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1.
Gut Microbes ; 16(1): 2313769, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-38353638

RESUMO

Melatonin has various physiological effects, such as the maintenance of circadian rhythms, anti-inflammatory functions, and regulation of intestinal barriers. The regulatory functions of melatonin in gut microbiota remodeling have also been well clarified; however, the role of gut microbiota in regulating host melatonin production remains poorly understood. To address this, we studied the contribution of gut microbiota to host melatonin production using gut microbiota-perturbed models. We demonstrated that antibiotic-treated and germ-free mice possessed diminished melatonin levels in the serum and elevated melatonin levels in the colon. The influence of the intestinal microbiota on host melatonin production was further confirmed by fecal microbiota transplantation. Notably, Lactobacillus reuteri (L. R) and Escherichia coli (E. coli) recapitulated the effects of gut microbiota on host melatonin production. Mechanistically, L. R and E. coli activated the TLR2/4/MyD88/NF-κB signaling pathway to promote expression of arylalkylamine N-acetyltransferase (AANAT, a rate-limiting enzyme for melatonin production), and MyD88 deficiency in colonic epithelial cells abolished the influence of intestinal microbiota on colonic melatonin production. Collectively, we revealed a specific underlying mechanism of gut microbiota to modulate host melatonin production, which might provide novel therapeutic ideas for melatonin-related diseases.


Assuntos
Microbioma Gastrointestinal , Melatonina , Animais , Camundongos , Escherichia coli , Fator 88 de Diferenciação Mieloide/genética , Proteínas Adaptadoras de Transdução de Sinal , Células Epiteliais
2.
Cells ; 12(8)2023 04 07.
Artigo em Inglês | MEDLINE | ID: mdl-37190016

RESUMO

Skeletal muscle formation is an extremely important step in animal growth and development. Recent studies have found that TMEM8c (also known as Myomaker, MYMK), a muscle-specific transmembrane protein, can promote myoblast fusion and plays a key role in the normal development of skeletal muscle. However, the effect of Myomaker on porcine (Sus scrofa) myoblast fusion and the underlying regulatory mechanisms remain largely unknown. Therefore, in this study, we focused on the role and corresponding regulatory mechanism of the Myomaker gene during skeletal muscle development, cell differentiation, and muscle injury repair in pigs. We obtained the entire 3' UTR sequence of porcine Myomaker using the 3' RACE approach and found that miR-205 inhibited porcine myoblast fusion by targeting the 3' UTR of Myomaker. In addition, based on a constructed porcine acute muscle injury model, we discovered that both the mRNA and protein expression of Myomaker were activated in the injured muscle, while miR-205 expression was significantly inhibited during skeletal muscle regeneration. The negative regulatory relationship between miR-205 and Myomaker was further confirmed in vivo. Taken together, the present study reveals that Myomaker plays a role during porcine myoblast fusion and skeletal muscle regeneration and demonstrates that miR-205 inhibits myoblast fusion through targeted regulation of the expression of Myomaker.


Assuntos
MicroRNAs , Doenças Musculares , Animais , Suínos , Regiões 3' não Traduzidas/genética , Mioblastos/metabolismo , Músculo Esquelético/metabolismo , Proteínas de Membrana/metabolismo , Doenças Musculares/genética , MicroRNAs/genética , MicroRNAs/metabolismo
3.
Front Immunol ; 13: 967576, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-36045669

RESUMO

Hypoxia is a common hallmark of healthy tissues in physiological states or chronically inflamed tissues in pathological states. Mammalian cells sense and adapt to hypoxia mainly through hypoxia-inducible factor (HIF) signaling. Many studies have shown that hypoxia and HIF signaling play an important regulatory role in development and function of innate immune cells and T cells, but their role in B cell biology is still controversial. B cells experience a complex life cycle (including hematopoietic stem cells, pro-B cells, pre-B cells, immature B cells, mature naïve B cells, activated B cells, plasma cells, and memory B cells), and the partial pressure of oxygen (PO2) in the corresponding developmental niche of stage-specific B cells is highly dynamic, which suggests that hypoxia and HIF signaling may play an indispensable role in B cell biology. Based on the fact that hypoxia niches exist in the B cell life cycle, this review focuses on recent discoveries about how hypoxia and HIF signaling regulate the development, metabolism, and function of B cells, to facilitate a deep understanding of the role of hypoxia in B cell-mediated adaptive immunity and to provide novel strategies for vaccine adjuvant research and the treatment of immunity-related or infectious diseases.


Assuntos
Linfócitos B , Hipóxia , Imunidade Adaptativa , Animais , Linfócitos B/metabolismo , Hipóxia/metabolismo , Mamíferos/metabolismo , Oxigênio/metabolismo , Linfócitos T
4.
Bioengineering (Basel) ; 9(9)2022 Sep 07.
Artigo em Inglês | MEDLINE | ID: mdl-36134996

RESUMO

As the initial antibody technology, the preparation of hybridoma cells has been widely used in discovering antibody drugs and is still in use. Various antibody drugs obtained through this technology have been approved for treating human diseases. However, the key to producing hybridoma cells is efficient cell fusion. High-voltage microsecond pulsed electric fields (µsHVPEFs) are currently one of the most common methods used for cell electrofusion. Nevertheless, the membrane potential induced by the external microsecond pulse is proportional to the diameter of the cell, making it difficult to fuse cells of different sizes. Although nanosecond pulsed electric fields (nsPEFs) can achieve the fusion of cells of different sizes, due to the limitation of pore size, deoxyribonucleic acid (DNA) cannot efficiently pass through the cell pores produced by nsPEFs. This directly causes the significant loss of the target gene and reduces the proportion of positive cells after fusion. To achieve an electric field environment independent of cell size and enable efficient cell fusion, we propose a combination of nanosecond pulsed electric fields and low-voltage microsecond pulsed electric fields (ns/µsLVPEFs) to balance the advantages and disadvantages of the two techniques. The results of fluorescence experiments and hybridoma culture experiments showed that after lymphocytes and myeloma cells were stimulated by a pulse (ns/µsLVPEF, µsHVPEF, and control), compared with µsHVPEF, applying ns/µsLVPEF at the same energy could increase the cell fusion efficiency by 1.5-3.0 times. Thus far, we have combined nanosecond and microsecond pulses and provided a practical solution that can significantly increase cell fusion efficiency. This efficient cell fusion method may contribute to the further development of hybridoma technology in electrofusion.

5.
Front Microbiol ; 13: 1065668, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-36620044

RESUMO

Methionine (Met) metabolism provides methyl groups for many important physiological processes and is implicated in multiple inflammatory diseases associated with the disrupted intestinal microbiota; nevertheless, whether intestinal microbiota determines Met metabolism in the host remains largely unknown. Here, we found that gut microbiota is responsible for host Met metabolism by using various animal models, including germ-free (GF) pigs and mice. Specifically, the Met levels are elevated in both GF pigs and GF mice that mainly metabolized to S-adenosine methionine (SAM) in the liver. Furthermore, antibiotic clearance experiments demonstrate that the loss of certain ampicillin- or neomycin-sensitive gut microbiota causes decreased Met in murine colon. Overall, our study suggests that gut microbiota mediates Met metabolism in the host and is a prospective target for the treatment of Met metabolism-related diseases.

6.
Front Nutr ; 8: 713256, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34490327

RESUMO

Post-weaning diarrhea of piglets is associated with gut microbiota dysbiosis and intestinal pathogen infection. Recent studies have shown that Lactococcus lactis (L.lactis) could help suppress pathogen infection. This study aimed to investigate the effects of L.lactis on various factors related to growth and immunity in weaning piglets. The results showed that L.lactis improved the growth performance, regulated the amino acid profile (for example, increasing serum tryptophan and ileal mucosal cystine) and the intestinal GABAergic system (including inhibiting ileal gene expression of SLC6A13, GABAAρ1, π, θ, and γ1, and promoting ileal GABAAα5 expression). L.lactis also modulated intestinal immunity by promoting jejunal interleukin 17, 18, 22, ileal toll-like receptor 2, 5, 6, and myeloid differentiation primary response protein 88 gene expression while inhibiting jejunal interferon-γ and ileal interleukin 22 expressions. L.lactis highly affected the intestinal microbiota by improving the beta diversity of gut microbiota and the relative abundance of Halomonas and Shewanella. In conclusion, L.lactis improved the growth performance and regulated amino acid profiles, intestinal immunity and microbiota in weaning piglets.

7.
J Biol Eng ; 15(1): 18, 2021 Jun 29.
Artigo em Inglês | MEDLINE | ID: mdl-34187511

RESUMO

Since the advances in protein engineering and manufacture, over the last 30 years, antibody-based immunotherapeutic has become a powerful strategy to treat diseases. The T-cell engaging bispecific antibody (BsAb) by combining the Fab binding domain of tumor antigens and Fab or single-chain variable fragments (scFvs) binding domain of CD3 molecules, could redirect cytotoxic T cells to kill tumor cells. The IgG-scFv format of BsAb is a dual bivalent and asymmetrical design, which adds the benefit of potent cytotoxicity and less complicated for manufacture but limits the stability and production. Here, we engineered a series of interchain disulfide bonds in the Fab region of IgG-svFv BsAbs and evaluated its biophysical and biological properties. We found that simultaneously replaced the position of VH44-VL100 and CH1126-CL121 residues with cysteine, to form two additional disulfide bonds, could markedly increase monomeric BsAb formation and yield. The thermostability and stability against aggregation and degradation also performed better than BsAbs without extra disulfide bonds introduction. Besides, the affinity of engineered BsAbs was maintained, and the h8B-BsAb antibody had a slight enhancement in an inhibitory effect on target cells.

8.
Sci Rep ; 11(1): 11237, 2021 05 27.
Artigo em Inglês | MEDLINE | ID: mdl-34045661

RESUMO

Although the importance of the intestinal microbiota in host growth and health is well known, the relationship between microbiota colonization and muscle development is unclear. In this study, the direct causal effects of the colonization of gut microorganisms on the muscle tissue of piglets were investigated. The body weight and lean mass of germ-free (GF) piglets were approximately 40% lower than those of normal piglets. The deletion of the intestinal microbiota led to weakened muscle function and a reduction in myogenic regulatory proteins, such as MyoG and MyoD, in GF piglets. In addition, the blinded IGF1/AKT/mTOR pathway in GF piglets caused muscle atrophy and autophagy, which were characterized by the high expression of Murf-1 and KLF15. Gut microbiota introduced to GF piglets via fecal microbiota transplantation not only colonized the gut but also partially restored muscle growth and development. Furthermore, the proportion of slow-twitch muscle fibers was lower in the muscle of GF piglets, which was caused by the reduced short-chain fatty acid content in the circulation and impaired mitochondrial function in muscle. Collectively, these findings suggest that the growth, development and function of skeletal muscle in animals are mediated by the intestinal microbiota.


Assuntos
Microbioma Gastrointestinal/fisiologia , Músculo Esquelético/fisiologia , Animais , Perfilação da Expressão Gênica , Fator de Crescimento Insulin-Like I/metabolismo , Proteínas Proto-Oncogênicas c-akt/metabolismo , Transdução de Sinais/fisiologia , Suínos , Serina-Treonina Quinases TOR/metabolismo
9.
J Immunother ; 44(3): 106-113, 2021 04 01.
Artigo em Inglês | MEDLINE | ID: mdl-33239522

RESUMO

Cancer therapies benefit from accelerated development of biotechnology, and many immunotherapeutic strategies spring up including vaccines, the immune checkpoint blockade, chimeric antigen receptor T cells, and bispecific antibodies (BsAbs). Glypican-3 (GPC3) is a member of the heparan sulfate proteoglycan family of proteins and is highly expressed in hepatocellular carcinoma (HCC) cell membranes. Here, the authors describe a new tetravalent BsAb h8B-BsAb targeting GPC3 and CD3 antigens and studied its antitumor activities against HCC. h8B-BsAb was designed based on immunoglobulin G with a fragment variable fused to the light chain, whose biophysical stabilities including degradation resistance and thermostability were improved by introducing disulfide bonds. In vitro activity of h8B-BsAb showed potent T-cell recruitment and activation for HCC cell lysis by the presence of peripheral blood mononuclear cells, but no specific killing in GPC3-negative cells. In HCC xenograft mouse studies, h8B-BsAb induced robust regression of tumors. In summary, we engineered a highly stable and efficacious BsAb as a potential candidate for HCC treatment.


Assuntos
Anticorpos Biespecíficos/imunologia , Carcinoma Hepatocelular/imunologia , Glipicanas/imunologia , Neoplasias Hepáticas/imunologia , Linfócitos T/imunologia , Animais , Complexo CD3/imunologia , Linhagem Celular , Linhagem Celular Tumoral , Feminino , Células HEK293 , Células Hep G2 , Humanos , Leucócitos Mononucleares/imunologia , Camundongos , Ensaios Antitumorais Modelo de Xenoenxerto/métodos
10.
Invest New Drugs ; 39(3): 615-626, 2021 06.
Artigo em Inglês | MEDLINE | ID: mdl-33215325

RESUMO

The acceleration of therapeutic antibody development has been motivated by the benefit to and their demand for human health. In particular, humanized transgenic antibody discovery platforms, combined with immunization, hybridoma fusion and/or single cell DNA sequencing are the most reliable and rapid methods for mining the human monoclonal antibodies. Human GPC3 protein is an oncofetal antigen, and it is highly expressed in most hepatocellular carcinomas and some types of squamous cell carcinomas. Currently, no fully human anti-GPC3 therapeutic antibodies have been reported and evaluated in extensive tumor tissues. Here, we utilized a new humanized transgenic mouse antibody discovery platform (CAMouse) that contains large V(D)J -regions and human gamma-constant regions of human immunoglobulin in authentic configurations to generate fully human anti-GPC3 antibodies. Our experiments resulted in four anti-GPC3 antibodies with high-specific binding and cytotoxicity to GPC3 positive cancer cells, and the antibody affinities are in the nanomolar range. Immunohistochemistry analysis demonstrated that these antibodies can recognize GPC3 protein on many types of solid tumors. In summary, the human anti-human GPC3 monoclonal antibodies described here are leading candidates for further preclinical studies of cancer therapy, further, the CAMouse platform is a robust tool for human therapeutic antibody discovery.


Assuntos
Anticorpos Monoclonais/farmacologia , Glipicanas/antagonistas & inibidores , Neoplasias/metabolismo , Animais , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Feminino , Glipicanas/imunologia , Glipicanas/metabolismo , Hepatite/metabolismo , Humanos , Intestino Delgado/metabolismo , Pulmão/metabolismo , Masculino , Camundongos Transgênicos , Placenta/metabolismo , Gravidez
11.
J Anim Sci ; 98(12)2020 Dec 01.
Artigo em Inglês | MEDLINE | ID: mdl-33205812

RESUMO

The present experiment was conducted to investigate the effects of exogenously infused short-chain fatty acids (SCFAs) on the growth development and intestinal functions in a germ-free (GF) pig model. Twelve hysterectomy-derived newborn piglets were reared in six sterile isolators. All piglets were hand-fed Co60-γ-irradiated sterile milk powder for 21 d and then were switched to sterile feed for another 21 d. During the second 21-d period, GF piglets (n = 6) were orally infused with 25 mL/kg sterile saline per day, and SCFA piglets (n = 6) were orally infused with 25 mL/kg SCFAs mixture (acetic, propionic, and butyric acids, 45, 15, and 11 mM, respectively) per day. We observed the concentrations of SCFAs in serum and intestine, and the messenger ribonucleic acid (mRNA) abundance of G-protein-coupled receptor-43 in the ileum was increased (P < 0.05) in the SCFA group. Meanwhile, oral infusion of SCFAs enhanced (P < 0.05) the contents of glucagon-like peptide-2 in the jejunum and serum and tended to increase the villi height in the ileum (P < 0.10). Besides, the activities of lipase, trypsin, sucrase, lactase, Na+-K+-adenosine triphosphatase ([ATPase] P < 0.05), and Ca2+-Mg2+-ATPase (P < 0.10) were stimulated and the mRNA expressions of solute carrier family 7 (SLC7A1) and regeneration protein (REG)-ΙΙΙ Î³ in the jejunum (P < 0.05) were upregulated in the SCFA group. Additionally, SCFAs infusion downregulated the mRNA abundances of interleukin (IL)-1ß and IL-6 in the jejunum, ileum, or colon (P < 0.05) and increased the counts of white blood cell, neutrophils, and lymphocyte in the blood (P < 0.05). Collectively, exogenous infusion of SCFAs might improve intestinal health through promoting intestinal development and absorption function, and enhancing intestinal immune function, and these effects were occur independently of the gut microbiota.


Assuntos
Microbioma Gastrointestinal , Animais , Ácidos Graxos Voláteis , Feminino , Íleo , Intestinos , Jejuno , Suínos
12.
Am J Physiol Gastrointest Liver Physiol ; 318(3): G542-G553, 2020 03 01.
Artigo em Inglês | MEDLINE | ID: mdl-31984787

RESUMO

A lack of sunlight exposure, residence in the northern latitudes, and dietary vitamin D insufficiency are coprevalent with metabolic syndrome (MetS), Type 2 diabetes (T2D), and nonalcoholic fatty liver diseases (NAFLD), implying a potential causality and underlying mechanism. Whether vitamin D supplementation or treatment can improve these disorders is controversial, in part, because of the absence of large-scale trials. Experimental investigations, on the other hand, have uncovered novel biological functions of vitamin D in development, tumor suppression, and immune regulation, far beyond its original role as a vitamin that maintained calcium homeostasis. While the large intestine harbors massive numbers of microbes, the small intestine has a minimal quantity of bacteria, indicating the existence of a gating system located in the distal region of the small intestine that may restrain bacterial translocation to the small intestine. Vitamin D receptor (VDR) was found to be highly expressed at the distal region of small intestine, where the vitamin D signaling promotes innate immunity, including the expression of α-defensins by Paneth cells, and maintains the intestinal tight junctions. Thus, a new hypothesis is emerging, indicating that vitamin D deficiency may impair the intestinal innate immunity, including downregulation of Paneth cell defensins, leading to bacterial translocation, endotoxemia, systemic inflammation, insulin resistance, and hepatic steatosis. Here, we review the studies for vitamin D for innate immunity and metabolic homeostasis, and we outline the clinical trials of vitamin D for mitigating MetS, T2D, and NAFLD.


Assuntos
Diabetes Mellitus Tipo 2/metabolismo , Microbioma Gastrointestinal , Imunidade Inata , Imunidade nas Mucosas , Mucosa Intestinal/metabolismo , Síndrome Metabólica/metabolismo , Hepatopatia Gordurosa não Alcoólica/tratamento farmacológico , Vitamina D/metabolismo , Diabetes Mellitus Tipo 2/tratamento farmacológico , Diabetes Mellitus Tipo 2/imunologia , Diabetes Mellitus Tipo 2/microbiologia , Microbioma Gastrointestinal/efeitos dos fármacos , Interações Hospedeiro-Patógeno , Humanos , Imunidade Inata/efeitos dos fármacos , Imunidade nas Mucosas/efeitos dos fármacos , Mucosa Intestinal/efeitos dos fármacos , Mucosa Intestinal/imunologia , Mucosa Intestinal/microbiologia , Síndrome Metabólica/tratamento farmacológico , Síndrome Metabólica/imunologia , Síndrome Metabólica/microbiologia , Hepatopatia Gordurosa não Alcoólica/imunologia , Hepatopatia Gordurosa não Alcoólica/microbiologia , Receptores de Calcitriol/metabolismo , Transdução de Sinais , Vitamina D/uso terapêutico
13.
Bioelectrochemistry ; 127: 171-179, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-30831355

RESUMO

The excessive cell death rate caused by electrofusion with unipolar pulses (UPs) has been a bottleneck to increasing cell fusion efficiency in monoclonal antibody technology. Several studies have confirmed that compared with UPs, bipolar pulses (BPs) with microsecond pulse widths can increase electropermeabilization while reducing cell death. Given these characteristics, BPs were used to increase cell fusion efficiency in this study. Cell staining and hybridoma culture experiments were performed using SP2/0 mouse myeloma cells and lymphocytes. Based on the equal energy principle, UPs and BPs were delivered to electrodes at a distance of 3.81 mm, with electric field intensities ranging from 2 kV/cm to 3 kV/cm and pulse duration of 40 µs for the UPs and 20-20 µs for the BPs. The results of cell staining experiments showed that cell fusion efficiency was 3-fold greater with BPs than with UPs. Similarly, the results of the hybridoma culture experiments showed that the hybridoma yields were 0.26‰ and 0.23‰ (2.5 kV/cm and 3 kV/cm, respectively) in the UP groups and increased to 0.46‰ and 0.35‰ in the BP groups. Taken together, the results show that the efficiency of heterologous cell fusion can be greatly increased if BPs are used instead of the commonly applied UPs. This study may provide a promising method for monoclonal antibody technology.


Assuntos
Anticorpos Monoclonais/metabolismo , Fusão Celular/métodos , Hibridomas/citologia , Linfócitos/citologia , Animais , Fusão Celular/instrumentação , Células Cultivadas , Eletricidade , Eletroporação/instrumentação , Eletroporação/métodos , Desenho de Equipamento , Hibridomas/metabolismo , Linfócitos/metabolismo , Masculino , Camundongos
14.
IEEE Trans Biomed Eng ; 66(5): 1353-1360, 2019 05.
Artigo em Inglês | MEDLINE | ID: mdl-30281431

RESUMO

OBJECTIVE: Unipolar pulses have been used in cell electrofusion over the last decades. However, the problem of high mortality with unipolar pulses has not been solved effectively. The cell fusion rate is restricted by cell mortality. By using the advantages of bipolar pulses which cause less cell damage, this paper attempts to use bipolar pulses to increase the cell fusion rate. METHODS: the transmembrane voltage and pore density of cells subjected to unipolar/bipolar pulses were simulated in COMSOL software. In an experiment, two 40 µs unipolar and two 20-20 µs bipolar pulses with electric fields of 2, 2.5, and 3 kV/cm were applied to SP2/0 murine myeloma cells. To determine the cell fusion rate and cell mortality, cells were stained with Hoechst 33342 and propidium iodide. RESULTS: the simulation in this paper showed that a high transmembrane voltage and a high pores density were concentrated only at the contact area of cells when bipolar pulses were used. The results of the cell staining experiment verified the simulation analysis. When bipolar pulses were applied, the cell mortality was significantly reduced. In addition, the cell fusion rate with bipolar pulses was almost two times higher than that with unipolar pulses. CONCLUSION: for cell electrofusion, compared with unipolar pulses, bipolar pulses can not only reduce the cell mortality remarkably but also improve the cell fusion rate obviously. SIGNIFICANCE: this paper introduces a novel way to increase the fusion rate of cells.


Assuntos
Fusão Celular/métodos , Membrana Celular/efeitos da radiação , Eletroporação/métodos , Animais , Morte Celular/efeitos da radiação , Linhagem Celular Tumoral , Simulação por Computador , Camundongos , Porosidade
15.
Macromol Biosci ; 16(9): 1381-90, 2016 09.
Artigo em Inglês | MEDLINE | ID: mdl-27280860

RESUMO

Photo-crosslinking and self-healing have received considerable attention for the design of intelligent materials. A novel photostimulated, self-healing, and cytocompatible hydrogel system is reported. A coumarin methacrylate crosslinker is synthesized to modify the polyacrylamide-based hydrogels. With the [2+2] cyclo-addition of coumarin moieties, the hydrogels exhibit excellent self-healing capacity when they are exposed to light with wavelengths at 280 and 365 nm, respectively. To enhance cell compatibility, a poly (amidoamine) crosslinker is also synthesized. Variations in light exposure times and irradiation wavelengths are found to alter the self-healing property of the hydrogels. The hydrogels are shown to induce a regular cellular pattern. The hydrogels are used to regulate bone marrow stromal cells differentiation. The relative mRNA expressions are recorded to monitor the osteogenic differentiation of the cells.


Assuntos
Diferenciação Celular/efeitos dos fármacos , Hidrogéis/farmacologia , Luz , Teste de Materiais/métodos , Células-Tronco Mesenquimais/citologia , Animais , Morte Celular/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Reagentes de Ligações Cruzadas/química , Regulação da Expressão Gênica/efeitos dos fármacos , Processamento de Imagem Assistida por Computador , Células-Tronco Mesenquimais/efeitos dos fármacos , Células-Tronco Mesenquimais/metabolismo , Metacrilatos/síntese química , Metacrilatos/química , Camundongos , Microscopia de Fluorescência , Osteogênese/efeitos dos fármacos , Osteogênese/genética , Poliaminas/síntese química , Poliaminas/química , Espectroscopia de Prótons por Ressonância Magnética , Reação em Cadeia da Polimerase em Tempo Real
16.
J Biomater Appl ; 30(9): 1385-91, 2016 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-26801474

RESUMO

Pelvic organ prolapse (POP) is a serious health issue that affects many adult women. Surgical treatments for POP patients comprise a common strategy in which scaffold materials are used to reconstruct the prolapsed pelvic. However, the existing materials for pelvic reconstruction cannot meet clinical requirements in terms of biocompatibility, mechanics and immunological rejection. To address these concerns, polypropylene (PP) mesh was selected because of its strong mechanical properties. Small intestinal submucosa (SIS) was used to modify the PP mesh via a mussel-inspired polydopamine coating to enhance its biocompatibility. The scanning electron microscopy (SEM) and atomic force microscopy (AFM) results demonstrated that SIS was successfully conjugated on the surface of the PP mesh. Moreover, the cytotoxicity results indicated that the PP mesh and SIS-modified PP mesh were safe to use. Furthermore, in vivo tests demonstrated that the fibroplasia around the implanted site in the SIS-modified PP mesh group was significantly less than the fibroplasia around the PP mesh group. In addition, the immunohistochemistry staining results indicated that the expression of pro-inflammatory macrophages (M1) was substantially lower and that the expression of pro-healing macrophages (M2) was higher in the SIS-modified PP mesh group. Furthermore, ELISA detection indicated that the expression of IL-1ß and IL-6 in the SIS-modified PP mesh group was reduced compared with the PP mesh group. These findings suggest that a SIS-modified polypropylene hybrid mesh via a mussel-inspired polydopamine coating is a promising approach in pelvic reconstruction.


Assuntos
Materiais Revestidos Biocompatíveis/química , Indóis/química , Mucosa Intestinal/química , Polímeros/química , Polipropilenos/química , Telas Cirúrgicas , Alicerces Teciduais/química , Animais , Materiais Biomiméticos/química , Materiais Biomiméticos/toxicidade , Bivalves/química , Linhagem Celular , Materiais Revestidos Biocompatíveis/toxicidade , Feminino , Indóis/imunologia , Indóis/toxicidade , Interleucina-1beta/análise , Interleucina-1beta/imunologia , Interleucina-6/análise , Interleucina-6/imunologia , Mucosa Intestinal/imunologia , Teste de Materiais , Camundongos , Polímeros/toxicidade , Polipropilenos/imunologia , Polipropilenos/toxicidade , Ratos Sprague-Dawley , Telas Cirúrgicas/efeitos adversos , Suínos
17.
Tissue Eng Part C Methods ; 21(10): 1015-24, 2015 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-25951851

RESUMO

A persistent challenge in tissue engineering is the fabrication of manipulatable scaffolds for implantation in clinical treatments and use in disease models for drug screening. Electrospinning of nanofibrous membranes is an emerging technology in artificial extracellular matrix (ECM) design that can offer precisely tunable microenvironments upon assembly into three-dimensional (3D) scaffolds that mimic the in vivo ECM structure. In this study, we report a facile and versatile strategy for preparing 3D multilayered constructs from Fe3O4/polycaprolactone (PCL)/gelatin nanofibrous membranes. This method combines membrane assembly with noncontact magnetic force to preserve the mechanical integrity and interconnectivity of the 3D scaffolds. An ordered layer structure can be achieved using a magnetic control technique through the addition of magnetic nanoparticles into the PCL/gelatin nanofibers. We first verified the magnetic properties and structures of magnetic nanofibers according to X-ray diffraction, hysteresis, scanning electron microscopy, and transmission electron microscopy. We tested the potential toxicity and osteogenic differentiation of mesenchymal stem cells seeded on the layered scaffolds. To add further functionality to the scaffolds, the membranes were coated with silver nanoparticles and shown to inhibit the growth of Escherichia coli and Staphylococcus aureus, which are responsible for most cases of infection-related implant failure. Finally, we tested the utility of magnetic membranes implanted in an animal model as a contrast agent for magnetic resonance imaging. Scaffolds formed using the presented magnetically guided fabrication strategy have the potential to mimic the structure and function of human tissues and also may be applied in disease models to study cell-cell interactions.


Assuntos
Compostos Férricos/química , Gelatina/química , Campos Magnéticos , Células-Tronco Mesenquimais/metabolismo , Osteogênese , Poliésteres/química , Alicerces Teciduais/química , Animais , Matriz Extracelular/química , Humanos , Células-Tronco Mesenquimais/citologia , Camundongos , Nanomedicina Teranóstica , Engenharia Tecidual
18.
Int J Nanomedicine ; 10: 3193-201, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-25995629

RESUMO

Pelvic organ prolapse (POP) is a serious health issue affecting many adult women. Complications of POP include pelvic pressure, pelvic pain, and problems in emptying their bowels or bladder. Sometimes, POP may even cause urinary outflow obstruction and lead to bladder or kidney infections. Currently, synthetic and naturally derived materials have been chosen for treatment of POP to reduce the high recurrence rates after surgical interventions. However, existing materials for POP treatment cannot meet the clinical requirements in terms of biocompatibility, mechanics, and minimal risk of rejection. Especially, erosion in synthetic polymers and rapid degradation in natural polymers limit their further applications in clinics. To address these concerns, we report a novel POP replacement using core-sheath polystyrene/gelatin electrospun nanofiber mesh. The outside gelatin sheath provides a hydrophilic surface and implantable integrity between host and guest, while the inner PS core offers the necessary mechanical support. The composite mesh shows graft accommodation in pelvic submucosa after implantation in vivo, as shown in hematoxylin-eosin staining and T helper cell phenotype and macrophage phenotype stainings. Qualitative analysis of inducible nitric oxide synthase, arginase, interferon-γ, and interleukin-10 gene expressions also indicates that the implanted composite mesh switches to accommodation mode 2 weeks postimplantation. Thus, these novel core-sheath polystyrene/gelatin nanofibrous membranes are promising in pelvic reconstruction.


Assuntos
Materiais Biocompatíveis , Gelatina , Nanofibras/química , Procedimentos de Cirurgia Plástica/instrumentação , Poliestirenos , Telas Cirúrgicas , Animais , Materiais Biocompatíveis/química , Materiais Biocompatíveis/farmacologia , Adesão Celular/efeitos dos fármacos , Células Cultivadas , Feminino , Gelatina/química , Gelatina/farmacologia , Masculino , Camundongos , Prolapso de Órgão Pélvico , Poliestirenos/química , Poliestirenos/farmacologia , Ratos Sprague-Dawley , Vagina/cirurgia
19.
Eur J Contracept Reprod Health Care ; 20(2): 136-40, 2015 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-25666811

RESUMO

OBJECTIVE: The study evaluated the efficacy of removing retained intrauterine devices (IUDs) under direct vision using a novel hysteroscopic hook. METHODS: In a retrospective observational study, 83 patients (group 1) underwent IUD extraction using a hysteroscopic IUD removal hook (HIRH) and 60 patients (group 2) underwent traditional hysteroscopic IUD extraction. We recorded the blood loss, operation time and success rate. RESULTS: The operation time was shorter (10.7 vs. 17.7 min; p < 0.001) and the success rate higher in group 1 compared with group 2 (odds ratio 1.09; p = 0.027). CONCLUSIONS: The HIRH is an effective, simple, inexpensive and durable tool for the direct visual removal of IUDs partially embedded in the endometrium and for damaged IUDs.


Assuntos
Remoção de Dispositivo/métodos , Histeroscopia/instrumentação , Dispositivos Intrauterinos , Adulto , Idoso , Feminino , Humanos , Migração de Dispositivo Intrauterino , Pessoa de Meia-Idade , Duração da Cirurgia , Estudos Retrospectivos , Resultado do Tratamento
20.
Eur J Obstet Gynecol Reprod Biol ; 183: 109-13, 2014 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-25461362

RESUMO

OBJECTIVE: To compare a new method using fallopian tube stripping forceps (FTSF) for salpingostomy in laparoscopic tubal pregnancy management. STUDY DESIGN: Comparative observational study. A total of 102 patients with ampullary tubal pregnancy were treated as follows: 56 patients (Group 1) underwent stripping by FTSF, and 46 patients (Group 2) underwent salpingostomy. The bleeding, operation time, persistent ectopic pregnancy (EP) rate, and the first reproductive performance were investigated. RESULTS: We found less intraoperative bleeding, shorter operation times, and lower rates of EP recurrence in Group 1 compared with Group 2. In contrast, we observed no significant differences in the persistent EP rate, the occurrences of spontaneous intrauterine pregnancy and miscarriage, and the rates of successful IVF between the two groups. CONCLUSION: For distal tubal pregnancy with an ectopic mass ≤30mm, laparoscopic fallopian tube stripping assisted by FTSF may be an easy, less-damaging, conservative operational modality with lower recurrent EP compared with salpingostomy for patients who desire future pregnancy.


Assuntos
Laparoscopia , Gravidez Tubária/cirurgia , Salpingostomia/instrumentação , Adulto , Feminino , Seguimentos , Humanos , Gravidez , Salpingostomia/métodos , Resultado do Tratamento
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