Cellular Scale Curvature in Bioceramic Scaffolds Enhanced Bone Regeneration by Regulating Skeletal Stem Cells and Vascularization.

Critical-sized segmental bone defects cannot heal spontaneously, leading to disability and significant increase in mortality. However, current treatments utilizing bone grafts face a variety of challenges from donor availability to poor osseointegration. Drugs such as growth factors increase cancer risk and are very costly. Here, a porous bioceramic scaffold that promotes bone regeneration via solely mechanobiological design is reported. Two types of scaffolds with high versus low pore curvatures are created using high-precision 3D printing technology to fabricate pore curvatures radius in the 100s of micrometers. While both are able to support bone formation, the high-curvature pores induce higher ectopic bone formation and increased vessel invasion. Scaffolds with high-curvature pores also promote faster regeneration of critical-sized segmental bone defects by activating mechanosensitive pathways. High-curvature pore recruits skeletal stem cells and type H vessels from both the periosteum and the marrow during the early phase of repair. High-curvature pores have increased survival of transplanted GFP-labeled skeletal stem cells (SSCs) and recruit more host SSCs. Taken together, the bioceramic scaffolds with defined micrometer-scale pore curvatures demonstrate a mechanobiological approach for orthopedic scaffold design.

Hyperbaric oxygen therapy enhances osteointegration of reimplanted cranial flap by regulating osteogenesis-angiogenesis coupling.

Craniectomy is a lifesaving procedure to alleviate dangerously high intracranial pressure by removing a bone flap from the calvarium. However, the osteointegration of reimplanted bone flap with the existing bone tissue is still a clinical challenge. Hyperbaric oxygen (HBO) therapy has shown efficacy in promoting bone repair and could be a promising treatment for accelerating postoperative recovery. However, the specific cell types that are responsive to HBO treatment are not well understood. In this study, we created a murine model of craniectomy, with reimplantation of the cranial flap after 1 week. The effects of HBO treatment on bone formation and blood vessel formation around reimplanted bone were examined by micro-computed tomography, histological staining, and immunofluorescence staining. Single-cell RNA sequencing (scRNAseq) was utilized to identify key cell subtypes and signaling pathways after HBO treatment. We found that HBO treatment increased bone volume around reimplanted cranial flaps. HBO also increased the volume of Osterix-expressing cells and type H vessels. scRNAseq data showed more mature osteoblasts and endothelial cells, with higher expressions of adhesion and migration-related genes after HBO treatment. Cell-cell interaction analysis revealed a higher expression level of genes between mature osteoblasts and endothelial cells from the angiopoietin 2-integrin α5ß1 pathway. Taken together, HBO therapy promotes the healing process of craniectomy by regulating the crosstalk between vascular endothelial cells and osteogenic cells. These findings provide evidence in a preclinical model that HBO therapy enhances osteointegration by regulating angiogenesis-osteogenesis coupling, providing a scientific basis for utilizing HBO therapy for accelerating postoperative recovery after craniectomy.

Axial Compressive Loading Attenuates Early Osteoarthritis by Reducing Subchondral Bone Remodeling.

BACKGROUND: Mechanical loading and alendronate (ALN) can be used as noninvasive physical therapy methods for osteoarthritis (OA). However, the timing and efficacy for treatments are unknown. PURPOSE: To determine whether the timing of mechanical loading and ALN influences the pathobiological changes of OA. STUDY DESIGN: Controlled laboratory study. METHODS: Mice with OA induced by anterior cruciate ligament transection were subjected to early (1-3 weeks) or late (5-7 weeks) axial compressive dynamic load or intraperitoneal injection of ALN. Changes in gait were analyzed using gait analysis system, pathobiological changes in subchondral bone, cartilage, osteophyte, and synovitis were assessed using micro-computed tomography, tartrate-resistant acid phosphatase staining, pathologic section staining, and immunohistochemistry at 1, 2, 4, and 8 weeks. RESULTS: At 1, 2, and 4 weeks, the OA limb had lower mean footprint pressure intensity, lower bone volume per tissue volume (BV/TV) in the subchondral bone, and more osteoclasts. At 4 weeks, the early loading, ALN, and load + ALN treatments induced less cartilage destruction, with a corresponding reduction in Osteoarthritis Research Society International score and increased hyaline cartilage thickness. The treatments also resulted in fewer osteoclasts and higher BV/TV and bone mineral density of subchondral bone and suppressed inflammation and interleukin 1ß- and tumor necrosis factor α-positive cells in synovium. At 8 weeks, early loading or load + ALN improved the mean footprint pressure intensity and knee flexion. At 8 weeks, early load + ALN had a synergistic effect on protecting hyaline cartilage and proteoglycans. Footprint pressure intensity and cartilage destruction were worse in late loading limbs, and no differences in BV/TV, bone mineral density, osteophyte formation, and synovium inflammation were found between the late load, ALN, and load + ALN groups and the anterior cruciate ligament transection group. CONCLUSION: Dynamic axial mechanical loading or ALN in the early stages of knee trauma protected against OA by suppressing subchondral bone remodeling. However, late loading promoted cartilage degeneration in advanced OA, indicating that reduced loading should be performed in the late stages of OA to avoid the acceleration of OA. CLINICAL RELEVANCE: Early low-level functional exercise or antiosteoporotic drugs could clearly slow or prevent the progression of early OA. For patients with mild to severe OA, loading reduction via brace protection or maintenance of joint stability via early ligament reconstruction surgery may ameliorate OA exacerbation.

Genome-wide identification of the CPK gene family in wheat (Triticum aestivum L.) and characterization of TaCPK40 associated with seed dormancy and germination.

Calcium-dependent protein kinases (CPKs), important sensors of calcium signals, play an essential role in plant growth, development, and stress responses. Although the CPK gene family has been characterized in many plants, the functions of the CPK gene family in wheat, including their relationship to seed dormancy and germination, remain unclear. In this study, we identified 84 TaCPK genes in wheat (TaCPK1-84). According to their phylogenetic relationship, they were divided into four groups (I-IV). TaCPK genes in the same group were found to have similar gene structures and motifs. Chromosomal localization indicated that TaCPK genes were unevenly distributed across 21 wheat chromosomes. TaCPK gene expansion occurred through segmental duplication events and underwent strong negative selection. A large number of cis-regulatory elements related to light response, phytohormone response, and abiotic stress response were identified in the upstream promoter sequences of TaCPK genes. TaCPK gene expression was found to be tissue- and growth-stage-diverse. Analysis of the expression patterns of several wheat varieties with contrasting seed dormancy and germination phenotypes resulted in the identification of 11 candidate genes (TaCPK38/-40/-43/-47/-50/-60/-67/-70/-75/-78/-80) which are likely associated with seed dormancy and germination. The ectopic expression of TaCPK40 in Arabidopsis promoted seed germination and reduced abscisic acid (ABA) sensitivity during germination, indicating that TaCPK40 negatively regulates seed dormancy and positively regulates seed germination. These findings advance our understanding of the multifaceted functions of CPK genes in seed dormancy and germination, and provide potential candidate genes for controlling wheat seed dormancy and germination.

Feasibility of one breath-hold cardiovascular magnetic resonance compressed sensing cine for left ventricular strain analysis.

Objective: To investigate the feasibility of 3D left ventricular global and regional strain by using one breath-hold (BH) compressed sensing cine (CSC) protocol and determine the agreement between CSC and conventional cine (CC) protocols. Methods: A total of 30 volunteers were enrolled in this study. Cardiovascular magnetic resonance (CMR) images were acquired using a 1.436 T magnetic resonance imaging (MRI) system. The CSC protocols included one BH CSC and the shortest BH CSC protocols with different parameters and were only performed in short-axis (SA) view following CC protocols. Left ventricular (LV) end-diastole volume (EDV), end-systole volume (ESV), stroke volume (SV), and ejection fraction (EF) global and regional strain were calculated by CC, one BH CSC, and shortest BH CSC protocols. The intraclass correlation coefficient (ICC) and coefficient of variance (CV) of these parameters were used to determine the agreement between different acquisitions. Results: The agreement of all volumetric variables and EF between the CC protocol and one BH CSC protocol was excellent (ICC > 0.9). EDV, ESV, and SV between CC and shortest BH CSC protocols also had a remarkable coherence (ICC > 0.9). The agreement of 3D LV global strain assessment between CC protocol and one BH CSC protocol was good (ICC > 0.8). Most CVs of variables were also good (CV < 15%). ICCs of all variables were lower than 0.8. CVs of all parameters were higher than 15% except global longitudinal strain (GLS) between CC and shortest BH CSC protocols. The agreement of regional strain between CC and BH CSC protocols was heterogeneous (-0.2 < ICC < 0.7). Many variables of CVs were poor. Conclusion: Notably, one BH CSC protocol can be used for 3D global strain analysis, along with a good correlation with the CC protocol. The regional strain should continue to be computed by the CC protocol due to poor agreement and a remarkable variation between the protocols. The shortest BH CSC protocol was insufficient to replace the CC protocol for 3D global and regional strain.

Prognostic value of high-resolution magnetic resonance imaging in evaluating carotid atherosclerotic plaque in patients with ischemic stroke.

BACKGROUND: Ischemic stroke (IS) is a devastating occurrence affecting millions worldwide. This study aimed to evaluate the prognostic value of high-resolution magnetic resonance imaging (HRMRI) in assessing carotid atherosclerotic plaque in IS patients. METHODS: Between January 2013 and March 2015, 338 IS patients were recruited for the investigative purposes of the study. All participants of the study underwent an HRMRI inspection procedure after being admitted into the hospital. During this study, we systematically analyzed and measured various types of fibrous caps, lipid compositions, and plaque lipid ratios. Univariate and multivariate logistic regression analyses were performed for predicting prognosis of IS patients. A receiver-operating characteristic (ROC) curve was employed to determine the accuracy of the IS prognosis. RESULTS: The percentage of type I fibrous caps exhibited significant decrease, while the percentage of type III fibrous caps, lipid compositions, and lipid ratios all displayed increase. The results of the univariate analysis indicated that age, hypertension, hyperlipidemia, treatment regimens, fibrous cap type, plaque type, lipid composition, and lipid ratio shared a correlation in regards to the poor prognosis of IS patients. Multivariate logistic regression analysis demonstrated that the prognosis of IS patients was not necessarily dependent on fibrous cap type, plaque type, or age. ROC curves revealed that the HRMRI possessed a strong predicative ability in relation to the identification of the prognosis of IS patients through factors such as type of plaque and fibrous caps determination. CONCLUSION: Our study conclusively intimated the promise of HRMRI as an evaluative tool for the determination of carotid atherosclerotic plaques in patients with IS.