluated using dye penetration and tensile tests. Results showed that cement fracture affected surface strain behavior on the crown and was detected using the surface strain measurement. The dye penetration test and the tensile test could not be used to detect the cement fracture. It is suggested that the three-dimensional relative positioning between the crown and the die affected the development of the cement fracture.

A comparison of in vivo and in vitro strain with posterior fixed partial dentures.

PURPOSE: Deformation of fixed partial dentures during function has been related to clinical failures. However, such deformation has not been measured quantitatively, and the effect on deformation of the viscoelastic behavior of peridental structures is not well understood. The purpose of this study was to determine the strain distribution of fixed partial dentures during function and to compare the biomechanical behavior of fixed partial dentures in vivo and in vitro. MATERIAL AND METHODS: Three conventional posterior fixed partial dentures and two complete crowns were fabricated. Both in vivo and in vitro strain measurements of each of the five prostheses were recorded by a strain gauge method. A static load was produced through occlusal force in vivo and with a universal testing machine in vitro. RESULTS: The resulting difference between the strain distribution for the two crowns in vivo and in vitro was negligible, but the whole fixed partial denture demonstrated a substantial amount of strain in vivo. CONCLUSIONS: The results of this study suggest that serious problems with strain may occur in a long-span mandibular posterior fixed partial denture.

Intercondylar notch width and its relation to the configuration and cross-sectional area of the anterior cruciate ligament. A cadaveric knee study.

If a narrow intercondylar notch contains a smaller anterior cruciate ligament, that may explain why people with narrow notches have a higher incidence of anterior cruciate ligament injuries. To investigate the significance of notch width measurement, we used 16 embalmed cadaveric knees. A positive mold of the entire anterior cruciate ligament, including its femoral and tibial insertions, was created with silicone rubber and plaster commonly used for dental molding. We had two hypotheses to test from this study. One was that the dimensions of the anterior cruciate ligament can be predicted by the notch width. The other was that the size of a person's anterior cruciate ligament can be predicted by a caliper measurement of the intercondylar notch. The width, sagittal length, and cross-sectional area of the midsubstance and the femoral and tibial insertions of the anterior cruciate ligament were measured. The notch width index, the ratio of notch width to width of the femoral condyle, showed a positive correlation only to the ratio of width to sagittal length of the tibial insertion. None of the parameters showed any differences between the knees with a notch width index less than or equal to 0.2 and those with a notch width index greater than 0.2. The knees with small notch width indexes did not have thinner anterior cruciate ligaments in them. These findings may not be applicable to knees obtained from other races, i.e., not Japanese.

A cadaveric study on the stability of lumbar segment after partial laminotomy and facetectomy with intact posterior ligaments.

This study aimed to biomechanically investigate a change in the stability of decompressed segments in the degenerative lumbar spine. Cyclic loading tests to the cadaveric spinal unit were conducted in compressive and bending directions with a stepwise resection of the posterior elements such as wide fenestration (partial laminotomy), unilateral and bilateral facetectomy with the intact supra- and interspinous ligaments and spinous process. Flexion stability did not decrease significantly, and may reflect the fact that the aforementioned ligamentous structures were preserved throughout the resection process. After wide fenestration, the loss of compression stiffness amounted to only half that of bilateral facetectomy, and extension stiffness dropped to this same level. Lateral bending stiffness was maintained until unilateral facetectomy was performed. We recommend restricted decompression surgeries for a single-level central stenosis in the degenerative lumbar spine.

Magnetic resonance imaging and pathologic studies on lateral fluid percussion injury as a model of focal brain injury in rats.

In this study, morphologic changes in brain lesions initiated by moderate lateral fluid percussion injury in rats were investigated chronologically using high-resolution magnetic resonance imaging (MRI) and histopathologic methods. Rats were subjected to moderate fluid percussion injury (average 2.80 +/- 0.48 atmospheres) over the exposed dura overlying the right parietal cortex. MRI obtained in vivo were compared with corresponding pathologic findings at 1, 6, and 24 h and at 3, 6, 14 and 80 days after injury. T2-weighted images showed scattered low-signal intensity in the injured cortex within a few hours after injury, whereas histologic findings revealed intraparenchymal hemorrhages. T2-weighted images of the ipsilateral cerebral cortex and/or corpus callosum showed a high-signal-intensity area 4 h after injury. The high-signal-intensity area became largest in size between 6 and 24 h, then declined gradually, and almost disappeared 14 days after injury. Histologic examination revealed pyknosis, retraction of the cell body of neurons with vacuolated neuropil in the corresponding regions 6 and 24 h after injury, and cystic necrosis 14 days after injury. The location and extent of these pathologic changes were depicted accurately by MRI in vivo. In the hippocampus, pyknosis and retraction of the cell body of pyramidal neurons were observed on the injured side 24 h after injury, and the number of neurons in the CA1 and CA2-CA3 regions decreased significantly on the same side by 14 days after injury. It is concluded that morphologic changes in the brain following experimental traumatic brain injury in rats are detectable in vivo by high-resolution MRI, and that MRI may be useful for the evaluation of treatment effects in experimental brain injury.

Tensile behaviour of fibroblasts cultured in collagen gel.

An in vitro morphological study on the relation of the stress field and the orientation of cells with the use of the time lapse video recording system was carried out. Specimens of thin collagen gel membrane, within which cells are proliferating, were examined under various mechanical conditions. Cell stretching, cell orientation, cell migration, cell proliferation and generation of tensile stress in collagen gel were recorded. It was demonstrated that fibroblasts generate tension and change their orientation along the tensile direction. A hypothetical mechanism for such phenomena is proposed, that is, fibroblasts generate tension and make tense collagen fibres, then cells stretch themselves along the tense fibres and increase tension in this direction. Thus, this mechanism works as a positive feedback loop which enables cells to make tensile stresses and align along them in accordance with the mechanical environment.

Changes in lCBF, morphology and related parameters by fluid percussion injury.

We investigated the pathophysiological and morphological responses of anaesthetized rats to fluid percussion brain injury generated by an original midline fluid percussion injury device. Following different grades of trauma, lCBF was measured continuously in the right parietal cortex through a burr hole using laser Doppler flowmeter, and physiological parameters were monitored. Pathological changes also were evaluated microscopically. During the first 2 hours following trauma, we found four patterns of cerebral circulatory responses. Little measurable pathophysiological response occurred after percussion pulses of less than 1.33 atmospheres (atm). In animals subjected to pulses of greater than 4.30 atm, lCBF increased synchronously with blood pressure, and then both parameters decreased continuously until death. In animals subjected to pulses of 1.53 to 2.33 atm, trauma produced a transient increase in lCBF with no synchronous rise in blood pressure. In animals subjected to pulses of 2.70 to 3.87 atm, lCBF increased synchronously with blood pressure immediately following the injury, but had decreased markedly by 60 seconds and remained below the pre-injury baseline. Blood pressure recovered to baseline within 4 minutes of the injury. The transient increase in lCBF occurred within 5 seconds following percussion pulses of greater than 1.53 atm and appeared to be independent of the rise in systemic blood pressure. Apnoea occurred in animals subjected to pulses of greater than 1.53 atm, and the duration of apnoea and mortality rate correlated with the magnitude of the applied injury. A power decrease in the electroencephalogram post-injury and a delay in its recovery, both depended on the magnitude of the injury with few regional differences in the beta-2 band power. The distribution and extent of blood-brain barrier disruption and small haemorrhages also correlated with the magnitude of the injury. The number of neurons decreased significantly in both hippocampi by 2 weeks following moderate trauma. The four patterns of lCBF changes demonstrated in the present study, as well as the other responses to injury, may be useful for studying graded models of various diffuse brain injuries.

An approach to computer aided porcelain forming system.

To examine a new concept for computerized production of dental prostheses, an experimental porcelain forming system was devised. Unlike other dental CAM systems, the system is intended to form porcelain much as human technicians do. The system builds up an object by alternately applying water and droplets of porcelain powder. Test specimens were made by forming porcelain powder into square plates under different forming conditions. Surface characteristics and shapes of formed porcelain were found to be affected by conditions such as quantity of water, forming area and forming thickness. The proper quantity of water for forming was influenced by evaporation. It was concluded that investigation into feedback control of the water supply in response to varying forming conditions is required.

Fuzzy and conventional control of high-frequency ventilation.

A high-frequency ventilator was developed, consisting of a single-phase induction motor, an unbalanced mass and a mechanical vibration system. Intermittent positive pressure respiration was combined with high-frequency ventilation to measure end-tidal pCO2. Hysteresis was observed between the rotational frequency of the high-frequency ventilator and end-tidal pCO2. A fuzzy proportional plus integral control system, designed on the basis of the static characteristics of the controlled system and a knowledge of respiratory physiology, successfully regulated end-tidal pCO2. The characteristics of gas exchange under high-frequency ventilation was approximated by a first-order linear model. A conventional PI control system, designed on the basis of the approximated model, regulated end-tidal pCO2 with a performance similar to that of the fuzzy PI control system. The design of the fuzzy control system required less knowledge about the controlled system than that of the conventional control system.

Effects of postoperative immobilization on the reconstructed anterior cruciate ligament. An experimental study in rabbits.

To investigate the effects of postoperative immobilization and limited motion on reconstructed anterior cruciate ligaments, 28 rabbits received an anterior cruciate ligament reconstruction using autogenous Achilles tendon and were then divided into three groups: fully immobilized, 4 weeks immobilized, and limited motion. Two rabbits from each group were evaluated macroscopically, histologically and microangiographically at 4-week intervals until 12 weeks postoperatively. An additional six rabbits in the 4 weeks immobilized and limited motion groups were studied biomechanically at 12 weeks postoperatively. Macroscopically, both immobilized groups showed more proliferation of the infrapatellar fat pad, which was adherent to the reconstructed anterior cruciate ligament. Histology revealed more rapid regeneration of reconstructed anterior cruciate ligaments in the limited motion group, with no findings of necrosis in the mid-substance. Microangiography indicated faster normalization of vascularity in the limited motion group. The biomechanical study showed no significant difference in laxity between the 4 weeks immobilized and limited motion groups. The graft stiffness and maximum load to failure were greater for the limited motion group, although the increase was not statistically significant. The histologic and microangiographic results from the limited number of animals in this study support limited postoperative motion in the anterior cruciate ligament reconstructed knee. However, there were no differences in terms of the biomechanical parameters at 12 weeks postoperatively between the immobilized and limited motion treatment modes.