Polydopamine-modified konjac glucomannan scaffold with sustained release of vascular endothelial growth factor to promote angiogenesis.

The fabrication of scaffolds capable of the sustained release of the vascular endothelial growth factor (VEGF) to promote angiogenesis for a long time remains a challenge in tissue engineering. Here, we report a facile approach for effectively fabricating a bioactive scaffold that gradually releases VEGF to promote angiogenesis. The scaffold was fabricated by coating polydopamine (PDA) on a konjac glucomannan (KGM) scaffold, followed by the surface immobilization of VEGF with PDA. The resulting VEGF-PDA/KGM scaffold, with a porous and interconnected microstructure (392 µm pore size with 84.80 porosity), combined the features of long-term biodegradability (10 weeks with 51 % degradation rate), excellent biocompatibility, and sustained VEGF release for up to 21 days. The bioactive VEGF-PDA/KGM scaffold exhibited multiple angiogenic activities over time, as confirmed by in vivo and in vitro experiments. For example, the scaffold significantly promoted the attachment and proliferation of human umbilical vein endothelial cells and the formation of vascular tubes in vitro. Moreover, the in vivo results demonstrated the formation and maturation of blood vessels after subcutaneous implantation in rats for four weeks. This promising strategy is a feasible approach for producing bioactive materials that can induce angiogenesis in vivo. These findings provide a new avenue for designing and fabricating biocompatible and long-term biodegradable scaffolds for sustained VEGF release to facilitate angiogenesis.

Effect of HER2 expression status on the prognosis of patients with HR+/HER2­ advanced breast cancer undergoing advanced first­line endocrine therapy.

The present study aimed to retrospectively assess the effects of human epidermal growth factor receptor 2 (HER2) expression on the diagnosis of patients with hormone receptor (HR)+/HER2- late-stage breast cancer undergoing advanced first-line endocrine-based treatment. A total of 72 late-stage breast tumor cases from June 2017 to June 2019 were selected from the Department of Surgical Oncology, Shaanxi Provincial People's Hospital (Xi'an, China) and included in the present study. The expression of estrogen receptor, progesterone receptor and HER2 was detected by immunohistochemistry. The subjects were divided into two groups: the HER2-negative (0) cohort (n=31) and the HER2 low expression cohort (n=41). The age, BMI, Karnofsky Performance Status (KPS) score, tumor size, lymph node metastasis, pathological type, Ki-67 expression and menopausal status of the patients were obtained through the electronic medical record system of Shaanxi Provincial People's Hospital. Progression-free survival (PFS) and overall survival (OS) were evaluated for all patients. The median PFS and OS of the HER2(0) cohort were longer than those of the HER2 low expression cohort (all P<0.05). It was shown that age (hazard ratio, 6.000 and 5.465), KPS score (hazard ratio, 4.000 and 3.865), lymph node metastasis (hazard ratio, 3.143; 2.983) and HER2 status (hazard ratio, 3.167 and 2.996) were independent influencing factors of the prognosis of patients with HR+/HER2- advanced breast cancer (ABC) (all P<0.05). Three models (model 1, no parameters adjusted; model 2, BMI, tumor size, pathological type, Ki-67 and menopausal status adjusted; and model 3, age, KPS functional status score and lymph node metastasis adjusted based on model 2) were established within the HER2(0) cohort as the reference for statistical analysis using the multivariate Cox's regression test. In models 2 and 3, the risk of poor prognosis of ABC within the HER2 low expression cohort was significantly higher compared with that in the HER2(0) cohort (hazard ratio, 3.558 and 4.477; 95% CI, 1.349-9.996 and 1.933-11.586; P=0.003 and P<0.001). The HER2 expression status of patients with HR+/HER2- ABC receiving advanced first-line endocrine therapy may affect PFS and OS.

Nanocellulose-based hydrogels as versatile drug delivery vehicles: A review.

Hydrogels designed with nanocellulose (i.e. cellulose nanocrystals (CNC), cellulose nanofibrils (CNF), and bacterial cellulose (BC)) have significant advantages as drug carriers due to their environmentally-benign features and excellent properties. Nanocellulose hydrogels have been demonstrated to sustainably deliver various kinds of drugs via different routes of administration, in which nanocellulose significantly improves the hydrogel properties and tunes the drug releasing profile. This article comprehensively summarizes the recent research progress on nanocellulose hydrogels in drug delivery. We carefully assessed the gelation methods for nanocellulose hydrogel design and highlighted the influence of nanocellulose on hydrogel properties and drug release behaviors. In particular, it is the first time to summarize the research on nanocellulose hydrogel-based drug carriers regarding specific routes of administration. This work provides a critical review of nanocellulose-based hydrogels as drug delivery vehicles, and also underlines the outlook in this field, with the objective to inspire/prompt future work, especially the practical applications of nanocellulose hydrogels in designing controlled drug delivery systems.

Biological Application of Novel Biodegradable Cellulose Composite as a Hemostatic Material.

Degradable hemostatic materials have unique advantages in reducing the amount of bleeding, shortening the surgical operation time, and improving patient prognosis. However, none of the current hemostatic materials are ideal and have disadvantages. Therefore, a novel biodegradable cellulose-based composite hemostatic material was prepared by crosslinking sodium carboxymethyl cellulose (CCNa) and hydroxyethyl cellulose (HEC), following an improved vacuum freeze-drying method. The resulting cellulose composite material was neutral in pH and spongy with a density of 0.042 g/cm3, a porosity of 77.68%, and an average pore size of 13.45 µm. The composite's compressive and tensile strengths were 0.1 MPa and 15.2 MPa, respectively. Under in vitro conditions, the composites were degraded gradually through petite molecule stripping and dissolution, reaching 96.8% after 14 days and 100% degradation rate at 21 days. When implanted into rats, the degradation rate of the composite was slightly faster, reaching 99.7% in 14 days and 100% in 21 days. Histology showed a stable inflammatory response and no evidence of cell degeneration, necrosis, or abnormal hyperplasia in the tissues around the embedded material, indicating good biocompatibility. In the hemorrhagic liver model, the time to hemostasis and the total blood loss in the cellulose composite group was significantly lower than in the medical gauze group and the blank control group (P < 0.05). These data indicate that the novel cellulose composite is a promising implantable hemostatic material in clinical settings.

Construction of a Prognostic Nomogram Model for Patients with Mucinous Breast Cancer.

Objective: The objective of the study is to develop a nomogram for estimating three- and five-year survival rates in mucinous breast cancer patients. Methods: Between 2010 and 2016, the National Cancer Institute's Surveillance, Epidemiology, and End Results (SEER) were searched as a data source for patients associated with mucinous breast cancer (MBC). A total of 3964 patients were recruited after screening. The multivariate Cox model and the univariate Kaplan-Meier (KM) approach were employed to evaluate the independent prognostic markers, followed by developing a nomogram for estimating three- and five-year survival rates in MBC patients. Consequently, the consistency index (C-index) was employed to assess the predictive accuracy of the generated nomogram. Results: Age, race, T stage, M stage, surgery, and radiotherapy were all independent predictive biomarkers for the MBC patients (P < 0.05). The nomogram was finally developed based on the underlined factors. Furthermore, the C-index of 0.803 and reliable calibration curves were obtained in the nomogram's assessment. Conclusions: In patients with mucinous breast cancer, the proposed nomogram provides a viable tool for accurate prognostic prediction. In clinical practice, it could serve as a personalized diagnosis tool, estimate prognosis, and help in suggesting treatment plans for patients with MBC.

Immunomodulatory role of mesenchymal stem cell therapy in liver fibrosis.

Liver fibrosis is a fibrogenic and inflammatory process that results from hepatocyte injury and is characterized by hepatic architectural distortion and resultant loss of liver function. There is no effective treatment for advanced fibrosis other than liver transplantation, but it is limited by expensive costs, immune rejection, and postoperative complications. With the development of regenerative medicine in recent years, mesenchymal stem cell (MSCs) transplantation has become the most promising treatment for liver fibrosis. The underlying mechanisms of MSC anti-fibrotic effects include hepatocyte differentiation, paracrine, and immunomodulation, with immunomodulation playing a central role. This review discusses the immune cells involved in liver fibrosis, the immunomodulatory properties of MSCs, and the immunomodulation mechanisms of MSC-based strategies to attenuate liver fibrosis. Meanwhile, we discuss the current challenges and future directions as well.

Mesenchymal Stem Cells (MSCs) in Targeted Drug Delivery: Literature Review and Exploratory Data on Migrating and Differentiation Capacities of Bone MSCs into Hepatic Progenitor Cells.

BACKGROUND AND OBJECTIVE: Mesenchymal stem cells (MSCs), particularly bone MSCs (BMSCs) offer great potentials for targeted therapeutic applications owing to their migratory and differentiation capacities. Significant advances have been achieved in the differentiation of hepatocyte or hepatocyte-like cells both in vitro and in vivo. However, there is limited knowledge on the differentiation of BMSCs into bipotential hepatic progenitor cells or cholangiocyte. This study reviews the potentials and advances in using MSCs as vehicles for targeted drug delivery and proposes a new method for the induction of differentiation in rat BMSCs into hepatic progenitor cells in vitro and assesses the differential and migratory capacities. METHODS: The BMSCs of Sprague Dawley (SD) rats were harvested from the femur and the tibiae of the rats. After isolation and culturing, BMSCs from Passage 1 were used for the study. The in vitro differentiation of the hepatic progenitor cells was performed using a 2-step induction approach after 5-day serum deprivation from the BMSCs and culturing in Dulbecco's modified eagle medium. Spontaneous in vitro differentiation of BMSCs was examined in the absence of growth factors for 15 days as control treatment. Hepatocytes differentiation was achieved by exposing the culture to collagen type I-coated plates. Cholangiocytes differentiation was achieved with replating the BMC-HepPCs on a layer of Matrigel. Immunofluorescence was conducted on twelve-well plates to determine cell differentiations. Real-Time Quantitative Reverse Transcription PCR (qRTPCR) was used to determine the total RNA extracted using the Trizol LS reagent. In the hepatocyte differentiation group, after periodic acid-schiff (PAS) staining for glycogen, inverted microscope was used to determine differentiations and undifferentiated BMC-HepPCs served as controls. The amount of low-density lipoprotein (LDL) uptake by the BMSCs-derived hepatocytes was assessed using fluorescence microscopy. The secretion of rat albumin was quantified using a quantitative ELISA kit. RESULTS: Differentiation induction is indicative of the sequential supplementation of sodium butyrate and cytokines, which are involved in the embryonic development of the mammalian liver. Hepatic progenitor cells, derived from bone marrow, can be differentiated bidirectionally in vitro into both hepatocyte and cholangiocyte cell-lines. The differentiated cells, including hepatic progenitor cells, hepatocytes, and bile duct-like cells, were identified and analyzed at mRNA and protein levels. CONCLUSION: Our findings show that BMSCs can be utilized as novel bipotential hepatic progenitor cells and thereby for hepatobiliary disease treatment or hepatobiliary tissue-engineering.

Doxorubicin-loaded Fe3O4-ZIF-8 nano-composites for hepatocellular carcinoma therapy.

Hepatocellular carcinoma (HCC) is one of the most common and malignant cancers and has no effective therapeutic approaches. Chemotherapeutic drug doxorubicin (DOX) is widely used for HCC therapy, but its application is limited by the clinical toxicity. In the present study, an Fe3O4-ZIF-8 magnetic nano-composite was fabricated and used for DOX delivery for HCC therapy. The morphology, structure and property of Fe3O4-ZIF-8 nano-composites were evaluated by scanning electron microscopy, transmission electron microscopy and N2 adsorption-desorption isotherms studies. The drug release from DOX@Fe3O4-ZIF-8 was measured in pH 7.4 phosphate-buffered saline. The cellular uptake ability of DOX@Fe3O4-ZIF-8 into hepatocarcinoma cell line (MHCC97H) was visualized with a confocal laser scanning microscope. The effects of Fe3O4-ZIF-8, DOX and DOX@Fe3O4-ZIF-8 against MHCC97H cells were evaluated by CCK-8 assay and flow cytometry assay. Fe3O4-ZIF-8 nano-composites were synthesized and used as a nano-carrier for the delivery of DOX. Because of high drug loading property of ZIF-8, 1 mg Fe3O4-ZIF-8 nano-composites loaded 120 µg DOX when DOX@Fe3O4-ZIF-8 was synthesized in 30 mg/mL DOX solution. The cumulative DOX release curve showed a slow and sustained release pattern over time. The results of CCK-8 assay showed that Fe3O4-ZIF-8 was nontoxic to MHCC97H cells, and DOX@Fe3O4-ZIF-8 presented effective inhibiting effect on cell viability of MHCC97H cells. Cellular uptake assay showed that DOX@Fe3O4-ZIF-8 accumulated in both cytoplasm and nuclei. Moreover, because of valid drug accumulation, DOX@Fe3O4-ZIF-8 exhibited a good inducing effect on cell apoptosis of MHCC97H cells. In conclusion, based on the nontoxic and high drug loading capability of Fe3O4-ZIF-8, DOX@Fe3O4-ZIF-8 presented enhanced effects on HCC cells compared to free DOX, indicating its potential for the chemotherapy of HCC.

Association analysis of telomere length related gene ACYP2 with the gastric cancer risk in the northwest Chinese Han population.

Gastric cancer (GC) is a complex multifactorial disease, and genetic factors are believed the predominant cause to the occurrence of GC. We sought to investigate the associations between single nucleotide polymorphisms (SNPs) in ACYP2 gene and the risk of GC in the Northwest Chinese Han population. We recruited 302 GC cases and 300 controls from northwest China and selected 13 SNPs from ACYP2 gene. SNPs were genotyped using Sequenom Mass-ARRAY technology. Odds ratios (ORs) and 95 % confidence intervals (CIs) were used to assess the association. Bonferroni's multiple adjustment was applied to the level of significance, which was set at P < 0.00078 (0.05/65). We found that the minor alleles of rs6713088, rs11125529, rs12615793, rs843711, rs11896604, rs843706 and rs17045754 significantly stimulated the risk of GC, and homozygous alleles of above SNPs except rs6713088 were also found increasing the GC risk (P < 0.05). Under additive model and recessive model, rs11125529, rs12615793, rs843711, rs11896604, and rs17045754 also activated the risk of GC (P < 0.05). However, after Bonferroni's multiple adjusted was applied to our data, no SNP in our study was significantly related to GC risk. Further results of haplotype analysis founds that the haplotypes "TTCTAATG" (rs1682111, rs843752, rs10439478, rs843645, rs11125529, rs12615793, rs843711, and rs11896604) and "AC" (rs843706 and rs17045754) were more frequency among patients with GC, on the contrary, the haplotypes "CG" had a protective role in the GC risk (P < 0.05). Our results indicate that ACYP2 polymorphisms may influence the GC risk and may serve as a new precursory biomarker in the northwest Chinese Han population.

[Strategies to choose scaffold materials for tissue engineering].

Current therapies of organ failure or a wide range of tissue defect are often not ideal. Transplantation is the only effective way for long time survival. But it is hard to meet huge patients demands because of donor shortage, immune rejection and other problems. Tissue engineering could be a potential option. Choosing a suitable scaffold material is an essential part of it. According to different sources, tissue engineering scaffold materials could be divided into three types which are natural and its modified materials, artificial and composite ones. The purpose of tissue engineering scaffold is to repair the tissues or organs damage, so could reach the ideal recovery in its function and structure aspect. Therefore, tissue engineering scaffold should even be as close as much to the original tissue or organs in function and structure. We call it "organic scaffold" and this strategy might be the drastic perfect substitute for the tissues or organs in concern. Optimized organization with each kind scaffold materials could make up for biomimetic structure and function of the tissue or organs. Scaffold material surface modification, optimized preparation procedure and cytosine sustained-release microsphere addition should be considered together. This strategy is expected to open new perspectives for tissue engineering. Multidisciplinary approach including material science, molecular biology, and engineering might find the most ideal tissue engineering scaffold. Using the strategy of drawing on each other strength and optimized organization with each kind scaffold material to prepare a multifunctional biomimetic tissue engineering scaffold might be a good method for choosing tissue engineering scaffold materials. Our research group had differentiated bone marrow mesenchymal stem cells into bile canaliculi like cells. We prepared poly(L-lactic acid)/poly(ε-caprolactone) biliary stent. The scaffold's internal played a part in the long-term release of cytokines which mixed with sustained-release nano-microsphere containing growth factors. What's more, the stent internal surface coated with glue/collagen matrix mixing layer containing bFGF and EGF so could supplying the early release of the two cytokines. Finally, combining the poly(L-lactic acid)/poly(ε-caprolactone) biliary stent with the induced cells was the last step for preparing tissue-engineered bile duct. This literature reviewed a variety of the existing tissue engineering scaffold materials and briefly introduced the impact factors on the characteristics of tissue engineering scaffold materials such as preparation procedure, surface modification of scaffold, and so on. We explored the choosing strategy of desired tissue engineering scaffold materials.

Liver regeneration using decellularized splenic scaffold: a novel approach in tissue engineering.

BACKGROUND: The potential application of decellularized liver scaffold for liver regeneration is limited by severe shortage of donor organs. Attempt of using heterograft scaffold is accompanied with high risks of zoonosis and immunological rejection. We proposed that the spleen, which procured more extensively than the liver, could be an ideal source of decellularized scaffold for liver regeneration. METHODS: After harvested from donor rat, the spleen was processed by 12-hour freezing/thawing x 2 cycles, then circulation perfusion of 0.02% trypsin and 3% Triton X-100 sequentially through the splenic artery for 32 hours in total to prepare decellularized scaffold. The structure and component characteristics of the scaffold were determined by hematoxylin and eosin and immumohistochemical staining, scanning electron microscope, DNA detection, porosity measurement, biocompatibility and cytocompatibility test. Recellularization of scaffold by 5 x 10(6) bone marrow mesenchymal stem cells (BMSCs) was carried out to preliminarily evaluate the feasibility of liver regeneration by BMSCs reseeding and differentiation in decellularized splenic scaffold. RESULTS: After decellularization, a translucent scaffold, which retained the gross shape of the spleen, was generated. Histological evaluation and residual DNA quantitation revealed the remaining of extracellular matrix without nucleus and cytoplasm residue. Immunohistochemical study proved the existence of collagens I, IV, fibronectin, laminin and elastin in decellularized splenic scaffold, which showed a similarity with decellularized liver. A scanning electron microscope presented the remaining three-dimensional porous structure of extracellular matrix and small blood vessels. The porosity of scaffold, aperture of 45.36 +/- 4.87 µm and pore rate of 80.14% +/- 2.99% was suitable for cell engraftment. Subcutaneous implantation of decellularized scaffold presented good histocompatibility, and recellularization of the splenic scaffold demonstrated that BMSCs could locate and survive in the decellularized matrix. CONCLUSION: Considering the more extensive organ source and satisfying biocompatibility, the present study indicated that the three-dimensional decellularized splenic scaffold might have considerable potential for liver regeneration when combined with BMSCs reseeding and differentiation.

[Optimization of the protocols for in vitro culture and induction of hepatic differentiation of rat mesenchymal stem cells].

OBJECTIVE: To optimize the protocols for isolation, in vitro culture, identification and induction of hepatic differentiation of rat bone marrow mesenchymal stem cells (BMSCs). METHODS: Rat BMSCs were separated and purified by differential adherent culture for 1.5 h with the first medium change at 12 h. The surface markers of BMSCs were detected by flow cytometry. The cells were induced to differentiate into adipogenic, osteogenic, and chondrogenesis lineages. A 3-step protocol including sequential addition of growth factors, cytokines and hormones was used to induce the BMSCs to differentiate into hepatocyte-like cells. RESULTS: The cells isolated using this protocol were positive for CD29, CD44, and CD90 and negative for CD29 and CD45. The adipogenic, osteogenic, and chondrogenic differentiation of the BMSCs were verified by Oil red, Alizarin red, and toluidine blue staining. The BMSCs induced with the 3-step protocol differentiated into hepatic-like cells that expressed hepatocyte-specific proteins (ALB and AFP) and genes. CONCLUSION: The optimized protocol allows simple and efficient isolation of highly purified populations of BMSCs, which can be induced into hepatic lineages in specific microenvironment.

[Optimization of the method for isolating and culturing rat mesenchymal stem cells].

OBJECTIVE: To optimize the protocols for isolation and culture of mesenchymal stem cells from rat bone marrow (BMSCs). METHODS: BMSCs were isolated by adherence to plastic with frequent medium change and reduced trypsinization time. The cell growth curves were drawn and the surface markers of BMSCs were detected by flow cytometry. The cells were induced to differentiate into osteogenic, adipogenic, hepatic and cholic lineages. RESULTS: The cells isolated using this method were positive for CD29, CD44, and CD90 and negative for the hematopoietic surface markers CD45. The osteogenic and adipogenic differentiation of the BMSCs was verified by alkaline phosphatase staining, Alizarin red staining and Oil red staining. The cell subcultures up to passage 10 maintained capacities of differentiation into osteogenic and adipogenic lineages. The BMSCs induced with sequential addition of growth factors, cytokines and hormones differentiated into cells expressing hepatocyte- and cholangiocyte-specific markers. CONCLUSION: The optimized method allows efficient isolation of homogenous populations of MSCs from rat bone marrow, which can be induced into multiple cell lineages.