Where and what damage occurs at the acromial undersurface in patients with rotator cuff tears?

BACKGROUND: The gross pathology of the acromial undersurface in shoulders with rotator cuff tears with subacromial impingement is not completely understood. Many researchers have focused on damage to the anterior one-third area of the acromial undersurface, but few have studied the middle and posterior one-third areas. The purpose of this study was to clarify where and what damage occurs at the acromial undersurface in patients with rotator cuff tears. METHODS: We performed arthroscopic shoulder (n = 182, all with rotator cuff tears; mean age, 64.9 ± 8.4 years) and cadaveric shoulder (n = 23, 14 intact cuffs and 9 rotator cuff tears; mean age, 74.8 years) evaluations to observe the extent and degree of damage to the acromial undersurface. We statistically analyzed the association between the severity of the damage to the acromial undersurface (assessed using the Copeland-Levy classification as A0, normal; A1, minor scuffing; A2, major damage; or A3, visualization of bare bone area) and rotator cuff tear size (assessed using the classification of DeOrio and Cofield as partial; small, <1 cm; medium, 1-3 cm; or large or massive, >3 cm). RESULTS: The anterior, middle, and posterior one-thirds of the acromial undersurface were somewhat damaged (class A1-A3) in 92.6%, 90.1%, and 78.6% of shoulders with rotator cuff tears, respectively, according to arthroscopic evaluation. Increasing cuff tear size was significantly associated with worsening degree of damage to the acromial undersurface (P < .001). In the 9 cadaveric shoulders with rotator cuff tears, class A1-A3 damage was identified in the anterior one-third area in 100%, in the middle one-third area in 88.9%, and in the posterior one-third area in 33.3%. In the 14 cadaveric shoulders with a normal rotator cuff, class A1-A3 damage was identified in the anterior one-third area in 35.7%, in the middle one-third area in 14.3%, and in the posterior one-third area in 0.71%. CONCLUSION: Damage to the acromial undersurface in patients with rotator cuff tears occurred at the middle, posterior, and anterior one-third areas, and the degree of damage was related to cuff tear size. Surgeons should evaluate the entire acromial undersurface to check for subacromial impingement damage at the middle and posterior one-third areas as well as the anterior one-third area of the acromial undersurface; this might aid in the treatment of patients with rotator cuff disease or subacromial impingement syndrome.

Influence of Nanobubble Size Distribution on Ultrasound-Mediated Plasmid DNA and Messenger RNA Gene Delivery.

The use of nanobubbles (NBs) for ultrasound-mediated gene therapy has recently attracted much attention. However, few studies have evaluated the effect of different NB size distribution to the efficiency of gene delivery into cells. In this study, various size of albumin stabilized sub-micron bubbles were examined in an in vitro ultrasound (1 MHz) irradiation setup in the aim to compare and optimize gene transfer efficiency. Results with pDNA showed that gene transfer efficiency in the presence of NB size of 254.7 ± 3.8 nm was 2.5 fold greater than those with 187.3 ± 4.8 nm. Similarly, carrier-free mRNA transfer efficiency increased in the same conditions. It is suggested that NB size greater than 200 nm contributed more to the delivery of genes into the cytoplasm with ultrasound. Although further experiments are needed to understand the underlying mechanism for this phenomenon, the present results offer valuable information in optimizing of NB for future ultrasound-mediate gene therapy.

Hypotonia-induced cell swelling enhances ultrasound-induced mechanical damage to cancer cells.

INTRODUCTION: It has been shown that killing of suspended cells by low-intensity ultrasound (0.08-0.11 W/cm(2)) can be enhanced by a mild non-lethal hypotonic (146 mOsm) medium. PURPOSE: In this study we wished to determine whether hypotonia-induced cell swelling of suspension cells was directly related to enhancement of ultrasound-mediated cell killing, and to verify whether similar effects could be observed on circulating and attached cells. METHODS: U937 cells under mild hypotonia were exposed to ultrasound for different times with real-time monitoring of cell size using a particle-size-distribution analyzer. To study the effect on attached cells, HeLa cells were exposed to ultrasound while under hypotonia in an in vivo-simulated set-up. RESULTS: The result showed that the enhanced cell killing (up to more than twice) was directly proportional to hypotonia-induced cell swelling. Similar membrane damage based on PI staining could be observed on HeLa cells treated with hypotonia. An in vivo-simulated circulating system also showed similar findings for hypotonia-enhanced ultrasound cell killing. CONCLUSION: These findings showed that mild hypotonia can be used to augment the effect of ultrasound in the treatment of cancers, particularly leukemia. The results showing that such enhancement is related to cell swelling could guide us toward proper timing of sonication while under hypotonic treatment.

Nanobubble Mediated Gene Delivery in Conjunction With a Hand-Held Ultrasound Scanner.

Recent research has revealed that nanobubbles (NBs) can be an effective tool for gene transfection in conjunction with therapeutic ultrasound (US). However, an approach to apply commercially available hand-held diagnostic US scanners for this purpose has not been evaluated as of now. In the present study, we first compared in vitro, the efficiency of gene transfer (pCMV-Luciferase) with lipid-based and albumin-based NBs irradiated by therapeutic US (1MHz, 5.0 W/cm2) in oral squamous carcinoma cell line HSC-2. Secondly, we similarly examined if gene transfer in mice is possible using a clinical hand-held US scanner (2.3MHz, MI 1.0). Results showed that lipid-based NBs induced more gene transfection compared to albumin-based NBs, in vitro. Furthermore, significant gene transfer was also obtained in mice liver with lipid-based NBs. Sub-micro sized bubbles proved to be a powerful gene transfer reagent in combination with conventional hand-held ultrasonic diagnostic device.

Growth inhibition of neurofibroma by ultrasound-mediated interferon γ transfection.

PURPOSE: We have previously shown that ultrasound-mediated transfection (sonotransfection) can be optimized using a concept based on the ultrasound-induced apoptosis produced in our in vitro experiments. At optimized conditions, we have shown, using five cancer cell lines, that sonotransfection is superior to other conventional nonviral methods. Interferon gamma (IFN-γ) transfection using lipofection has been found to markedly inhibit the proliferation of neurofibroma cell lines. In this study, we investigated whether sonotransfection of IFN-γ to neurofibroma cell lines can suppress cell proliferation. METHODS: The ultrasound device used was the SonoPore KTAC-4000, which is capable of various acoustic settings. Ultrasound transducers at an oscillation frequency of 1.011 MHz were used; the potential ideal conditions were an intensity of 0.17 W/cm(2) at a burst frequency of 0.5 Hz, 25% duty factor, and 30-s sonication duration. Cells were assayed at 3 and 5 days after sonication. RESULTS: The transfection efficiency was found to be 12%. The ultrasound-treated cells were successfully transfected with IFN-γ genes as detected by enzyme-linked immunosorbent assay, and the cell growth ratio in the IFN-γ sonotransfection group tended to be lower than that in the other experimental groups. CONCLUSION: These results suggested that IFN-γ sonotransfection could potentially become a nonsurgical method for treating skin lesions such as neurofibromas.

Echographic and physical characterization of albumin-stabilized nanobubbles.

There has been increasing interest in using nanobubbles (NBs) for ultrasound mediated drug delivery as well as for ultrasound imaging. Albumin NBs are especially attractive for its potential of becoming a versatile platform for drug carriers and molecular targeted therapy agents. However, physical characterization of NBs is generally considered to be difficult due to various technical issues, such as concentration limitations, nanoparticle contamination, etc. In the present study, we measured the size distribution, concentration and weight density of albumin stabilized NBs by means of multiple nanoscale measurement modalities. Laser nanoparticle tracking analysis, multicolor flow cytometry, resonance mass evaluation showed consistent measurement results of the NBs with low mass weight density and diameter size ranging from 100 nm to 400 nm. Furthermore, the NB solution showed excellent images by high frequency ultrasound (30-50 MHz) in flow model acoustic phantoms. The NBs also induced acute cell disruption by low intensity ultrasound (0.8 W/cm2) irradiation. We successfully fabricated and characterized albumin stabilized NBs which could serve as an effective platform for future theranositic agents.

Predictors of safety margin for coracoid transfer: a cadaveric morphometric analysis.

BACKGROUND: The purpose of this study was to investigate the relationship between the bone length available for coracoid transfer without coracoclavicular ligament injury and the distance from the coracoid tip to the attachments of the coracoacromial ligament or pectoralis minor. We hypothesized that cadaver height and the soft tissue attachments on the coracoid process were predictive factors for sufficient bone length for coracoid transfer. METHODS: This study included 28 shoulders from Japanese cadavers: 19 male and 9 female. The distance from the coracoid tip to the distal attachment of the coracoclavicular ligament and the anterior and posterior margins of the coracoacromial ligament or pectoralis minor on the coracoid process were measured. RESULTS: The mean available length for coracoid transfer was 24.8 ± 3.4 mm. There was a significant difference in length between male and female subjects, being 26.0 ± 2.9 mm and 22.2 ± 3.0 mm, respectively (p = 0.004). High positive correlations were found between the length of the coracoid transfer and cadaver's height (r = 0.48, p = 0.009) and the distance from the coracoid tip to the anterior coracoacromial ligament attachment (r = 0.63, p < 0.001). The receiver operating characteristic curve area under the curve for cadaver height was 0.72 while that for distance from coracoid tip to anterior coracoacromial ligament was 0.88 when predicted for a sufficient length for coracoid transfer > 25 mm. CONCLUSIONS: Our findings will aid surgeons in preoperative planning and performing of osteotomy of the coracoid safely by predicting the available length of coracoid bone graft.

Anatomical study of the position and orientation of the coracoclavicular ligaments: Differences in bone tunnel position by gender.

BACKGROUND: Reconstructing both coracoclavicular ligaments following acromioclavicular dislocation has recently been reported to restore the function of the acromioclavicular joint better than traditional procedures. Knowing the appropriate position and orientation of the bone tunnels and the potential risks of neurovascular injuries leads to safe reconstruction. We aimed to answer the following questions: what is the difference in the accurate clavicular bone tunnel positions (BTPs) during coracoclavicular ligament reconstruction between sex, and what are the potential risks for neurovascular injuries? HYPOTHESIS: The BTPs differ by sex at the site of coracoclavicular ligament reconstruction. PATIENTS AND METHODS: We introduced two Kirschner wires into 25 cadaver shoulders (17 male, 8 female), one through the insertion center of the trapezoid ligament and one through the conoid ligament, and measured the distance from the respective Kirschner wire insertion points to the bony landmarks of the clavicle and the oblique angle of each Kirschner wire. The shortest distance from the insertion point of each Kirschner wire to the suprascapular nerve and artery was also measured. RESULTS: While the distance from the acromioclavicular joint to the respective Kirschner wire insertion points tended to be longer in males, the ratio of these insertion points to total clavicle length was constant. Other measurements for respective Kirschner wire insertions to the bony landmarks and neurovascular structures were comparable, as were abduction and retroversion angles. The distance from the suprascapular nerve to the insertion point of the conoid ligament at the coracoid process was 13.8±4.0mm, while the distance from the suprascapular artery was 7.1±3.3mm. DISCUSSION: Appropriate position and orientation of the bone tunnels, and the ratio of the BTPs to the total clavicular length, aid surgeons in performing the reconstruction. The conoid ligament insertion on the coracoid was just proximal to the suprascapular artery, so surgeons should be careful with conoid insertion. LEVEL OF EVIDENCE: V, cadaver study.

Therapeutic potential of low-intensity ultrasound (part 1): thermal and sonomechanical effects.

In this first part of the review, we will focus on and discuss various aspects of low-intensity ultrasound (US), with emphasis on mild thermal effects, apoptosis induction, and sonomechanical effects. Mild thermal effects of US have been commonly applied to physical therapy. Though US has clear beneficial effects, the advantage of using US over other heating modalities remains unclear. US has also been used in vivo and clinically in the treatment of wounds and fractures, with promising results. On the biomolecular level, studies have shown that US can induce apoptosis and that certain conditions can provide optimal apoptosis induction. As to potential therapeutic applications, in addition to the thermal and other physical effects, apoptosis induction by US may offer direct and rapid treatment of tumors or cancer tissues. Technological advances and rapidly accelerating research in this field are providing an ever-increasing array of therapeutic options for lowintensity US.

Enhanced cell killing and apoptosis of oral squamous cell carcinoma cells with ultrasound in combination with cetuximab coated albumin microbubbles.

Targeted microbubbles have the potential to be used for ultrasound (US) therapy and diagnosis of various cancers. In the present study, US was irradiated to oral squamous cell carcinoma cells (HSC-2) in the presence of cetuximab-coated albumin microbubbles (CCAM). Cell killing rate with US treatment at 0.9 W/cm2 and 1.0 W/cm2 in the presence of CCAM was greater compared to non-targeted albumin microbubbles (p < .05). On the other hand, selective cell killing was not observed in human myelomonocytic lymphoma cell line (U937) that had no affinity to cetuximab. Furthermore, US irradiation in the presence of CCAM showed a fivefold increase of cell apoptotic rate for HSC-2 cells (21.0 ± 3.8%) as compared to U937 cells (4.0 ± 0.8%). Time-signal intensity curve in a tissue phantom demonstrated clear visualisation of CCAM with conventional US imaging device. Our experiment verifies the hypothesis that CCAM was selective to HSC-2 cells and may be applied as a novel therapeutic/diagnostic microbubble for oral squamous cell carcinoma.

Topographical relationship between the human lateral circumflex femral artery and saphenous nerve.

This study was performed to investigate causes of various types of topographical relationship between the lateral circumflex femoral artery (L) and the saphenous nerve (S). Femoral artery (F), deep femoral artery (P), L and S of 186 legs of 93 Japanese adult cadavers were submitted to anatomy. Further, the levels of origin of L in thigh were measured. L were classified into nine types by the origins of L and topographical relationship between L and S. The incidence of various types of L is different among researchers. Our findings proved that these differences were caused by the differences in evaluations of twig from ascending branch (AB) or descending branch (DB) of L. In cases of L originating from F, incidence of L positioned in front of S is significantly higher than L originating from P (p < 0.01). In cases of L originating from F, L positioned in front of S originates from F at the significantly more proximal level compared to L positioned posterior to S (p < 0.001). Furthermore, also in cases of L originating from P, L positioned in front of S originates from P at the significantly more proximal level compared to L positioned posterior to S (p < 0.001). It is supposed that the topographical relationship between L and S changes depending on the artery where L originates and the level of origin of L.

A morphological study on the human obturator externus muscle with reference to anomalous muscle and anomalous fasciculus originating from the obturator externus muscle.

To elucidate the essential nature of the human obturator externus muscle (OE), the gross anatomy of the anomalous muscle and anomalous fasciculus originating from this muscle as well as the posterior division of the obturator nerve (P) were examined in 73 thighs of 45 Japanese adult cadavers. The investigation of anomalous muscle and anomalous fasciculus was effected by observation of the supernumerary muscular branch and supernumerary muscular twig from the P. The anomalous muscle occurring between the adductor brevis muscle (AB) and adductor minimus muscle (AMi) (37/73 thighs; 50.7%) and the anomalous fasciculus fused to the posterior surface of the AMi (18/73 thighs; 24.7%) were detected. It was demonstrated by the course of the P and the suppling nerve, that the OE was divided into its superior fasciculus (or fasciculi) (SF) and main belly by the P, moreover the anomalous muscle originated from the former and the anomalous fasciculus from the latter. Presumably, in the process of ontogeny, the SF of the original OE retained its original morphology (23/73 thighs; 31.5%), a part of the SF was converted to anomalous muscle (27/73 thighs; 37.0%), the entire SF was converted to anomalous muscle (10/73 thighs; 13.7%), and the entire SF underwent retrogression and disappeared (13/73 thighs; 17.8%). The presence (50/73 thighs; 68.5%) or absence (23/73 thighs; 31.5%) of piercing of the OE by the P seemed to depend upon the circumstances surrounding the morphological change in the SF. The original OE was considered to occur as an OE pierced by the P not possessing any supernumerary muscular branch or twig. Actually, the original OE was confirmed to occur with a low frequency (15/73 thighs; 20.5%). It was inferred that the original OE readily underwent variation in the process of ontogeny due to the influence of the obturator nerve. The segmental composition of the obturator nerve was considered not to be involved in the morphological change in the OE.

Ultrasound activation of TiO2 in melanoma tumors.

Sonodynamic therapy (SDT) is a new modality using ultrasound (US) to activate certain chemical sensitizers for cancer therapy. In this study, the effect of US combined with a nanoparticle titanium dioxide (TiO(2)) on melanoma cell was investigated in vitro and in vivo. Melanoma cells (C32) were irradiated with US in the presence and/or absence of TiO(2). Cell viability was measured immediately after US irradiation (1MHz, 0.5 and 1.0W/cm(2) for 10s). The effect of the combination of TiO(2) and US exposure (1MHz, 1.0W/cm(2), 2 min duration) on subcutaneously implanted C32 solid tumors in mice were investigated by measuring tumor volume regression. The cell viability was significantly decreased only after US irradiation in the presence of TiO(2). In vivo results showed significant inhibition of tumor growth in groups treated with TiO(2) and US. To our knowledge, this is the first report to demonstrate the cell killing effect of TiO(2) nanoparticles under the irradiation US in vitro and in vivo.

Reconstruction of cartilage tissue using scaffold-free organoid culture technique.

We applied the scaffold-free culture method to chondrocytes and attempted to reconstitute articular cartilage grafts. Primary rat costal chondrocytes were immobilized into hollow fibers by centrifugation at a density of 3 x 10(8) cells/cm(3) to induce the formation of cylindrical-shaped multicellular aggregates (organoids) and cultured for one month. The organoids were evaluated by histological and gene expression analyses. Chondrocytes formed cylindrical organoids in hollow fibers (HFs). Histochemical analysis revealed the accumulation of a cartilage extracellular matrix (collagen and proteoglycan) around cells in the lumen of HFs with culture time, forming a low-cellular-density tissue similar to native cartilage by day 28. Furthermore, in contrast to that in traditional monolayer culture, the organoid maintained the gene expression of the cartilage extracellular matrix (type II collagen, aggrecan) for one month of culture. In conclusion, our organoid formation method was effective in producing a cartilage-like tissue. This result suggests that the technique may be applicable to the development of an articular cartilage graft.