Chenglong Wang, Jinlei Dong, Fanxiao Liu, Nan Liu, Lianxin Li
{"title":"3D-printed PCL@BG scaffold integrated with SDF-1α-loaded hydrogel for enhancing local treatment of bone defects","authors":"Chenglong Wang, Jinlei Dong, Fanxiao Liu, Nan Liu, Lianxin Li","doi":"10.1186/s13036-023-00401-4","DOIUrl":"https://doi.org/10.1186/s13036-023-00401-4","url":null,"abstract":"The long-term nonunion of bone defects is always a difficult problem in orthopaedics treatment. Artificial bone implants made of polymeric materials are expected to solve this problem due to their suitable degradation rate and good biocompatibility. However, the lack of mechanical strength, low osteogenic induction ability and poor hydrophilicity of these synthetic polymeric materials limit their large-scale clinical application. In this study, we used bioactive glass (BG) (20%, W/W) and polycaprolactone (PCL, 80%, W/W) as raw materials to prepare a bone repair scaffold (PCL@BG20) using fused deposition modelling (FDM) three-dimensional (3D) printing technology. Subsequently, stromal cell-derived factor-1α (SDF-1α) chemokines were loaded into the PCL@BG20 scaffold pores with gelatine methacryloyl (GelMA) hydrogel. The experimental results showed that the prepared scaffold had a porous biomimetic structure mimicking that of cancellous bone, and the compressive strength (44.89 ± 3.45 MPa) of the scaffold was similar to that of cancellous bone. Transwell experiments showed that scaffolds loaded with SDF-1α could promote the recruitment of bone marrow stromal cells (BMSCs). In vivo data showed that treatment with scaffolds containing SDF-1α and BG (PCL@BG-GelMA/SDF-1α) had the best effect on bone defect repair compared to the other groups, with a large amount of new bone and mature collagen forming at the bone defect site. No significant organ toxicity or inflammatory reactions were observed in any of the experimental groups. The results show that this kind of scaffold containing BG and SDF-1α serves the dual functions of recruiting stem cell migration in vivo and promoting bone repair in situ. We envision that this scaffold may become a new strategy for the clinical treatment of bone defects.\u0000","PeriodicalId":15053,"journal":{"name":"Journal of Biological Engineering","volume":"15 1","pages":""},"PeriodicalIF":5.6,"publicationDate":"2024-01-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139084451","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Guanyu Su, Yushi Zhang, Chunyu Jin, Qiyue Zhang, Jiarui Lu, Zengqian Liu, Qiang Wang, Xue Zhang, Jia Ma
{"title":"3D printed zirconia used as dental materials: a critical review","authors":"Guanyu Su, Yushi Zhang, Chunyu Jin, Qiyue Zhang, Jiarui Lu, Zengqian Liu, Qiang Wang, Xue Zhang, Jia Ma","doi":"10.1186/s13036-023-00396-y","DOIUrl":"https://doi.org/10.1186/s13036-023-00396-y","url":null,"abstract":"In view of its high mechanical performance, outstanding aesthetic qualities, and biological stability, zirconia has been widely used in the fields of dentistry. Due to its potential to produce suitable advanced configurations and structures for a number of medical applications, especially personalized created devices, ceramic additive manufacturing (AM) has been attracting a great deal of attention in recent years. AM zirconia hews out infinite possibilities that are otherwise barely possible with traditional processes thanks to its freedom and efficiency. In the review, AM zirconia’s physical and adhesive characteristics, accuracy, biocompatibility, as well as their clinical applications have been reviewed. Here, we highlight the accuracy and biocompatibility of 3D printed zirconia. Also, current obstacles and a forecast of AM zirconia for its development and improvement have been covered. In summary, this review offers a description of the basic characteristics of AM zirconia materials intended for oral medicine. Furthermore, it provides a generally novel and fundamental basis for the utilization of 3D printed zirconia in dentistry.","PeriodicalId":15053,"journal":{"name":"Journal of Biological Engineering","volume":"31 1","pages":""},"PeriodicalIF":5.6,"publicationDate":"2023-12-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"138823604","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Michael Killinger, Adéla Kratochvilová, Eva Ingeborg Reihs, Eva Matalová, Karel Klepárník, Mario Rothbauer
{"title":"Microfluidic device for enhancement and analysis of osteoblast differentiation in three-dimensional cell cultures","authors":"Michael Killinger, Adéla Kratochvilová, Eva Ingeborg Reihs, Eva Matalová, Karel Klepárník, Mario Rothbauer","doi":"10.1186/s13036-023-00395-z","DOIUrl":"https://doi.org/10.1186/s13036-023-00395-z","url":null,"abstract":"Three-dimensional (3D) cell cultures are to date the gold standard in biomedical research fields due to their enhanced biological functions compared to conventional two-dimensional (2D) cultures. 3D cell spheroids, as well as organoids, are better suited to replicate tissue functions, which enables their use both as in vitro models for basic research and toxicology, as well as building blocks used in tissue/organ biofabrication approaches. Culturing 3D spheroids from bone-derived cells is an emerging technology for both disease modelling and drug screening applications. Bone tissue models are mainly limited by the implementation of sophisticated devices and procedures that can foster a tissue-specific 3D cell microenvironment along with a dynamic cultivation regime. In this study, we consequently developed, optimized and characterized an advanced perfused microfluidic platform to improve the reliability of 3D bone cell cultivation and to enhance aspects of bone tissue maturation in vitro. Moreover, biomechanical stimulation generated by fluid flow inside the arrayed chamber, was used to mimic a more dynamic cell environment emulating a highly vascularized bone we expected to improve the osteogenic 3D microenvironment in the developed multifunctional spheroid-array platform. The optimized 3D cell culture protocols in our murine bone-on-a-chip spheroid model exhibited increased mineralization and viability compared to static conditions. As a proof-of-concept, we successfully confirmed on the beneficial effects of a dynamic culture environment on osteogenesis and used our platform for analysis of bone-derived spheroids produced from primary human pre-osteoblasts. To conclude, the newly developed system represents a powerful tool for studying human bone patho/physiology in vitro under more relevant and dynamic culture conditions converging the advantages of microfluidic platforms with multi-spheroid array technologies. ","PeriodicalId":15053,"journal":{"name":"Journal of Biological Engineering","volume":"12 1","pages":""},"PeriodicalIF":5.6,"publicationDate":"2023-12-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"138631369","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Tyler Kozisek, Luke Samuelson, Andrew Hamann, Angela K. Pannier
{"title":"Systematic comparison of nonviral gene delivery strategies for efficient co-expression of two transgenes in human mesenchymal stem cells","authors":"Tyler Kozisek, Luke Samuelson, Andrew Hamann, Angela K. Pannier","doi":"10.1186/s13036-023-00394-0","DOIUrl":"https://doi.org/10.1186/s13036-023-00394-0","url":null,"abstract":"Human mesenchymal stem cells (hMSCs) are being researched for cell-based therapies due to a host of unique properties, however, genetic modification of hMSCs, accomplished through nonviral gene delivery, could greatly advance their therapeutic potential. Furthermore, expression of multiple transgenes in hMSCs could greatly advance their clinical significance for treatment of multifaceted diseases, as individual transgenes could be expressed that target separate pathogenic drivers of complex diseases. Expressing multiple transgenes can be accomplished by delivering multiple DNA vectors encoding for each transgene, or by delivering a single poly-cistronic vector that encodes for each transgene and accomplishes expression through either use of multiple promoters, an internal ribosome entry site (IRES), or a 2A peptide sequence. These different transgene expression strategies have been used to express multiple transgenes in various mammalian cells, however, they have not been fully evaluated in difficult-to-transfect primary cells, like hMSCs. This study systematically compared four transgene expression and delivery strategies for expression of two reporter transgenes in four donors of hMSCs from two tissue sources using lipid- and polymer-mediate transfection, as follows: (i) delivery of separate DNA vectors in separate nanoparticles; (ii) delivery of separate DNA vectors combined in the same nanoparticle; (iii) delivery of a bi-cistronic DNA vector with an IRES sequence via nanoparticles; and (iv) delivery of a bi-cistronic DNA vector with a dual 2A peptide sequence via nanoparticles. Our results indicate that expression of two transgenes in hMSCs, independent of expression or delivery strategy, is inefficient compared to expressing a single transgene. However, delivery of separate DNA vectors complexed in the same nanoparticle, or delivery of a bi-cistronic DNA vector with a dual 2A peptide sequence, significantly increased the number of hMSCs expressing both transgenes compared to other conditions tested. Separate DNA vectors delivered in the same nanoparticle and bi-cistronic DNA vectors with dual 2A peptide sequences are highly efficient at simultaneously expressing two transgenes in multiple donors of hMSCs from different tissue sources. The data presented in this work can guide the development of hMSC transfection systems for delivery of multiple transgenes, with the goal of producing clinically relevant, genetically modified hMSCs.\u0000","PeriodicalId":15053,"journal":{"name":"Journal of Biological Engineering","volume":"16 1","pages":""},"PeriodicalIF":5.6,"publicationDate":"2023-12-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"138554439","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Nan Zhou, TingWei Gu, Yang Xu, Yuda Liu, LiHua Peng
{"title":"Challenges and progress of neurodrug: bioactivities, production and delivery strategies of nerve growth factor protein.","authors":"Nan Zhou, TingWei Gu, Yang Xu, Yuda Liu, LiHua Peng","doi":"10.1186/s13036-023-00392-2","DOIUrl":"10.1186/s13036-023-00392-2","url":null,"abstract":"<p><p>Nerve growth factor (NGF) is a vital cytokine that plays a crucial role in the development and regeneration of the nervous system. It has been extensively studied for its potential therapeutic applications in various neural diseases. However, as a protein drug, limited natural source seriously hinders its translation and clinical applications. Conventional extraction of NGF from mouse submandibular glands has a very high cost and potentially induces immunogenicity; total synthesis and semi-synthesis methods are alternatives, but have difficulty in obtaining correct protein structure; gene engineering of plant cells is thought to be non-immunogenic, bioactive and economical. Meanwhile, large molecular weight, high polarity, and negative electrical charge make it difficult for NGF to cross the blood brain barrier to reach therapeutic targets. Current delivery strategies mainly depend on the adenovirus and cell biodelivery, but the safety and efficacy remain to be improved. New materials are widely investigated for the controllable, safe and precise delivery of NGF. This review illustrates physiological and therapeutic effects of NGF for various diseases. Moreover, new progress in production and delivery technologies for NGF are summarized. Bottlenecks encountered in the development of NGF as therapeutics are also discussed with the countermeasures proposed.</p>","PeriodicalId":15053,"journal":{"name":"Journal of Biological Engineering","volume":"17 1","pages":"75"},"PeriodicalIF":5.6,"publicationDate":"2023-12-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10696794/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"138482391","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Chih-Yun Lee, Swathi Nedunchezian, Sung-Yen Lin, Yu-Feng Su, Che-Wei Wu, Shun-Cheng Wu, Chung-Hwan Chen, Chih-Kuang Wang
{"title":"Bilayer osteochondral graft in rabbit xenogeneic transplantation model comprising sintered 3D-printed bioceramic and human adipose-derived stem cells laden biohydrogel.","authors":"Chih-Yun Lee, Swathi Nedunchezian, Sung-Yen Lin, Yu-Feng Su, Che-Wei Wu, Shun-Cheng Wu, Chung-Hwan Chen, Chih-Kuang Wang","doi":"10.1186/s13036-023-00389-x","DOIUrl":"10.1186/s13036-023-00389-x","url":null,"abstract":"<p><p>Reconstruction of severe osteochondral defects in articular cartilage and subchondral trabecular bone remains a challenging problem. The well-integrated bilayer osteochondral graft design expects to be guided the chondrogenic and osteogenic differentiation for stem cells and provides a promising solution for osteochondral tissue repair in this study. The subchondral bone scaffold approach is based on the developed finer and denser 3D β-tricalcium phosphate (β-TCP) bioceramic scaffold process, which is made using a digital light processing (DLP) technology and the novel photocurable negative thermo-responsive (NTR) bioceramic slurry. Then, the concave-top disc sintered 3D-printed bioceramic incorporates the human adipose-derived stem cells (hADSCs) laden photo-cured hybrid biohydrogel (HG + 0.5AFnSi) comprised of hyaluronic acid methacryloyl (HAMA), gelatin methacryloyl (GelMA), and 0.5% (w/v) acrylate-functionalized nano-silica (AFnSi) crosslinker. The 3D β-TCP bioceramic compartment is used to provide essential mechanical support for cartilage regeneration in the long term and slow biodegradation. However, the apparent density and compressive strength of the 3D β-TCP bioceramics can be obtained for ~ 94.8% theoretical density and 11.38 ± 1.72 MPa, respectively. In addition, the in vivo results demonstrated that the hADSC + HG + 0.5AFnSi/3D β-TCP of the bilayer osteochondral graft showed a much better osteochondral defect repair outcome in a rabbit model. The other word, the subchondral bone scaffold of 3D β-TCP bioceramic could accelerate the bone formation and integration with the adjacent host cancellous tissue at 12 weeks after surgery. And then, a thicker cartilage layer with a smooth surface and uniformly aligned chondrocytes were observed by providing enough steady mechanical support of the 3D β-TCP bioceramic scaffold.</p>","PeriodicalId":15053,"journal":{"name":"Journal of Biological Engineering","volume":"17 1","pages":"74"},"PeriodicalIF":5.6,"publicationDate":"2023-11-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10680339/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"138444742","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Negar Karimi Hajishoreh, Mehdi Dadashpour, Abolfazl Akbarzadeh
{"title":"Preparation and in vitro evaluation of biological agents based on Zinc-laponite- curcumin incorporated in alginate hydrogel.","authors":"Negar Karimi Hajishoreh, Mehdi Dadashpour, Abolfazl Akbarzadeh","doi":"10.1186/s13036-023-00391-3","DOIUrl":"10.1186/s13036-023-00391-3","url":null,"abstract":"<p><p>Due to their outstanding structures and properties, three-dimensional (3D) hydrogels and nanoparticles have been widely studied and indicated a very high potential for medical, therapeutic, and diagnostic applications. However, hydrogels and nanoparticles systems have particular drawbacks that limit their widespread applications. In recent years, the incorporation of nanostructured systems into hydrogel has been developed as a novel way for the formation of new biomaterials with various functions to solve biomedical challenges. In this study, alginate-loaded Zinc- laponite-curcumin (Zn/La/Cur) nanocomposites were fabricated via ionic cross-linking. The prepared nanocomposite hydrogels were characterized via FTIR and FE-SEM. Moreover, energy dispersive x-ray spectroscopy (EDX) was used to study the elements of the Zn/La/Cur nanocomposite. The NIH3T3 fibroblast cell line was utilized for the MTT assay to determine the cell viability of the fabricated alginate-loaded Zn/La/Cur nanocomposites. MTT results demonstrated that there was no evidence of toxicity in the samples. These outcomes suggest that applying Al/Zn/La/Cur nanocomposite as a biological agent could be a novel tissue engineering strategy for treating soft tissue disorders.</p>","PeriodicalId":15053,"journal":{"name":"Journal of Biological Engineering","volume":"17 1","pages":"73"},"PeriodicalIF":5.6,"publicationDate":"2023-11-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10675890/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"138434051","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Meike Bielfeldt, Kai Budde-Sagert, Nikolai Weis, Maren Buenning, Susanne Staehlke, Julius Zimmermann, Nils Arbeiter, Sahba Mobini, María Ujué González, Henrike Rebl, Adelinde Uhrmacher, Ursula van Rienen, Barbara Nebe
{"title":"Discrimination between the effects of pulsed electrical stimulation and electrochemically conditioned medium on human osteoblasts.","authors":"Meike Bielfeldt, Kai Budde-Sagert, Nikolai Weis, Maren Buenning, Susanne Staehlke, Julius Zimmermann, Nils Arbeiter, Sahba Mobini, María Ujué González, Henrike Rebl, Adelinde Uhrmacher, Ursula van Rienen, Barbara Nebe","doi":"10.1186/s13036-023-00393-1","DOIUrl":"10.1186/s13036-023-00393-1","url":null,"abstract":"<p><strong>Background: </strong>Electrical stimulation is used for enhanced bone fracture healing. Electrochemical processes occur during the electrical stimulation at the electrodes and influence cellular reactions. Our approach aimed to distinguish between electrochemical and electric field effects on osteoblast-like MG-63 cells. We applied 20 Hz biphasic pulses via platinum electrodes for 2 h. The electrical stimulation of the cell culture medium and subsequent application to cells was compared to directly stimulated cells. The electric field distribution was predicted using a digital twin.</p><p><strong>Results: </strong>Cyclic voltammetry and electrochemical impedance spectroscopy revealed partial electrolysis at the electrodes, which was confirmed by increased concentrations of hydrogen peroxide in the medium. While both direct stimulation and AC-conditioned medium decreased cell adhesion and spreading, only the direct stimulation enhanced the intracellular calcium ions and reactive oxygen species.</p><p><strong>Conclusion: </strong>The electrochemical by-product hydrogen peroxide is not the main contributor to the cellular effects of electrical stimulation. However, undesired effects like decreased adhesion are mediated through electrochemical products in stimulated medium. Detailed characterisation and monitoring of the stimulation set up and electrochemical reactions are necessary to find safe electrical stimulation protocols.</p>","PeriodicalId":15053,"journal":{"name":"Journal of Biological Engineering","volume":"17 1","pages":"71"},"PeriodicalIF":5.6,"publicationDate":"2023-11-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10668359/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"138299145","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Xinyu Luo, Jiayue Niu, Guanyu Su, Linxi Zhou, Xue Zhang, Ying Liu, Qiang Wang, Ningning Sun
{"title":"Research progress of biomimetic materials in oral medicine.","authors":"Xinyu Luo, Jiayue Niu, Guanyu Su, Linxi Zhou, Xue Zhang, Ying Liu, Qiang Wang, Ningning Sun","doi":"10.1186/s13036-023-00382-4","DOIUrl":"10.1186/s13036-023-00382-4","url":null,"abstract":"<p><p>Biomimetic materials are able to mimic the structure and functional properties of native tissues especially natural oral tissues. They have attracted growing attention for their potential to achieve configurable and functional reconstruction in oral medicine. Though tremendous progress has been made regarding biomimetic materials, significant challenges still remain in terms of controversy on the mechanism of tooth tissue regeneration, lack of options for manufacturing such materials and insufficiency of in vivo experimental tests in related fields. In this review, the biomimetic materials used in oral medicine are summarized systematically, including tooth defect, tooth loss, periodontal diseases and maxillofacial bone defect. Various theoretical foundations of biomimetic materials research are reviewed, introducing the current and pertinent results. The benefits and limitations of these materials are summed up at the same time. Finally, challenges and potential of this field are discussed. This review provides the framework and support for further research in addition to giving a generally novel and fundamental basis for the utilization of biomimetic materials in the future.</p>","PeriodicalId":15053,"journal":{"name":"Journal of Biological Engineering","volume":"17 1","pages":"72"},"PeriodicalIF":5.7,"publicationDate":"2023-11-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10668381/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"138299146","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Follicular reconstruction and neo-oogenesis in xenotransplantation of human ovarian isolated cells derived from chemotherapy-induced POF patients.","authors":"Sara Khaleghi, Farideh Eivazkhani, Somayeh Tavana, Ashraf Moini, Marefat Ghaffari Novin, Petkov Stoyan, Hamid Nazarian, Rouhollah Fathi","doi":"10.1186/s13036-023-00384-2","DOIUrl":"10.1186/s13036-023-00384-2","url":null,"abstract":"<p><strong>Background: </strong>Developing new strategies to restore fertility in patients with chemotherapy-induced Premature Ovarian Failure (Chemo-POF) is important. We aimed to construct an Artificial Ovary (AO) by seeding Human Ovarian Cortical Cells (HOCCs) into Human ovarian Decellularized Cortical Tissue (DCT). We assessed the AO's ability to produce new ovarian follicles following xenotransplantation to NMRI mice.</p><p><strong>Material and methods: </strong>The DCTs were prepared, and cell removal was confirmed through DNA content, MTT assay, DAPI and H&E staining. Next, HOCCs were isolated from both Chemo-POF and Trans (as a control group) ovarian patients. The HOCCs were characterized using immunostaining (FRAGILIS, Vimentin, and Inhibin α) and real time PCR (DDX4, STELLA, FRAGILIS, Vimentin, FSH-R, KI67) assays. The HOCCs were then seeded into the DCTs and cultured for one week to construct an AO, which was subsequently xenotransplanted into the mice. The existence of active follicles within the AO was studied with H&E and immunofluorescence (GDF9) staining, Real-time PCR (GDF9, ZP3) and hormone analysis (Estradiol, FSH and AMH).</p><p><strong>Results: </strong>The results of gene expression and immunostaining showed that 85-86% of the isolated cells from both Trans and Chemo-POF groups were positive for vimentin, while 2-5% were granulosa cells and OSCs were less than 3%. After xenotransplantation, histological study confirmed the presence of morphologically healthy reconstructed human ovarian follicles. Additionally, immunofluorescence staining of GDF9 and hormonal assay confirmed the presence of secretory-active follicles on the AO.</p><p><strong>Conclusion: </strong>Our findings demonstrate that an artificial ovary produced by seeding HOCCs on DCT can support HOCCs proliferation as well as neo-oogenesis, and enable sex hormone secretion following xenotransplantation.</p>","PeriodicalId":15053,"journal":{"name":"Journal of Biological Engineering","volume":"17 1","pages":"70"},"PeriodicalIF":5.7,"publicationDate":"2023-11-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10662631/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"138176262","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}