Pub Date : 2022-06-09DOI: 10.1088/1748-605X/ac7763
Anurag Kumar Pandey, R. Gautam, C. Behera
Titanium-based biomedical alloys are susceptible as they are used as a substitute for human bone. In this study, titanium alloy, Ti–5Cu–x%Nb (x = 0, 5, 10, 15) (%wt) was developed by powder metallurgy route. The effect of alloying niobium with Ti–5Cu alloy and its effect on the microstructure, mechanical strength, corrosion resistance, and antibacterial properties have been evaluated. The results show that the sintered alloy has both α-Ti and Ti2Cu phases. With increasing niobium content in the alloy, β-Ti was also detected. Additionally, it was found that the micro-hardness and compressive strength of the studied alloy was better than commercially pure titanium (cpTi), while the Young’s modulus was lower than cpTi. These properties are highly favorable for using this alloy to replicate the human cortical bone. The alloy was also tested for anticorrosive property in Ringer’s solution. The antibacterial activity was also examined for Staphylococcus aureus and Escherichia coli bacteria. The alloy showed promising anticorrosive and antibacterial ability.
{"title":"Microstructure, mechanical strength, chemical resistance, and antibacterial behavior of Ti–5Cu–x%Nb biomedical alloy","authors":"Anurag Kumar Pandey, R. Gautam, C. Behera","doi":"10.1088/1748-605X/ac7763","DOIUrl":"https://doi.org/10.1088/1748-605X/ac7763","url":null,"abstract":"Titanium-based biomedical alloys are susceptible as they are used as a substitute for human bone. In this study, titanium alloy, Ti–5Cu–x%Nb (x = 0, 5, 10, 15) (%wt) was developed by powder metallurgy route. The effect of alloying niobium with Ti–5Cu alloy and its effect on the microstructure, mechanical strength, corrosion resistance, and antibacterial properties have been evaluated. The results show that the sintered alloy has both α-Ti and Ti2Cu phases. With increasing niobium content in the alloy, β-Ti was also detected. Additionally, it was found that the micro-hardness and compressive strength of the studied alloy was better than commercially pure titanium (cpTi), while the Young’s modulus was lower than cpTi. These properties are highly favorable for using this alloy to replicate the human cortical bone. The alloy was also tested for anticorrosive property in Ringer’s solution. The antibacterial activity was also examined for Staphylococcus aureus and Escherichia coli bacteria. The alloy showed promising anticorrosive and antibacterial ability.","PeriodicalId":9016,"journal":{"name":"Biomedical materials","volume":" ","pages":""},"PeriodicalIF":4.0,"publicationDate":"2022-06-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"45299079","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2022-06-08DOI: 10.1088/1748-605X/ac76fb
Alba Herrero-Gómez, Marcelo Azagra, Irene Marco-Rius
Technologies to cryogenically preserve (a.k.a. cryopreserve) living tissue, cell lines and primary cells have matured greatly for both clinicians and researchers since their first demonstration in the 1950s and are widely used in storage and transport applications. Currently, however, there remains an absence of viable cryopreservation and thawing methods for bioengineered, three-dimensional (3D) cell models, including patients’ samples. As a first step towards addressing this gap, we demonstrate a viable protocol for spheroid cryopreservation and survival based on a 3D carboxymethyl cellulose scaffold and precise conditions for freezing and thawing. The protocol is tested using hepatocytes, for which the scaffold provides both the 3D structure for cells to self-arrange into spheroids and to support cells during freezing for optimal post-thaw viability. Cell viability after thawing is improved compared to conventional pellet models where cells settle under gravity to form a pseudo-tissue before freezing. The technique may advance cryobiology and other applications that demand high-integrity transport of pre-assembled 3D models (from cell lines and in future cells from patients) between facilities, for example between medical practice, research and testing facilities.
{"title":"A cryopreservation method for bioengineered 3D cell culture models","authors":"Alba Herrero-Gómez, Marcelo Azagra, Irene Marco-Rius","doi":"10.1088/1748-605X/ac76fb","DOIUrl":"https://doi.org/10.1088/1748-605X/ac76fb","url":null,"abstract":"Technologies to cryogenically preserve (a.k.a. cryopreserve) living tissue, cell lines and primary cells have matured greatly for both clinicians and researchers since their first demonstration in the 1950s and are widely used in storage and transport applications. Currently, however, there remains an absence of viable cryopreservation and thawing methods for bioengineered, three-dimensional (3D) cell models, including patients’ samples. As a first step towards addressing this gap, we demonstrate a viable protocol for spheroid cryopreservation and survival based on a 3D carboxymethyl cellulose scaffold and precise conditions for freezing and thawing. The protocol is tested using hepatocytes, for which the scaffold provides both the 3D structure for cells to self-arrange into spheroids and to support cells during freezing for optimal post-thaw viability. Cell viability after thawing is improved compared to conventional pellet models where cells settle under gravity to form a pseudo-tissue before freezing. The technique may advance cryobiology and other applications that demand high-integrity transport of pre-assembled 3D models (from cell lines and in future cells from patients) between facilities, for example between medical practice, research and testing facilities.","PeriodicalId":9016,"journal":{"name":"Biomedical materials","volume":" ","pages":""},"PeriodicalIF":4.0,"publicationDate":"2022-06-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"44877481","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2022-06-02DOI: 10.1088/1748-605X/ac759f
Anusha Dravid, Amy McCaughey-Chapman, B. Raos, S. O'Carroll, B. Connor, D. Svirskis
Three-dimensional bioprinting continues to advance as an attractive biofabrication technique to employ cell-laden hydrogel scaffolds in the creation of precise, user-defined constructs that can recapitulate the native tissue environment. Development and characterisation of new bioinks to expand the existing library helps to open avenues that can support a diversity of tissue engineering purposes and fulfil requirements in terms of both printability and supporting cell attachment. In this paper, we report the development and characterisation of agarose–gelatin (AG–Gel) hydrogel blends as a bioink for extrusion-based bioprinting. Four different AG–Gel hydrogel blend formulations with varying gelatin concentration were systematically characterised to evaluate suitability as a potential bioink for extrusion-based bioprinting. Additionally, autoclave and filter sterilisation methods were compared to evaluate their effect on bioink properties. Finally, the ability of the AG–Gel bioink to support cell viability and culture after printing was evaluated using SH-SY5Y cells encapsulated in bioprinted droplets of the AG–Gel. All bioink formulations demonstrate rheological, mechanical and swelling properties suitable for bioprinting and cell encapsulation. Autoclave sterilisation significantly affected the rheological properties of the AG–Gel bioinks compared to filter sterilisation. SH-SY5Y cells printed and differentiated into neuronal-like cells using the developed AG–Gel bioinks demonstrated high viability (>90%) after 23 d in culture. This study demonstrates the properties of AG–Gel as a printable and biocompatible material applicable for use as a bioink.
{"title":"Development of agarose–gelatin bioinks for extrusion-based bioprinting and cell encapsulation","authors":"Anusha Dravid, Amy McCaughey-Chapman, B. Raos, S. O'Carroll, B. Connor, D. Svirskis","doi":"10.1088/1748-605X/ac759f","DOIUrl":"https://doi.org/10.1088/1748-605X/ac759f","url":null,"abstract":"Three-dimensional bioprinting continues to advance as an attractive biofabrication technique to employ cell-laden hydrogel scaffolds in the creation of precise, user-defined constructs that can recapitulate the native tissue environment. Development and characterisation of new bioinks to expand the existing library helps to open avenues that can support a diversity of tissue engineering purposes and fulfil requirements in terms of both printability and supporting cell attachment. In this paper, we report the development and characterisation of agarose–gelatin (AG–Gel) hydrogel blends as a bioink for extrusion-based bioprinting. Four different AG–Gel hydrogel blend formulations with varying gelatin concentration were systematically characterised to evaluate suitability as a potential bioink for extrusion-based bioprinting. Additionally, autoclave and filter sterilisation methods were compared to evaluate their effect on bioink properties. Finally, the ability of the AG–Gel bioink to support cell viability and culture after printing was evaluated using SH-SY5Y cells encapsulated in bioprinted droplets of the AG–Gel. All bioink formulations demonstrate rheological, mechanical and swelling properties suitable for bioprinting and cell encapsulation. Autoclave sterilisation significantly affected the rheological properties of the AG–Gel bioinks compared to filter sterilisation. SH-SY5Y cells printed and differentiated into neuronal-like cells using the developed AG–Gel bioinks demonstrated high viability (>90%) after 23 d in culture. This study demonstrates the properties of AG–Gel as a printable and biocompatible material applicable for use as a bioink.","PeriodicalId":9016,"journal":{"name":"Biomedical materials","volume":" ","pages":""},"PeriodicalIF":4.0,"publicationDate":"2022-06-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"44759298","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2022-05-26DOI: 10.1088/1748-605X/ac73cd
E. Saygili, U. Devamoglu, B. Goker-Bagca, O. Goksel, C. Biray-Avci, T. Goksel, O. Yesil‐Celiktas
Associated with a high mortality rate, pulmonary fibrosis (PF) is the end stage of several interstitial lung diseases. Although many factors are linked to PF progression, initiation of the fibrotic process remains to be studied. Current research focused on generating new strategies to gain a better understanding of the underlying disease mechanism as the animal models remain insufficient to reflect human physiology. Herein, to account complex cellular interactions within the fibrotic tissue, a multicellular spheroid model where human bronchial epithelial cells incorporated with human lung fibroblasts was generated and treated with bleomycin (BLM) to emulate drug-induced PF. Recapitulating the epithelial-interstitial microenvironment, the findings successfully reflected the PF disease, where excessive alpha smooth muscle actin and collagen type I secretion were noted along with the morphological changes in response to BLM. Moreover, increased levels of fibrotic linked COL13A1, MMP2, WNT3 and decreased expression level of CDH1 provide evidence for the model reliability on fibrosis modelling. Subsequent administration of the Food and Drug Administration approved nintedanib and pirfenidone anti-fibrotic drugs proved the drug-responsiveness of the model.
{"title":"A drug-responsive multicellular human spheroid model to recapitulate drug-induced pulmonary fibrosis","authors":"E. Saygili, U. Devamoglu, B. Goker-Bagca, O. Goksel, C. Biray-Avci, T. Goksel, O. Yesil‐Celiktas","doi":"10.1088/1748-605X/ac73cd","DOIUrl":"https://doi.org/10.1088/1748-605X/ac73cd","url":null,"abstract":"Associated with a high mortality rate, pulmonary fibrosis (PF) is the end stage of several interstitial lung diseases. Although many factors are linked to PF progression, initiation of the fibrotic process remains to be studied. Current research focused on generating new strategies to gain a better understanding of the underlying disease mechanism as the animal models remain insufficient to reflect human physiology. Herein, to account complex cellular interactions within the fibrotic tissue, a multicellular spheroid model where human bronchial epithelial cells incorporated with human lung fibroblasts was generated and treated with bleomycin (BLM) to emulate drug-induced PF. Recapitulating the epithelial-interstitial microenvironment, the findings successfully reflected the PF disease, where excessive alpha smooth muscle actin and collagen type I secretion were noted along with the morphological changes in response to BLM. Moreover, increased levels of fibrotic linked COL13A1, MMP2, WNT3 and decreased expression level of CDH1 provide evidence for the model reliability on fibrosis modelling. Subsequent administration of the Food and Drug Administration approved nintedanib and pirfenidone anti-fibrotic drugs proved the drug-responsiveness of the model.","PeriodicalId":9016,"journal":{"name":"Biomedical materials","volume":" ","pages":""},"PeriodicalIF":4.0,"publicationDate":"2022-05-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"43100037","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2022-05-24DOI: 10.1088/1748-605X/ac7308
R. Noroozi, M. Shamekhi, R. Mahmoudi, A. Zolfagharian, Fatemeh Asgari, A. Mousavizadeh, M. Bodaghi, Amin Hadi, N. Haghighipour
The aim of this paper was to design and fabricate a novel composite scaffold based on the combination of 3D-printed polylactic acid-based triply periodic minimal surfaces (TPMSs) and cell-laden alginate hydrogel. This novel scaffold improves the low mechanical properties of alginate hydrogel and can also provide a scaffold with a suitable pore size, which can be used in bone regeneration applications. In this regard, an implicit function was used to generate some gyroid TPMS scaffolds. Then the fused deposition modeling process was employed to print the scaffolds. Moreover, the micro computed tomography technique was employed to assess the microstructure of 3D-printed TPMS scaffolds and obtain the real geometries of printed scaffolds. The mechanical properties of composite scaffolds were investigated under compression tests experimentally. It was shown that different mechanical behaviors could be obtained for different implicit function parameters. In this research, to assess the mechanical behavior of printed scaffolds in terms of the strain–stress curves on, two approaches were presented: equivalent volume and finite element-based volume. Results of strain–stress curves showed that the finite-element based approach predicts a higher level of stress. Moreover, the biological response of composite scaffolds in terms of cell viability, cell proliferation, and cell attachment was investigated. In this vein, a dynamic cell culture system was designed and fabricated, which improves mass transport through the composite scaffolds and applies mechanical loading to the cells, which helps cell proliferation. Moreover, the results of the novel composite scaffolds were compared to those without alginate, and it was shown that the composite scaffold could create more viability and cell proliferation in both dynamic and static cultures. Also, it was shown that scaffolds in dynamic cell culture have a better biological response than in static culture. In addition, scanning electron microscopy was employed to study the cell adhesion on the composite scaffolds, which showed excellent attachment between the scaffolds and cells.
{"title":"In vitro static and dynamic cell culture study of novel bone scaffolds based on 3D-printed PLA and cell-laden alginate hydrogel","authors":"R. Noroozi, M. Shamekhi, R. Mahmoudi, A. Zolfagharian, Fatemeh Asgari, A. Mousavizadeh, M. Bodaghi, Amin Hadi, N. Haghighipour","doi":"10.1088/1748-605X/ac7308","DOIUrl":"https://doi.org/10.1088/1748-605X/ac7308","url":null,"abstract":"The aim of this paper was to design and fabricate a novel composite scaffold based on the combination of 3D-printed polylactic acid-based triply periodic minimal surfaces (TPMSs) and cell-laden alginate hydrogel. This novel scaffold improves the low mechanical properties of alginate hydrogel and can also provide a scaffold with a suitable pore size, which can be used in bone regeneration applications. In this regard, an implicit function was used to generate some gyroid TPMS scaffolds. Then the fused deposition modeling process was employed to print the scaffolds. Moreover, the micro computed tomography technique was employed to assess the microstructure of 3D-printed TPMS scaffolds and obtain the real geometries of printed scaffolds. The mechanical properties of composite scaffolds were investigated under compression tests experimentally. It was shown that different mechanical behaviors could be obtained for different implicit function parameters. In this research, to assess the mechanical behavior of printed scaffolds in terms of the strain–stress curves on, two approaches were presented: equivalent volume and finite element-based volume. Results of strain–stress curves showed that the finite-element based approach predicts a higher level of stress. Moreover, the biological response of composite scaffolds in terms of cell viability, cell proliferation, and cell attachment was investigated. In this vein, a dynamic cell culture system was designed and fabricated, which improves mass transport through the composite scaffolds and applies mechanical loading to the cells, which helps cell proliferation. Moreover, the results of the novel composite scaffolds were compared to those without alginate, and it was shown that the composite scaffold could create more viability and cell proliferation in both dynamic and static cultures. Also, it was shown that scaffolds in dynamic cell culture have a better biological response than in static culture. In addition, scanning electron microscopy was employed to study the cell adhesion on the composite scaffolds, which showed excellent attachment between the scaffolds and cells.","PeriodicalId":9016,"journal":{"name":"Biomedical materials","volume":" ","pages":""},"PeriodicalIF":4.0,"publicationDate":"2022-05-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"46254470","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Artificial tissue replacement is a promising strategy for better healing outcomes for tendon and ligament injuries, due to the very limited self-regeneration capacity of these tissues in mammals, including humans. Because clinically available synthetic and biological scaffolds for tendon repair have performed more poorly than autografts, both biological and mechanical compatibility need to be improved. Here we propose a rapid fabrication method for tendon-like structure from collagen hydrogel, simultaneously achieving collagen fibre alignment and intermolecular cross-linking. Collagen gel, 24 h after polymerization, was subjected to mechanical loading in the presence of the chemical cross-linker, genipin, for 24 or 48 h. Mechanical loading during gel incubation oriented collagen fibres in the loading direction and made chemical cross-linking highly effective in a loading magnitude-dependent manner. Gel incubated with 4 g loading in the presence of genipin for 48 h possessed tensile strength of 4 MPa and tangent modulus of 60 MPa, respectively, which could fulfill the minimum biomechanical requirement for artificial tendon. Although mechanical properties of gels fabricated using the present method can be improved by using a larger amount of collagen in the starting material and through optimisation of mechanical loading and cross-linking, the method is a simple and effective for producing highly aligned collagen fibrils with excellent mechanical properties.
{"title":"Rapid fabrication of tendon-like collagen gel via simultaneous fibre alignment and intermolecular cross-linking under mechanical loading","authors":"Eijiro Maeda, Ryota Kawamura, Takashi Suzuki, Takeo Matsumoto","doi":"10.1088/1748-605X/ac7305","DOIUrl":"https://doi.org/10.1088/1748-605X/ac7305","url":null,"abstract":"Artificial tissue replacement is a promising strategy for better healing outcomes for tendon and ligament injuries, due to the very limited self-regeneration capacity of these tissues in mammals, including humans. Because clinically available synthetic and biological scaffolds for tendon repair have performed more poorly than autografts, both biological and mechanical compatibility need to be improved. Here we propose a rapid fabrication method for tendon-like structure from collagen hydrogel, simultaneously achieving collagen fibre alignment and intermolecular cross-linking. Collagen gel, 24 h after polymerization, was subjected to mechanical loading in the presence of the chemical cross-linker, genipin, for 24 or 48 h. Mechanical loading during gel incubation oriented collagen fibres in the loading direction and made chemical cross-linking highly effective in a loading magnitude-dependent manner. Gel incubated with 4 g loading in the presence of genipin for 48 h possessed tensile strength of 4 MPa and tangent modulus of 60 MPa, respectively, which could fulfill the minimum biomechanical requirement for artificial tendon. Although mechanical properties of gels fabricated using the present method can be improved by using a larger amount of collagen in the starting material and through optimisation of mechanical loading and cross-linking, the method is a simple and effective for producing highly aligned collagen fibrils with excellent mechanical properties.","PeriodicalId":9016,"journal":{"name":"Biomedical materials","volume":" ","pages":""},"PeriodicalIF":4.0,"publicationDate":"2022-05-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"47674709","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2022-05-24DOI: 10.1088/1748-605X/ac7307
Mohammad Asadi Tokmedash, Elham Seyyedi Zadeh, Elham Nezami Balouchi, Z. Salehi, M. Ardestani
Multifunctional nanocarriers, specifically for tumor targeting and traceable features, have been increasingly considered in cancer therapies. Herein, a novel targeting agent (TA), tryptophan (TRP), was proposed for the synthesis of functionalized (3-aminopropyl) triethoxysilane-iron oxide nanoparticles using two methods, creating a smart drug delivery system (DDS). In one method, two-step, glutaraldehyde (GA) as a linker, bonded TRP and amino-functionalized magnetite, and in the second method, one step, TRP binding was carried out by (3-dimethyl aminopropyl)-N’-ethyl carbodiimide hydrochloride (EDC)/N-hydroxysuccinimide ester. The synthesis yield of the second method was 7% higher than the first method. After synthesizing DDS, 5-fluorouracil (5-FU) was loaded on nanocarriers and was observed TRP functionalized nanoparticles by GA have better loading efficiency, which was 50% greater than the product from the one-step method. A pH-sensitive release profile was also studied for 5-FU/DDS with the release of almost 75% and 50% at pH 5.5 and 7.4, respectively. To analyze the biological aspects of nanocarriers, human breast cancer, MCF-7, and embryonic kidney, HEK293, cell lines were used for cellular uptake and 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assays. In vitro studies confirmed that TRP can act as a TA as its cellular uptake through cancerous cells was 40% greater than normal cells, and the MTT assay confirmed that using DDS can increase and decrease the cell viability of normal cells and cancerous cells, respectively, compared to free drug. Therefore, it was concluded that advanced nano-assembly is a great candidate for breast cancer cell-targeted delivery.
多功能纳米载体，特别是具有肿瘤靶向性和可追溯性的特点，在癌症治疗中越来越受到重视。本文提出了一种新的靶向剂色氨酸(TA)，用于两种方法合成功能化(3-氨基丙基)三乙氧基硅烷-氧化铁纳米颗粒，创建了一个智能给药系统(DDS)。方法一是以戊二醛(GA)为连接剂，两步结合TRP与氨基功能化磁铁矿;方法二是以(3-二甲基氨基丙基)-N′-乙基碳二亚胺盐酸盐(EDC)/ n -羟基琥珀酰亚胺酯一步结合TRP。第二种方法的合成收率比第一种方法高7%。合成DDS后，将5-氟尿嘧啶(5-FU)负载在纳米载体上，通过GA观察到TRP功能化纳米颗粒的负载效率更高，比一步法的产物高50%。研究了5-FU/DDS的pH敏感性释放曲线，在pH为5.5和7.4时，5-FU/DDS的释放率分别接近75%和50%。为了分析纳米载体、人乳腺癌、MCF-7和胚胎肾、HEK293的生物学特性，采用细胞系进行细胞摄取和3-(4,5 -二甲基噻唑-2-基)- 2,5 -二苯基溴化四唑(MTT)测定。体外研究证实，TRP可以作为TA，因为其通过癌细胞的细胞摄取比正常细胞高40%，MTT试验证实，与游离药物相比，使用DDS可以分别增加和降低正常细胞和癌细胞的细胞活力。因此，我们得出结论，先进的纳米组装是乳腺癌细胞靶向递送的一个很好的候选。
{"title":"Synthesis of smart carriers based on tryptophan-functionalized magnetic nanoparticles and its application in 5-fluorouracil delivery","authors":"Mohammad Asadi Tokmedash, Elham Seyyedi Zadeh, Elham Nezami Balouchi, Z. Salehi, M. Ardestani","doi":"10.1088/1748-605X/ac7307","DOIUrl":"https://doi.org/10.1088/1748-605X/ac7307","url":null,"abstract":"Multifunctional nanocarriers, specifically for tumor targeting and traceable features, have been increasingly considered in cancer therapies. Herein, a novel targeting agent (TA), tryptophan (TRP), was proposed for the synthesis of functionalized (3-aminopropyl) triethoxysilane-iron oxide nanoparticles using two methods, creating a smart drug delivery system (DDS). In one method, two-step, glutaraldehyde (GA) as a linker, bonded TRP and amino-functionalized magnetite, and in the second method, one step, TRP binding was carried out by (3-dimethyl aminopropyl)-N’-ethyl carbodiimide hydrochloride (EDC)/N-hydroxysuccinimide ester. The synthesis yield of the second method was 7% higher than the first method. After synthesizing DDS, 5-fluorouracil (5-FU) was loaded on nanocarriers and was observed TRP functionalized nanoparticles by GA have better loading efficiency, which was 50% greater than the product from the one-step method. A pH-sensitive release profile was also studied for 5-FU/DDS with the release of almost 75% and 50% at pH 5.5 and 7.4, respectively. To analyze the biological aspects of nanocarriers, human breast cancer, MCF-7, and embryonic kidney, HEK293, cell lines were used for cellular uptake and 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assays. In vitro studies confirmed that TRP can act as a TA as its cellular uptake through cancerous cells was 40% greater than normal cells, and the MTT assay confirmed that using DDS can increase and decrease the cell viability of normal cells and cancerous cells, respectively, compared to free drug. Therefore, it was concluded that advanced nano-assembly is a great candidate for breast cancer cell-targeted delivery.","PeriodicalId":9016,"journal":{"name":"Biomedical materials","volume":" ","pages":""},"PeriodicalIF":4.0,"publicationDate":"2022-05-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"43040997","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2022-05-24DOI: 10.1088/1748-605X/ac7304
Junqiang Xue, Wei Yang, Xinping Wang, Peiyan Wang, Xinyue Meng, Tengbo Yu, C. Fan
The macro-porous hydrogel scaffolds can not only enhance the proliferation of laden chondrocytes but also favor the deposition of hyaline cartilaginous extracellular matrix, however, the underlying molecular mechanism is still unclear. Herein, the global gene expression of human cartilage chondrocytes (HCCs) encapsulated in traditional hydrogel (Gel) constructs and micro-cavitary gel (MCG) constructs are investigated by using high-throughput RNA sequencing (RNA-seq). The differentially expressed genes (DEGs) between the HCCs cultured in Gel and MCG constructs have been identified via bioinformatics analysis. Significantly, the DEGs that promote cell proliferation (e.g. POSTN, MKI67, KIF20A) or neo-cartilage formation (e.g. COL2, ASPN, COMP, FMOD, FN1), are more highly expressed in MCG constructs than in Gel constructs, while the expressions of the DEGs associated with chondrocyte hypertrophy (e.g. EGR1, IBSP) are upregulated in Gel constructs. The expression of representative DEGs is verified at both mRNA and protein levels. Besides, cellular viability and morphology as well as the enriched signaling pathway of DEGs are studied in detail. These results of this work may provide data for functional tissue engineering of cartilage.
{"title":"A transcriptome sequencing study on the effect of macro-pores in hydrogel scaffolds on global gene expression of laden human cartilage chondrocytes","authors":"Junqiang Xue, Wei Yang, Xinping Wang, Peiyan Wang, Xinyue Meng, Tengbo Yu, C. Fan","doi":"10.1088/1748-605X/ac7304","DOIUrl":"https://doi.org/10.1088/1748-605X/ac7304","url":null,"abstract":"The macro-porous hydrogel scaffolds can not only enhance the proliferation of laden chondrocytes but also favor the deposition of hyaline cartilaginous extracellular matrix, however, the underlying molecular mechanism is still unclear. Herein, the global gene expression of human cartilage chondrocytes (HCCs) encapsulated in traditional hydrogel (Gel) constructs and micro-cavitary gel (MCG) constructs are investigated by using high-throughput RNA sequencing (RNA-seq). The differentially expressed genes (DEGs) between the HCCs cultured in Gel and MCG constructs have been identified via bioinformatics analysis. Significantly, the DEGs that promote cell proliferation (e.g. POSTN, MKI67, KIF20A) or neo-cartilage formation (e.g. COL2, ASPN, COMP, FMOD, FN1), are more highly expressed in MCG constructs than in Gel constructs, while the expressions of the DEGs associated with chondrocyte hypertrophy (e.g. EGR1, IBSP) are upregulated in Gel constructs. The expression of representative DEGs is verified at both mRNA and protein levels. Besides, cellular viability and morphology as well as the enriched signaling pathway of DEGs are studied in detail. These results of this work may provide data for functional tissue engineering of cartilage.","PeriodicalId":9016,"journal":{"name":"Biomedical materials","volume":" ","pages":""},"PeriodicalIF":4.0,"publicationDate":"2022-05-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"42685756","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2022-05-17DOI: 10.1088/1748-605X/ac6b06
P. Brumm, Anna Fritschen, Lara Doß, E. Dörsam, A. Blaeser
Mammalian tissue comprises a plethora of hierarchically organized channel networks that serve as routes for the exchange of liquids, nutrients, bio-chemical cues or electrical signals, such as blood vessels, nerve fibers, or lymphatic conduits. Despite differences in function and size, the networks exhibit a similar, highly branched morphology with dendritic extensions. Mimicking such hierarchical networks represents a milestone in the biofabrication of tissues and organs. Work to date has focused primarily on the replication of the vasculature. Despite initial progress, reproducing such structures across scales and increasing biofabrication efficiency remain a challenge. In this work, we present a new biofabrication method that takes advantage of the viscous fingering phenomenon. Using flexographic printing, highly branched, inter-connective channel structures with stochastic, biomimetic distribution and dendritic extensions can be fabricated with unprecedented efficiency. Using gelatin (5%–35%) as resolvable sacrificial material, the feasability of the proposed method is demonstrated on the example of a vascular network. By selectively adjusting the printing velocity (0.2–1.5 m s−1), the anilox roller dip volume (4.5–24 ml m−2) as well as the shear viscosity of the printing material used (10–900 mPas), the width of the structures produced (30–400 µm) as well as their distance (200–600 µm) can be specifically determined. In addition to the flexible morphology, the high scalability (2500–25 000 mm2) and speed (1.5 m s−1) of the biofabrication process represents an important unique selling point. Printing parameters and hydrogel formulations are investigated and tuned towards a process window for controlled fabrication of channels that mimic the morphology of small blood vessels and capillaries. Subsequently, the resolvable structures were casted in a hydrogel matrix enabling bulk environments with integrated channels. The perfusability of the branched, inter-connective structures was successfully demonstrated. The fabricated networks hold great potential to enable nutrient supply in thick vascularized tissues or perfused organ-on-a-chip systems. In the future, the concept can be further optimized and expanded towards large-scale and cost-efficient biofabrication of vascular, lymphatic or neural networks for tissue engineering and regenerative medicine.
哺乳动物组织包括大量分层组织的通道网络，这些通道网络充当液体、营养物质、生化线索或电信号（如血管、神经纤维或淋巴管）交换的途径。尽管在功能和大小上存在差异，但网络表现出相似的、高度分支的形态，并具有树枝状延伸。模仿这样的层级网络代表着组织和器官生物制造的一个里程碑。迄今为止的工作主要集中在血管系统的复制上。尽管取得了初步进展，但跨规模复制这种结构和提高生物制造效率仍然是一个挑战。在这项工作中，我们提出了一种新的生物制造方法，利用粘性指法现象。使用柔性版印刷，可以以前所未有的效率制造具有随机、仿生分布和树枝状延伸的高度分支、互连通道结构。使用明胶（5%-35%）作为可溶解的牺牲材料，以血管网络为例证明了该方法的可行性。通过选择性地调整印刷速度（0.2–1.5 m s−1）、网纹辊浸渍体积（4.5–24 ml m−2）以及所用印刷材料的剪切粘度（10–900 mPas）、所产生结构的宽度（30–400µm）及其距离（200–600µm），可以具体确定。除了灵活的形态外，高可扩展性（2500–25 000 mm2）和生物制造过程的速度（1.5 m s−1）代表了一个重要的独特卖点。研究了印刷参数和水凝胶配方，并将其调整为模拟小血管和毛细血管形态的通道的受控制造的工艺窗口。随后，将可分解结构浇铸在水凝胶基质中，从而实现具有集成通道的本体环境。分支结缔结构的可灌注性得到了成功的证明。所制造的网络具有巨大的潜力，可以在厚血管组织或灌注芯片上的器官系统中提供营养。未来，该概念可以进一步优化和扩展，用于组织工程和再生医学的血管、淋巴或神经网络的大规模和成本效益高的生物制造。
{"title":"Fabrication of biomimetic networks using viscous fingering in flexographic printing","authors":"P. Brumm, Anna Fritschen, Lara Doß, E. Dörsam, A. Blaeser","doi":"10.1088/1748-605X/ac6b06","DOIUrl":"https://doi.org/10.1088/1748-605X/ac6b06","url":null,"abstract":"Mammalian tissue comprises a plethora of hierarchically organized channel networks that serve as routes for the exchange of liquids, nutrients, bio-chemical cues or electrical signals, such as blood vessels, nerve fibers, or lymphatic conduits. Despite differences in function and size, the networks exhibit a similar, highly branched morphology with dendritic extensions. Mimicking such hierarchical networks represents a milestone in the biofabrication of tissues and organs. Work to date has focused primarily on the replication of the vasculature. Despite initial progress, reproducing such structures across scales and increasing biofabrication efficiency remain a challenge. In this work, we present a new biofabrication method that takes advantage of the viscous fingering phenomenon. Using flexographic printing, highly branched, inter-connective channel structures with stochastic, biomimetic distribution and dendritic extensions can be fabricated with unprecedented efficiency. Using gelatin (5%–35%) as resolvable sacrificial material, the feasability of the proposed method is demonstrated on the example of a vascular network. By selectively adjusting the printing velocity (0.2–1.5 m s−1), the anilox roller dip volume (4.5–24 ml m−2) as well as the shear viscosity of the printing material used (10–900 mPas), the width of the structures produced (30–400 µm) as well as their distance (200–600 µm) can be specifically determined. In addition to the flexible morphology, the high scalability (2500–25 000 mm2) and speed (1.5 m s−1) of the biofabrication process represents an important unique selling point. Printing parameters and hydrogel formulations are investigated and tuned towards a process window for controlled fabrication of channels that mimic the morphology of small blood vessels and capillaries. Subsequently, the resolvable structures were casted in a hydrogel matrix enabling bulk environments with integrated channels. The perfusability of the branched, inter-connective structures was successfully demonstrated. The fabricated networks hold great potential to enable nutrient supply in thick vascularized tissues or perfused organ-on-a-chip systems. In the future, the concept can be further optimized and expanded towards large-scale and cost-efficient biofabrication of vascular, lymphatic or neural networks for tissue engineering and regenerative medicine.","PeriodicalId":9016,"journal":{"name":"Biomedical materials","volume":"294 3","pages":""},"PeriodicalIF":4.0,"publicationDate":"2022-05-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"41330839","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2022-05-12DOI: 10.1088/1748-605X/ac6f38
Zichang Zhang, Fan Zhou, Jianwei Zheng, J. Mu, P. Bo, Bin You
To provide better treatment of myocardial infarction, DiI-labeled bone marrow mesenchymal stem cells (BMSCs) were contact co-cultured with cardiomyocytes (CMs) on polycaprolactone (PCL) film to prepare myocardial patches. BMSCs from Sprague Dawley rats were isolated, cultured, and characterized for expression of surface markers by flow cytometry. CMs were isolated from suckling rats. After BMSCs were cultured for three generations, they were labeled with DiI dye. DiI-labeled BMSCs were co-cultured with CMs on PCL film in the experimental group, while CMs were replaced with the same amount of unlabeled BMSCs in the control group. After 24 h, cell growth was observed by light microscopy and cells were fixed for scanning electron microscopy (SEM). After 7 d of co-culture, cells were stained for immunofluorescence detection of myocardial markers cardiac troponin T (cTnT) and α-actin. Differentiation of BMSCs on PCL was observed by fluorescence microscopy. The efficiency of BMSC differentiation into CMs was analyzed by flow cytometry on the first and seventh days of co-culture. CMs were stained with calcein alone and contact co-cultured with DiI-labeled BMSCs on PCL film to observe intercellular dye transfer. Finally, cells were stained for immunofluorescence detection of connexin 43 (Cx43) expression and to observe the relationship between gap junctions and contact co-culture. BMSCs were identified by flow cytometry as strongly positive for CD90 and CD44H, and negative for CD11b/c and CD45. After co-culture for 24 h, cells were observed to have attached to PCL by light microscopy. Upon appropriate excitation, DiI-labeled BMSCs exhibited red fluorescence, while unlabeled CMs did not. SEM revealed a large number of cells on the PCL membrane and their cell state appeared normal. On the seventh day, some DiI-labeled BMSCs expressed cTnT and α-actin. Flow cytometry showed that the rate of stem cell differentiation in the experimental group was significantly higher than the control group on the seventh day (20.12% > 3.49%, P < 0.05). From the second day of co-culture, immunofluorescence staining for Cx43 revealed green fluorescent puncta in some BMSCs; from the third day of co-culture, a portion of BMSCs exhibited green fluorescence in dye transfer tests. Contact co-culture of DiI-labeled BMSCs and CMs on PCL film generated primary myocardial patches. The mechanism by which contact co-culture promoted differentiation of the myocardial patch may be related to gap junctions and gap junction-mediated intercellular signaling pathways.
{"title":"Preparation of myocardial patches from DiI-labeled rat bone marrow mesenchymal stem cells and neonatal rat cardiomyocytes contact co-cultured on polycaprolactone film","authors":"Zichang Zhang, Fan Zhou, Jianwei Zheng, J. Mu, P. Bo, Bin You","doi":"10.1088/1748-605X/ac6f38","DOIUrl":"https://doi.org/10.1088/1748-605X/ac6f38","url":null,"abstract":"To provide better treatment of myocardial infarction, DiI-labeled bone marrow mesenchymal stem cells (BMSCs) were contact co-cultured with cardiomyocytes (CMs) on polycaprolactone (PCL) film to prepare myocardial patches. BMSCs from Sprague Dawley rats were isolated, cultured, and characterized for expression of surface markers by flow cytometry. CMs were isolated from suckling rats. After BMSCs were cultured for three generations, they were labeled with DiI dye. DiI-labeled BMSCs were co-cultured with CMs on PCL film in the experimental group, while CMs were replaced with the same amount of unlabeled BMSCs in the control group. After 24 h, cell growth was observed by light microscopy and cells were fixed for scanning electron microscopy (SEM). After 7 d of co-culture, cells were stained for immunofluorescence detection of myocardial markers cardiac troponin T (cTnT) and α-actin. Differentiation of BMSCs on PCL was observed by fluorescence microscopy. The efficiency of BMSC differentiation into CMs was analyzed by flow cytometry on the first and seventh days of co-culture. CMs were stained with calcein alone and contact co-cultured with DiI-labeled BMSCs on PCL film to observe intercellular dye transfer. Finally, cells were stained for immunofluorescence detection of connexin 43 (Cx43) expression and to observe the relationship between gap junctions and contact co-culture. BMSCs were identified by flow cytometry as strongly positive for CD90 and CD44H, and negative for CD11b/c and CD45. After co-culture for 24 h, cells were observed to have attached to PCL by light microscopy. Upon appropriate excitation, DiI-labeled BMSCs exhibited red fluorescence, while unlabeled CMs did not. SEM revealed a large number of cells on the PCL membrane and their cell state appeared normal. On the seventh day, some DiI-labeled BMSCs expressed cTnT and α-actin. Flow cytometry showed that the rate of stem cell differentiation in the experimental group was significantly higher than the control group on the seventh day (20.12% > 3.49%, P < 0.05). From the second day of co-culture, immunofluorescence staining for Cx43 revealed green fluorescent puncta in some BMSCs; from the third day of co-culture, a portion of BMSCs exhibited green fluorescence in dye transfer tests. Contact co-culture of DiI-labeled BMSCs and CMs on PCL film generated primary myocardial patches. The mechanism by which contact co-culture promoted differentiation of the myocardial patch may be related to gap junctions and gap junction-mediated intercellular signaling pathways.","PeriodicalId":9016,"journal":{"name":"Biomedical materials","volume":" ","pages":""},"PeriodicalIF":4.0,"publicationDate":"2022-05-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"45942437","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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