Soft Tissue Stabilization of the Hinge Position for Lateral Closing-Wedge Distal Femoral Osteotomy: An Anatomic Study.

Background: Soft tissue plays an important role in stabilizing the hinge point for osteotomy around the knee. However, insufficient data are available on the anatomic features of the soft tissue around the hinge position for lateral closing-wedge distal femoral osteotomy (LCWDFO). Purpose: To (1) anatomically analyze the soft tissue around the hinge position for LCWDFO, (2) histologically analyze the soft tissue based on the anatomic analysis results, and (3) radiologically define the appropriate hinge point to prevent unstable hinge fracture based on the results of the anatomic and histological analyses. Study Design: Descriptive laboratory study. Methods: In 20 cadaveric knees (age, 82.7 ± 7.8 years; range, 60-96 years), the soft tissue of the distal medial side of the femur was anatomically analyzed. The thicknesses of the periosteum and direct insertion of the adductor tendon (AT) were histologically examined and measured using an electron microscope. The thickness of the periosteum was visualized graphically, and the graph of the periosteum and radiograph of the knee were overlaid using image editing software. The appropriate hinge position was determined based on the periosteal thickness and attachment of the AT. Results: The mean thickness of the periosteum of the metaphysis was 352.7 ± 58.6 µm (range, 213.6-503.4 µm). The overlaid graph and radiograph revealed that the thickness of the periosteum changed at the part corresponding to the transition between the diaphyseal and metaphyseal ends of the femur. The mean width of the AT attached to the distal medial femur from the adductor tubercle toward the distal direction was 7.9 ± 1.3 mm (range, 6.3-9.7 mm). Conclusion: Results indicated that the periosteum and AT support the hinge for LCWDFO within the area surrounded by the apex of the adductor tubercle and the upper border of the posterior part of the lateral femoral condyle. Clinical Relevance: When the hinge point is located within the area surrounded by the apex of the adductor tubercle and the upper border of the posterior part of the lateral femoral condyle, these soft tissues work as stabilizers, and there is no risk of cutting into the joint space.

Intracranial Gene Delivery Mediated by Albumin-Based Nanobubbles and Low-Frequency Ultrasound.

Research in the field of high-intensity focused ultrasound (HIFU) for intracranial gene therapy has greatly progressed over the years. However, limitations of conventional HIFU still remain. That is, genes are required to cross the blood-brain barrier (BBB) in order to reach the neurological disordered lesion. In this study, we introduce a novel direct intracranial gene delivery method, bypassing the BBB using human serum albumin-based nanobubbles (NBs) injected through a less invasive intrathecal route via lumbar puncture, followed by intracranial irradiation with low-frequency ultrasound (LoFreqUS). Focusing on both plasmid DNA (pDNA) and messenger RNA (mRNA), our approach utilizes LoFreqUS for deeper tissue acoustic penetration and enhancing gene transfer efficiency. This drug delivery method could be dubbed as the "Spinal Back-Door Approach", an alternative to the "front door" BBB opening method. Experiments showed that NBs effectively responded to LoFreqUS, significantly improving gene transfer in vitro using U-87 MG cell lines. In vivo experiments in mice demonstrated significantly increased gene expression with pDNA; however, we were unable to obtain conclusive results using mRNA. This novel technique, combining albumin-based NBs and LoFreqUS offers a promising, efficient, targeted, and non-invasive solution for central nervous system gene therapy, potentially transforming the treatment landscape for neurological disorders.

Efficient mRNA Delivery with Lyophilized Human Serum Albumin-Based Nanobubbles.

In this study, we developed an efficient mRNA delivery vehicle by optimizing a lyophilization method for preserving human serum albumin-based nanobubbles (HSA-NBs), bypassing the need for artificial stabilizers. The morphology of the lyophilized material was verified using scanning electron microscopy, and the concentration, size, and mass of regenerated HSA-NBs were verified using flow cytometry, nanoparticle tracking analysis, and resonance mass measurements, and compared to those before lyophilization. The study also evaluated the response of HSA-NBs to 1 MHz ultrasound irradiation and their ultrasound (US) contrast effect. The functionality of the regenerated HSA-NBs was confirmed by an increased expression of intracellularly transferred Gluc mRNA, with increasing intensity of US irradiation. The results indicated that HSA-NBs retained their structural and functional integrity markedly, post-lyophilization. These findings support the potential of lyophilized HSA-NBs, as efficient imaging, and drug delivery systems for various medical applications.

Soft tissue stabilization of the hinge position in medial closed wedge distal femoral osteotomy: an anatomical study.

BACKGROUND: Soft tissue has an important role in stabilizing the hinge point of medial closed wedge distal femoral osteotomy (MCWDFO). However, there are conflicting data on the soft tissue anatomy around the hinge point of MCWDFO and, therefore, further anatomical data are needed. The purposes of the study were to: 1) anatomically analyze the soft tissue around the hinge point of MCWDFO; 2) radiologically define the appropriate hinge point to prevent an unstable hinge fracture based on the result of the anatomical analysis; and 3) histologically analyze the soft tissue based on the result of the anatomical analysis. METHODS: In 20 cadaveric knees, the capsule attachment of the distal lateral side of the femur was marked with a radiopaque ball bearing. A digital planning tool was used to calculate the area of the marked capsule attachment around the ideal hinge point of MCWDFO on radiographs. The soft tissue around the hinge point was histologically examined and the periosteal thickness was measured and visualized graphically. The graph and radiograph were overlayed using image editing software, and the appropriate hinge position was determined based on the periosteal thickness. RESULTS: As a result, the periosteal thickness of the distal lateral femur tended to rapidly decrease from the metaphyseal region toward the diaphyseal region. The overlayed graph and radiograph revealed that the periosteal thickness changed in the region corresponding to the apex of the turning point of the femoral metaphysis in all cases. CONCLUSIONS: In conclusion, the periosteum might support the hinge of MCWDFO within the area surrounded by the apex of the turning point of the femoral metaphysis and the upper border of the posterior part of the lateral femoral condyle.

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.

Ultrasound-Induced DNA Damage and Cellular Response: Historical Review, Mechanisms Analysis, and Therapeutic Implications.

The biological effects of ultrasound may be classified into thermal and nonthermal mechanisms. The nonthermal effects may be further classified into cavitational and noncavitational mechanisms. DNA damage induced by ultrasound is considered to be related to nonthermal cavitations. For this aspect, many in vitro studies on DNA have been conducted for evaluating the safety of diagnostic ultrasound, particularly in fetal imaging. Technological advancement in detecting DNA damage both in vitro and in vivo have elucidated the mechanism of DNA damage formation and their cellular response. Damage to DNA, and the residual damages after DNA repair are implicated in the biological effects. Here, we discuss the historical evidence of ultrasound on DNA damage and the mechanism of DNA damage formation both in vitro and in vivo, compared with those induced by ionizing radiation. We also offer a commentary on the safety of ultrasound over X-ray-based imaging. Also, understanding the various mechanisms involved in the bioeffects of ultrasound will lead us to alternative strategies for use of ultrasound for therapy.

Low-intensity ultrasound inhibits melanoma cell proliferation in vitro and tumor growth in vivo.

PURPOSE: To determine the effect of low-intensity ultrasound on cancer cell proliferation in vitro and tumor growth in vivo. METHODS: In vitro, several cancer cell lines were exposed to low-intensity ultrasound at 0.11 W/cm2 for 2 min. Of the cell lines screened, melanoma C32 is one of the cell lines that showed sensitivity to growth inhibition by ultrasound and was therefore used in succeeding experiments. In vivo, under the same ultrasound conditions used in vitro, C32 tumors in mice were exposed to ultrasound daily for 2 weeks, and the tumor volumes were monitored weekly using sonography. RESULTS: In vitro, C32 cell growth was inhibited, attaining 43.2% inhibition on the 3rd day. In vivo, tumor growth was significantly inhibited, with the treated tumors exhibiting 2.7-fold slowed tumor growth vs. untreated tumors at week 2. Such inhibition was not associated with increased cell death. Several genes related to the cell cycle and proliferation were among those significantly regulated. CONCLUSION: These findings highlight the potential of low-intensity ultrasound to inhibit tumor growth in a noninvasive, safe, and easy-to-administer way. In addition, this may suggest that the mechanical stress induced by ultrasound on C32 cells may have affected the intrinsic biomolecular mechanism related to the cell growth of this particular cell line. Further research is needed to identify which of the regulated genes played key roles in growth inhibition.

High-Intensity Focused Ultrasound in the Treatment of Breast Cancer.

High-intensity focused ultrasound (HIFU) is a non-invasive method of ablating malignant tumors. This paper will review the current clinical application of HIFU specially in the treatment of breast cancer. In addition to clinical studies, this review will also look into some basic studies that could address the technical issues related to this modality. In general, HIFU is considered to be safe being non-invasive and non-ionizing. The complication occurrence rate is low and repeated treatment is possible, making it an attractive option for some patients. However, for more than two decades since it was first used to treat breast cancer, clinical studies on HIFU still remain at the investigative stage and are only available in several centers. Reasons contributing to such few studies on HIFU include lack of specialized medical team and bioengineering technical staff, and breast cancer-dedicated imaging-HIFU platform to attain positive outcomes. Despite these circumstances, we believe that HIFU will eventually become the treatment of choice for most breast cancer patients in the near future.

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.

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.

Enhanced effect of recombinant human soluble thrombomodulin by ultrasound irradiation in acute liver failure.

The administration of recombinant human soluble thrombomodulin (rhsTM) significantly improves liver inflammation and increases the survival rate of patients with acute liver failure (ALF). However, rhsTM is dose-dependently correlated to the risk of bleeding. Recently, ultrasound (US) was found to enhance the effect of various drugs. Thus, the present study aimed to determine the enhancement effect of US irradiation on rhsTM in ALF. rhsTM (1 mg/kg) and US (1 MHz, 0.3 W/cm2) were irradiated to the liver of lipopolysaccharide/D-galactosamine-induced ALF mice model. The post-treatment aspartate aminotransferase, alanine aminotransferase, and high-mobility group box 1 levels were significantly lower in the rhsTM + US group than in the rhsTM alone group. Histopathological findings revealed significantly reduced liver injury and apoptosis in the rhsTM + US group. By contrast, US irradiation had no effect on rhsTM and TNF-α concentration in the liver tissue. In conclusion, US irradiation enhanced the effect of rhsTM in the ALF mice model. However, further studies must be conducted to determine the exact mechanism of such enhancement effect.

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.

Cavitation-threshold Determination and Rheological-parameters Estimation of Albumin-stabilized Nanobubbles.

Nanobubbles (NBs) are of high interest for ultrasound (US) imaging as contrast agents and therapy as cavitation nuclei. Because of their instability (Laplace pressure bubble catastrophe) and low sensitivity to US, reducing the size of commonly used microbubbles to submicron-size is not trivial. We introduce stabilized NBs in the 100-250-nm size range, manufactured by agitating human serum albumin and perfluoro-propane. These NBs were exposed to 3.34- and 5.39-MHz US, and their sensitivity to US was proven by detecting inertial cavitation. The cavitation-threshold information was used to run a numerical parametric study based on a modified Rayleigh-Plesset equation (with a Newtonian rheology model). The determined values of surface tension ranged from 0 N/m to 0.06 N/m. The corresponding values of dilatational viscosity ranged from 5.10-10 Ns/m to 1.10-9 Ns/m. These parameters were reported to be 0.6 N/m and 1.10-8 Ns/m for the reference microbubble contrast agent. This result suggests the possibility of using albumin as a stabilizer for the nanobubbles that could be maintained in circulation and presenting satisfying US sensitivity, even in the 3-5-MHz range.

Effects of polyphenols on doxorubicin-induced oral keratinocyte cytotoxicity and anticancer potency against oral cancer cells.

BACKGROUND: Normal human oral keratinocytes are highly sensitive to anticancer drugs including doxorubicin. Resveratrol, epigallocatechin gallate, and tannic acid are polyphenolic compounds that were reported to have cardioprotective effect when combined with doxorubicin. However, it is unknown whether these polyphenols could protect normal human oral keratinocytes against doxorubicin-induced cytotoxicity without weakening its cytotoxic potential against oral cancer cells. Here, we examined the effects of the 3 polyphenolic compounds on doxorubicin-induced cytotoxicity in normal human oral keratinocytes and also investigated their effects on doxorubicin potency in HSC-2 human oral squamous cell carcinoma cells. METHODS: Cell viability was evaluated, followed by the analysis of apoptosis and necrosis. The changes in intracellular reactive oxygen species at the early stage after treatment were also examined. RESULTS: The results revealed that resveratrol in combination with doxorubicin additively augmented doxorubicin cytotoxicity in both types of cells. However, epigallocatechin gallate and tannic acid at a certain concentration mitigated the doxorubicin-induced keratinocyte toxicity mainly due to reduced doxorubicin-induced necrosis in normal human oral keratinocytes without weaken doxorubicin anticancer efficacy. The exact mechanism is still unknown but intracellular reactive oxygen species might be not the sole factor. CONCLUSIONS: This study for the first time reported the effects of resveratrol, epigallocatechin gallate, and tannic acid on doxorubicin-induced cytotoxicity in normal oral keratinocytes and oral cancer cells. The combined use of epigallocatechin gallate or tannic acid with doxorubicin at a certain concentration could mitigate doxorubicin-induced keratinocyte cytotoxicity without weakening doxorubicin anticancer efficacy.

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.

Apoptotic and genotoxic effects of low-intensity ultrasound on healthy and leukemic human peripheral mononuclear blood cells.

PURPOSE: To scrutinize the apoptotic and genotoxic effects of low-intensity ultrasound and an ultrasound contrast agent (SonoVue; Bracco Diagnostics Inc., EU) on human peripheral mononuclear blood cells (PMBCs). METHODS: PMBCs were subjected to a low-intensity ultrasound field (1-MHz frequency; spatial peak temporal average intensity 0.18 W/cm2) followed by analysis for apoptosis and DNA damage (single-strand breaks + double-strand breaks). The comet assay was then repeated after 2 h to examine the ability of cells to repair DNA breaks. RESULTS: The results demonstrated that low-intensity ultrasound was capable of selectively inducing apoptosis in leukemic PMBCs, but not in healthy cells. The introduction of ultrasound contrast agent SonoVue resulted in an increase in apoptosis in both groups. DNA analysis after ultrasound exposure indicated that ultrasound triggered DNA damage in leukemic PMBCs (66.05 ± 13.36%), while the damage was minimal (7.01 ± 0.89%) in control PMBCs. However, both cell lines demonstrated an ability to repair DNA single- and double-strand breaks 2 h after sonication. CONCLUSIONS: The study demonstrated that low-intensity ultrasound selectively induced apoptosis in cancer PMBCs. Ultrasound-induced DNA damage was observed primarily in leukemic PMBCs. Nevertheless, both cell lines were able to repair ultrasound-mediated DNA strand breaks.

Efficient delivery of signal-responsive gene carriers for disease-specific gene expression via bubble liposomes and sonoporation.

Sonoporation is a promising method to intracellularly deliver synthetic gene carriers that have lower endocytotic uptake than viral carriers. Here, we applied sonoporation to deliver genes via polyethylene glycol (PEG)-grafted polymeric carriers that specifically respond to hyperactivated protein kinase A (PKA). PEG-grafted polymeric carrier/DNA polyplexes were not efficiently delivered into cells via the endocytotic pathway because of the hydrophilic PEG layer surrounding the polyplexes. However, the delivery of polyplexes into cells was significantly increased by sonoporation. The delivered polyplexes exhibited PKA-responsive transgene expression in PKA-overexpressing cells, but not in cells with low PKA activation. These results show that the sonoporation-mediated delivery of PEG-modified PKA-responsive polyplexes is a promising approach for safely applying gene therapy to abnormal cells with hyperactivated PKA.

Hyperthermia enhances bortezomib-induced apoptosis in human white blood cancer cells.

At present, the current therapeutic strategy for apoptosis induction mainly relies on the administration of pharmacological apoptotic modulators. Apart from that, apoptosis can be induced by various external stimuli such as hyperthermia, ionizing radiation, and electric fields. Despite advantages, both physical and pharmacological approaches bear some limitations as well. The rationale of this study was to overcome the limitations by combining hyperthermia and apoptotic modulator 'bortezomib' (Velcade). Two types of human blood cancer cell lines were utilized: human leukemic monocyte lymphoma cell U937 line and peripheral blood mononuclear cells (PMBCs) derived from the patient diagnosed with acute myeloid leukemia. Prior to apoptosis experiments, cytotoxicity tests were performed at three types of temperature regimes (40°, 42° and 44°C). We observed a gradual inhibition of cell viability correlating with an increase of temperature and drug concentration in both cell lines. However, there was no significant difference between sham group and groups of leukemic PMBCs treated by high temperature (44°C) and bortezomib. In U937 cells, combined treatment by heat shock and bortezomib led to an increase the number of cells underwent the late apoptosis stage. At the same time, similar treatment of PMBCs resulted in the stimulation of early apoptosis. Our data suggest that combination of bortezomib and hyperthermia enhances apoptosis induction in human cancer white blood cells, indicating a therapeutic potential for blood cancer therapy.