Combination Of Growth Factors Research Articles

MP33-07 ANTI-PROGRAMMED DEATH RECEPTOR 1 PLUS ANTI-EPIDERMAL GROWTH FACTOR RECEPTOR ANTIBODIES AND COMBINATION CHEMOTHERAPY FOR RELAPSED LOCALLY ADVANCED AND/OR METASTATIC PENILE SQUAMOUS CELL CARCINOMA

You have accessJournal of UrologyCME1 Apr 2023MP33-07 ANTI-PROGRAMMED DEATH RECEPTOR 1 PLUS ANTI-EPIDERMAL GROWTH FACTOR RECEPTOR ANTIBODIES AND COMBINATION CHEMOTHERAPY FOR RELAPSED LOCALLY ADVANCED AND/OR METASTATIC PENILE SQUAMOUS CELL CARCINOMA Longbin Xiong, Lichao Wei, Ting Xue, Ru Yan, Tingyu Liu, Lijuan Jiang, Huali Ma, Yanxia Shi, Xin An, Yijun Zhang, Shengjie Guo, Kai Yao, Fangjian Zhou, and Hui Han Longbin XiongLongbin Xiong More articles by this author , Lichao WeiLichao Wei More articles by this author , Ting XueTing Xue More articles by this author , Ru YanRu Yan More articles by this author , Tingyu LiuTingyu Liu More articles by this author , Lijuan JiangLijuan Jiang More articles by this author , Huali MaHuali Ma More articles by this author , Yanxia ShiYanxia Shi More articles by this author , Xin AnXin An More articles by this author , Yijun ZhangYijun Zhang More articles by this author , Shengjie GuoShengjie Guo More articles by this author , Kai YaoKai Yao More articles by this author , Fangjian ZhouFangjian Zhou More articles by this author , and Hui HanHui Han More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000003266.07AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: Relapsed locally advanced and/or metastatic penile cancer has an extremely poor prognosis with a need for effective therapy. This study retrospectively estimated the efficacy and tolerability of anti-programmed death receptor-1 (PD1) plus anti-epidermal growth factor receptor (EGFR) antibodies and combination chemotherapy in relapsed locally advanced and/or metastatic penile squamous cell carcinoma. METHODS: A retrospective review of 30 relapsed locally advanced and/or metastatic (T4/N2-3/M1) penile squamous cell carcinoma received anti-PD1 plus anti-EGFR antibodies and cisplatin-based chemotherapy in Sun Yat-sen University Cancer Center from Jan 2018 to Aug 2022. Of 15 were chemotherapy-pretreated patients, and the remaining 15 were chemotherapy-naïve patients. Clinical responses were assessed using the Response Evaluation Criteria in Solid Tumours (RECIST v1.1) in 27 evaluable patients. Adverse events (AEs) were graded according to Common Terminology Criteria for Adverse Events (CTCAE v5.0). Progression-free survival (PFS) and overall survival (OS) were estimated using the Kaplan-Meier method. RESULTS: Partial responses were observed in 63% (17/27) patients, and 14 were chemotherapy-naïve patients. The objective response rate (ORR) was 100% (14/14) and 23% (3/13) in chemotherapy-naïve and chemotherapy-pretreated patients, respectively. Fourteen patients subsequently received consolidated surgery. Eight patients achieved pathological complete response, and six were chemotherapy-naïve patients. The median follow-up time was 11.4 (range: 2.3-36.2) months. The median PFS and OS were 15.6 (1.0-32.6) and 20.9 (10.5-30.3) months in chemotherapy-pretreated patients, respectively. However, these were not reached in overall cohort and chemotherapy-naïve patients. 2-year PFS (OS) was 53% (62%) in overall cohort, whereas 75% (79%) and 35% (47%) in chemotherapy-naïve and chemotherapy-pretreated patients, respectively. Treatment-related AEs were experienced by all patients, with 60% (18/30) experiencing a grade 3 or 4 AEs. The most common grade 3-4 treatment-related AEs were lymphopenia (40%), anemia (23%), and neutropenia (13%). CONCLUSIONS: Anti-PD1 antibody plus anti-EGFR antibody and cisplatin-based combination chemotherapy were effective, well-tolerated, and satisfying long-term survival outcome for relapsed locally advanced and/or metastatic penile squamous cell carcinoma. However, prolonged follow-up and large prospective clinical trials are needed to validate the findings on this important issue. Source of Funding: This study was supported by the National Natural Science Foundation of China (No. 81772755). © 2023 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 209Issue Supplement 4April 2023Page: e452 Advertisement Copyright & Permissions© 2023 by American Urological Association Education and Research, Inc.MetricsAuthor Information Longbin Xiong More articles by this author Lichao Wei More articles by this author Ting Xue More articles by this author Ru Yan More articles by this author Tingyu Liu More articles by this author Lijuan Jiang More articles by this author Huali Ma More articles by this author Yanxia Shi More articles by this author Xin An More articles by this author Yijun Zhang More articles by this author Shengjie Guo More articles by this author Kai Yao More articles by this author Fangjian Zhou More articles by this author Hui Han More articles by this author Expand All Advertisement PDF downloadLoading ...

Regulation of hypoxic stress and oxidative stress in bone grafting: Current trends and future perspectives

Tissue engineering aims to offer large-scale replacement of damaged organs using implants with the combination of cells, growth factors and scaffolds. However, the intra/peri‑implant region is exposed to severe hypoxic stress and oxidative stress during the early stage of implantation with bone graft materials, which endangers the survival, proliferation and differentiation of seed cells within the implants as well as the host cells surrounding the implants. If the bone graft material could spontaneously and intelligently regulate the hypoxic stress and oxidative stress to a moderate level, it will facilitate the vascularization of the implants and the rapid regeneration of the bone tissue. In this review, we will first introduce the signaling pathways of cellular response under hypoxic stress and oxidative stress, then present the classical material designs and examples in response to hypoxic stress and oxidative stress. And finally, we will address the important role of epigenetic mechanisms in the regulation of hypoxic stress and oxidative stress and describe the potential applications and prospective smart bone graft materials based on novel epigenetic factors against hypoxic stress and oxidative stress in bone repair. The main content of this review is summarized in the following graphical abstract.

Ayurvedic Management of Polypoidal Choroidal Vasculopathy (PCV) -A Case Report

Polypoidal Choroidal Vasculopathy (PCV), a subtype of neovascular Age-related Macular Degenerationn (AMD) is a disease characterized by persistent, recurrent serous leakage and hemorrhage in the macular area, mostly seen in the elderly population. While contemporary science offers anti Vasculo Endothelial Growth Factor (VEG-F) monotherapy and combination with Photodynamic Therapy (PDT), the outcomes of these remain doubtful. Repeated anti-VEG-F injections not only cause a financial and mental burden on the patients with no or very little visual outcome. Chronicity paves the way to complete loss of vision due to exudative retinal detachment. Symptoms like Raktavatdristi (to visualize things as if through a column of blood), Pashyatiaasyamanaasikam (unable to recognize the faces of people around) and aaviladarshanam (blurred vision) were suggestive of Raktaja Timira (an eye disease caused by Raktadosa) associated with Kaphadushti), according to Ayurveda. A 77-year-old male patient diagnosed with PCV sought Ayurvedic treatment after being advised to undergo anti-VEG-F therapy for the 3rd time in a row. His treatment comprised internal medicines consisting of Mahatiktakakashaya, Guduchyadikashaya, Chandraprabhavati and Avipathichurna; external treatments like Pratimarshanasya with Anu tailam and Gandusha with Triphalakashaya; Kriyakalpa (eye treatments) like Seka, Vidalaka, Aschotana and Anjana karma. At the end of 3 months of Ayurvedic treatment, his visual acuity in the right eye improved to 6/6, Optical Coherence Tomography (OCT) affirmed complete normalcy of the macula and Fundus examination confirmed the absence of neovascularization. Ayurvedic treatment is effective in reversing the process of neovascularization, thereby bringing about the marked visual outcome in PCV. Such patients should be encouraged to adopt Ayurvedic treatment instead of expensive and invasive treatment procedures like intra-vitreal injections even when the absolute effects of these remain doubtful.

Osteochondrogenesis by TGF-β3, BMP-2 and noggin growth factor combinations in an ex vivo muscle tissue model: Temporal function changes affecting tissue morphogenesis.

In the absence of clear molecular insight, the biological mechanism behind the use of growth factors applied in osteochondral regeneration is still unresolved. The present study aimed to resolve whether multiple growth factors applied to muscle tissue in vitro, such as TGF-β3, BMP-2 and Noggin, can lead to appropriate tissue morphogenesis with a specific osteochondrogenic nature, thereby revealing the underlying molecular interaction mechanisms during the differentiation process. Interestingly, although the results showed the typical modulatory effect of BMP-2 and TGF-β3 on the osteochondral process, and Noggin seemingly downregulated specific signals such as BMP-2 activity, we also discovered a synergistic effect between TGF-β3 and Noggin that positively influenced tissue morphogenesis. Noggin was observed to upregulate BMP-2 and OCN at specific time windows of culture in the presence of TGF-β3, suggesting a temporal time switch causing functional changes in the signaling protein. This implies that signals change their functions throughout the process of new tissue formation, which may depend on the presence or absence of specific singular or multiple signaling cues. If this is the case, the signaling cascade is far more intricate and complex than originally believed, warranting intensive future investigations so that regenerative therapies of a critical clinical nature can function properly.

Ophthalmic Solution of Smart Supramolecular Peptides to Capture Semaphorin 4D against Diabetic Retinopathy (Adv. Sci. 3/2023)

Smart Supramolecular Peptides In article number 2203351, Lei Wang, Hao Wang, Bo Hu, and co-workers report the formulation of a smart supramolecular peptide (SSP) eye drop for diabetic retinopathy treatment that is effective via specifically identifying and capturing Semaphorin4D (Sema 4D), a strongly pro-angiogenesis and exudates factor. The combination of SSP eye drop and anti-vascular endothelial growth factor (VEGF) injection showed approximately two times better therapeutic effect over anti-VEGF treatment alone. This work provides an alternative and effective method for noninvasive treatment for diabetic retinopathy.

In Vitro Generation of Human Cross-Presenting Type 1 Conventional Dendritic Cells (cDC1s) and Plasmacytoid Dendritic Cells (pDCs).

Dendritic cells (DCs) represent one of the most important immune cell subsets in preventing the host from pathogen invasion by promoting both innate and adaptive immunity. Most research on human dendritic cells has focused on the easy-to-obtain dendritic cells derived in vitro from monocytes (MoDCs). However, many questions remain unanswered regarding the role of different dendritic cell types. The investigation of their roles in human immunity is hampered by their rarity and fragility, which especially holds true for type 1 conventional dendritic cells (cDC1s) and for plasmacytoid dendritic cells (pDCs). In vitro differentiation from hematopoietic progenitors emerged as a common way to produce different DC types, but the efficiency and reproducibility of these protocols needed to be improved and the extent to which the DCs generated in vitro resembled their in vivo counterparts required a more rigorous and global assessment. Here, we describe a cost-effective and robust in vitro differentiation system for the production of cDC1s and pDCs equivalent to their blood counterparts, from cord blood CD34+ hematopoietic stem cells (HSCs) cultured on a stromal feeder layer with a combination of cytokines and growth factors.

LNK/SH2B3 As a Novel Driver in Juvenile Myelomonocytic Leukemia

Introduction Juvenile myelomonocytic leukemia (JMML) is a rare and aggressive myelodysplastic/myeloproliferative disorder in toddlers. Ninety-five percent of patients have mutations detected in the Ras/MAPK signaling pathway including CBL, KRAS, NF1, NRAS, RRAS, RRAS2, and PTPN11. Oncogenic fusion proteins leading to hyperactive Ras signaling and other mutations in genes encoding for proteins upstream the Ras pathway have also been described. One of these upstream proteins is LNK which is encoded by SH2B3. Loss of function mutations in SH2B3 result in abnormal proliferation of hematopoietic cells due to cytokine hypersensitivity and activation of JAK/STAT signaling. Mutations in SH2B3 have been detected in a variety of hematologic malignancies, specifically in myeloproliferative neoplasms in adults. We previously identified secondary mutations in SH2B3 in JMML. Here we report a case series of four JMML patients with initiating mutations in SH2B3. We also show that SH2B3 mutant induced pluripotent stem cell (iPSC)-derived JMML like hematopoietic progenitor cells (HPC) are sensitive to JAK inhibitors including ruxolitinib. Patients and Methods Genomic DNA from peripheral blood, bone marrow or buccal swabs was extracted using standard protocols. DNA samples were sequenced using a custom amplicon-based targeted sequencing approach. Ficoll-purified mononuclear cells from bone marrow of JMML patients and control samples were reprogrammed by using Sendai virus expressing doxycycline-regulated OCT4, KLF4, MYC, and SOX2. Control and JMML iPSCs were differentiated to HPCs by culturing in serum-free media with sequential combinations of growth factors to support multipotent hematopoietic progenitor formation. The half-maximal inhibitory concentration (IC50) for each kinase inhibitor was determined by performing Cell Titer Glo assays (Promega). Results Clinical characteristics of patients with SH2B3 mutant JMML described in this study are highlighted in Table 1. iPSC-derived JMML-like HPCs with different mutational backgrounds were analyzed in cell proliferation assays after exposure to multiple different JAK inhibitors (ruxolitinib, momelotinib, tofacitinib). HPCs with alterations in SH2B3 were more sensitive to chemical JAK inhibition compared to HPCs not harboring mutations in SH2B3. This finding was observed for all JAK inhibitors but was most prominent with ruxolitinib (Figure 1). Patient 3 received ruxolitinib monotherapy after being refractory to 6-mercaptopurine. Treatment resulted in resolution of splenomegaly but no reduction in the variant allele frequency of SH2B3. The patient is now alive more than one year after hematopoietic stem cell transplantation (HSCT). Discussion In a previous study of 100 patients with JMML we first identified seven patients with secondary SH2B3 mutations. Here, we report four additional patients with JMML that appear to be driven by initiating mutations in SH2B3 (Table 1). The mutations identified here result in a truncated protein (patient 1, 3 and 4) or affect the SH2 domain (patient 2). Interestingly, copy neutral loss of heterozygosity of SH2B3 in patient 1, also known as uniparental isodisomy, is a mechanism that has been observed in other genes (e.g. CBL and NF1) in JMML. Since in vitro data showed that loss of LNK results in increased JAK/STAT signaling, we hypothesized that this cohort of patients may benefit from JAK inhibitor therapy. Our data from iPSC-derived JMML like HPCs shows, that those cells with an additional SH2B3 mutation are more sensitive to JAK inhibitors, specifically ruxolitinib, a JAK1/2 inhibitor that is already approved or under investigation in multiple clinical trials. In summary, we expand the list of initiating mutations in JMML to include SH2B3 and raise the possibility of targeting the JAK pathway in these patients. Figure 1View largeDownload PPTFigure 1View largeDownload PPT Close modal

Integrated Transcriptomic and Proteomic Analysis Identifies Plasma Biomarkers of Hepatocellular Failure in Alcohol-Associated Hepatitis

Alcohol-associated hepatitis (AH) is a form of liver failure with high short-term mortality. Recent studies have shown that defective function of hepatocyte nuclear factor 4 alpha (HNF4a) and systemic inflammation are major disease drivers of AH. Plasma biomarkers of hepatocyte function could be useful for diagnostic and prognostic purposes. Herein, an integrative analysis of hepatic RNA sequencing and liquid chromatography-tandem mass spectrometry was performed to identify plasma protein signatures for patients with mild and severe AH. Alcohol-related liver disease cirrhosis, nonalcoholic fatty liver disease, and healthy subjects were used as comparator groups. Levels of identified proteins primarily involved in hepatocellular function were decreased in patients with AH, which included hepatokines, clotting factors, complement cascade components, and hepatocyte growth activators. A protein signature of AH disease severity was identified, including thrombin, hepatocyte growth factor α, clusterin, human serum factor H-related protein, and kallistatin, which exhibited large abundance shifts between severe and nonsevere AH. The combination of thrombin and hepatocyte growth factor α discriminated between severe and nonsevere AH with high sensitivity and specificity. These findings were correlated with the liver expression of genes encoding secreted proteins in a similar cohort, finding a highly consistent plasma protein signature reflecting HNF4A and HNF1A functions. This unbiased proteomic-transcriptome analysis identified plasma protein signatures and pathways associated with disease severity, reflecting HNF4A/1A activity useful for diagnostic assessment in AH.

TGFβ superfamily signaling regulates the state of human stem cell pluripotency and capacity to create well-structured telencephalic organoids.

SummaryTelencephalic organoids generated from human pluripotent stem cells (hPSCs) are a promising system for studying the distinct features of the developing human brain and the underlying causes of many neurological disorders. While organoid technology is steadily advancing, many challenges remain, including potential batch-to-batch and cell-line-to-cell-line variability, and structural inconsistency. Here, we demonstrate that a major contributor to cortical organoid quality is the way hPSCs are maintained prior to differentiation. Optimal results were achieved using particular fibroblast-feeder-supported hPSCs rather than feeder-independent cells, differences that were reflected in their transcriptomic states at the outset. Feeder-supported hPSCs displayed activation of diverse transforming growth factor β (TGFβ) superfamily signaling pathways and increased expression of genes connected to naive pluripotency. We further identified combinations of TGFβ-related growth factors that are necessary and together sufficient to impart broad telencephalic organoid competency to feeder-free hPSCs and enhance the formation of well-structured brain tissues suitable for disease modeling.

Effect of the combination of composite skin grafting and recombinant human basic fibroblast growth factor on plastic surgery for extensive sc ars after burns

Purpose: To investigate the effect of combined use of composite skin grafting and recombinant human basic fibroblast growth factor (rh-bFGF) on plastic surgery for extensive burn scars, and its influence on wound healing and inflammatory responses in patients.
 Methods: Hospital records of 120 patients who underwent extensive scar plastic surgery after burns in Hanchuan People’s Hospital from February 2019 to February 2020 were retrospectively analyzed. The patients were equally assigned to study group (STG) and control group (COG), based on the order of admission. All patients received composite skin grafting surgery and post-surgery topical silver sulphadiazine cream regularly, while those in study group (STG) received additional treatment with rh-bFGF spray on the wound surfaces. Scar scores, degree of wound healing, populations of fibrocytes and capillaries, levels of vascular endothelial growth factor (VEGF) and transforming growth factor-β1 (TGF-β1), and activities of lactate dehydrogenase (LDH) and succinate dehydrogenase (SDH) were evaluated.
 Results: Patients in STG had significantly better scar scores, shorter mean wound healing time, higher mean degree of wound healing, higher scores on inflammatory reactions near the wounds, and higher populations of fibrocytes and capillaries, when compared with patients in COG group (p < 0.001). After surgery, there were lower levels of VEGF and TGF-β1, and lower levels of activities of LDH and SDH in STG than in COG group (p < 0.001).
 Conclusion: Rh-bFGF accelerates the proliferation of wound tissue, improves local microcirculation, regulates inflammatory response, and enhances wound healing in burn scar patients when used in combination with composite skin grafting. Therefore, Rh-bFGF has a potential for enhanced management of burn scars.

THE EFFECT OF COMBINATIONS OF GROWTH FACTORS AND HYALURONIC ACID ON THE PROLIFERATION AND CHONDROGENIC DIFFERENTIATION OF HUMAN SYNOVIUM DERIVED MESENCHYMAL STEM CELLS

Growth factors and hyaluronic acid (HA) have been shown to stimulate functional activity of mesenchymal stem cells (MSCs) and thus may influence damaged tissue healing, including cartilage. In this study we investigated the effects of individual and combinatorial application of HA, platelet derived growth factor–BB (PDGF-BB), transforming growth factor β1 (TGF-β1), bone morphogenetic protein-4 (BMP-4) and fibroblast growth factor-2 (FGF-2) on the proliferation and chondrogenic differentiation of human synovium derived MSCs (SD MSCs). Isolated SD MSCs were expanded in α-MEM medium and characterized using immunocytochemistry, multilineage differentiation assay and CFU-assay. Proliferation activity of MSCs was examined with MTT-assay. Chondrogenic differentiation and glycosaminoglycan level in micro-pellet culture of MSCs were evaluated with histochemical and biochemical analysis. Our results showed that the individual use of FGF-2 and PDGF-BB significantly increased the rate of MSC proliferation compared to the control. At the same time, TGF-β1 did not affect cell growth. Combinations of growth factors: TGF-β1+FGF-2, TGF-β1+PDGF-BB, and FGF-2+PDGF-BB were able to significantly increase the proliferative activity of MSCs. The greatest synergistic effect was found with combination of all three factors TGF-β1, FGF-2, and PDGF-BB. Histochemical and biochemical analysis revealed that combination of TGF-β1 and BMP-4 have a significant effect on chondrogenic differentiation and the synthesis of glycosaminoglycans in micro-pellet culture of MSCs. The addition of HA to growth factors has minor effect on the proliferation and does not affect the chondrogenic differentiation of MSCs. This study demonstrated that combination of growth factors can promote proliferation and chondrogenic differentiation of SD MSCs and may be effective in enhancing cartilage healing.

The pathogenesis of cryptoglandular anal fistula: New insight into the immunological profile.

The aetiology of cryptoglandular anal fistula (AF) is poorly understood. Evidence suggests that persistence and/or recurrence of the disease is more related to inflammatory than infectious factors. The aim of this study was to investigate the immune profile of cryptoglandular AF and to perform a histopathological characterization. Fistulectomy was performed in all patients; healthy ischioanal fat from the same patients was used as a control. Samples were evaluated by the Luminex xMAP system for the detection of 27 analytes. AF tissues were analysed using immunofluorescence. Staining was performed using primary antibodies to identify M1 inflammatory and M2 anti-inflammatory macrophages. Selective staining of total T lymphocytes and different T lymphocyte subsets was performed. Twenty patients with AF underwent a fistulectomy. Specific cytokine pathways differentiated AF from healthy tissue: pro-inflammatory cytokines interleukin (IL)-1β, IL-4, IL-8 and IL-17 and the anti-inflammatory cytokine IL-10 were overexpressed in AF compared with controls. Chemokines involved in macrophage recruitment (CCL2, CCL3, CCL4) were higher in AF than in healthy fatty tissue. Moreover, we showed that Tc17 cells characterize AF patients, thus confirming the enzyme-linked immunosorbent assay data. Furthermore, elevated infiltration of CD68+ myeloid cells and a reduction of the M1/M2 ratio characterize AF patients. A combination of inflammatory cytokines, chemokines and growth factors reside in the wound microenvironment of AF patients. For the first time an important prevalence of Tc17 cells and a reduction in the M1/M2 ratio was observed, thus suggesting new insights into the immunological characterization of AF patients.

Novel Approaches Used in Ovarian Tissue Transplantation for Fertility Preservation: Focus on Tissue Engineering Approaches and Angiogenesis Capacity.

Due to the impact of the modern lifestyle, female infertility has been reduced because of different reasons. For example, in combined chemotherapeutic therapies, a small fraction of cancer survivors has faced different post-complications and side effects such as infertility. Besides, in modern society, delayed age of childbearing has also affected fertility. Nowadays, ovarian tissue cryopreservation and transplantation (OTC/T) is considered one of the appropriate strategies for the restoration of ovarian tissue and bioactivity in patients with the loss of reproductive function. In this regard, several procedures have been considered to improve the efficacy and safety of OTT. Among them, a surgical approach is used to transplant ovaries into the optimal sites, but the existence of ischemic changes and lack of appropriate revascularization can lead to bulk follicular atresia. Besides, the role of OTC/T is limited in women of advanced maternal age undergoing lifesaving chemo-radiation. As a correlate, the development of de novo approaches with efficacious regenerative outcomes is highly welcomed. Tissue engineering shows high therapeutic potentialities to restore fertility in males and females using the combination of biomaterials, cells, and growth factors. Unfortunately, most synthetic and natural materials are at the experimental stage and only the efficacy has been properly evaluated in limited cases. Along with these descriptions, strategies associated with the induction of angiogenesis in transplanted ovaries can diminish the injuries associated with ischemic changes. In this review, the authors tried to summarize recent techniques, especially tissue engineering approaches for improving ovarian function and fertility by focusing on angiogenesis and neovascularization.

Abstract P1142: Engineering Vascular Therapies With Angiogenic Factors For Cardiac Regeneration And Tissue Remodeling

The limited regenerative capacity of the adult human heart presents a critical need to stabilize and support injured heart muscle to mitigate the progression of heart failure. The effectiveness of remuscularization therapies using human induced pluripotent stem cell derived cardiomyocytes depends host mediated angiogenesis to direct the formation of stable, perfused vascular networks within the engineered cardiac implant. We hypothesize that potent combinations of growth factors will stimulate angiogenesis in the myocardium and increase vascular development in engineered cardiac tissue. In a fractional factorial experiment, we assessed the effect of ten growth factors, cytokines and enzymes: VEGF, bFGF, Shh, PDGF, IGF-1, EGF, TGF-β1, Ang-1, MCP-1 and MMP-9 on in vivo angiogenesis in a subcutaneous Matrigel plug assay in the rat. Five factors were highly potent by metrics of vessel density and vascular cell recruitment in explants and warranted further investigation. We then designed a two-level, fractional factorial experiment using a defined collagen-alginate gel to deliver high/low doses of these 5 factors subcutaneously to elucidate main effects and two-way interactions that regulate vessel development in vivo . After 7 days, Griffonia Simplicfolia lectin I was administered to label perfused vasculature then explants were removed and processed for immunohistochemistry. Our data show that 3 unique cocktails yielded a 10-fold increase in lumen density, a 3.3-fold increase in lumen size, and a 3.7-fold increase in endothelial cell recruitment relative to unloaded controls. Ongoing analyses will determine which angiogenic cocktail to deliver in a customizable biomaterial-based vascular therapy for applications in the heart. This novel therapy will be delivered with an engineered cardiac tissue onto the epicardium in a rat I/R MI model to quantify vascular remodeling in the implant and underlying host tissue and whole heart function over 4 weeks. In conclusion, we have identified five potent angiogenic factors for modulating revascularization responses and our ongoing work aims to understand the potential synergy between revascularization and remuscularization therapies for heart regeneration.

The initial delay to mitotic activity in primary cultures of equine satellite cells is reduced by combinations of growth factors.

Satellite cell (SC) activation is defined as the time frame during which the stem cell becomes poised to reenter G1 of the cell cycle. The growth factors and events leading to full mitotic activation in equine SCs remain largely unknown. Insulin-like growth factor I (IGF-I), hepatocyte growth factor (HGF), and fibroblast growth factor 2 (FGF2) are sequentially transcribed during the muscle repair and recovery period following strenuous exercise in adult horses. Expression of IGF-I occurs within 24 h of the postexercise recovery period suggesting it may affect early SC actions. As a first step, gluteus medius muscle cryosections from adult horses (n = 9) were examined for the presence of central nuclei (CN), a marker of SC addition to the fiber. Results demonstrate few CN fibers prior to exercise with a 3-fold increase (P = 0.05) 24 h postexercise. Cultures of SC (n = 4 isolates) were treated with 100 ng/mL IGF-I for varying times prior to measurement of myogenic events. Results demonstrate that IGF-I does not affect the initial lag period, proliferation, or subsequent differentiation of equine SC in vitro (P > 0.05). However, media containing a combination of IGF-I and 10 ng/mL FGF2 and 25 ng/mL HGF hastens (P < 0.05) the time to S-phase entry in fresh isolates of SCs. Media supplementation with optimal concentrations of FGF2, HGF, or a combination of HGF and FGF2 suppresses (P < 0.05) the percentage of myogenin immunopositive SCs to levels below that found in control- or IGF-I-treated SCs. These results provide new insight into the combinatorial roles growth factors play during equine SC myogenesis.

Extracellular Matrix Regulation of Vascular Morphogenesis, Maturation, and Stabilization.

The extracellular matrix represents a critical regulator of tissue vascularization during embryonic development and postnatal life. In this perspective, we present key information and concepts that focus on how the extracellular matrix controls capillary assembly, maturation, and stabilization, and, in addition, contributes to tissue stability and health. In particular, we present and discuss mechanistic details underlying (1) the role of the extracellular matrix in controlling different steps of vascular morphogenesis, (2) the ability of endothelial cells (ECs) and pericytes to coassemble into elongated and narrow capillary EC-lined tubes with associated pericytes and basement membrane matrices, and (3) the identification of specific growth factor combinations ("factors") and peptides as well as coordinated "factor" and extracellular matrix receptor signaling pathways that are required to form stabilized capillary networks.

Polysaccharides, as biological macromolecule-based platforms in skeletal muscle tissue engineering: a systematic review

Traumatic injuries to skeletal muscles caused by accidents, surgeries, and strains result in a significant disability. Currently, the golden standard of treatment for skeletal muscle injuries is tissue grafting. However, this treatment strategy is associated with several challenges. Therefore, developing alternative strategies is highly demanded. In this context, skeletal muscle tissue engineering (SMTE) aims to regenerate defective muscle tissue using combinations of cells, growth factors, and biomaterials. Although significant progress has been made in SMTE, the inherent complexity of skeletal muscle tissue requires a versatile approach to develop scaffolds with high similarity to native muscle tissue. These scaffolds can be produced using synthetic or natural polymers. Among different biomaterial options, polysaccharides are ideal scaffolding platforms with potential applicability in different biomedical fields, such as drug delivery, guided tissue regeneration, and driving stem cells fate. Due to their versatile properties, biocompatibility, low cost, high availability, low immunogenicity, surface modification capabilities, and processability, the appeal of polysaccharides in SMTE is on the rise. Although different polysaccharides can be potentially used in SMTE, only alginate, chitosan, cellulose, and hyaluronic acid have been studied. In the current review, the recent progress, applications, and challenges of polysaccharides in SMTE will be highlighted.

A streamlined CRISPR workflow to introduce mutations and generate isogenic iPSCs for modeling amyotrophic lateral sclerosis.

Amyotrophic lateral sclerosis (ALS) represents a complex neurodegenerative disorder with significant genetic heterogeneity. To date, both the genetic etiology and the underlying molecular mechanisms driving this disease remain poorly understood, although in recent years several studies have highlighted a number of genetic mutations causative for ALS. With these mutations pointing to potential pathways that may be affected within individuals with ALS, having the ability to generate human neurons and other disease relevant cells containing these mutations becomes even more critical if new therapies are to emerge. Recent developments with the advent of induced pluripotent stem cells (iPSCs) and clustered regularly interspaced short palindromic repeats (CRISPR) gene editing fields gave us the tools to introduce or correct a specific mutation at any site within the genome of an iPSC, and thus model the specific contribution of risk mutations. In this study we describe a rapid and efficient way to either introduce a mutation into a control line, or to correct an allele-specific mutation, generating an isogenic control line from patient-derived iPSCs with a given mutation. The mutations introduced were the G94A (also known as G93A) mutation into SOD1 or H517Q into FUS, and the mutation corrected was a patient iPSC line with I114T mutation in SOD1. A combination of small molecules and growth factors were used to guide a stepwise differentiation of the edited cells into motor neurons in order to demonstrate that disease-relevant cells could be generated for downstream applications. Through a combination of iPSCs and CRISPR editing, the cells generated here will provide fundamental insights into the molecular mechanisms underlying neuron degeneration in ALS.

P-466 Decellularization of whole organ human cervix: Physio and biochemical aspects of decellularized extracellular matrix

Abstract Study question Can the whole organ-cervix be decellularized while maintaining its fiber ultrastructure and extracellular matrix (ECM) functionality? Summary answer Cervix after decellularization could maintain the physical properties, ECM was found to be intact and functional. What is known already ECM-rich bioscaffolds are obtained from the process that utilizes the decellularization of tissues and organs. Cervical bioengineering is needed for clinical cervicovaginal reconstruction and tissue constructs can help women with cervical abnormalities. Study design, size, duration Human cervices were obtained from hysterectomized samples from 3 women undergoing surgery for complete hysterectomy during January 2021 to December 2021 Participants/materials, setting, methods Human cervices were obtained from hysterectomized samples from 3 women undergoing surgery and were subjected to whole organ decellularization. Various physical, chemical methods were employed to remove cellular remnants for a period of 10 days via perfusion of ionic and detergent solutions. Validation of complete decellularization was done via hematoxylin and eosin (H&amp;E) staining, DNA quantification. ECM functionality was observed. Main results and the role of chance Well-organized ECM was obtained. Effective complete decellularization was dictated and verified by cervical tissue density, histopathological studies, DNA quantification and biochemical tests. H&amp;E staining reflected the absence of any nuclear material in the endocervical and ectocervical tissues. Tissue microarchitecture and descending uterine artery were intact and could be helpful in the nutrient and gas exchange. DNA was found to be around 70 ng/mg tissue weight. Cell-free tissue-derived scaffolds were obtained along with a combination of growth factors and proteins. Limitations, reasons for caution Acellular cervical scaffold as a whole organ with vasculature intact for site-specific transplantation is a bigger question and still unaddressed. Wider implications of the findings Our study creates numerous prospects for in-vitro and in-vivo whole cervix tissue engineering. Site-specific tissue holds a promising avenue for the reconstruction of cervical abnormalities. Trial registration number Not applicable

Effect of natural-based biological hydrogels combined with growth factors on skin wound healing

Abstract Skin wound healing is a continuous and complex process affected by many factors. Growth factors play an important role in the process of wound healing. Local application of growth factors can significantly promote wound healing. However, the degradation and time dependence of growth factors require appropriate delivery systems to help them play a role in wound healing. In recent years, wound dressing products with hydrogels as matrix materials or main components have shown obvious advantages in promoting wound healing. By modifying the hydrogel or combining it with other factors or materials that are beneficial to wound healing, the healing effect can be further enhanced. This review will introduce the research status of growth factors and hydrogels based on natural biological materials in skin wound repair and review the effects and research progress of the combination of growth factors and hydrogels in skin wound healing.