Isabel Haferland, Andreas Pinter, Tanja Rossmanith, Sandra Diehl, Claudia Buerger, Tanja Ickelsheimer, Roland Kaufmann, Anke Koenig
Hidradenitis suppurativa (HS) is a chronic inflammatory skin disease. Patients can present with inflammatory nodules, abscesses up to fistulas, or sinus tracts in intertriginous body parts. Occlusion of the sebaceous gland unit leads to its rupture, with a subsequent exuberant immune response. Given there is still no causative therapy, to better understand HS and develop novel therapeutic concepts, research activities in the HS field are constantly growing. Primary skin cells, blood cells, and ex vivo explant cultures from HS patients have been previously used as HS cell culture models. In vitro reconstituted epidermal models are established to study inflammatory dermatoses, such as psoriasis or atopic dermatitis. For HS, the exploration of epidermis models would be an excellent addition, e.g., biomarkers or barrier function in testing new topic treatment options. We therefore established a stratified in vitro HS epidermis model based on primary cells from HS lesions. After isolating keratinocytes from lesional skin, we cultured them submerged in a transwell system. To induce differentiation, we then lifted them to the air-liquid interface. Immunohistochemical staining demonstrated that our HS-epidermis model meets the expected differentiation pattern. In addition, we detected the secretion of the inflammatory cytokines interleukin-1β and TNF-α.
{"title":"A Novel Epidermis Model Using Primary Hidradenitis Suppurativa Keratinocytes","authors":"Isabel Haferland, Andreas Pinter, Tanja Rossmanith, Sandra Diehl, Claudia Buerger, Tanja Ickelsheimer, Roland Kaufmann, Anke Koenig","doi":"10.1155/2024/4363876","DOIUrl":"10.1155/2024/4363876","url":null,"abstract":"<div>\u0000 <p>Hidradenitis suppurativa (HS) is a chronic inflammatory skin disease. Patients can present with inflammatory nodules, abscesses up to fistulas, or sinus tracts in intertriginous body parts. Occlusion of the sebaceous gland unit leads to its rupture, with a subsequent exuberant immune response. Given there is still no causative therapy, to better understand HS and develop novel therapeutic concepts, research activities in the HS field are constantly growing. Primary skin cells, blood cells, and <i>ex vivo</i> explant cultures from HS patients have been previously used as HS cell culture models. <i>In vitro</i> reconstituted epidermal models are established to study inflammatory dermatoses, such as psoriasis or atopic dermatitis. For HS, the exploration of epidermis models would be an excellent addition, e.g., biomarkers or barrier function in testing new topic treatment options. We therefore established a stratified <i>in vitro</i> HS epidermis model based on primary cells from HS lesions. After isolating keratinocytes from lesional skin, we cultured them submerged in a transwell system. To induce differentiation, we then lifted them to the air-liquid interface. Immunohistochemical staining demonstrated that our HS-epidermis model meets the expected differentiation pattern. In addition, we detected the secretion of the inflammatory cytokines interleukin-1<i>β</i> and TNF-<i>α</i>.</p>\u0000 </div>","PeriodicalId":202,"journal":{"name":"Journal of Tissue Engineering and Regenerative Medicine","volume":"2024 1","pages":""},"PeriodicalIF":3.3,"publicationDate":"2024-02-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1155/2024/4363876","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140426706","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}
In rheumatoid arthritis, dysregulated cytokine signaling has been implicated as a primary factor in chronic inflammation. Many antirheumatic and biological therapies are used to suppress joint inflammation, but despite these advances, effectiveness is not universal, and delivery is often at high doses, which can predispose patients to significant off-target effects. During chronic inflammation, the inappropriate regulation of signaling factors by macrophages accelerates the progression of disease by driving an imbalance of inflammatory cytokines, making macrophages an ideal cellular target. To develop a macrophage-based therapy to treat chronic inflammation, we engineered a novel induced pluripotent stem cell (iPSC)-derived macrophage capable of delivering soluble TNF receptor 1 (sTNFR1), an anti-inflammatory biologic inhibitor of tumor necrosis factor alpha (TNF-α), in an autoregulated manner in response to TNF-α. Murine iPSCs were differentiated into macrophages (iMACs) over a 17-day optimized protocol with continued successful differentiation confirmed at key timepoints. Varying inflammatory and immunomodulatory stimuli demonstrated traditional macrophage function and phenotypes. In response to TNF-α, therapeutic iMACs produced high levels of sTNFR1 in an autoregulated manner, which inhibited inflammatory signaling. This self-regulating iMAC system demonstrated the potential for macrophage-based drug delivery as a novel therapeutic approach for a variety of chronic inflammatory diseases.
{"title":"Genetically Engineered Macrophages Derived from iPSCs for Self-Regulating Delivery of Anti-Inflammatory Biologic Drugs","authors":"Molly Klimak, Farshid Guilak","doi":"10.1155/2024/6201728","DOIUrl":"10.1155/2024/6201728","url":null,"abstract":"<div>\u0000 <p>In rheumatoid arthritis, dysregulated cytokine signaling has been implicated as a primary factor in chronic inflammation. Many antirheumatic and biological therapies are used to suppress joint inflammation, but despite these advances, effectiveness is not universal, and delivery is often at high doses, which can predispose patients to significant off-target effects. During chronic inflammation, the inappropriate regulation of signaling factors by macrophages accelerates the progression of disease by driving an imbalance of inflammatory cytokines, making macrophages an ideal cellular target. To develop a macrophage-based therapy to treat chronic inflammation, we engineered a novel induced pluripotent stem cell (iPSC)-derived macrophage capable of delivering soluble TNF receptor 1 (sTNFR1), an anti-inflammatory biologic inhibitor of tumor necrosis factor alpha (TNF-<i>α</i>), in an autoregulated manner in response to TNF-<i>α</i>. Murine iPSCs were differentiated into macrophages (iMACs) over a 17-day optimized protocol with continued successful differentiation confirmed at key timepoints. Varying inflammatory and immunomodulatory stimuli demonstrated traditional macrophage function and phenotypes. In response to TNF-<i>α</i>, therapeutic iMACs produced high levels of sTNFR1 in an autoregulated manner, which inhibited inflammatory signaling. This self-regulating iMAC system demonstrated the potential for macrophage-based drug delivery as a novel therapeutic approach for a variety of chronic inflammatory diseases.</p>\u0000 </div>","PeriodicalId":202,"journal":{"name":"Journal of Tissue Engineering and Regenerative Medicine","volume":"2024 1","pages":""},"PeriodicalIF":3.3,"publicationDate":"2024-01-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1155/2024/6201728","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139380725","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}
Liza A Bruk, Xin Fan, Jayde L. Resnick, Morgan V. DiLeo
Chronic tympanic membrane (TM) perforation increases patient susceptibility to infection, hearing loss, and other side effects. Current clinical treatment, surgical grafting, can result in detrimental side effects including nerve damage, dizziness, or hearing loss. Therefore, it is essential to develop novel therapeutic procedures that can induce or accelerate healing in minimally or noninvasive approaches. Cell-free therapies have safety advantages over stem cells and are logistically favorable for clinical use. The regenerative potential by mesenchymal stem cell-conditioned media (CM) has been promising. In this study, poly(lactic-co-glycolic acid) (PLGA) microspheres with CM encapsulated have been developed as a cell-free alternative regenerative treatment for TM perforation. The results suggest that the PLGA microspheres were capable of encapsulating and releasing CM for up to 21 days. The in vitro scratch wound proliferation assays showed increased wound healing ability of CM-loaded microspheres. In vivo guinea pig models treated with CM drops and CM-loaded microspheres using a thermoresponsive gel carrier demonstrated potential for wound healing in TM perforation. These studies provide a basis for further examination of the delivery of stem cell CM and investigation of time-dependent wound healing, long-term ototoxicity, and hearing restoration.
慢性鼓膜（TM）穿孔会增加患者感染、听力损失和其他副作用的风险。目前的临床治疗方法--手术移植可能会导致有害的副作用，包括神经损伤、头晕或听力下降。因此，有必要开发新型治疗程序，以微创或无创方式诱导或加速愈合。与干细胞相比，无细胞疗法具有安全优势，而且在临床应用上也很方便。间充质干细胞调理介质（CM）的再生潜力很有希望。本研究开发了包裹间充质干细胞的聚乳酸-共聚乙醇酸（PLGA）微球，作为治疗颞下颌关节穿孔的无细胞替代再生疗法。结果表明，PLGA 微球能够包裹和释放 CM 长达 21 天。体外划痕伤口增殖试验表明，含有 CM 的微球能增强伤口愈合能力。使用 CM 滴剂和使用热致伸缩凝胶载体的 CM 负载微球治疗的体内豚鼠模型显示了 TM 穿孔伤口愈合的潜力。这些研究为进一步研究干细胞CM的输送、调查随时间变化的伤口愈合、长期耳毒性和听力恢复提供了基础。
{"title":"Controlled Release of Mesenchymal Stem Cell-Conditioned Media from a Microsphere/Gel-Based Drug Delivery System for Wound Healing of Tympanic Membrane Perforations","authors":"Liza A Bruk, Xin Fan, Jayde L. Resnick, Morgan V. DiLeo","doi":"10.1155/2023/6039254","DOIUrl":"https://doi.org/10.1155/2023/6039254","url":null,"abstract":"Chronic tympanic membrane (TM) perforation increases patient susceptibility to infection, hearing loss, and other side effects. Current clinical treatment, surgical grafting, can result in detrimental side effects including nerve damage, dizziness, or hearing loss. Therefore, it is essential to develop novel therapeutic procedures that can induce or accelerate healing in minimally or noninvasive approaches. Cell-free therapies have safety advantages over stem cells and are logistically favorable for clinical use. The regenerative potential by mesenchymal stem cell-conditioned media (CM) has been promising. In this study, poly(lactic-co-glycolic acid) (PLGA) microspheres with CM encapsulated have been developed as a cell-free alternative regenerative treatment for TM perforation. The results suggest that the PLGA microspheres were capable of encapsulating and releasing CM for up to 21 days. The in vitro scratch wound proliferation assays showed increased wound healing ability of CM-loaded microspheres. In vivo guinea pig models treated with CM drops and CM-loaded microspheres using a thermoresponsive gel carrier demonstrated potential for wound healing in TM perforation. These studies provide a basis for further examination of the delivery of stem cell CM and investigation of time-dependent wound healing, long-term ototoxicity, and hearing restoration.","PeriodicalId":202,"journal":{"name":"Journal of Tissue Engineering and Regenerative Medicine","volume":"32 4","pages":""},"PeriodicalIF":3.3,"publicationDate":"2023-12-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139132491","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}
Anyesha Sarkar, Shanta M. Messerli, Md Moin Uddin Talukder, M. Messerli
Therapeutic electric fields (EFs) are applied to the epidermis to accelerate the healing of chronic epidermal wounds and promote skin transplantation. While research has emphasized understanding the role of EFs in polarizing the migration of superficial epidermal cells, there are no reports describing the effect of EFs on polarization of the underlying vasculature. We explored the effects of EFs on the growth of endothelial sprouts, precursors to functional blood vessels. We discovered that DC EFs of the same magnitude near wounded epidermis polarize initiation, growth, and turning of endothelial sprouts toward the anode. While EFs polarize sprouts, they do not change the frequency of primary sprout or branch formation. Unidirectional electrical pulses also polarize sprouts based on their time-averaged EF magnitude. Sprout polarization occurs antiparallel to the direction of electrically driven water flow (electro-osmosis) and is consistent with the direction of sprout polarization induced by pressure-driven flow. These results support the role of EFs in controlling the direction of neovascularization during the healing of soft tissues and tissue engineering.
治疗性电场（EF）被应用于表皮，以加速慢性表皮伤口的愈合并促进皮肤移植。虽然研究强调了解电场在极化表皮细胞迁移中的作用，但还没有报告描述电场对底层血管极化的影响。我们探讨了 EFs 对内皮芽（功能性血管的前体）生长的影响。我们发现，受伤表皮附近同等强度的直流环流可极化内皮萌芽的启动、生长和转向阳极。虽然直流电极化了萌芽，但并没有改变初级萌芽或分支形成的频率。单向电脉冲也会根据其时间平均 EF 幅值极化萌芽。萌芽极化与电驱动水流（电渗）的方向相反，与压力驱动水流诱导的萌芽极化方向一致。这些结果支持 EF 在软组织愈合和组织工程中控制新生血管方向的作用。
{"title":"Applied Electric Fields Polarize Initiation and Growth of Endothelial Sprouts","authors":"Anyesha Sarkar, Shanta M. Messerli, Md Moin Uddin Talukder, M. Messerli","doi":"10.1155/2023/6331148","DOIUrl":"https://doi.org/10.1155/2023/6331148","url":null,"abstract":"Therapeutic electric fields (EFs) are applied to the epidermis to accelerate the healing of chronic epidermal wounds and promote skin transplantation. While research has emphasized understanding the role of EFs in polarizing the migration of superficial epidermal cells, there are no reports describing the effect of EFs on polarization of the underlying vasculature. We explored the effects of EFs on the growth of endothelial sprouts, precursors to functional blood vessels. We discovered that DC EFs of the same magnitude near wounded epidermis polarize initiation, growth, and turning of endothelial sprouts toward the anode. While EFs polarize sprouts, they do not change the frequency of primary sprout or branch formation. Unidirectional electrical pulses also polarize sprouts based on their time-averaged EF magnitude. Sprout polarization occurs antiparallel to the direction of electrically driven water flow (electro-osmosis) and is consistent with the direction of sprout polarization induced by pressure-driven flow. These results support the role of EFs in controlling the direction of neovascularization during the healing of soft tissues and tissue engineering.","PeriodicalId":202,"journal":{"name":"Journal of Tissue Engineering and Regenerative Medicine","volume":"55 1","pages":""},"PeriodicalIF":3.3,"publicationDate":"2023-12-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139162934","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}
Femke Bellen, Elisa Carbone, Pieter Baatsen, E. A. Jones, F. Kabirian, Ruth Heying
Polycaprolactone (PCL) is a promising material for the fabrication of alternatives to autologous grafts used in coronary bypass surgery. PCL biodegrades over time, allowing cells to infiltrate the polymeric matrix, replacing the biodegrading graft, and creating a fully functional vessel constituted of autologous tissue. However, the high hydrophobicity of PCL is associated with poor cell affinity. Surface modification of PCL can increase this cell affinity, making PCL an improved scaffold material for acellular vascular grafts. In this study, the surface of PCL films was modified by hydrolysis, aminolysis, and the combination thereof to introduce carboxyl, hydroxyl, and amino groups on the surface. Only the hydrolyzed films exhibited a significant increase in their hydrophilicity, although further testing showed that all aminolysis conditions had amino groups on the surface. Furthermore, in vitro experiments with human umbilical endothelial cells (HUVECs) were performed to assess changes in cell affinity for PCL due to the surface treatments. PCL treated with sodium hydroxide (NaOH), a hydrolysis reaction, showed a significant increase in endothelial cell adhesion after 24 hours with a significant increase in cell survival after 72 hours. Thus, NaOH treatment improves the biocompatibility and endothelialization of PCL, creating a competent candidate for artificial, acellular, biodegradable vascular grafts.
{"title":"Improvement of Endothelial Cell-Polycaprolactone Interaction through Surface Modification via Aminolysis, Hydrolysis, and a Combined Approach","authors":"Femke Bellen, Elisa Carbone, Pieter Baatsen, E. A. Jones, F. Kabirian, Ruth Heying","doi":"10.1155/2023/5590725","DOIUrl":"https://doi.org/10.1155/2023/5590725","url":null,"abstract":"Polycaprolactone (PCL) is a promising material for the fabrication of alternatives to autologous grafts used in coronary bypass surgery. PCL biodegrades over time, allowing cells to infiltrate the polymeric matrix, replacing the biodegrading graft, and creating a fully functional vessel constituted of autologous tissue. However, the high hydrophobicity of PCL is associated with poor cell affinity. Surface modification of PCL can increase this cell affinity, making PCL an improved scaffold material for acellular vascular grafts. In this study, the surface of PCL films was modified by hydrolysis, aminolysis, and the combination thereof to introduce carboxyl, hydroxyl, and amino groups on the surface. Only the hydrolyzed films exhibited a significant increase in their hydrophilicity, although further testing showed that all aminolysis conditions had amino groups on the surface. Furthermore, in vitro experiments with human umbilical endothelial cells (HUVECs) were performed to assess changes in cell affinity for PCL due to the surface treatments. PCL treated with sodium hydroxide (NaOH), a hydrolysis reaction, showed a significant increase in endothelial cell adhesion after 24 hours with a significant increase in cell survival after 72 hours. Thus, NaOH treatment improves the biocompatibility and endothelialization of PCL, creating a competent candidate for artificial, acellular, biodegradable vascular grafts.","PeriodicalId":202,"journal":{"name":"Journal of Tissue Engineering and Regenerative Medicine","volume":"125 S179","pages":""},"PeriodicalIF":3.3,"publicationDate":"2023-12-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139006347","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}
Minae Kawasaki, Gerald D. Dykstra, C. McConnel, C. Burbick, Y. Ambrosini
Recent progress in bovine intestinal organoid research has expanded opportunities for creating improved in vitro models to study intestinal physiology and pathology. However, the establishment of a culture condition capable of generating organoids from all segments of the cattle intestine has remained elusive. Although previous research has described the development of bovine jejunal, ileal, and colonic organoids, this study marks the first report of successful bovine duodenal and rectal organoid development. Maintenance of these organoids through serial passages and cryopreservation was achieved, with higher success rates observed in large intestinal organoids compared to their small intestinal counterparts. A novel approach involving the use of biopsy forceps during initial tissue sampling streamlined the subsequent tissue processing, simplifying the procedure compared to previously established protocols in cattle. In addition, our study introduced a more cost-effective culture medium based on advanced DMEM/F12, diverging from frequently used commercially available organoid culture media. This enhancement improves the accessibility to organoid technology by reducing culture costs. Crucially, the derived organoids from the jejunum, ileum, colon, and rectum faithfully preserved the structural, cellular, and genetic characteristics of the in vivo intestinal tissue. This research underscores the significant potential of adult bovine intestinal organoids as a physiologically and morphologically relevant in vitro model. Such organoids provide a renewable and sustainable resource for a broad spectrum of studies, encompassing investigations into normal intestinal physiology in cattle and the intricate host-pathogen interactions of clinically and economically significant enteric pathogens.
{"title":"Adult Bovine-Derived Small and Large Intestinal Organoids: In Vitro Development and Maintenance","authors":"Minae Kawasaki, Gerald D. Dykstra, C. McConnel, C. Burbick, Y. Ambrosini","doi":"10.1155/2023/3095002","DOIUrl":"https://doi.org/10.1155/2023/3095002","url":null,"abstract":"Recent progress in bovine intestinal organoid research has expanded opportunities for creating improved in vitro models to study intestinal physiology and pathology. However, the establishment of a culture condition capable of generating organoids from all segments of the cattle intestine has remained elusive. Although previous research has described the development of bovine jejunal, ileal, and colonic organoids, this study marks the first report of successful bovine duodenal and rectal organoid development. Maintenance of these organoids through serial passages and cryopreservation was achieved, with higher success rates observed in large intestinal organoids compared to their small intestinal counterparts. A novel approach involving the use of biopsy forceps during initial tissue sampling streamlined the subsequent tissue processing, simplifying the procedure compared to previously established protocols in cattle. In addition, our study introduced a more cost-effective culture medium based on advanced DMEM/F12, diverging from frequently used commercially available organoid culture media. This enhancement improves the accessibility to organoid technology by reducing culture costs. Crucially, the derived organoids from the jejunum, ileum, colon, and rectum faithfully preserved the structural, cellular, and genetic characteristics of the in vivo intestinal tissue. This research underscores the significant potential of adult bovine intestinal organoids as a physiologically and morphologically relevant in vitro model. Such organoids provide a renewable and sustainable resource for a broad spectrum of studies, encompassing investigations into normal intestinal physiology in cattle and the intricate host-pathogen interactions of clinically and economically significant enteric pathogens.","PeriodicalId":202,"journal":{"name":"Journal of Tissue Engineering and Regenerative Medicine","volume":"38 1","pages":""},"PeriodicalIF":3.3,"publicationDate":"2023-11-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139232648","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}
Anna-Laura Nelson, Kelsey M. O’Hara, Philip C. Nolte, N. Fukase, Yoichi Murata, Anna-Katharina Nolte, Johnny Huard, David L. Bernholt, Peter J. Millett, C. Bahney
Rotator cuff tears are a common soft tissue injury that can significantly decrease function of the shoulder and cause severe pain. Despite progress in surgical technique, rotator cuff repairs (RCRs) do not always heal efficiently. Many failures occur at the bone-tendon interface as a result of poor healing capacity of the tendon and failure to regenerate the native histological anatomy of the enthesis. While allografts are commercially available, clinical use is limited as they do not stimulate tissue regeneration and are associated with a structural failure of up to 40% in re-tear cases. Novel tissue engineering strategies are being developed with promise, but most involve addition of cells and/or growth factors which extends the timeline for clinical translation. Thus, there exists a significant unmet clinical need for easily translatable surgical augmentation approaches that can improve healing in RCR. Here we describe the development of a decellularized tendon matrix (DTM) putty that preserves native tendon bioactivity using a novel processing technique. In vitro, DTM promoted proliferation of tenocytes and adipose-derived stem cells with an increase in expression-specific transcription factors seen during enthesis development, Scleraxis and Sox9. When placed in a rabbit model of a chronic rotator cuff tear, DTM improved histological tissue repair by promoting calcification at the bone-tendon interface more similar to the normal fibrocartilaginous enthesis. Taken together, these data indicate that the engineered DTM putty retains a pro-regenerative bioactivity that presents a promising translational strategy for improving healing at the enthesis.
{"title":"Engineered Decellularized Tendon Matrix Putty Preserves Native Tendon Bioactivity to Promote Cell Proliferation and Enthesis Repair","authors":"Anna-Laura Nelson, Kelsey M. O’Hara, Philip C. Nolte, N. Fukase, Yoichi Murata, Anna-Katharina Nolte, Johnny Huard, David L. Bernholt, Peter J. Millett, C. Bahney","doi":"10.1155/2023/4665795","DOIUrl":"https://doi.org/10.1155/2023/4665795","url":null,"abstract":"Rotator cuff tears are a common soft tissue injury that can significantly decrease function of the shoulder and cause severe pain. Despite progress in surgical technique, rotator cuff repairs (RCRs) do not always heal efficiently. Many failures occur at the bone-tendon interface as a result of poor healing capacity of the tendon and failure to regenerate the native histological anatomy of the enthesis. While allografts are commercially available, clinical use is limited as they do not stimulate tissue regeneration and are associated with a structural failure of up to 40% in re-tear cases. Novel tissue engineering strategies are being developed with promise, but most involve addition of cells and/or growth factors which extends the timeline for clinical translation. Thus, there exists a significant unmet clinical need for easily translatable surgical augmentation approaches that can improve healing in RCR. Here we describe the development of a decellularized tendon matrix (DTM) putty that preserves native tendon bioactivity using a novel processing technique. In vitro, DTM promoted proliferation of tenocytes and adipose-derived stem cells with an increase in expression-specific transcription factors seen during enthesis development, Scleraxis and Sox9. When placed in a rabbit model of a chronic rotator cuff tear, DTM improved histological tissue repair by promoting calcification at the bone-tendon interface more similar to the normal fibrocartilaginous enthesis. Taken together, these data indicate that the engineered DTM putty retains a pro-regenerative bioactivity that presents a promising translational strategy for improving healing at the enthesis.","PeriodicalId":202,"journal":{"name":"Journal of Tissue Engineering and Regenerative Medicine","volume":"14 3","pages":""},"PeriodicalIF":3.3,"publicationDate":"2023-11-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139269084","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}
Regenerative medicine using lymphatic vascular engineering is a promising approach for treating lymphedema. However, its development lags behind that of artificial blood vascular tissue for ischemic diseases. In this study, we constructed artificial 3D lymphatic vascular tissue, termed ASCLT, by co-cultivation of ECM-nanofilm-coated human adipose tissue-derived mesenchymal stromal cells (hASCs) and human dermal lymphatic endothelial cells (HDLECs). The effect of hASCs in lymphatic vessel network formation was evaluated by comparison with the tissue based on fibroblasts, termed FbLT. Our results showed that the density of lymphatic vascular network in ASCLT was higher than that in FbLT, demonstrating a promoting effect of hASCs on lymphatic vascular formation. This result was also supported by higher levels of lymphangiogenesis-promoting factors, such as bFGF, HGF, and VEGF-A in ASCLT than in FbLT. To evaluate the therapeutic effects, FbLTs and ASCLTs were subcutaneously transplanted to mouse hindlimb lymphatic drainage interruption models by removal of popliteal and subiliac lymph nodes. Despite the restricted engraftment of lymphatic vessels, ASCLT promoted regeneration of irregular and diverse lymphatic drainage in the skin, as visualized by indocyanine green imaging. Moreover, transplantation of ASCLT to the popliteal lymph node resection area also resulted in lymphatic drainage regeneration. Histological analysis of the generated drainage visualized by FITC-dextran injection revealed that the drainage was localized in the subcutaneous area shallower than the dermal muscle. These findings demonstrate that ASCLT promotes lymphatic drainage in vivo and that hASCs can serve as an autologous source for treatment of secondary lymphedema by tissue engineering.
{"title":"Lymphatic Drainage-Promoting Effects by Engraftment of Artificial Lymphatic Vascular Tissue Based on Human Adipose Tissue-Derived Mesenchymal Stromal Cells in Mice","authors":"Yoshiya Asano, Hiroshi Shimoda, Daisuke Okano, Michiya Matsusaki, Mitsuru Akashi","doi":"10.1155/2023/7626767","DOIUrl":"https://doi.org/10.1155/2023/7626767","url":null,"abstract":"Regenerative medicine using lymphatic vascular engineering is a promising approach for treating lymphedema. However, its development lags behind that of artificial blood vascular tissue for ischemic diseases. In this study, we constructed artificial 3D lymphatic vascular tissue, termed ASCLT, by co-cultivation of ECM-nanofilm-coated human adipose tissue-derived mesenchymal stromal cells (hASCs) and human dermal lymphatic endothelial cells (HDLECs). The effect of hASCs in lymphatic vessel network formation was evaluated by comparison with the tissue based on fibroblasts, termed FbLT. Our results showed that the density of lymphatic vascular network in ASCLT was higher than that in FbLT, demonstrating a promoting effect of hASCs on lymphatic vascular formation. This result was also supported by higher levels of lymphangiogenesis-promoting factors, such as bFGF, HGF, and VEGF-A in ASCLT than in FbLT. To evaluate the therapeutic effects, FbLTs and ASCLTs were subcutaneously transplanted to mouse hindlimb lymphatic drainage interruption models by removal of popliteal and subiliac lymph nodes. Despite the restricted engraftment of lymphatic vessels, ASCLT promoted regeneration of irregular and diverse lymphatic drainage in the skin, as visualized by indocyanine green imaging. Moreover, transplantation of ASCLT to the popliteal lymph node resection area also resulted in lymphatic drainage regeneration. Histological analysis of the generated drainage visualized by FITC-dextran injection revealed that the drainage was localized in the subcutaneous area shallower than the dermal muscle. These findings demonstrate that ASCLT promotes lymphatic drainage in vivo and that hASCs can serve as an autologous source for treatment of secondary lymphedema by tissue engineering.","PeriodicalId":202,"journal":{"name":"Journal of Tissue Engineering and Regenerative Medicine","volume":"27 6","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2023-11-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"135584787","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}
Aishi Song, Wei Wang, Yuying Zhang, Peng Zhou, Jiaxing Li, Jean de Dieu Habimana, Omar Mukama, Wei Xie, Sihao Deng, Shusheng Zhang, Ming Li, Bin Ni, Yabing Tang, Xiao-Xin Yan, Jufang Huang, Zhiyuan Li
Background. Periodontitis is characterized by bone resorption and periodontal tissue destruction owing to oral microbiota, mechanical stress, and systemic diseases such as diabetes mellitus. Human dental pulp mesenchymal stem cells (hDPMSCs) were analyzed as potential candidates for periodontal tissue regeneration. Acetylsalicylic acid (ASA), also known as aspirin, has been shown to promote osteogenic differentiation of mesenchymal stem cells. We investigated the effect of ASA pretreatment on periodontitis in order to achieve a more appealing prognosis of bone resorption. Methods. The effect of ASA on cell proliferation was detected by the CCK-8 assay, and alkaline phosphatase (ALP) staining, alizarin red staining (ARS), and western blot were used to investigate the effect of different ASA concentrations on hDPMSCs’ osteogenic differentiation and possible signaling pathways. Periodontitis was induced for 4 weeks. Stem cells pretreated with 50 µg/mL of ASA were transplanted into six-week-old male Sprague-Dawley rats by local and systemic injection once a week for two weeks. Four weeks after cell therapy, the rats were sacrificed for sampling to complete the molecular and morphological experiments. Results. In vitro experiments revealed that 50 µg/mL of ASA had a significant effect on cell osteogenic differentiation. That is, when ASA was administered, the MAPK signaling pathway was activated. Notably, further vivo experiments revealed that ASA-hDPMSCs increased the area of bone regeneration and the OPG/RANKL ratio, suppressed TNF-α and IL-1 expression, and promote alveolar bone repair. Conclusion. Our study extends the findings of previous research, firstly demonstrating that the use of ASA-pretreated hDPMSCs offers a novel therapy for the treatment of periodontitis for future clinical application.
{"title":"Acetylsalicylic Acid Promotes Osteogenic Differentiation of Human Dental Pulp Mesenchymal Stem Cells and Regeneration of Alveolar Bone in Experimental Periodontitis Rats","authors":"Aishi Song, Wei Wang, Yuying Zhang, Peng Zhou, Jiaxing Li, Jean de Dieu Habimana, Omar Mukama, Wei Xie, Sihao Deng, Shusheng Zhang, Ming Li, Bin Ni, Yabing Tang, Xiao-Xin Yan, Jufang Huang, Zhiyuan Li","doi":"10.1155/2023/3077814","DOIUrl":"https://doi.org/10.1155/2023/3077814","url":null,"abstract":"Background. Periodontitis is characterized by bone resorption and periodontal tissue destruction owing to oral microbiota, mechanical stress, and systemic diseases such as diabetes mellitus. Human dental pulp mesenchymal stem cells (hDPMSCs) were analyzed as potential candidates for periodontal tissue regeneration. Acetylsalicylic acid (ASA), also known as aspirin, has been shown to promote osteogenic differentiation of mesenchymal stem cells. We investigated the effect of ASA pretreatment on periodontitis in order to achieve a more appealing prognosis of bone resorption. Methods. The effect of ASA on cell proliferation was detected by the CCK-8 assay, and alkaline phosphatase (ALP) staining, alizarin red staining (ARS), and western blot were used to investigate the effect of different ASA concentrations on hDPMSCs’ osteogenic differentiation and possible signaling pathways. Periodontitis was induced for 4 weeks. Stem cells pretreated with 50 µg/mL of ASA were transplanted into six-week-old male Sprague-Dawley rats by local and systemic injection once a week for two weeks. Four weeks after cell therapy, the rats were sacrificed for sampling to complete the molecular and morphological experiments. Results. In vitro experiments revealed that 50 µg/mL of ASA had a significant effect on cell osteogenic differentiation. That is, when ASA was administered, the MAPK signaling pathway was activated. Notably, further vivo experiments revealed that ASA-hDPMSCs increased the area of bone regeneration and the OPG/RANKL ratio, suppressed TNF-α and IL-1 expression, and promote alveolar bone repair. Conclusion. Our study extends the findings of previous research, firstly demonstrating that the use of ASA-pretreated hDPMSCs offers a novel therapy for the treatment of periodontitis for future clinical application.","PeriodicalId":202,"journal":{"name":"Journal of Tissue Engineering and Regenerative Medicine","volume":"12 9","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2023-11-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"135818342","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}
Anna Hillenmayer, Laura D. Strehle, Christina Hilterhaus, Andreas Ohlmann, Christian M. Wertheimer, Armin Wolf
Proliferative vitreoretinopathy (PVR) as a rare fibrotic ocular disease is the main reason for failure of retinal detachment surgery and a reduced prognosis following surgery. Amniotic membrane (AM) is a versatile surgical tool for tissue stabilization, antifibrotic properties, and regeneration. Initial clinical case studies now demonstrated intravitreal tolerance as well as good anatomical and functional results for degenerative retinal diseases. Due to its diverse wound healing properties, AM could have promoting, suppressive, or no effects on PVR. To illuminate the potential of epiretinal AM transplantation in complex retinal detachment cases, we investigated its influence on human primary PVR (hPVR) cells in vitro. In our cell culture study, hPVR cells were isolated from surgically removed PVR membranes. Following incubation with AM for 48 h, AM-incubated hPVR showed significantly reduced proliferation (BrdU-ELISA; ), migration (Boyden chamber, scratch assay; = 0.003 and ), and cell adhesion ( = 0.005). Collagen contraction was nearly unaffected ( = 0.04), and toxicity (histone-complexed DNA ELISA, WST-1 assay, and life/dead staining) was excluded. Next, immunofluorescence showed a myofibroblastic phenotype with reduced expression of fibrosis markers in AM-incubated cells, which was confirmed by Western blot analysis. In the proteomics assay, AM significantly regulated proteins by a more than 2-fold increase in expression which were related to the cytoskeleton, lipid metabolism, cell-matrix contraction, and protein folding. In conclusion, this in vitro work suggests no induction of fibrosis and other key steps in the pathogenesis of PVR through AM but rather inhibiting properties of profibrotic cell behavior, making it a possible candidate for suppression of PVR. Further clinical studies are necessary to evaluate the therapeutic relevance.
{"title":"Epiretinal Amniotic Membrane Influences the Cellular Behavior of Profibrotic Dedifferentiated Cells of Proliferative Vitreoretinopathy In Vitro","authors":"Anna Hillenmayer, Laura D. Strehle, Christina Hilterhaus, Andreas Ohlmann, Christian M. Wertheimer, Armin Wolf","doi":"10.1155/2023/8820844","DOIUrl":"https://doi.org/10.1155/2023/8820844","url":null,"abstract":"Proliferative vitreoretinopathy (PVR) as a rare fibrotic ocular disease is the main reason for failure of retinal detachment surgery and a reduced prognosis following surgery. Amniotic membrane (AM) is a versatile surgical tool for tissue stabilization, antifibrotic properties, and regeneration. Initial clinical case studies now demonstrated intravitreal tolerance as well as good anatomical and functional results for degenerative retinal diseases. Due to its diverse wound healing properties, AM could have promoting, suppressive, or no effects on PVR. To illuminate the potential of epiretinal AM transplantation in complex retinal detachment cases, we investigated its influence on human primary PVR (hPVR) cells in vitro. In our cell culture study, hPVR cells were isolated from surgically removed PVR membranes. Following incubation with AM for 48 h, AM-incubated hPVR showed significantly reduced proliferation (BrdU-ELISA; <math xmlns=\"http://www.w3.org/1998/Math/MathML\" id=\"M1\"> <mi>p</mi> <mo><</mo> <mn>0.001</mn> </math> ), migration (Boyden chamber, scratch assay; <math xmlns=\"http://www.w3.org/1998/Math/MathML\" id=\"M2\"> <mi>p</mi> </math> = 0.003 and <math xmlns=\"http://www.w3.org/1998/Math/MathML\" id=\"M3\"> <mi>p</mi> <mo><</mo> <mn>0.001</mn> </math> ), and cell adhesion ( <math xmlns=\"http://www.w3.org/1998/Math/MathML\" id=\"M4\"> <mi>p</mi> </math> = 0.005). Collagen contraction was nearly unaffected ( <math xmlns=\"http://www.w3.org/1998/Math/MathML\" id=\"M5\"> <mi>p</mi> </math> = 0.04), and toxicity (histone-complexed DNA ELISA, WST-1 assay, and life/dead staining) was excluded. Next, immunofluorescence showed a myofibroblastic phenotype with reduced expression of fibrosis markers in AM-incubated cells, which was confirmed by Western blot analysis. In the proteomics assay, AM significantly regulated proteins by a more than 2-fold increase in expression which were related to the cytoskeleton, lipid metabolism, cell-matrix contraction, and protein folding. In conclusion, this in vitro work suggests no induction of fibrosis and other key steps in the pathogenesis of PVR through AM but rather inhibiting properties of profibrotic cell behavior, making it a possible candidate for suppression of PVR. Further clinical studies are necessary to evaluate the therapeutic relevance.","PeriodicalId":202,"journal":{"name":"Journal of Tissue Engineering and Regenerative Medicine","volume":"30 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2023-10-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"135825128","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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