Regenerative Therapy最新文献

Introduction: Exploring techniques for differentiating and culturing canine hepatocytes serves as a means to establish systems for liver transplantation and drug metabolism testing. However, establishing consistent methods for culturing stable hepatocytes remains a challenge. Recently, several investigations have shown the reprogramming of mature hepatocytes into hepatic progenitor cells by applying specific small molecule compounds, including Y-27632, (a ROCK inhibitor), A-83-01 (a TGFβ inhibitor), and CHIR99021 (a GSK3 inhibitor) (termed YAC) in rat, mouse, and humans, respectively. However, reports or evidence of successful reprogramming using these small-molecule compounds in dogs are absent. This study aimed to induce the differentiation of mature canine hepatocytes into progenitor cells.

Methods: Cryopreserved canine hepatocytes (cHep) were cultured for 14 d in a YAC-supplemented hepatocyte growth medium. Subsequently, an assessment was conducted involving morphological observations, quantitative real-time polymerase chain reaction (qRT-PCR), and immunocytochemistry.

Results: Notably, cryopreserved cHep cells emerged and exhibited ongoing proliferation and concurrently developed colonies within the YAC-enriched culture. These observations indicated that the mature hepatocytes reprogrammed into hepatic progenitor cells. Moreover, qRT-PCR analysis revealed a notable enhancement in gene expression levels. Specifically, the genes encoding α-fetoprotein (AFP), epithelial cell adhesion molecule (EpCAM), Cytokeratin 19 (CK19) and SRY-box9 (Sox9) displayed approximately 12-, 2.2-, 517- and 2.9- increases in hepatic progenitor cells, respectively, on day 14 as compared to their state before induction of differentiation. Hepatocyte-related protein expression of AFP, EPCAM, SOX9 and CK19 was confirmed via immunocytochemistry on day 21. In contrast, ALB and MRP2, which are highly expressed in mature hepatocytes, were decreased compared to those before YAC addition, which is consistent with the characteristics of undifferentiated hepatocytes.

Conclusions: Herein, we effectively promoted the reprogramming of cryopreserved cHep cells into hepatic progenitor cells using three small-molecule compounds. The mRNA and protein expression analyses demonstrated increased levels of hepatic progenitor cells-specific markers, whereas markers related to mature hepatocytes decreased, suggesting that reprogramming cryopreserved cHep cells to hepatic progenitor cells was achieved using YAC. Therefore, cultivating liver progenitor cells holds the potential to offer valuable insights into the development of artificial livers for drug discovery research and transplantation therapy aimed at addressing liver diseases in dogs.

The introduction of three-dimensional (3D) printing scaffolds has emerged as an effective approach to achieving satisfactory revascularization for bone tissue engineering (BTE). However, there is a notable absence of analytical and descriptive investigations concerning the trajectory, essential research directions, current research scenario, pivotal investigative focuses, and forthcoming perspectives. Hence, the objective of this research is to offer a thorough overview of the advancements achieved in 3D printing structures for vascularized BTE within the last 10 years. Information extracted from the Web of Science repository spans from January 1, 2014, to April 1, 2024. Utilizing advanced analytical instruments, we conducted comprehensive scientometric and visual analyses. The findings underscore the predominant influence of China, representing 59.62 % of the overall publications and playing a pivotal role in shaping research within this field. Notable productivity was evident at various institutions, including Shanghai Jiao Tong University, Chinese Academy of Sciences, and Sichuan University. Wang Jinwu and Wu Chengtie stand out as the most prolific contributors in this domain. The highest number of publications in this area was contributed by the journal Advanced Healthcare Materials. In this study, osteogenesis imperfecta, osteosarcoma, fractures, osteonecrosis, and cartilage diseases were identified as the most significant disorders investigated in this research area. By providing a comprehensive scientometric assessment, this study benefits both experienced researchers and newcomers alike, offering prompt access to essential information and fostering the extraction of innovative concepts within this specific field.

Introduction: Cirrhosis remains a significant clinical challenge due to its poor prognosis and limited treatment options, creating a high unmet medical need for the development of novel therapies. In this study, we analyzed the effects of a novel approach to treat cirrhosis using platelet-rich plasma-derived extracellular vesicles (PRPEV) in mice.

Methods: PRPEV were collected from platelet-rich plasma using ultrafiltration, and their proteomes were analyzed. The carbon tetrachloride (CCl₄)-induced cirrhosis model of mice was used to evaluate the effect of PRPEV administration and compared with the control group (n = 8). In vitro and in vivo mechanistic analyses of PRPEV administration were confirmed using real time-PCR and immunostaining.

Results: Gene ontology analysis based on the proteome revealed that PRPEV contain many factors associated with EV and immune responses. In vitro, PRPEV polarize macrophages into an anti-inflammatory phenotype. Following PRPEV administration, there was a decrease in serum alanine aminotransferase levels and reduction in liver fibrosis, while mRNA levels of regenerative factors were upregulated and transforming growth factor β-1 was downregulated. Furthermore, the number of anti-inflammatory macrophages in the liver increased.

Conclusions: PRPEV may contribute to hepatocyte proliferation, anti-inflammation, and anti-fibrogenesis in the liver. This novel concept paves the way for cirrhosis treatment.

People still hold the concept of using cell-based treatments to regenerate missing neurons in high esteem. CD117⁺ cells are considered favorable stem cells for regenerative medicine. The objective of this research was to examine the impact of Alginate-Gelatin (Alg-Gel) hydrogel on the process of neurogenic differentiation of CD117⁺ cells utilizing a cytokines secretion test conducted in a laboratory setting. To achieve this objective, bone marrow-CD117⁺ cells were isolated using the MACS technique and then transformed into neuron cells using a neurogenic differentiation medium. The characterization of enriched CD117⁺ cells has been done with flow cytometry as well as immunocytochemistry. Next, the cells underwent western blotting assay to evaluate the signaling pathways. Subsequently, the culture media was obtained from both groups in order to determine cytokine levels. The study revealed that the Alg-Gel hydrogel had a notable impact on enhancing the protein expression of neuron markers such as β-tubulin and Wnt/catenin signaling pathway components in CD117⁺ neurogenic differentiated cells. Furthermore, the cultured medium from the experimental group exhibited a notable abundance of IL-6 and IL-10 in comparison to the control group. The observed in vitro effects of Alg-Gel hydrogel on neurogenic differentiation of CD117⁺ cells are likely to be caused by the cytokines that are released.

Extracellular vesicles (EVs) have gainedsignificant attention due totheir crucialroles invarious biological systems. This review aims to explore the functions of EVs in both in physiological and pathological states of the liver, with a specific focus on the potential mechanisms and concrete evidence of EVs in liver regeneration processes. The review begins by emphasizing the importance of EVs in maintaining liver health and their involvement in different pathological conditions, starting from the liver's own EVs. Reviewing the role of EVs in liver diseases to reveal the impact of EVs in pathological processes (e.g., hepatitis, liver fibrosis, and cirrhosis) and elucidate their signaling functions at the molecular level. Subsequently, the work concentrates on the functions of EVs in liver regeneration, revealing their key role in repair and regeneration following liver injury by carrying growth factors, nucleic acids, and other bioactive molecules. This part not only theoretically clarifies the mechanisms of EVs in liver regeneration but also experimentally demonstrates their role in promoting liver cell proliferation, inhibiting apoptosis, regulating immune responses, and fostering angiogenesis, laying the groundwork for future clinical applications. Moreover, this work provides a comprehensive analysis of the challenges faced by existing EV-based therapies in liver regeneration and offers prospects for future research directions. It highlights that despite the tremendous potential of EVs in treating liver diseases, there are still technical challenges (e.g., EV isolation and purification, dosage control, and targeted delivery). To overcome these challenges, the review suggests improvements to current technologies and the development of new methods to realize the clinical application of EVs in treating liver diseases.

Generally, diabetic wounds heal very slowly and inefficiently with an increasing risk of infections. Recent nanotechnology and biomaterial advances elaborate developed multi-functional hydrogels and nanoparticles offer promising solutions to accelerate wound healing for diabetic patients. This research work demonstrates to use of solvent diffusion method to develop hydrogel nanocomposites composed of chitosan (CS), hyaluronic acid (HA), gold (Au), and fibroblast growth factors (FGF). The biological analysis of nanocomposites exhibited enhanced wound healing efficiency by incorporating bioactive molecules like FGF and bioactive Au nanoparticles. In vitro, cell compatibility analysis (MTT assay) of prepared hydrogel nanocomposites was studied on fibroblast cell lines NIH-3T3-L1 and L929 and exhibited greater cell survival ability (>90 %), cell proliferation and migration ability, which demonstrated the suitability of nanocomposite for wound healing treatment. In vitro, anti-bacterial analyses established that FGF-Au@CS/HA has strong antibacterial effectiveness against gram-positive and gram-negative pathogens. The observation of the present research revealed that prepared FGF-Au@CS/HA hydrogel composites could be a suitable biomaterial for diabetic wound care, potentially improving its antibacterial and healing efficacies.

Introduction: Self-assembling peptide nanofibers have emerged as promising biomaterials in the realm of bone tissue engineering due to their biocompatibility, biodegradability, and ability to mimic the native extracellular matrix. This study delved into the comparative efficacy of two distinct self-assembling peptide nanofibers, RADA-BMHP1 and KSL-BMHP1, both incorporating the biological motif of BMHP1, but differing in their core peptide sequences.

Methods: Cell viability and osteogenic differentiation in rat mesenchymal stem cells (rMSCs), and bone regeneration in rat were compared.

Results: In vitro assays revealed that KSL-BMHP1 promoted enhanced cell viability, and nitric oxide production than RADA-BMHP1, an effect potentially attributable to its lower hydrophobicity and higher net charge at physiological pH. Conversely, RADA-BMHP1 induced superior osteogenic differentiation, evidenced by upregulation of key osteogenic genes, increased alkaline phosphatase activity (ALP), and enhanced matrix mineralization which may be attributed to its higher protein-binding potential and grand hydropathy, facilitating interactions between the peptide nanofibers and proteins involved in osteogenesis. In vivo experiments utilizing a rat bone defect model demonstrated that both peptide nanofibers improved bone regeneration at the genes level and ALP activity, with RADA-BMHP1 exhibiting a more pronounced increase in bone formation compared to KSL-BMHP1. Histological evaluation using H&E, Masson's trichrome and Wright-Giemsa staining confirmed the biocompatibility of both nanofibers.

Conclusion: These findings underscore the pivotal role of the core structure of self-assembling peptide nanofibers, beyond their biological motif, in the fate of tissue regeneration. Further research is warranted to optimize the physicochemical properties and functionalization of these nanofibers to enhance their efficacy in bone regeneration applications.

Neurodegenerative diseases are central or peripheral nervous system disorders associated with progressive brain cell degeneration. Common neurodegenerative diseases such as Alzheimer's disease, Parkinson's disease, Huntington's disease, and amyotrophic lateral sclerosis have been widely studied. However, current therapeutics only reduce the symptoms and do not ameliorate the pathogenesis of these diseases. Recent studies suggested the roles of neuroinflammation, apoptosis, and oxidative stress in neurodegenerative diseases. Mesenchymal stem cells (MSCs) exert anti-apoptotic, anti-inflammatory, and antioxidative effects. Therefore, investigating the effects of MSCs and their applications may lead to the discovery of more effective therapies for neurodegenerative diseases. In this study, we review different approaches used to identify therapies for neurodegenerative diseases using MSCs.

Background: Breast cancer is the most common cancer among women. Partial mastectomy is an alternative to mastectomy in early-stage breast cancer to restore a poor quality of life (QOL). However, the aesthetic satisfaction with this procedure is inadequate. The standard methods for breast reconstruction have certain limitations. We developed bioabsorbable implants consisting of an outer mesh composed of poly L-lactic acid (PLLA) and an inner component filled with a collagen sponge (CS). These implants were designed to promote and sustain adipogenesis in vivo, without the addition of exogenous cells or growth factors. In this study, we used PLLA mesh implants to investigate the effects of irradiation on fat formation, which is important in partial mastectomy.

Methods: The implants were inserted into both the inguinal regions of the rats. One month after the implantation, a dose of 13 Gy was delivered to the left-side implants. We compared adipose tissue formation in the non-irradiated and irradiated groups at 6 and 12 months after irradiation.

Results: Irradiation of implants did not lead to malignant tumor formation. The newly formed tissues and adipose tissue were not significantly different between the two groups at 6 and 12 months after irradiation.

Conclusions: PLLA mesh implants containing CS are desirable bioabsorbable implants that can be replaced with autologous adipose tissue after in vivo implantation under irradiation. These implants serve as an effective material for partial mastectomy and have the potential to improve the QOL of patients after mastectomy.

Autologous fat grafting technology has become a new method for breast reconstruction after breast surgery due to its advantages of simple operation, low immunogenicity, fewer complications, high patient acceptance, and natural filling effect. However, the unpredictable fate of transplanted fat limits its widespread application. Currently, many studies have made certain progress in improving the survival rate of fat grafts. This article provides an overview of autologous fat grafting technology, including the mechanisms of fat graft survival, techniques for obtaining and transplanting adipose tissue, methods for enhancing graft survival, and complications associated with fat grafting.