Cellular and molecular bioengineering最新文献

{"title":"Causative Role of Anoxic Environment in Bacterial Regulation of Human Intestinal Function.","authors":"Chengyao Wang,&nbsp;Andrea Cancino,&nbsp;Jasmine Baste,&nbsp;Daniel Marten,&nbsp;Advait Anil Joshi,&nbsp;Amreen Nasreen,&nbsp;Abhinav Bhushan","doi":"10.1007/s12195-022-00735-x","DOIUrl":"https://doi.org/10.1007/s12195-022-00735-x","url":null,"abstract":"<p><strong>Introduction: </strong>Life on Earth depends on oxygen; human tissues require oxygen signaling, whereas many microorganisms, including bacteria, thrive in anoxic environments. Despite these differences, human tissues and bacteria coexist in close proximity to each other such as in the intestine. How oxygen governs intestinal-bacterial interactions remains poorly understood.</p><p><strong>Methods: </strong>To address to this gap, we created a dual-oxygen environment in a microfluidic device to study the role of oxygen in regulating the regulation of intestinal enzymes and proteins by gut bacteria. Two-layer microfluidic devices were designed using a fluid transport model and fabricated using soft lithography. An oxygen-sensitive material was integrated to determine the oxygen levels. The intestinal cells were cultured in the upper chamber of the device. The cells were differentiated, upon which bacterial strains, a facultative anaerobe, <i>Escherichia coli</i> Nissle 1917, and an obligate anaerobe, <i>Bifidobacterium Adolescentis</i>, were cultured with the intestinal cells.</p><p><strong>Results: </strong>The microfluidic device successfully established a dual-oxygen environment. Of particular importance in our findings was that both strains significantly upregulated mucin proteins and modulated several intestinal transporters and transcription factors but only under the anoxic-oxic oxygen gradient, thus providing evidence of the role of oxygen on bacterial-epithelial signaling.</p><p><strong>Conclusions: </strong>Our work that integrates cell and molecular biology with bioengineering presents a novel strategy to engineer an accessible experimental system to provide tailored oxygenated environments. The work could provide new avenues to study intestine-microbiome signaling and intestinal tissue engineering, as well as a novel perspective on the indirect effects of gut bacteria on tissues including tumors.</p><p><strong>Supplementary information: </strong>The online version contains supplementary material available at 10.1007/s12195-022-00735-x.</p>","PeriodicalId":9687,"journal":{"name":"Cellular and molecular bioengineering","volume":"15 5","pages":"493-504"},"PeriodicalIF":2.8,"publicationDate":"2022-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9700550/pdf/12195_2022_Article_735.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10616026","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Myosin II Adjusts Motility Properties and Regulates Force Production Based on Motor Environment.","authors":"Omayma Y Al Azzam,&nbsp;Janie C Watts,&nbsp;Justin E Reynolds,&nbsp;Juliana E Davis,&nbsp;Dana N Reinemann","doi":"10.1007/s12195-022-00731-1","DOIUrl":"https://doi.org/10.1007/s12195-022-00731-1","url":null,"abstract":"<p><strong>Introduction: </strong>Myosin II has been investigated with optical trapping, but single motor-filament assay arrangements are not reflective of the complex cellular environment. To understand how myosin interactions propagate up in scale to accomplish system force generation, we devised a novel actomyosin ensemble optical trapping assay that reflects the hierarchy and compliancy of a physiological environment and is modular for interrogating force effectors.</p><p><strong>Methods: </strong>Hierarchical actomyosin bundles were formed <i>in vitro</i>. Fluorescent template and cargo actin filaments (AF) were assembled in a flow cell and bundled by myosin. Beads were added in the presence of ATP to bind the cargo AF and activate myosin force generation to be measured by optical tweezers.</p><p><strong>Results: </strong>Three force profiles resulted across a range of myosin concentrations: high force with a ramp-plateau, moderate force with sawtooth movement, and baseline. The three force profiles, as well as high force output, were recovered even at low solution concentration, suggesting that myosins self-optimize within AFs. Individual myosin steps were detected in the ensemble traces, indicating motors are taking one step at a time while others remain engaged in order to sustain productive force generation.</p><p><strong>Conclusions: </strong>Motor communication and system compliancy are significant contributors to force output. Environmental conditions, motors taking individual steps to sustain force, the ability to backslip, and non-linear concentration dependence of force indicate that the actomyosin system contains a force-feedback mechanism that senses the local cytoskeletal environment and communicates to the individual motors whether to be in a high or low duty ratio mode.</p><p><strong>Supplementary information: </strong>The online version contains supplementary material available at 10.1007/s12195-022-00731-1.</p>","PeriodicalId":9687,"journal":{"name":"Cellular and molecular bioengineering","volume":"15 5","pages":"451-465"},"PeriodicalIF":2.8,"publicationDate":"2022-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9700534/pdf/12195_2022_Article_731.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9999863","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"A Versatile Micromanipulation Apparatus for Biophysical Assays of the Cell Nucleus.","authors":"Marilena L Currey, Viswajit Kandula, Ronald Biggs, John F Marko, Andrew D Stephens","doi":"10.1007/s12195-022-00734-y","DOIUrl":"10.1007/s12195-022-00734-y","url":null,"abstract":"<p><strong>Intro: </strong>Force measurements of the nucleus, the strongest organelle, have propelled the field of mechanobiology to understand the basic mechanical components of the nucleus and how these components properly support nuclear morphology and function. Micromanipulation force measurement provides separation of the relative roles of nuclear mechanical components chromatin and lamin A.</p><p><strong>Methods: </strong>To provide access to this technique, we have developed a universal micromanipulation apparatus for inverted microscopes. We outline how to engineer and utilize this apparatus through dual micromanipulators, fashion and calibrate micropipettes, and flow systems to isolate a nucleus and provide force vs. extensions measurements. This force measurement approach provides the unique ability to measure the separate contributions of chromatin at short extensions and lamin A strain stiffening at long extensions. We then investigated the apparatus' controllable and programmable micromanipulators through compression, isolation, and extension in conjunction with fluorescence to develop new assays for nuclear mechanobiology.</p><p><strong>Results: </strong>Using this methodology, we provide the first rebuilding of the micromanipulation setup outside of its lab of origin and recapitulate many key findings including spring constant of the nucleus and strain stiffening across many cell types. Furthermore, we have developed new micromanipulation-based techniques to compress nuclei inducing nuclear deformation and/or rupture, track nuclear shape post-isolation, and fluorescence imaging during micromanipulation force measurements.</p><p><strong>Conclusion: </strong>We provide the workflow to build and use a micromanipulation apparatus with any inverted microscope to perform nucleus isolation, force measurements, and various other biophysical techniques.</p><p><strong>Supplementary information: </strong>The online version contains supplementary material available at 10.1007/s12195-022-00734-y.</p>","PeriodicalId":9687,"journal":{"name":"Cellular and molecular bioengineering","volume":"15 4","pages":"303-312"},"PeriodicalIF":2.3,"publicationDate":"2022-09-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9474788/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9319373","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"How to Thrive as a First-Generation College Student in Engineering.","authors":"Emily D Fabiano","doi":"10.1007/s12195-022-00736-w","DOIUrl":"https://doi.org/10.1007/s12195-022-00736-w","url":null,"abstract":"<p><p>First-generation college students face unique challenges compared to non-first-generation college students, especially in STEM fields. First-generation STEM students drop out of their major at higher rates than non-first-generation students. This may be due to a lack of role models or mentorship in the college environment or in the field, a challenging curriculum, and difficulty balancing personal and academic commitments. There has been a lack of significant attention given to studying first-generation college students in undergraduate engineering, and therefore, there is limited understanding of how to navigate post-secondary education as a first-generation college student to succeed in undergraduate engineering. Here, I lay out tips for success based on my own experience as a first-generation student in engineering. This includes how to find the right major for you, adjust to college, have a solid support system, seek out research opportunities, become involved in outreach, experience inclusivity, balance courses with other commitments, and apply for scholarships. This article also discusses considerations in pursuing graduate education. With more support, mentoring and guidance, a greater percentage of first-generation students will succeed in pursuing undergraduate engineering degrees.</p>","PeriodicalId":9687,"journal":{"name":"Cellular and molecular bioengineering","volume":"15 4","pages":"293-299"},"PeriodicalIF":2.8,"publicationDate":"2022-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9474774/pdf/12195_2022_Article_736.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10492934","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Mechanotransduction in Endothelial Cells in Vicinity of Cancer Cells.","authors":"Alessandra Ebben,&nbsp;Mahsa Dabagh","doi":"10.1007/s12195-022-00728-w","DOIUrl":"https://doi.org/10.1007/s12195-022-00728-w","url":null,"abstract":"<p><p><i>Introduction</i>-Local hemodynamics impact the mechanotransduction in endothelial cells (ECs) lining the vascular network. On the other hand, cancer cells are shown to influence the local hemodynamics in their vicinity, in microvasculature. The first objective of present study is to explore how cancer cell-induced changes in local hemodynamics can impact the forces experienced by intra/inter-cellular organelles of ECs that are believed to play important roles in mechanotransduction. Moreover, extracellular matrix (ECM) stiffening has been shown to correlate with progression of most cancer types. However, it is still not well understood how ECM stiffness impacts ECs mechanosensors. The second objective of this study is to elucidate the role of ECM stiffness on mechanotransduction in ECs. <i>Methods</i>-A three-dimensional, multiscale, multicomponent, viscoelastic model of focally adhered ECs is developed to simulate the force transmission through ECs mechanosensors [actin cortical layer, nucleus, cytoskeleton, focal adhesions (FAs), and adherens junctions (ADJs)]. <i>Results</i>-Our results show that cancer cell-altered hemodynamics results in significantly high forces transmitted to subcellular organelles of ECs which are in vicinity of cancer cells. This impact is more drastic on stress fibers (SFs) both centrally located and peripheral ones. Furthermore, we demonstrate that ADJs, FAs, and SFs experience higher stresses in ECs attached to stiffer ECM. Impact of ECM stiffness is particularly significant in ECs exposed to fluid shear stresses of 2 Pa or lower. This finding reveals the role of organ-specific stiffness in promoting cancer cell transmigration even in capillaries larger than cancer cell diameter. <i>Conclusions</i>-ÊCancer cell-induced-changes in ECs mechanotransduction represents an important potential mechanism for cancer cell transmigration in the microvasculature particularly with stiffer ECM. The identification of ECs mechanosensors involved in early stages of EC-cancer cell interaction will help with developing more efficient therapeutic interventions to suppress cancer cell transmigration in the microvasculature.</p>","PeriodicalId":9687,"journal":{"name":"Cellular and molecular bioengineering","volume":"15 4","pages":"313-330"},"PeriodicalIF":2.8,"publicationDate":"2022-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9474981/pdf/12195_2022_Article_728.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9808877","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Structural Mapping of BMP Conformational Epitopes and Bioengineering Design of Osteogenic Peptides to Specifically Target the Epitope-Binding Sites.","authors":"Hao Chen,&nbsp;Yaodong Zhou,&nbsp;Qirong Dong","doi":"10.1007/s12195-022-00725-z","DOIUrl":"https://doi.org/10.1007/s12195-022-00725-z","url":null,"abstract":"<p><strong>Introduction: </strong>Human bone morphogenetic proteins (BMPs) constitute a large family of cytokines related to members of the transforming growth factor-β superfamily, which fulfill biological functions by specificity binding to their cognate type I (BRI) and type II (BRII) receptors through conformational wrist and linear knuckle epitopes, respectively.</p><p><strong>Methods and results: </strong>We systematically examined the intermolecular recognition and interaction between the BMP proteins and BRI receptor at structural, energetic and dynamic levels. The BRI-binding site consists of three hotspot regions on BMP surface, which totally contribute ~70% potency to the BMP-BRI binding events and represent the core sections of BMP conformational wrist epitope; the contribution increases in the order: hotspot 2 (~ 8%) < hotspot 3 (~ 20%) < hotspot 1 (~ 40%). Multiple sequence alignment and structural superposition revealed a consensus sequence pattern and a similar binding mode of the three hotspots shared by most BMP members, indicating a high conservation of wrist epitope in BMP family. The three hotspots are natively folded into wellstructured <i>U</i>-shaped,, loop and double-stranded conformations in BMP proteins, which, however, would become largely disordered when splitting from the protein context to derive osteogenic peptides in free state, thus largely impairing their rebinding capability to BRI receptor. In this respect, cyclization strategy was employed to constrain hotspot 1/3-derived peptides into a native-like conformation, which was conducted by adding a disulfide bond across the ending arms of linear peptides based on their native conformations. Fluorescence-based assays substantiated that the cyclization can effectively improve the binding affinities of osteogenic peptides to BRI receptor by 3-6-fold. The cyclic peptides also exhibit a good selectivity for BRI over BRII (> 5-fold), confirming that they can specifically target the wrist epitope-binding site of BRI receptor.</p><p><strong>Conclusion: </strong>The rationally designed cyclic peptides can be regarded as the promising lead entities that should be further chemically modified to enhance their in vivo biological stability for further bioengineering therapeutic osteogenic peptides against chondrocyte senescence and bone disorder.</p>","PeriodicalId":9687,"journal":{"name":"Cellular and molecular bioengineering","volume":"15 4","pages":"341-352"},"PeriodicalIF":2.8,"publicationDate":"2022-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9474794/pdf/12195_2022_Article_725.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9416442","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Leadership Lessons Learned in Biomedical Engineering.","authors":"Michael R King","doi":"10.1007/s12195-022-00732-0","DOIUrl":"https://doi.org/10.1007/s12195-022-00732-0","url":null,"abstract":"","PeriodicalId":9687,"journal":{"name":"Cellular and molecular bioengineering","volume":"15 4","pages":"301-302"},"PeriodicalIF":2.8,"publicationDate":"2022-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9474766/pdf/12195_2022_Article_732.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10401493","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"OTUB2 Promotes Proliferation and Migration of Hepatocellular Carcinoma Cells by PJA1 Deubiquitylation.","authors":"Gang Hu,&nbsp;Jianwu Yang,&nbsp;Hongwen Zhang,&nbsp;Zhen Huang,&nbsp;Heming Yang","doi":"10.1007/s12195-022-00720-4","DOIUrl":"https://doi.org/10.1007/s12195-022-00720-4","url":null,"abstract":"<p><strong>Introduction: </strong>Recent studies have revealed that several deubiquitinating enzymes (DUBs) play important roles in hepatocellular carcinoma (HCC) progression, but the roles of Otubain 2 (OTUB2) in HCC remain obscure.</p><p><strong>Methods: </strong>In this study, we investigated the expression of OTUB2 in HCC based on clinical samples and a public online database (ENCORI), and its roles and working mechanisms were further explored by <i>in vitro</i> experiments.</p><p><strong>Results: </strong>It was found that the expression of OTUB2 was significantly up-regulated in HCC tissues, and correlated with poor prognosis of HCC patients. Functionally, the overexpression of OTUB2 could promote malignant proliferation and metastasis of HCC cells, while knockdown of OTUB2 exerted the opposite results. Using two bioinformatics tools, PJA1 was identified as a potential gene regulated by OTUB2. Mechanistically, it was found that OTUB2 promoted the stabilization of PJA1 by deubiquitylation, based on immunoprecipitation (IP) and cycloheximide (CHX) assays. Moreover, the suppressive effects of OTUB2 depletion on the malignant phenotypes of HCC cells could be reversed by overexpressing PJA1.</p><p><strong>Conclusion: </strong>In conclusion, our study indicated that OTUB2 could promote the malignant proliferation and migration of HCC cells by increasing the stability of PJA1 <i>via</i> deubiquitylation.</p>","PeriodicalId":9687,"journal":{"name":"Cellular and molecular bioengineering","volume":"15 3","pages":"281-292"},"PeriodicalIF":2.8,"publicationDate":"2022-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9124278/pdf/12195_2022_Article_720.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10603455","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Ibrutinib Inhibits BMX-Dependent Endothelial VCAM-1 Expression <i>In Vitro</i> and Pro-Atherosclerotic Endothelial Activation and Platelet Adhesion <i>In Vivo</i>.","authors":"Tia C L Kohs,&nbsp;Sven R Olson,&nbsp;Jiaqing Pang,&nbsp;Kelley R Jordan,&nbsp;Tony J Zheng,&nbsp;Aris Xie,&nbsp;James Hodovan,&nbsp;Matthew Muller,&nbsp;Carrie McArthur,&nbsp;Jennifer Johnson,&nbsp;Bárbara B Sousa,&nbsp;Michael Wallisch,&nbsp;Paul Kievit,&nbsp;Joseph E Aslan,&nbsp;João D Seixas,&nbsp;Gonçalo J L Bernardes,&nbsp;Monica T Hinds,&nbsp;Jonathan R Lindner,&nbsp;Owen J T McCarty,&nbsp;Cristina Puy,&nbsp;Joseph J Shatzel","doi":"10.1007/s12195-022-00723-1","DOIUrl":"https://doi.org/10.1007/s12195-022-00723-1","url":null,"abstract":"<p><strong>Introduction: </strong>Inflammatory activation of the vascular endothelium leads to overexpression of adhesion molecules such as vascular cell adhesion molecule-1 (VCAM-1), contributing to the pro-thrombotic state underpinning atherogenesis. While the role of TEC family kinases (TFKs) in mediating inflammatory cell and platelet activation is well defined, the role of TFKs in vascular endothelial activation remains unclear. We investigated the role of TFKs in endothelial cell activation <i>in vitro</i> and in a nonhuman primate model of diet-induced atherosclerosis <i>in vivo</i>.</p><p><strong>Methods and results: </strong><i>In vitro</i>, we found that ibrutinib blocked activation of the TFK member, BMX, by vascular endothelial growth factors (VEGF)-A in human aortic endothelial cells (HAECs). Blockade of BMX activation with ibrutinib or pharmacologically distinct BMX inhibitors eliminated the ability of VEGF-A to stimulate VCAM-1 expression in HAECs. We validated that treatment with ibrutinib inhibited TFK-mediated platelet activation and aggregation in both human and primate samples as measured using flow cytometry and light transmission aggregometry. We utilized contrast-enhanced ultrasound molecular imaging to measure platelet GPIbα and endothelial VCAM-1 expression in atherosclerosis-prone carotid arteries of obese nonhuman primates. We observed that the TFK inhibitor, ibrutinib, inhibited platelet deposition and endothelial cell activation <i>in vivo</i>.</p><p><strong>Conclusion: </strong>Herein we found that VEGF-A signals through BMX to induce VCAM-1 expression in endothelial cells, and that VCAM-1 expression is sensitive to ibrutinib <i>in vitro</i> and in atherosclerosis-prone carotid arteries <i>in vivo</i>. These findings suggest that TFKs may contribute to the pathogenesis of atherosclerosis and could represent a novel therapeutic target.</p>","PeriodicalId":9687,"journal":{"name":"Cellular and molecular bioengineering","volume":"15 3","pages":"231-243"},"PeriodicalIF":2.8,"publicationDate":"2022-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9124262/pdf/12195_2022_Article_723.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9418435","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Amelotin Promotes Mineralization and Adhesion in Collagen-Based Systems.","authors":"Yuichi Ikeda,&nbsp;James Holcroft,&nbsp;Eri Ikeda,&nbsp;Bernhard Ganss","doi":"10.1007/s12195-022-00722-2","DOIUrl":"https://doi.org/10.1007/s12195-022-00722-2","url":null,"abstract":"<p><strong>Introduction: </strong>Periodontitis is characterized by the destruction of tooth-supporting tissues including the alveolar bone. Barrier membranes are used in dentistry for tissue regenerative therapy. Nevertheless, conventional membranes have issues related to membrane stability and direct induction of bone mineralization. Amelotin (AMTN), an enamel matrix protein, regulates hydroxyapatite crystal nucleation and growth. To apply an AMTN membrane in clinical practice, we investigated the mineralizing and adhesive effects of recombinant human (rh) AMTN <i>in vitro</i> using a collagen-based system.</p><p><strong>Methods: </strong>Collagen hydrogel incorporated with rhAMTN (AMTN gel) and rhAMTN-coated dentin slices were prepared. AMTN gel was then applied on a commercial membrane (AMTN membrane). Samples were incubated for up to 24 h in mineralization buffer, and the structures were observed. The peak adhesive tensile strength between the dentin and AMTN membrane was measured. Using an enzyme-linked immunosorbent assay, the release kinetics of rhAMTN from the membrane were investigated.</p><p><strong>Results: </strong>The AMTN gel resulted in the formation of hydroxyapatite deposits both onto and within the collagen matrix. Furthermore, coating the dentin surface with rhAMTN promoted the precipitation of mineral deposits on the surface. Interestingly, site-specific mineralization was observed in the AMTN membrane. Only 1% of rhAMTN was released from the membrane. Hence, the AMTN membrane adhered to the dentin surface with more than twofold greater tensile strength than that detected for a rhAMTN-free barrier membrane.</p><p><strong>Conclusions: </strong>RhAMTN can accelerate mineralization and adhesion in collagen-based systems. Furthermore, the AMTN membrane could inform the optimal design of calcified tissue regenerative materials.</p><p><strong>Supplementary information: </strong>The online version contains supplementary material available at 10.1007/s12195-022-00722-2.</p>","PeriodicalId":9687,"journal":{"name":"Cellular and molecular bioengineering","volume":"15 3","pages":"245-254"},"PeriodicalIF":2.8,"publicationDate":"2022-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9124263/pdf/12195_2022_Article_722.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9531472","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}