Endothelial cell membrane-based biosurface for targeted delivery to acute injury: analysis of leukocyte-mediated nanoparticle transportation.

Mimicking and leveraging biological structures and materials provide important approaches to develop functional vehicles for drug delivery. Taking advantage of the affinity and adhesion between the activated endothelial cells and innate immune cells during inflammatory responses, hybrid polyester nanoparticles coated with endothelial cell membranes (EM-P) containing adhesion molecules were fabricated and their capability as vehicles to travel to the acute injury sites through leukocyte-mediated processes was investigated. The in vivo studies and quantitative analyses performed through the lung-inflammation mouse models demonstrated that the EM-Ps preferentially interacted with the neutrophils and monocytes in the circulation and the cellular membrane-based biosurface improved the nanoparticle transportation to the inflamed lung possibly via the motility of neutrophils. Utilizing the transgenic zebrafish model, the leukocyte-mediated transportation and biodistribution of EM-Ps were further visualized in real time at the whole-organism level. Endothelial membranes provided a new biosurface for developing biomimetic vehicles to allow the immune cell-mediated transportation and may enable advanced systems for active and highly efficient drug delivery.

Feasibility of microwave ablation of the vertebral growth plate for spine growth regulation: a preliminary study.

PURPOSE: To explore the feasibility of microwave ablation (MWA) of the vertebral growth plate as a minimally invasive treatment for early-onset scoliosis. MATERIALS AND METHODS: One side of the L1-L3 vertebral growth plates were ablated using different MWA powers. Ablation safety and size were examined. Subsequently, L1-L3 vertebral growth plates were ablated on one side for 40 s at 20 W. At 2, 4, and 6 weeks after the ablation, growth changes of the spine were observed. RESULTS: No piglets died during and after ablation, and all had modified Tarlov Grade 5. The safe MWA time (time for safely ablating the vertebral growth plate) was 17.0 ± 1.5 s at 50 W, 23.0 ± 2.3 s at 40 W, 31.0 ± 3.1 s at 30 W, 47.0 ± 3.7 s at 20 W, 70.0 ± 4.2 s at 15 W, and 158.0 ± 5.0 s at 10 W. With power <15 W, the vertebral growth plate could not be effectively ablated within the safe ablation time. Within the safe ablation times, the MWA size on hematoxylin and eosin slices on a transverse diameter was between 7 and 10 mm; and that on longitudinal diameter was mainly determined by the ablation needle length. Moreover, the growth plate and annulus fibrosus on the ablated side grew poorly over time, the vertebral body showed significant wedge-shaped changes, and the spine showed significant unbalanced growth. CONCLUSION: MWA of the vertebral growth plate can be performed safely when accompanied with appropriate thermometry, and could be a new minimally invasive strategy in regulating spine growth.

Cervical Abnormalities in Severe Spinal Deformity: A 10-year MRI Review.

OBJECTIVE: To investigate the incidence of cervical anomalies (CA), including cervical intraspinal neural axis abnormalities (CIINAA) and/or cervical osseous abnormalities (COA), and the clinical relevance in severe spinal deformities (SSD) at a single center. METHODS: A retrospective study of SSD admitted for spinal surgery from January 2003 to January 2015 was conducted at a single center. INCLUSION CRITERIA: patients who present with coronal Cobb over 90° (and/or sagittal cobb ≥90°); and patients with complete imaging and clinical data preoperatively. EXCLUSION CRITERIA: ankylosing spondylitis, adult onset scoliosis, scoliosis secondary to bone destruction. There were 108 SSD patients who fulfilled the criteria in this research (41 males and 67 females). The mean age of the patients was 18.1 ± 2.7 years (range, 10-45 years). The clinical and radiological data of these patients were reviewed to identify CA and to analyze the relationship between clinical and radiographic characteristics in the population of SSD. RESULTS: The major curves of scoliosis and segmental kyphosis were 109.1° ± 24.7° and 91.2° ± 29.1°. Cervical abnormalities were detected in 56 patients (51.85%) with 9 different CA, including 28 patients (25.9%) with 6 different COA, 21 patients (19.4%) with 3 different CIINAA, and 7 patients (6.5%) with a combination of COA and cervical intraspinal neural axis abnormalities (CINAA). Basilar invagination and Klippel-Feil syndrome were the most frequent COA. Syringomyelia was the most frequent CINAA. SSD with COA in upper vertebral levels (UVL) had a higher incidence of CINAA than those in subaxial vertebral levels (SVL) (P = 0.024) and SSD with multiple COA (mCOA) in UVL had a higher incidence of CINAA than those with single COA (sCOA) (P = 0.029). In the present study, 83.9% of the SSD with CA were asymptomatic. CONCLUSION: The incidence of CA in SSD was 51.85%, with most presenting with intact neurologic status. As the diversity of COA increased, we found a higher incidence of CINAA, especially in UVL.

Effect of Spinal Shortening for Protection of Spinal Cord Function in Canines with Spinal Cord Angulation.

BACKGROUND Posterior vertebral column resection (PVCR) has been widely used as a treatment for severe spinal deformity. By using the canine model of vertebral column resection, this study explored the effect of spinal shortening on blood flow and function of the spinal cord during spinal cord angulation. MATERIAL AND METHODS The canine model of L1 vertebral column resection was constructed with the PVCR technique. The canines were divided into 5 groups according to the degree of shortening: the 0/4 group, the 1/4 group, the 2/4 group, the 3/4 group, and the control group. Spinal cord blood flow, neuroelectrophysiology, HE staining, nitric oxide, and endothelin-1 were measured during the procedure of vertebral column resection and spinal cord angulation. RESULTS The results showed that, in the 1/4 group and the 2/4 group, the blood flow of the spinal cord decreased by 16.5% and 10.6%, respectively, with no obvious damage in the spinal cord; in the 0/4 group and the 3/4 group, the blood flow decreased by 23.5% and 23.1%, respectively, with significant damage in the spinal cord. CONCLUSIONS When the spinal cord is shortened by 1/4 to 2/4, the tolerance of the spinal cord can increase and spinal cord injury resulting from angulation can be avoided. However, when the shortening reaches 3/4, it is harmful to the spinal cord. Proper shortening of the spinal cord by 1/4 to 2/4 may increase the tolerance of the spinal cord to the damage caused by angulation during PVCR.

Nanoparticle-mediated delivery of siRNA into zebrafish heart: a cell-level investigation on the biodistribution and gene silencing effects.

Nanomaterials hold promise for the delivery of nucleic acids to facilitate gene therapy in cardiac diseases. However, as much of the in vivo study of nanomaterials was conducted via the "trial and error" method, the understanding of the nanomaterial-mediated delivery in cardiac tissue was limited to the gross efficiency in manipulating the gene expression while little was known about the delivery process and mechanism in particular at the cell level. In this study, small interfering RNA (siRNA) nanoparticles formulated with a polyamidoamine (PAMAM) nanomaterial were applied to the injured heart of zebrafish. The distribution of nanoparticles in cardiomyocytes, endothelial cells, macrophages and leukocytes was quantitatively analyzed with precision at the cell level by using transgenic models. Based on the distribution characteristics, gene silencing effects in a specific group of cells were analyzed to illustrate how siRNA nanoparticles could get potent gene silencing in different cells in vivo. The results elucidated the heterogeneous distribution of siRNA nanoparticles and how nanoparticles could be efficient despite the significant difference in cellular uptake efficiency in different cells. It demonstrated a paradigm and the need to decouple cellular processes to understand nanoparticle-mediated delivery in complex tissue and the investigation/methodology may lead to important information to guide the design of advanced targeted drug-delivery systems in heart.

Expert consensus for PVCR in severe, rigid and angular spinal deformity treatment: The Kunming consensus.

In the past decades, an increasing number of surgeons started using posterior vertebral column resection (PVCR) to treat severe, rigid and angular spinal deformities. Little high-level evidence is available to guide surgical treatment. The aims of our study were to identify important surgical strategies and key technical points of Chinese experts who have extensive experience in the management of severe, rigid and angular spinal deformities using PVCR, and to standardize and unify the current core concepts. Workgroups of consensus were formed by selecting nationwide representing experts and comprehensive consultations. Eight task forces for major issues were established, then retrieval of literature, collection of expert opinions and writing of review articles were carried out. A modified Delphi process was chosen in round-table forum with three face-to-face meetings. Consensus was reached with items graded more than seven points including: indications and contraindications of PVCR; review PVCR in the evolution of spinal osteotomies; The corrective mechanism and safety of spinal cord; monitoring and responses of spinal cord crisis; characteristics and therapeutic outcome of pulmonary function; management of bleeding during PVCR; relationship of pedicle screw insertion and spinal cord safety; and analysis of non-neurologic complications and prevention strategies. In conclusion, The essential properties regarding PVCR procedure are tightly linked with various factors such as medical and surgical indication, range and level of vertebral column resection, strategies of correction, corrective efficiency and control of neurological risk. PVCR is used mainly for severe, rigid spinal deformity that is not manageable by other osteotomy techniques.

A Neutralized Noncharged Polyethylenimine-Based System for Efficient Delivery of siRNA into Heart without Toxicity.

Cationic polymers constitute an important class of materials in development of delivery vehicles for nucleic acid-based therapeutics. Among them, polyethylenimine (PEI) has been a classical cationic carrier intensively studied for therapeutic delivery of DNA, RNA, and short RNA molecules to treat diseases. However, the development of PEI for in vivo applications has been hampered by the inherent problems associated with the material, particularly its cytotoxicity and the instability of the nucleic acid complexation systems formed via electrostatic interactions. Here, we demonstrate a strategy to modify PEI polymers via hydrazidation to create neutralized, stable, and multifunctional system for delivering siRNA molecules. Through substitution of the primary amino groups of PEI with neutral hydrazide groups, cross-linked nanoparticles with surface decorated with a model targeting ligands were generated. The neutral cross-linked siRNA nanoparticles not only showed favorable biocompatibility and cell internalization efficiency in vitro but also allowed for significant tissue uptake and gene silencing efficiency in zebrafish heart in vivo. Our study suggests transformation of conventional branched PEI into a neutral polymer that can lead to a new category of nonviral carriers, and the resulting functional delivery systems may be further explored for development of siRNA therapeutics for treating cardiovascular disease/injury.

The risk factors of neurologic deficits of one-stage posterior vertebral column resection for patients with severe and rigid spinal deformities.

PURPOSE: To determine the risk factors of neurologic deficits during PVCR correction, so as to help improve safety during and after surgery. METHODS: A consecutive series of 76 patients with severe and rigid spinal deformities who were treated with PVCR at a single institution between October 2004 and July 2011 were included in our study. Of the 76 patients, 37 were male and 39 female, with an average age of 17.5 years (range 10-48 years). There were 52 adolescent patients (with an age <18 years) and 24 adult patients (with an age ≥18 years). Preoperatively, postoperatively and 6 months after surgery, we performed systemically neurologic function evaluations of each patients through meticulous physical examination. Any new abnormality or deterioration in evaluation of neurologic function than preoperative is reckoned postoperative neurologic deficits. Ten variables that might affect the safety of neurologic deficits during PVCR procedures, including imaging factors, clinical factors and operational factors, were analyzed using univariate analysis. Then the variables with statistical difference were analyzed by using multi-factor unconditional logistic regression analysis. RESULTS: No patient in this series had permanent paraplegia and nerve root injury due to operation. Change of neurologic status was found in six patients after surgery. Results of single-factor comparison demonstrated that the following seven variables were statistically different (P < 0.05): location of apex at main curve (X 3), Cobb angle at the main curve at the coronal plane (X 4), scoliosis associated with thoracic hyperkyphosis (X 5), level of vertebral column resected (X 6), number of segmental vessels ligated (X 7), preexisting neurologic dysfunction (X 8), and associated with intraspinal and brain stem anomalies (X 9). The multi-factor unconditional logistic regression analysis revealed that X 8 (OR = 49.322), X 9 (OR = 18.423), X 5 (OR = 11.883), and X 6 (OR = 8.769) were independent and positively correlated with the neurologic deficit. CONCLUSIONS: Preexisting neurologic dysfunction, associated with intraspinal and brain stem anomalies, scoliosis associated with thoracic hyperkyphosis and level of vertebral column resected are independent risk factors for neurologic deficits during PVCR procedure.

A five-step remedial screw placement method to treat severe spinal deformity with free-hand transpedicular screw placement.

PURPOSE: Severe spinal deformity is a complex morphological deformation that occurs and develops in three-dimensional space combined with abnormal development and morphology of anatomical structures, which presents great difficulties in the process of transpedicular screw placement. This study tried to explore the methods of transpedicular screw placement in surgical correction of severe spinal deformities. METHODS: Surgical corrections through posterior approach were performed in all the 76 cases (mean age 20.4 years). The averaging preoperative Cobb's angle of scoliosis was 108.2° ± 33.6° (range 100°-170°). Among these patients, 34 cases were combined with kyphosis; the average Cobb's angle of kyphosis was 77.3° (range 63°-160°). During operation, the screw tract was first established with the regular free-hand pedicle screw placement method. When this failed, in order to adjust the screw trajectory, a five-step remedial method was performed in the following order: (1) the"funnel" method; (2) exploring the pedicle exterior edge through the costotransverse joint; (3) exploring the superior and inferior edges of pedicle through the nerve root canal; (4) the vertebral plate fenestration; and (5) hemilaminectomy. RESULTS: Among all 1,472 screws planned to be placed for the patients, 1,210 (82.2 %) were successfully placed after using the regular method, and 262 (17.8 %) failed in this stage. After applying the five-step remedial method, 256 of the failed 262 screws were successfully placed. Among them, 176 screws (68.8 %) were successfully placed after Step 1, 44 (17.2 %) after Step 2, 21 (8.2 %) after Step 3, 12 (4.7 %) after Step 4, and 3 (1.2 %) after Step 5. In only six, pedicles screws could not be placed eventually. No nerve or blood vessel damages occurred in all cases. All final screw positions were validated by CT. CONCLUSION: The five-step remedial method proved to be an effective supplementary method for transpedicular screw placement to treat patients with severe spinal deformities. The key points include a detailed preoperative plan, a meticulous hand drilling sensation, and an experienced probing technique for screw tract.