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This study was conducted to know the impact of cryopreservation on caprine fetal adnexa derived mesenchymal stem cells (MSCs) on the basic stem cell characteristics. Gravid caprine uteri (2-3 months) were collected from local abattoir to derive (amniotic fluid [cAF], amniotic sac [cAS], Wharton's jelly [cWJ], and cord blood [cCB]) MSCs and expanded in vitro. Cells were cryopreserved at 3rd passage (P3) using 10% DMSO. Post-thaw viability and cellular properties were assessed. Cells were expanded to determine growth kinetics, tri-lineage differentiation, localization, and molecular expression of MSCs and pluripotency markers; thereafter, these cells were transplanted in the full-thickness (2 × 2cm2 ) rat skin wound to determine their wound healing potential. The post-thaw (pt) growth kinetics study suggested that cWJ MSCs expanded more rapidly with faster population doubling time (PDT) than that of other fetal adnexa MSCs. The relative mRNA expression of surface antigens (CD73, CD90, and CD 105) and pluripotency markers (Oct4, KLF, and cMyc) was higher in cWJ MSCs in comparison to cAS, cAF, and cCB MSCs post-thaw. The percent wound contraction on 7th day was more than 50% for all the MSC-treated groups (pre and post-thaw), against 39.55% in the control group. On day 28th, 99% and more wound contraction was observed in cAF, cAF-pt, cAS-pt, cWJ, cWJ-pt, and cCB, MSCs with better scores for epithelization, neovascularization, and collagen characteristics at a non-significant level. It is concluded that these MSCs could be successfully cryopreserved without altering their stemness and wound healing properties. J. Cell. Physiol. 232: 2186-2200, 2017. © 2016 Wiley Periodicals, Inc.
Assuntos
Proliferação de Células/efeitos dos fármacos , Transplante de Células-Tronco de Sangue do Cordão Umbilical , Criopreservação , Crioprotetores/farmacologia , Dimetil Sulfóxido/farmacologia , Células-Tronco Fetais/efeitos dos fármacos , Células-Tronco Fetais/transplante , Transplante de Células-Tronco Mesenquimais , Células-Tronco Mesenquimais/efeitos dos fármacos , Ferida Cirúrgica/cirurgia , Cicatrização , Líquido Amniótico/citologia , Animais , Biomarcadores/metabolismo , Diferenciação Celular , Linhagem da Célula , Sobrevivência Celular/efeitos dos fármacos , Células Cultivadas , Modelos Animais de Doenças , Feminino , Sangue Fetal/citologia , Células-Tronco Fetais/metabolismo , Cabras , Xenoenxertos , Cinética , Masculino , Células-Tronco Mesenquimais/metabolismo , Fenótipo , Gravidez , Ratos Wistar , Ferida Cirúrgica/metabolismo , Ferida Cirúrgica/patologia , Geleia de Wharton/citologiaRESUMO
The widespread application of mesenchymal stem cells (MSCs) in veterinary regenerative medicine highlights their promising therapeutic potential. However, the lack of standardized characterization and reporting practices across studies poses a significant challenge, compromising the assessment of their safety and efficacy. While criteria established for human MSCs serve as a foundation, the unique characteristics of animal-derived MSCs warrant updated guidelines tailored to veterinary medicine. A recent position statement outlining minimal reporting criteria for MSCs in veterinary research reflects efforts to address this need, aiming to enhance research quality and reproducibility. Standardized reporting criteria ensure transparency, facilitate evidence synthesis, and promote best practices adoption in MSC isolation, characterization, and administration. Adherence to minimal reporting criteria is crucial for maintaining scientific rigor and advancing the field of veterinary regenerative medicine. Ongoing collaboration among stakeholders is essential for effective implementation and adherence to updated guidelines, fostering excellence and innovation in MSC-based therapies for animal patients.
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Células-Tronco Mesenquimais , Medicina Regenerativa , Animais , Medicina Regenerativa/normas , Células-Tronco Mesenquimais/citologia , Transplante de Células-Tronco Mesenquimais/veterinária , Transplante de Células-Tronco Mesenquimais/normas , Transplante de Células-Tronco Mesenquimais/métodos , Medicina Veterinária/normas , Medicina Veterinária/métodosRESUMO
BACKGROUND: Stem cell-based therapies display immense potential in regenerative medicine, highlighting the crucial significance of devising efficient delivery methods. This study centers on a pioneering approach that utilizes Pluronic F127 (PF127) as a thermoresponsive and injectable hydrogel designed for the encapsulation of adipose-derived mesenchymal stem cells (AdMSCs). METHODS: The degradation profile, gelation time, and microstructure of the PF127 hydrogel were thoroughly examined. AdMSCs were isolated, expanded, and characterized based on their multi-lineage differentiation potential. AdMSCs from the third passage were specifically employed for encapsulation within the PF127 hydrogel. Subsequently, the cytotoxicity of the AdMSC-loaded PF127 hydrogel was assessed using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and apoptosis assays. RESULTS: Characterized by scanning electron microscopy (SEM), the PF127 hydrogel exhibited a porous structure, indicating its suitability for accommodating AdMSCs and facilitating wound healing. The PF127 hydrogel demonstrated reversible phase transitions, rendering it suitable for in vivo applications. Studies on the gelation time of PF127 hydrogel unveiled a concentration-dependent decrease in gelation time, offering adaptability for diverse medical applications. Analysis of the degradation profile showcased a seven-day degradation period, leading to the decision for weekly topical applications. Cytotoxicity assessments confirmed that AdMSCs loaded into the PF127 hydrogel maintained heightened metabolic activity for up to one week, affirming the safety and appropriateness of the PF127 hydrogel for encapsulating cellular therapeutics. Furthermore, cell apoptosis assays consistently indicated low rates of apoptosis, emphasizing the viability and robust health of AdMSCs when delivered within the hydrogel. CONCLUSIONS: These findings underscore the vast potential of PF127 hydrogel as a versatile and biocompatible delivery system for AdMSCs in the realm of regenerative medicine. Boasting adjustable gelation properties and a remarkable capacity for cell encapsulation, this pioneering delivery system presents a promising path for applications in tissue engineering and wound healing. Ultimately, these advancements propel and elevate the landscape of regenerative medicine.
Assuntos
Hidrogéis , Células-Tronco Mesenquimais , Humanos , Hidrogéis/química , Poloxâmero/químicaRESUMO
Stem cell research has the transformative potential to revolutionize medicine. Language models like ChatGPT, which use artificial intelligence (AI) and natural language processing, generate human-like text that can aid researchers. However, it is vital to ensure the accuracy and reliability of AI-generated references. This study assesses Chat Generative Pre-Trained Transformer (ChatGPT)'s utility in stem cell research and evaluates the accuracy of its references. Of the 86 references analyzed, 15.12% were fabricated and 9.30% were erroneous. These errors were due to limitations such as no real-time internet access and reliance on preexisting data. Artificial hallucinations were also observed, where the text seems plausible but deviates from fact. Monitoring, diverse training, and expanding knowledge cut-off can help to reduce fabricated references and hallucinations. Researchers must verify references and consider the limitations of AI models. Further research is needed to enhance the accuracy of such language models. Despite these challenges, ChatGPT has the potential to be a valuable tool for stem cell research. It can help researchers to stay up-to-date on the latest developments in the field and to find relevant information.
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BACKGROUND AND AIM: Adipose tissue-derived stromal vascular fraction (SVF) contains a heterogeneous cell population comprising multipotent adipose-derived stem cells. Regenerative therapy using adipose-derived SVF has broad applications in bone tissue engineering due to the superior osteogenic potential of SVF. This study was designed to standardize and characterize adipose-derived SVF obtained from New Zealand white rabbits for bone tissue engineering and other potential applications. MATERIALS AND METHODS: Ten skeletally mature and clinically healthy adult New Zealand white rabbits were used in this study. The SVF was prepared using surgically resected interscapular adipose tissue following enzymatic digestion with 0.1% collagenase type I solution. The SVF pellet obtained after the final centrifugation step was suspended in a 0.5 mL control solution to obtain ready-to-use adipose-derived SVF. The freshly prepared SVF was characterized based on the total SVF cell count and cell yield per gram of adipose tissue. The SVF cells were enumerated using a hemocytometer. RESULTS: Interscapular adipose tissue depots are ideal for preparing autologous adipose-derived SVF due to the ease of access. The interscapular adipose-derived SVF prepared by enzymatic digestion had an average cell yield of 3.15±0.09×106 cells/g adipose tissue. Freshly prepared SVF had a total cell count of 3.15±0.09×104 cells/µL. CONCLUSION: The enzymatic digestion of adipose tissue using 0.1% collagenase resulted in better cell yield per gram than methods previously reported in rabbits. The use of adipose-derived SVF can preclude the requirement for an additional culture period. In addition, it may also reduce the risk of extensive cell contamination, which makes it a safe and cost-effective strategy for bone tissue engineering applications.
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Ivermectin is considered to be a wonder drug due to its broad-spectrum antiparasitic activity against both ectoparasites and endoparasites (under class of endectocide) and has multiple applications in both veterinary and human medicine. In particular, ivermectin is commonly used in the treatment of different kinds of infections and infestations. By altering the vehicles used in the formulations, the pharmacokinetic properties of different ivermectin preparations can be altered. Since its development, various vehicles have been evaluated to assess the efficacy, safety, and therapeutic systemic concentrations of ivermectin in different species. A subcutaneous route of administration is preferred over a topical or an oral route for ivermectin due to superior bioavailability. Different formulations of ivermectin have been developed over the years, such as stabilized aqueous formulations, osmotic pumps, controlled release capsules, silicone carriers, zein microspheres, biodegradable microparticulate drug delivery systems, lipid nanocapsules, solid lipid nanoparticles, sustained-release ivermectin varnish, sustained-release ivermectin-loaded solid dispersion suspension, and biodegradable subcutaneous implants. However, several reports of ivermectin resistance have been identified in different parts of the world over the past few years. Continuous use of suboptimal formulations or sub-therapeutic plasma concentrations may predispose an individual to resistance toward ivermectin. The current research trend is focused toward the need for developing ivermectin formulations that are stable, effective, and safe and that reduce the number of doses required for complete clinical cure in different parasitic diseases. Therefore, single-dose long-acting preparations of ivermectin that provide effective therapeutic drug concentrations need to be developed and commercialized, which may revolutionize drug therapy and prophylaxis against various parasitic diseases in the near future. The present review highlights the current advances in pharmacokinetic modulation of ivermectin formulations and their potent therapeutic applications, issues related to emergence of ivermectin resistance, and future trends of ivermectin usage.
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Irrespective of aetiology, infectious respiratory diseases of sheep and goats contribute to 5.6 percent of the total diseases of small ruminants. These infectious respiratory disorders are divided into two groups: the diseases of upper respiratory tract, namely, nasal myiasis and enzootic nasal tumors, and diseases of lower respiratory tract, namely, peste des petits ruminants (PPR), parainfluenza, Pasteurellosis, Ovine progressive pneumonia, mycoplasmosis, caprine arthritis encephalitis virus, caseous lymphadenitis, verminous pneumonia, and many others. Depending upon aetiology, many of them are acute and fatal in nature. Early, rapid, and specific diagnosis of such diseases holds great importance to reduce the losses. The advanced enzyme-linked immunosorbent assays (ELISAs) for the detection of antigen as well as antibodies directly from the samples and molecular diagnostic assays along with microsatellites comprehensively assist in diagnosis as well as treatment and epidemiological studies. The present review discusses the advancements made in the diagnosis of common infectious respiratory diseases of sheep and goats. It would update the knowledge and help in adapting and implementing appropriate, timely, and confirmatory diagnostic procedures. Moreover, it would assist in designing appropriate prevention protocols and devising suitable control strategies to overcome respiratory diseases and alleviate the economic losses.
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Mechanism of Mycobacterium avium subsp. paratuberculosis (Map) invasion through intestinal mucosa is not completely understood. In the present study, we developed an in vivo multiple-intestinal loop model in lambs to investigate (i) the type of cells involved in the bacterial uptake across the intestinal mucosa, (ii) the efficiency of bacterial uptake in different segments of the small intestine and (iii) the ability of different strains of Map to invade the various segments of the small intestine. Four loops on ileum and four loops each on Peyer's patch and non-Peyer's patch areas of jejunum were constructed by surgical procedure. The caprine, bovine, and vaccine strains of Map were used for infection. Map-infected intestinal loop tissues were collected at 1, 3, 6, 12, and 24 h post-infection and processed for electron microscopy, histology, bacterial culture and bacterial counting. All these parameters revealed that Map invaded through M cells and the enterocytes and bacterial translocation across M cells was greater than the enterocytes. Bacterial invasion was greater in ileal loops when compared to jejunal loops. Within the jejunal loops, bacterial uptake was higher in Peyer's patch areas than that of non-Peyer's patch areas. The caprine and bovine strains of Map showed greater ability for invasion into the small intestinal mucosa than that of the vaccine strain.