Enyu Mao, Yu Hu, Yinzi Xin, Zheyi Sun, Jun Zhang, Song Li
Mesenchymal stem cell (MSC)-based therapies for articular cartilage regeneration are effective mostly due to paracrine signals mediated by extracellular vesicles, especially small extracellular vesicles (sEV). However, it is unknown whether dental follicle cell-derived sEV (DFC-sEV) affect cartilage regeneration in temporomandibular joint osteoarthritis (TMJ-OA). In this study, the effects of DFC-sEV on IL-1β-induced mandibular condylar chondrocytes (MCCs) were determined using CCK8 assays, scratch assays, flow cytometry, and Western blot analysis of matrix synthesis and catabolic proteins. Furthermore, we used an abnormal occlusion-induced rat model and intra-articular injection of DFC-sEV, the pathological changes of which were observed by HE staining, safranin O staining, immunohistochemistry, and micro-CT analysis of subchondral bone loss. Gene set enrichment analysis (GSEA) was used to determine the related mechanism involved in the effect of DFC-sEV. Immunofluorescence analysis and Western blotting were used to evaluate the expression of HIF-1α, HIF-2α, MMP13, and VEGF in MCCs. Then, lentivirus-induced Epas1 overexpression and Western blot analysis of the downstream regulators of HIF-2α were performed. We found that DFC-sEV promoted MCCs proliferation and migration and protected against cartilage matrix destruction induced by IL-1β. In addition, DFC-sEV prevented cartilage destruction in an abnormal occlusion rat model. Furthermore, we found that DFC-sEV reduced the expression of HIF-1α and HIF-2α in vitro and in vivo and decreased the downstream regulators of HIF-2α, including MMP13 and VEGF. Our study indicated that DFC-sEV attenuated TMJ cartilage damage in vitro and in vivo, which might be involved in the regulation of HIF-2α.
{"title":"Human Dental Follicle Cell-Derived Small Extracellular Vesicles Attenuate Temporomandibular Joint Cartilage Damage through Inhibiting HIF-2α","authors":"Enyu Mao, Yu Hu, Yinzi Xin, Zheyi Sun, Jun Zhang, Song Li","doi":"10.1155/2023/6625123","DOIUrl":"https://doi.org/10.1155/2023/6625123","url":null,"abstract":"Mesenchymal stem cell (MSC)-based therapies for articular cartilage regeneration are effective mostly due to paracrine signals mediated by extracellular vesicles, especially small extracellular vesicles (sEV). However, it is unknown whether dental follicle cell-derived sEV (DFC-sEV) affect cartilage regeneration in temporomandibular joint osteoarthritis (TMJ-OA). In this study, the effects of DFC-sEV on IL-1β-induced mandibular condylar chondrocytes (MCCs) were determined using CCK8 assays, scratch assays, flow cytometry, and Western blot analysis of matrix synthesis and catabolic proteins. Furthermore, we used an abnormal occlusion-induced rat model and intra-articular injection of DFC-sEV, the pathological changes of which were observed by HE staining, safranin O staining, immunohistochemistry, and micro-CT analysis of subchondral bone loss. Gene set enrichment analysis (GSEA) was used to determine the related mechanism involved in the effect of DFC-sEV. Immunofluorescence analysis and Western blotting were used to evaluate the expression of HIF-1α, HIF-2α, MMP13, and VEGF in MCCs. Then, lentivirus-induced Epas1 overexpression and Western blot analysis of the downstream regulators of HIF-2α were performed. We found that DFC-sEV promoted MCCs proliferation and migration and protected against cartilage matrix destruction induced by IL-1β. In addition, DFC-sEV prevented cartilage destruction in an abnormal occlusion rat model. Furthermore, we found that DFC-sEV reduced the expression of HIF-1α and HIF-2α in vitro and in vivo and decreased the downstream regulators of HIF-2α, including MMP13 and VEGF. Our study indicated that DFC-sEV attenuated TMJ cartilage damage in vitro and in vivo, which might be involved in the regulation of HIF-2α.","PeriodicalId":202,"journal":{"name":"Journal of Tissue Engineering and Regenerative Medicine","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2023-09-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"135815053","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}
Meghan McGraw, Gabrielle Gilmer, Juliana Bergmann, Vishnu Seshan, Kai Wang, David Pekker, Michel Modo, Fabrisia Ambrosio
Magnetic field exposure is a well-established diagnostic tool. However, its use as a therapeutic in regenerative medicine is relatively new. To better understand how magnetic fields affect neural repair in vitro, we started by performing a systematic review of publications that studied neural repair responses to magnetic fields. The 38 included articles were highly heterogeneous, representing 13 cell types, magnetic field magnitudes of 0.0002–10,000 mT with frequencies of 0–150 Hz, and exposure times ranging from one hour to several weeks. Mathematical modeling based on data from the included manuscripts revealed higher magnetic field magnitudes enhance neural progenitor cell (NPC) viability. Finally, for those regenerative processes not influenced by magnitude, frequency, or time, we integrated the data by meta-analyses. Results revealed that magnetic field exposure increases NPC proliferation while decreasing astrocytic differentiation. Collectively, our approach identified neural repair processes that may be most responsive to magnetic field exposure.
{"title":"Mapping the Landscape of Magnetic Field Effects on Neural Regeneration and Repair: A Combined Systematic Review, Mathematical Model, and Meta-Analysis","authors":"Meghan McGraw, Gabrielle Gilmer, Juliana Bergmann, Vishnu Seshan, Kai Wang, David Pekker, Michel Modo, Fabrisia Ambrosio","doi":"10.1155/2023/5038317","DOIUrl":"https://doi.org/10.1155/2023/5038317","url":null,"abstract":"Magnetic field exposure is a well-established diagnostic tool. However, its use as a therapeutic in regenerative medicine is relatively new. To better understand how magnetic fields affect neural repair in vitro, we started by performing a systematic review of publications that studied neural repair responses to magnetic fields. The 38 included articles were highly heterogeneous, representing 13 cell types, magnetic field magnitudes of 0.0002–10,000 mT with frequencies of 0–150 Hz, and exposure times ranging from one hour to several weeks. Mathematical modeling based on data from the included manuscripts revealed higher magnetic field magnitudes enhance neural progenitor cell (NPC) viability. Finally, for those regenerative processes not influenced by magnitude, frequency, or time, we integrated the data by meta-analyses. Results revealed that magnetic field exposure increases NPC proliferation while decreasing astrocytic differentiation. Collectively, our approach identified neural repair processes that may be most responsive to magnetic field exposure.","PeriodicalId":202,"journal":{"name":"Journal of Tissue Engineering and Regenerative Medicine","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2023-09-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"136130785","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}
The rehabilitation of bone defects after radiotherapy requires the development of osteoinductive bone substitutes. MicroRNA could be used as an osteogenic factor to fabricate functional materials for bone regeneration. In this study, we used miR-34a to enhance bone regeneration after irradiation. We lyophilized lipofectamine-agomiR-34a lipoplexes on hydroxyapatite (HA) to develop miR-34a-functionalized hydroxyapatite (HA-agomiR-34a). The morphology was observed by scanning electron microscope and atomic force microscope. Fluorescence microscopy confirmed the retention of agomiR-34a on the surface of HA. HA-agomiR-34a showed high transfection efficiency and good biocompatibility. HA-agomiR-34a enhanced the osteoblastic differentiation of radiation-impaired bone marrow stromal cells (BMSCs). Implantation of HA-agomiR-34a promoted bone regeneration in irradiated bone defects. HA-agomiR-34a may be a novel and safe bone substitute to promote the reconstruction of bone defects after radiotherapy.
{"title":"MiR-34a-Functionalized Hydroxyapatite by Lyophilization Promoted Bone Regeneration in Irradiated Bone Defects","authors":"Xi Wu, Xiaoke Feng, Gang Zhang, Huan Liu","doi":"10.1155/2023/9946012","DOIUrl":"https://doi.org/10.1155/2023/9946012","url":null,"abstract":"The rehabilitation of bone defects after radiotherapy requires the development of osteoinductive bone substitutes. MicroRNA could be used as an osteogenic factor to fabricate functional materials for bone regeneration. In this study, we used miR-34a to enhance bone regeneration after irradiation. We lyophilized lipofectamine-agomiR-34a lipoplexes on hydroxyapatite (HA) to develop miR-34a-functionalized hydroxyapatite (HA-agomiR-34a). The morphology was observed by scanning electron microscope and atomic force microscope. Fluorescence microscopy confirmed the retention of agomiR-34a on the surface of HA. HA-agomiR-34a showed high transfection efficiency and good biocompatibility. HA-agomiR-34a enhanced the osteoblastic differentiation of radiation-impaired bone marrow stromal cells (BMSCs). Implantation of HA-agomiR-34a promoted bone regeneration in irradiated bone defects. HA-agomiR-34a may be a novel and safe bone substitute to promote the reconstruction of bone defects after radiotherapy.","PeriodicalId":202,"journal":{"name":"Journal of Tissue Engineering and Regenerative Medicine","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2023-09-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"135939249","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}
Studying the crosstalk between the muscle and tendon tissue is an important yet understudied area in musculoskeletal research. In vitro models can help elucidate the function and repair of the myotendinous junction (MTJ) under static and dynamic culture conditions using engineered muscle tissues. The goal of this study was to culture engineered muscle tissues in a novel bioreactor in both static and mechanically stimulated cultures and evaluate the expression of MTJ-specific proteins within the muscle-tendon unit(paxillin and type XXII collagen). C2C12 myoblasts were seeded in hydrogels made from type I collagen ortendon-derived extracellular matrix (tECM) and allowed to form around movable anchors. Engineered tissues were allowed to form and stabilize for 10 days. After 10 days in the culture, stimulated cultures were cyclically stimulated for 3 hours per day for 2 and 4 weeks alongside static cultures. Strain values at the maximum displacement of the anchors averaged about 0.10, a target that has been shown to induce myogenic phenotype in C2C12s. Protein expression of paxillin after 2 weeks did not differ between hydrogel materials in static cultures but increased by 62% in tECM when mechanically stimulated. These differences continued after 4 weeks, with 31% and 57% increases in tECM tissues relative to type I collagen. Expression of type XXII collagen was similarly influenced by hydrogel material and culture conditions. Overall, this research combined a relevant microenvironment to study muscle and tendon interactions with a novel bioreactor to apply mechanical strain, an important regulator of the formation and maintenance of the native MTJ.
{"title":"Tendon Extracellular Matrix Promotes Myotendinous Junction Protein Expression in Engineered Muscle Tissue under Both Static and Mechanically Stimulated Culture Conditions","authors":"Lewis S. Gaffney, M. Fisher, D. Freytes","doi":"10.1155/2023/6658543","DOIUrl":"https://doi.org/10.1155/2023/6658543","url":null,"abstract":"Studying the crosstalk between the muscle and tendon tissue is an important yet understudied area in musculoskeletal research. In vitro models can help elucidate the function and repair of the myotendinous junction (MTJ) under static and dynamic culture conditions using engineered muscle tissues. The goal of this study was to culture engineered muscle tissues in a novel bioreactor in both static and mechanically stimulated cultures and evaluate the expression of MTJ-specific proteins within the muscle-tendon unit(paxillin and type XXII collagen). C2C12 myoblasts were seeded in hydrogels made from type I collagen ortendon-derived extracellular matrix (tECM) and allowed to form around movable anchors. Engineered tissues were allowed to form and stabilize for 10 days. After 10 days in the culture, stimulated cultures were cyclically stimulated for 3 hours per day for 2 and 4 weeks alongside static cultures. Strain values at the maximum displacement of the anchors averaged about 0.10, a target that has been shown to induce myogenic phenotype in C2C12s. Protein expression of paxillin after 2 weeks did not differ between hydrogel materials in static cultures but increased by 62% in tECM when mechanically stimulated. These differences continued after 4 weeks, with 31% and 57% increases in tECM tissues relative to type I collagen. Expression of type XXII collagen was similarly influenced by hydrogel material and culture conditions. Overall, this research combined a relevant microenvironment to study muscle and tendon interactions with a novel bioreactor to apply mechanical strain, an important regulator of the formation and maintenance of the native MTJ.","PeriodicalId":202,"journal":{"name":"Journal of Tissue Engineering and Regenerative Medicine","volume":null,"pages":null},"PeriodicalIF":3.3,"publicationDate":"2023-08-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"48683730","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}
Miles Markey, Caroline D. Pena, T. Venkatesh, L. Cai, Maribel Vazquez
Retinal degeneration is an escalating public health challenge, as diseases such as age-related macular degeneration, diabetic retinopathy, and retinitis pigmentosa cause irreversible vision loss in millions of adults each year. Regenerative medicine has pioneered the development of stem cell replacement therapies, which treat degeneration by replacing damaged retinal neurons with transplanted stem-like cells (SCs). While the collective migration of SCs plays critical roles during retinal development, our understanding of collective SC behaviors within biomaterials transplanted into adult tissue remains understudied. This project examines the potential therapeutic impacts of collective SC migration during transplantation by correlating the expression of cadherin, cell-cell cohesion molecules that maintain intercellular communication during development, with receptor proteins of chemoattractant molecules prevalent in degenerated adult tissue. Experiments examine these well-conserved biomechanisms by using two different model organisms: Drosophila melanogaster, a seminal model for retinal development, and Mus, an important preclinical model for transplantation. Results indicate that SCs from both animal models significantly upregulate cadherin expression to achieve more directed collective migration towards species-specific chemoattractants and exhibit longer distance motility upon different extracellular matrix substrates.
{"title":"Retinal Progenitor Cells Exhibit Cadherin-Dependent Chemotaxis across Transplantable Extracellular Matrix of In Vitro Developmental and Adult Models","authors":"Miles Markey, Caroline D. Pena, T. Venkatesh, L. Cai, Maribel Vazquez","doi":"10.1155/2023/1381620","DOIUrl":"https://doi.org/10.1155/2023/1381620","url":null,"abstract":"Retinal degeneration is an escalating public health challenge, as diseases such as age-related macular degeneration, diabetic retinopathy, and retinitis pigmentosa cause irreversible vision loss in millions of adults each year. Regenerative medicine has pioneered the development of stem cell replacement therapies, which treat degeneration by replacing damaged retinal neurons with transplanted stem-like cells (SCs). While the collective migration of SCs plays critical roles during retinal development, our understanding of collective SC behaviors within biomaterials transplanted into adult tissue remains understudied. This project examines the potential therapeutic impacts of collective SC migration during transplantation by correlating the expression of cadherin, cell-cell cohesion molecules that maintain intercellular communication during development, with receptor proteins of chemoattractant molecules prevalent in degenerated adult tissue. Experiments examine these well-conserved biomechanisms by using two different model organisms: Drosophila melanogaster, a seminal model for retinal development, and Mus, an important preclinical model for transplantation. Results indicate that SCs from both animal models significantly upregulate cadherin expression to achieve more directed collective migration towards species-specific chemoattractants and exhibit longer distance motility upon different extracellular matrix substrates.","PeriodicalId":202,"journal":{"name":"Journal of Tissue Engineering and Regenerative Medicine","volume":null,"pages":null},"PeriodicalIF":3.3,"publicationDate":"2023-08-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"48303787","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}
Farhad Chariyev-Prinz, N. Neto, M. Monaghan, D. Kelly
It is generally accepted that the application of hydrostatic pressure (HP) is beneficial for MSC chondrogenesis. There is, however, evidence to suggest that the timing of application might determine its impact on cell fate and tissue development. Furthermore, understanding how the maturity of engineered cartilage affects its response to the application of HP can provide critical insight into determining when such a graft is ready for in vivo implantation into a mechanically loaded environment. In this study, we systematically examined chondrogenic maturation of hMSCs over 35 days in the presence of TGF-β3 in vitro. At specific timepoints, the response of hMSCs to the application of HP following the removal of TGF-β3 was assessed; this partially models conditions such grafts will experience in vivo upon implantation. In free swelling culture, the expression of chondrogenic (COL2A1 and ACAN) and hypertrophic (COL10A1) markers increased with time. At early timepoints, the expression of such markers continued to increase following TGF-β3 withdrawal; however, this was not observed after prolonged periods of chondrogenic priming (35 days). Interestingly, the application of HP was only beneficial after 35 days of chondrogenic priming, where it enhanced sGAG synthesis and improved key chondrogenic gene ratios. It was also found that HP can facilitate a metabolic shift towards oxidative phosphorylation, which can be viewed as a hallmark of successfully differentiating MSCs. These results point to the importance of mechanical loading as a key stimulus for maintaining a chondrogenic phenotype once MSCs are removed from chemically defined culture conditions.
{"title":"Time-Dependent Anabolic Response of hMSC-Derived Cartilage Grafts to Hydrostatic Pressure","authors":"Farhad Chariyev-Prinz, N. Neto, M. Monaghan, D. Kelly","doi":"10.1155/2023/9976121","DOIUrl":"https://doi.org/10.1155/2023/9976121","url":null,"abstract":"It is generally accepted that the application of hydrostatic pressure (HP) is beneficial for MSC chondrogenesis. There is, however, evidence to suggest that the timing of application might determine its impact on cell fate and tissue development. Furthermore, understanding how the maturity of engineered cartilage affects its response to the application of HP can provide critical insight into determining when such a graft is ready for in vivo implantation into a mechanically loaded environment. In this study, we systematically examined chondrogenic maturation of hMSCs over 35 days in the presence of TGF-β3 in vitro. At specific timepoints, the response of hMSCs to the application of HP following the removal of TGF-β3 was assessed; this partially models conditions such grafts will experience in vivo upon implantation. In free swelling culture, the expression of chondrogenic (COL2A1 and ACAN) and hypertrophic (COL10A1) markers increased with time. At early timepoints, the expression of such markers continued to increase following TGF-β3 withdrawal; however, this was not observed after prolonged periods of chondrogenic priming (35 days). Interestingly, the application of HP was only beneficial after 35 days of chondrogenic priming, where it enhanced sGAG synthesis and improved key chondrogenic gene ratios. It was also found that HP can facilitate a metabolic shift towards oxidative phosphorylation, which can be viewed as a hallmark of successfully differentiating MSCs. These results point to the importance of mechanical loading as a key stimulus for maintaining a chondrogenic phenotype once MSCs are removed from chemically defined culture conditions.","PeriodicalId":202,"journal":{"name":"Journal of Tissue Engineering and Regenerative Medicine","volume":null,"pages":null},"PeriodicalIF":3.3,"publicationDate":"2023-08-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"44449622","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}
Wei Liu, J. Venkatesan, Mahnaz Amini, T. Oláh, G. Schmitt, H. Madry, Magali Cucchiarini
Recombinant adeno-associated virus (rAAV) vectors have a strong potential to promote the healing of traumatic cartilage defects and osteoarthritic lesions upon delivery and overexpression of therapeutic genes from suitable biomaterials that support a controlled release of the candidate constructs. The goal of the present work is to examine whether the administration of chondrogenic rAAV sox9 and rAAV TGF-ß gene vehicles via alginate hydrogel-guided vector delivery stimulates the biological and chondroreparative activities of human bone marrow-derived mesenchymal stromal cells (hMSCs) as a source of improved reparative cells for future implantation in sites of cartilage damage. The delivery of rAAV using an alginate (AlgPH155) hydrogel system is successfully achieved in hMSCs over time (21 days), leading to the effective overexpression of sox9 and TGF-ß that significantly increases the proliferation and chondrogenic differentiation activities of the cells relative to control (marker lacZ) gene transfer while advantageously preventing premature hypertrophy, osteogenesis, and mineralization. This study reveals the potential of alginate hydrogel-based systems to control the delivery of rAAV (sox9 and TGF-ß) gene vectors to adeptly trigger the chondroreparative activities of hMSCs for future applications that aim at improving cartilage repair.
{"title":"Effects of rAAV-Mediated Overexpression of sox9 and TGF-ß via Alginate Hydrogel-Guided Vector Delivery on the Chondroreparative Activities of Human Bone Marrow-Derived Mesenchymal Stromal Cells","authors":"Wei Liu, J. Venkatesan, Mahnaz Amini, T. Oláh, G. Schmitt, H. Madry, Magali Cucchiarini","doi":"10.1155/2023/4495697","DOIUrl":"https://doi.org/10.1155/2023/4495697","url":null,"abstract":"Recombinant adeno-associated virus (rAAV) vectors have a strong potential to promote the healing of traumatic cartilage defects and osteoarthritic lesions upon delivery and overexpression of therapeutic genes from suitable biomaterials that support a controlled release of the candidate constructs. The goal of the present work is to examine whether the administration of chondrogenic rAAV sox9 and rAAV TGF-ß gene vehicles via alginate hydrogel-guided vector delivery stimulates the biological and chondroreparative activities of human bone marrow-derived mesenchymal stromal cells (hMSCs) as a source of improved reparative cells for future implantation in sites of cartilage damage. The delivery of rAAV using an alginate (AlgPH155) hydrogel system is successfully achieved in hMSCs over time (21 days), leading to the effective overexpression of sox9 and TGF-ß that significantly increases the proliferation and chondrogenic differentiation activities of the cells relative to control (marker lacZ) gene transfer while advantageously preventing premature hypertrophy, osteogenesis, and mineralization. This study reveals the potential of alginate hydrogel-based systems to control the delivery of rAAV (sox9 and TGF-ß) gene vectors to adeptly trigger the chondroreparative activities of hMSCs for future applications that aim at improving cartilage repair.","PeriodicalId":202,"journal":{"name":"Journal of Tissue Engineering and Regenerative Medicine","volume":null,"pages":null},"PeriodicalIF":3.3,"publicationDate":"2023-08-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"44149045","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}
Cory K. Mayfield, Elizabeth Lechtholz-Zey, M. Ayad, O. Sugiyama, Jay R. Lieberman
Novel treatment strategies for segmental bone loss in orthopaedic surgery remain under investigation. Regional gene therapy that involves transduction of mesenchymal stem cells with a lentiviral vector that expresses BMP-2 has gained particular interest as this strategy provides osteogenic and osteoinductive factors for bone growth. In particular, transduced adipose-derived stems cells (ASCs) and bone marrow-derived stem cells (BMSCs) have emerged as the leading candidates for the treatment of segmental defects in preclinical models. The aim of the present study was to evaluate the influence of demographic information on in vitro growth characteristics and bone morphogenetic protein-2 production following lentiviral transduction in a large cohort of human donors. We further sought to assess the effects of ASC harvest site on cell yield and growth characteristics. We evaluated a total of 187 human donors (124 adipose harvests and 63 bone marrow aspirates) in our cohort. We found that across all donors, ASCs demonstrated favorable growth characteristics and could be cultured in vitro more reliably than BMSCs regardless of patient-related factors. Furthermore, we noted that following lentiviral transduction, ASCs produced significantly higher levels of BMP-2 compared to BMSCs. Lastly, despite higher initial cell yields from lipoaspirate, posttransduction BMP-2 production was less than that of infrapatellar fat pad samples. These results support the continued investigation of ASCs as a cellular delivery vehicle for regional gene therapy to deliver osteoinductive proteins to specific anatomic bone repair sites.
{"title":"Human Bone Marrow versus Adipose-Derived Stem Cells: Influence of Donor Characteristics on Expandability and Implications for Osteogenic Ex Vivo BMP-2 Regional Gene Therapy","authors":"Cory K. Mayfield, Elizabeth Lechtholz-Zey, M. Ayad, O. Sugiyama, Jay R. Lieberman","doi":"10.1155/2023/8061890","DOIUrl":"https://doi.org/10.1155/2023/8061890","url":null,"abstract":"Novel treatment strategies for segmental bone loss in orthopaedic surgery remain under investigation. Regional gene therapy that involves transduction of mesenchymal stem cells with a lentiviral vector that expresses BMP-2 has gained particular interest as this strategy provides osteogenic and osteoinductive factors for bone growth. In particular, transduced adipose-derived stems cells (ASCs) and bone marrow-derived stem cells (BMSCs) have emerged as the leading candidates for the treatment of segmental defects in preclinical models. The aim of the present study was to evaluate the influence of demographic information on in vitro growth characteristics and bone morphogenetic protein-2 production following lentiviral transduction in a large cohort of human donors. We further sought to assess the effects of ASC harvest site on cell yield and growth characteristics. We evaluated a total of 187 human donors (124 adipose harvests and 63 bone marrow aspirates) in our cohort. We found that across all donors, ASCs demonstrated favorable growth characteristics and could be cultured in vitro more reliably than BMSCs regardless of patient-related factors. Furthermore, we noted that following lentiviral transduction, ASCs produced significantly higher levels of BMP-2 compared to BMSCs. Lastly, despite higher initial cell yields from lipoaspirate, posttransduction BMP-2 production was less than that of infrapatellar fat pad samples. These results support the continued investigation of ASCs as a cellular delivery vehicle for regional gene therapy to deliver osteoinductive proteins to specific anatomic bone repair sites.","PeriodicalId":202,"journal":{"name":"Journal of Tissue Engineering and Regenerative Medicine","volume":null,"pages":null},"PeriodicalIF":3.3,"publicationDate":"2023-08-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"47103066","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}
E. Dohle, Tatjana Fecht, Tobias Wolfram, F. Reinauer, Anke Wunder, Katja Heppe, R. Sader, C. Kirkpatrick, S. Ghanaati
During the present study, an in vitro coculture bone tissue mimic based on primary osteoblasts and primary endothelial cells was used for a complex and broad evaluation of a newly developed material for applications in pediatric maxillofacial traumatology. The biomaterial was composed of PDLLA (poly(D,L-lactide)) in various combinations with calcium carbonate (CC), magnesium (Mg), and chitosan (CH). Besides classical biocompatibility analyses, the present study evaluated material-dependent effects on fundamental processes that are essential for successful material integration and regeneration. Therefore, inflammation-associated factors such as E-selectin and interleukins were analyzed in the in vitro model system on gene expression and protein level depending on the different materials. Furthermore, in order to test the capability of vascularization of the material, the effect of the different materials on the formation of microvessel-like structures as well as the expression and release of proangiogenic factors was investigated in vitro in the bone coculture model. In addition, the mineralization capacity as well as the relative gene expression of osteogenic differentiation factors was analyzed in response to the different materials. As a result, the authors could assess the material combination PDLLA: CC CH as the most functionally tested material with regard to biocompatibility, inflammatory response, and microvessel-like structure formation as well as osteogenic differentiation in the in vitro coculture system. In conclusion, by using tissue-engineered human bone tissue equivalents as proposed here in an in vitro coculture model, biomaterial-mediated effects can be readily investigated. Moreover, it is proposed that these complex in vitro evaluations could contribute to the understanding and improvement of the development of novel materials for pediatric traumatological care for prospective clinical applications.
{"title":"In Vitro Coculture of Primary Human Cells to Analyze Angiogenesis, Osteogenesis, and the Inflammatory Response to Newly Developed Osteosynthesis Material for Pediatric Maxillofacial Traumatology: A Potential Pretesting Model before In Vivo Experiments","authors":"E. Dohle, Tatjana Fecht, Tobias Wolfram, F. Reinauer, Anke Wunder, Katja Heppe, R. Sader, C. Kirkpatrick, S. Ghanaati","doi":"10.1155/2023/4040504","DOIUrl":"https://doi.org/10.1155/2023/4040504","url":null,"abstract":"During the present study, an in vitro coculture bone tissue mimic based on primary osteoblasts and primary endothelial cells was used for a complex and broad evaluation of a newly developed material for applications in pediatric maxillofacial traumatology. The biomaterial was composed of PDLLA (poly(D,L-lactide)) in various combinations with calcium carbonate (CC), magnesium (Mg), and chitosan (CH). Besides classical biocompatibility analyses, the present study evaluated material-dependent effects on fundamental processes that are essential for successful material integration and regeneration. Therefore, inflammation-associated factors such as E-selectin and interleukins were analyzed in the in vitro model system on gene expression and protein level depending on the different materials. Furthermore, in order to test the capability of vascularization of the material, the effect of the different materials on the formation of microvessel-like structures as well as the expression and release of proangiogenic factors was investigated in vitro in the bone coculture model. In addition, the mineralization capacity as well as the relative gene expression of osteogenic differentiation factors was analyzed in response to the different materials. As a result, the authors could assess the material combination PDLLA: CC CH as the most functionally tested material with regard to biocompatibility, inflammatory response, and microvessel-like structure formation as well as osteogenic differentiation in the in vitro coculture system. In conclusion, by using tissue-engineered human bone tissue equivalents as proposed here in an in vitro coculture model, biomaterial-mediated effects can be readily investigated. Moreover, it is proposed that these complex in vitro evaluations could contribute to the understanding and improvement of the development of novel materials for pediatric traumatological care for prospective clinical applications.","PeriodicalId":202,"journal":{"name":"Journal of Tissue Engineering and Regenerative Medicine","volume":null,"pages":null},"PeriodicalIF":3.3,"publicationDate":"2023-08-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"46958080","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}
Yongwen Guo, Mengting He, Peiqi Wang, D. Bai, Jeong‐Hui Park, K. Dashnyam, Jung-Hwan Lee, O. Huck, N. Benkirane-Jessel, Hae-Won Kim, M. Ramalingam
While treated dentin matrix (TDM) has been used for regeneration of dental tissues, the quality and quantity of regenerated periodontal tissue structure are suboptimal. The present study was undertaken to test whether the combined use of the TDM with dental follicle cells (DFCs) and Hertwig’s epithelial root sheath (HERS) cells enhances the regeneration of periodontal structures in a nondental microenvironment. TDMs were fabricated from 3-month-old Sprague–Dawley (SD) rats. DFCs and HERS cells were isolated from postnatal 7-day SD rats. Purified DFCs and HERS cells, both in combination or alone, were seeded and cultured on TDM in vitro and characterized. The cell-seeded TDMs were subsequently implanted into a 3-month-old rat greater omentum for 6 weeks, and further histological evaluation was performed. The results showed that cells grew well on the surface of TDMs, and mineralized nodules could be seen, especially in the HERS + DFCs group. After transplantation in rat omentum, periodontal ligament-like fibers and cementum-like structures were observed around the TDM in 1/3 of the samples in both the HERS group and the DFCs group and in 2/3 of the samples in the HERS + DFCs group, while almost no attached tissue formation was found in the TDM only group. The formed cementum width and the periodontal ligament length were significantly larger in the HERS + DFCs group. The periodontal ligament-like fibers in the HERS + DFCs group were orderly arranged and attached to the cementum-like tissues, which resembled the cementum-periodontal structure. Therefore, the combined use of DFCs, TDM, and HERS cells may be a promising strategy for the regeneration of the periodontal structures, especially in the nondental microenvironment.
{"title":"A Combinatorial Approach to Regenerate the Periodontal Ligament and Cementum in a Nondental Microenvironment","authors":"Yongwen Guo, Mengting He, Peiqi Wang, D. Bai, Jeong‐Hui Park, K. Dashnyam, Jung-Hwan Lee, O. Huck, N. Benkirane-Jessel, Hae-Won Kim, M. Ramalingam","doi":"10.1155/2023/1277760","DOIUrl":"https://doi.org/10.1155/2023/1277760","url":null,"abstract":"While treated dentin matrix (TDM) has been used for regeneration of dental tissues, the quality and quantity of regenerated periodontal tissue structure are suboptimal. The present study was undertaken to test whether the combined use of the TDM with dental follicle cells (DFCs) and Hertwig’s epithelial root sheath (HERS) cells enhances the regeneration of periodontal structures in a nondental microenvironment. TDMs were fabricated from 3-month-old Sprague–Dawley (SD) rats. DFCs and HERS cells were isolated from postnatal 7-day SD rats. Purified DFCs and HERS cells, both in combination or alone, were seeded and cultured on TDM in vitro and characterized. The cell-seeded TDMs were subsequently implanted into a 3-month-old rat greater omentum for 6 weeks, and further histological evaluation was performed. The results showed that cells grew well on the surface of TDMs, and mineralized nodules could be seen, especially in the HERS + DFCs group. After transplantation in rat omentum, periodontal ligament-like fibers and cementum-like structures were observed around the TDM in 1/3 of the samples in both the HERS group and the DFCs group and in 2/3 of the samples in the HERS + DFCs group, while almost no attached tissue formation was found in the TDM only group. The formed cementum width and the periodontal ligament length were significantly larger in the HERS + DFCs group. The periodontal ligament-like fibers in the HERS + DFCs group were orderly arranged and attached to the cementum-like tissues, which resembled the cementum-periodontal structure. Therefore, the combined use of DFCs, TDM, and HERS cells may be a promising strategy for the regeneration of the periodontal structures, especially in the nondental microenvironment.","PeriodicalId":202,"journal":{"name":"Journal of Tissue Engineering and Regenerative Medicine","volume":null,"pages":null},"PeriodicalIF":3.3,"publicationDate":"2023-07-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"46841567","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}
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