Coadministration of adipose-derived stem cells and control-released basic fibroblast growth factor facilitates angiogenesis in a murine ischemic hind limb model.

OBJECTIVE: Adipose-derived stem cells (ASCs) have angiogenic potential owing to their differentiation into endothelial cells and their release of angiogenic growth factors to elicit paracrine effects. In addition, control-released basic fibroblast growth factor (bFGF) sustained with a gelatin hydrogel also supports effective angiogenesis. We sought to determine if coadministration of ASCs and control-released bFGF into murine ischemic limbs facilitates angiogenesis. METHODS: Levels of growth factors in the conditioned media of ASCs cultured with or without control-released bFGF were measured by enzyme-linked immunosorbent assays. A murine ischemic hind limb model was generated and intramuscularly injected with the following: gelatin hydrogel (group 1), a high number of ASCs (group 2), control-released bFGF (group 3), a small number of ASCs and control-released bFGF (group 4), and a high number of ASCs and control-released bFGF (group 5). Macroscopic and microscopic vascular changes were evaluated until day 7 by laser Doppler perfusion imaging and histologic analyses, respectively. RESULTS: Secretion of hepatocyte growth factor, vascular endothelial growth factor, and transforming growth factor-ß1 was enhanced by control-released bFGF. Vascular improvement was achieved in groups 4 and 5 according to laser Doppler perfusion imaging. Hematoxylin and eosin staining and CD31 immunohistochemical staining demonstrated an increase in the vascular density, vessel diameter, and thickness of vessel walls in groups 4 and 5. Cells positively stained for CD146, α-smooth muscle actin, and transforming growth factor-ß1 were observed around vessel walls in groups 4 and 5. CONCLUSIONS: These findings suggest that coadministration of ASCs and control-released bFGF facilitates angiogenesis in terms of vessel maturation in a murine ischemic hind limb model.

Proapoptotic effect of control-released basic fibroblast growth factor on skin wound healing in a diabetic mouse model.

The ability of basic fibroblast growth factor (bFGF) to improve wound healing is attenuated by its short half-life in free form. This study aimed to enhance skin wound healing in a diabetes mouse model while concomitantly decreasing scar formation using control-released bFGF together with acidic gelatin hydrogel microspheres (AGHMs). Bilateral full-thickness wounds (10 mm in diameter) were made on the backs of db/db mice. Forty-five mice were divided into three groups, and the base of the wound under the panniculus carnosus and the wound periphery were injected with phosphate-buffered saline (300 µL) containing (1) control-released bFGF (50 µg), (2) control-released bFGF (20 µg), or (3) AGHMs alone. The size of the wound area was recorded on each postoperative day (POD). Mice were sacrificed on postoperative day 4, 7, 10, 14, and 28, and skin wound specimens were obtained to assess the endothelium/angiogenesis index via cluster of differentiation 31 immunohistochemistry, the proliferation index via Ki-67 immunohistochemistry, and the myofibroblast and fibroblast apoptosis indices by terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling and alpha-smooth muscle actin or vimentin staining, respectively. Epithelialization rates and indices of proliferation and myofibroblast/fibroblast apoptosis were higher in the bFGF groups than in the AGHM group, mainly within 2 weeks of injury. No dose-effect relationship was found for control-released bFGF, although the actions of 50 µg bFGF seemed to last longer than those of 20 µg bFGF. Therefore, control-released bFGF may accelerate diabetic skin wound healing and induce myofibroblast/fibroblast apoptosis, thereby reducing scar formation.

Adverse events related to platelet-rich plasma therapy and future issues to be resolved.

Platelet-rich plasma (PRP) is the portion of plasma with a platelet concentration above baseline that is recovered through centrifugation of autologous blood. PRP therapy is currently used for wound healing and pain relief in diverse medical fields. Although there have been recent reports of adverse events (AEs) possibly related to PRP treatment, the safety profile of PRP treatment remains unclear. Therefore, this review discusses the risks inherent in PRP therapy and the current issues by surveying reports on AEs associated with PRP treatment within different fields. PubMed was searched for research articles referring to AEs associated with PRP therapy from inception to January 2024. Literature survey revealed that PRP therapy may involve several AEs, including postoperative infections, blindness, inflammation, allergic reactions, and nodule development. The most commonly reported AE was postoperative infections. Since PRP therapy generally proceeds in the process of blood collection, manufacturing, and administration to patients, it is conjectured that PRP may have been contaminated with microorganisms at some point in this series of processes, leading to bacterial infection. Additionally, because PRP cannot be sterilized like pharmaceuticals, it is important to prevent microbial contamination during each PRP treatment process. However, the specific process that involves the risk of microbial contamination remains unclear. To take measures to prevent microbial contamination of PRP, it may be necessary to elucidate the risk factors for microbial contamination during PRP treatment. It may be important to elucidate the effectiveness and risks of PRP therapy as well as to establish a follow-up system after PRP treatment. Currently, most reports of AEs related to PRP therapy are case reports; therefore, the accumulation of high-quality evidence and detailed verification are necessary to determine the causal relationship between PRP therapy and each AE.

Association of Social Network with Physical Function Among Community-Dwelling Older Adults in Rural Thailand: A Cross-Sectional Study.

Purpose: As the number of older adults in society increases, their social roles and networks, as well as their physical function, decrease. This study aimed to clarify the association between social networks and physical function among people aged ≥ 60 years in rural Thailand. Patients and Methods: This cross-sectional study was conducted in the Photharam District, Ratchaburi Province, Thailand. Participants were required to be at least 60 years old and be able to walk to the health center. Social networks were surveyed using the Thai version of Lubben Social Network Scores-6. Four physical function measures, namely hand grip strength, five-times-sit-to-stand test, timed up-and-go (TUG) test, and one-leg standing, were considered. Regression analysis was conducted with Lubben Social Network Scores-6 as the dependent variable and the four types of physical function as independent variables. Results: A total of 497 older adults aged 60 years or more were enrolled; 82 were males, and 412 were females. The mean Lubben Social Network Scores-6 was 14.9 ± 5.7. Only the TUG test was associated in a single and multiple regression analysis with the Lubben Social Network Scores-6 as the dependent variable and the four physical function assessments as independent variables. Conclusion: The TUG test assessed the smoothness of normal standing and walking, which are essential physical functions for maintaining a social network and meeting people. This suggests a relationship between physical function and social network.

Identification of risk factors for the onset of delirium associated with COVID-19 by mining nursing records.

COVID-19 has a range of complications, from no symptoms to severe pneumonia. It can also affect multiple organs including the nervous system. COVID-19 affects the brain, leading to neurological symptoms such as delirium. Delirium, a sudden change in consciousness, can increase the risk of death and prolong the hospital stay. However, research on delirium prediction in patients with COVID-19 is insufficient. This study aimed to identify new risk factors that could predict the onset of delirium in patients with COVID-19 using machine learning (ML) applied to nursing records. This retrospective cohort study used natural language processing and ML to develop a model for classifying the nursing records of patients with delirium. We extracted the features of each word from the model and grouped similar words. To evaluate the usefulness of word groups in predicting the occurrence of delirium in patients with COVID-19, we analyzed the temporal changes in the frequency of occurrence of these word groups before and after the onset of delirium. Moreover, the sensitivity, specificity, and odds ratios were calculated. We identified (1) elimination-related behaviors and conditions and (2) abnormal patient behavior and conditions as risk factors for delirium. Group 1 had the highest sensitivity (0.603), whereas group 2 had the highest specificity and odds ratio (0.938 and 6.903, respectively). These results suggest that these parameters may be useful in predicting delirium in these patients. The risk factors for COVID-19-associated delirium identified in this study were more specific but less sensitive than the ICDSC (Intensive Care Delirium Screening Checklist) and CAM-ICU (Confusion Assessment Method for the Intensive Care Unit). However, they are superior to the ICDSC and CAM-ICU because they can predict delirium without medical staff and at no cost.

Interleukin-10 induces TNF-driven apoptosis and ROS production in salivary gland cancer cells.

Treatment resistance after chemo-/immunotherapy occurs in patients with head and neck squamous cell cancers (HNSCs), including salivary gland cancers (SGCs). Interleukin-10 (IL-10), a cytokine with pro- and anti-cancer effects, has an unclear impact on HNSC/SGC cells. We show that HNSC patients exhibiting high expression of IL-10 and its receptor IL-10Rα experience have prolonged overall survival. Immunoreactive IL-10 was low in ductal cells of human SGC biopsies. Human (A253) and murine WR21-SGC cells expressed IL-10Rß, but only A253 cells expressed IL-10 and IL-10Rα. The addition of recombinant IL-10 impaired SGC cell proliferation and induced apoptosis in vitro. N-acetylcysteine restored IL-10-induced reactive oxygen species (ROS) production but did not prevent IL-10-mediated viability loss. Mechanistically, recIL-10 delayed cell cycle progression from G0/G1 to the S phase with cyclin D downregulation and upregulation of NF-kB. IL-10 increased tumor necrosis factor-α (TNF-α) in A253 and WR21 and FasL in WR21 cells. Neutralizing antibodies against TNF-α and NF-kB inhibition restored SGC proliferation after IL-10 treatment, emphasizing the critical role of TNF-α and NF-kB in IL-10-mediated anti-tumor effects. These findings underscore the potential of IL-10 to impede SGC cell growth through apoptosis induction, unraveling potential therapeutic targets for intervention in salivary gland carcinomas.

Concise review: Adipose-derived stem cells as a novel tool for future regenerative medicine.

The potential use of stem cell-based therapies for the repair and regeneration of various tissues and organs offers a paradigm shift that may provide alternative therapeutic solutions for a number of diseases. The use of either embryonic stem cells (ESCs) or induced pluripotent stem cells in clinical situations is limited due to cell regulations and to technical and ethical considerations involved in the genetic manipulation of human ESCs, even though these cells are, theoretically, highly beneficial. Mesenchymal stem cells seem to be an ideal population of stem cells for practical regenerative medicine, because they are not subjected to the same restrictions. In particular, large number of adipose-derived stem cells (ASCs) can be easily harvested from adipose tissue. Furthermore, recent basic research and preclinical studies have revealed that the use of ASCs in regenerative medicine is not limited to mesodermal tissue but extends to both ectodermal and endodermal tissues and organs, although ASCs originate from mesodermal lineages. Based on this background knowledge, the primary purpose of this concise review is to summarize and describe the underlying biology of ASCs and their proliferation and differentiation capacities, together with current preclinical and clinical data from a variety of medical fields regarding the use of ASCs in regenerative medicine. In addition, future directions for ASCs in terms of cell-based therapies and regenerative medicine are discussed.

Periodontal tissue regeneration by combined implantation of adipose tissue-derived stem cells and platelet-rich plasma in a canine model.

BACKGROUND AIMS: One goal of periodontal therapy is to regenerate periodontal tissues. Stem cells, growth factors and scaffolds and biomaterials are vital for the restoration of the architecture and function of complex tissues. Adipose tissue-derived stem cells (ASCs) are an ideal population of stem cells for practical regenerative medicine. In addition, platelet-rich plasma (PRP) can be useful for its ability to stimulate tissue regeneration. PRP contains various growth factors and may be useful as a cell carrier in stem cell therapies. The purpose of this study was to determine whether a mixture of ASCs and PRP promoted periodontal tissue regeneration in a canine model. METHODS: Autologous ASCs and PRP were implanted into areas with periodontal tissue defects. Periodontal tissue defects that received PRP alone or non-implantation were also examined. Histologic, immunohistologic and x-ray studies were performed 1 or 2 months after implantation. The amount of newly formed bone and the scale of newly formed cementum in the region of the periodontal tissue defect were analyzed on tissue sections. RESULTS: The areas of newly formed bone and cementum were greater 2 months after implantation of ASCs and PRP than at 1 month after implantation, and the radiopacity in the region of the periodontal tissue defect increased markedly by 2 months after implantation. The ASCs and PRP group exhibited periodontal tissue with the correct architecture, including alveolar bone, cementum-like structures and periodontal ligament-like structures, by 2 months after implantation. CONCLUSIONS: These findings suggest that a combination of autologous ASCs and PRP promotes periodontal tissue regeneration that develops the appropriate architecture for this complex tissue.

Preclinical efficacy of slow-release bFGF in ischemia-reperfusion injury in a Dorsal Island skin flap model.

The effect of slow-release basic fibroblast growth factor (bFGF) on ischemia-reperfusion injury was examined using an island skin flap model in rats. Paired rectangular island skin flaps were elevated on the dorsum of 30 Fischer rats. The flaps were subjected to 6 hours of ischemia. Before reperfusion the flaps were injected with acidic gelatin hydrogel microspheres + phosphate-buffered saline (PBS) (group I), 20 µg slow-release bFGF + PBS (group II), 50 µg slow-release bFGF + PBS (group III), and 150 µg slow-release bFGF + PBS (group IV). The mean percent flap survival area and the average number of vessels detected by microangiography were significantly higher in group IV (p < 0.05) than in groups I, II, and III. The immunohistochemical staining for vasculogenic growth factors was quantitatively higher in group IV (p < 0.01). In conclusion, slow-release bFGF prevents ischemia-reperfusion injury by upregulating the secretion of vasculogenic growth factors.

Current status of platelet-rich plasma therapy under the act on the safety of regenerative medicine in Japan.

In Japan, a legal framework has been established for the safe and effective application of regenerative medicine. After eight years of the Act on the Safety of Regenerative Medicine (RM Act), discussions have been underway in the Ministry of Health, Labor and Welfare of Japan to revise the law owing to numerous novel technologies and inappropriate case reports not anticipated when the law was enacted. Therefore, in this review article, we have reviewed the regenerative medicine provision plans and the contribution of platelet-rich plasma (PRP) therapy, a regenerative medicine technique widely used in Japan post RM Act implementation, to these plans. As of January 2022, 97.2% of the regenerative medicine provided under the RM Act had been for private practice, and most of them were Class Ⅲ regenerative medicine. Notably, PRP was the most used processed cell under the RM Act. PRP therapy accounted for approximately 66% of the regenerative medicine provision plans in clinical research or private practice and was the most provided regenerative medicine technology in Japan. PRP therapy was primarily used in dentistry to regenerate periodontal tissue (approximately 50%), followed by orthopedics, where it is used to treat osteoarthritis. We suggest that further discussion is essential to determine the factors that should be addressed by the RM act to evaluate the efficacy and safety of PRP therapy.

The Role of the Plasminogen/Plasmin System in Inflammation of the Oral Cavity.

The oral cavity is a unique environment that consists of teeth surrounded by periodontal tissues, oral mucosae with minor salivary glands, and terminal parts of major salivary glands that open into the oral cavity. The cavity is constantly exposed to viral and microbial pathogens. Recent studies indicate that components of the plasminogen (Plg)/plasmin (Pm) system are expressed in tissues of the oral cavity, such as the salivary gland, and contribute to microbial infection and inflammation, such as periodontitis. The Plg/Pm system fulfills two major functions: (a) the destruction of fibrin deposits in the bloodstream or damaged tissues, a process called fibrinolysis, and (b) non-fibrinolytic actions that include the proteolytic modulation of proteins. One can observe both functions during inflammation. The virus that causes the coronavirus disease 2019 (COVID-19) exploits the fibrinolytic and non-fibrinolytic functions of the Plg/Pm system in the oral cavity. During COVID-19, well-established coagulopathy with the development of microthrombi requires constant activation of the fibrinolytic function. Furthermore, viral entry is modulated by receptors such as TMPRSS2, which is necessary in the oral cavity, leading to a derailed immune response that peaks in cytokine storm syndrome. This paper outlines the significance of the Plg/Pm system for infectious and inflammatory diseases that start in the oral cavity.

Clinical Research on the Safety Evaluation of Platelet-rich Plasma Treatment in Oral Diseases: A Study Protocol.

Background: Platelet-rich plasma (PRP) is a biological product obtained from autologous blood that contains growth factors, promoting the healing and regeneration of human tissues. Several oral diseases require surgical intervention, producing residual wounds that undergo a healing process, accompanied by pain, swelling, superinfections, and bone remodeling. This protocol study aims to evaluate the safety of PRP use for the following dental procedures: post-extraction socket healing, periodontal tissue regeneration, maxillary sinus floor elevation, tooth transplantation, and intentional tooth replantation. Methods: Ten patients will be enrolled and subjected to the required treatment with the addition of PRP, after appropriate hematological and biochemical evaluations. The participants will then be subjected to an observation period of 4 weeks to monitor adverse events through clinical observation. Secondary outcomes will regard pain, and clinical evolution of the treated site. Among these, presence of infection, swelling, wound healing, stability of the transplanted tooth. Discussion: Safety of medical procedures represents the first requirement for their introduction in routine practice. A careful evaluation of clinical response during follow-up period and registration of adverse effects is fundamental for safety confirmation and subsequent use of PRP for the proposed dental procedures. Trial registration: Japan Registry of Clinical Trials (https://jrct.niph.go.jp/, registry number: jRCTc030190273, jRCTc030190274, jRCTc030190275, jRCTc030190276, jRCTc030190277; Date of registration: 31 March 2020).

Difficulties in ensuring review quality performed by committees under the Act on the Safety of Regenerative Medicine in Japan.

We outlined five studies regarding the quality of the review by committees based on the Act on the Safety of Regenerative Medicine. The findings raise serious concerns about the independence, integrity, and quality of reviews of therapeutic plans by these committees with inappropriately close relationships to medical institutions and companies.

The stem cell transcription factor ZFP296 transforms NIH3T3 cells and promotes anchorage-independent growth of cancer cells.

Cancer cells and embryonic stem (ES) cells share several biological properties, suggesting that some genes expressed in ES cells may play an important role in cancer cell growth. In this study, we investigated the possible role of zinc finger protein 296 (ZFP296), a transcription factor expressed in ES cells, in cancer development. First, we found that overexpression of Zfp296 in NIH3T3 mouse fibroblasts induced two phenomena indicative of cell transformation: enhanced proliferation under low-serum conditions and anchorage-independent growth. We also found that Zfp296 expression was upregulated in the tumor area of a mouse model of colon carcinogenesis. In addition, the expression levels of ZFP296 in various human cell lines were generally low in normal cells and relatively high in cancer cells. Finally, using a soft agar assay, we found that overexpression of ZFP296 promoted the anchorage-independent growth of cancer cells, while its knockdown had the opposite effect. Overall, these results suggest a possible role of the ES-specific transcription factor ZFP296 in cancer.

Reactogenicity and immunogenicity of BNT162b2 or mRNA-1273 COVID-19 booster vaccinations after two doses of BNT162b2 among healthcare workers in Japan: a prospective observational study.

BACKGROUND: This study evaluates the immunogenicity and reactogenicity of BNT162b2 and mRNA-1273 booster doses after the primary two-dose BNT162b2 series in Japan and is the first report from Western Pacific region. RESEARCH DESIGN AND METHODS: Healthcare workers receiving the two-dose BNT162b2 series and eligible for booster vaccination were enrolled. Self-reported adverse reactions were recorded for 8 days. Antibody titer was measured at baseline and on day 28. RESULTS: A total of 2,931 and 890 subjects received BNT162b2 (homologous) and mRNA-1273 (heterologous) booster vaccinations, respectively. The anti-SARS-CoV-2 spike protein IgG titer increased by 50.9- and 64.3-fold in the homologous and heterologous groups, respectively. Immunogenicity was greater with increasing age, regardless of sex. Adverse reactions were mild to moderate and decreased with age. The most common adverse reactions were injection-site pain (92.2%), fatigue (71.8%), headache (58.3%), and fever ≥37.5°C (46.5%). Two cases of non-severe myocarditis occurred in the heterologous group and resolved without clinical sequelae. CONCLUSION: Homologous booster schedules had fewer reported adverse reactions; heterologous boosters elicited greater immunogenicity. Among different age groups, subjects aged 60 or over had the lowest immunogenicity before the booster, and both homologous and heterologous boosters restored vaccine immunogenicity level comparable to those of younger age groups.

Study protocol for periodontal tissue regeneration with a mixture of autologous adipose-derived stem cells and platelet rich plasma: A multicenter, randomized, open-label clinical trial.

Introduction: Adipose-derived stem cells (ASCs) secrete various growth factors to promote wound healing and to regenerate various tissues, such as bone, cartilage, and fat tissue. Subcutaneous adipose tissue is a considerable cell source in clinical practice and can be collected relatively easily and safely under local anesthesia. Moreover, platelet-rich plasma (PRP), a plasma component containing many platelets purified by centrifuging the collected blood, also promotes wound healing. PRP can be easily gelled and is therefore attracting attention as a scaffolding material for transplanted cells. The usefulness of a mixture of ASCs and PRP for periodontal tissue regeneration has been in vitro demonstrated in our previous study. The aim of this study is to present the protocol of translation of tissue regeneration with ASCs and PRP into practical use, evaluating its efficacy. Methods: This study is a multicenter, randomized, open-label comparative clinical trial. Fifteen patients will be randomly assigned to the treatment with mixture of ASCs and PRP or enamel matrix derivate administration into periodontal tissue defects. Increase in height of new alveolar bone in the transplanted area will be evaluated. The evaluation will be performed using dental radiographs after 36 weeks of transplantation. Occurrence of adverse events will be evaluated as secondary outcome. Results: This clinical study was initiated after meeting the regulations to be complied with, including ethical review and regulatory notifications. Conclusions: If effective, this cell therapy using autologous mesenchymal stem cells can represent a useful medical technology for regeneration of periodontal defects.

Prefabrication of tissue engineered bone grafts: an experimental study.

The purpose of this study was to determine whether angiogenesis could successfully be induced into bone tissue that was engineered by cultured adipose-derived stem cells with porous beta-tricalcium phosphate and whether its biologic properties could be maintained by flap prefabrication technique.Adipose-derived stem cells with porous beta-tricalcium phosphate were implanted into the superficial inferior epigastric artery flap of the Fisher rats. After prefabrication for 8 weeks, the prefabricated flaps were elevated and the pedicles were clamped for 4 hours. The samples were harvested after 2 weeks for analyses.Angiogenesis was significantly increased in the prefabricated groups (P < 0.05). There was no significant difference between the prefabricated and nonprefabricated groups in terms of the osteogenic capacity (P > 0.5).The promising results obtained with prefabrication in tissue engineered bone grafts encourage the clinical application of this technology. Thus, prefabrication may be a useful technique in any engineered bone tissue transfer.

Current status of bone regeneration using adipose-derived stem cells.

Many bone regeneration therapies have been developed for clinical use and have variable outcomes and serious limitations. The goal of bone regeneration is to repair a bone defect in a stable and durable manner. Cellular strategies play an important role in bone tissue engineering. Clinical factors important for successful bone regeneration are the recruitment of cells to the defect site and the production of a suitable extracellular matrix consistent with bone tissues. Adipose-derived stem cells (ASCs) can be obtained in large quantities with little donor site morbidity or patient discomfort. They are multipotent somatic stem cells and have a strong potential to differentiate and secrete growth factors. In this review, we discuss the osteogenic potential of ASCs with/without several types of scaffolds in vivo and their clinical application for bone regeneration.

Bone Regeneration with a Combination of Adipose-Derived Stem Cells and Platelet-Rich Plasma.

Mesenchymal stem cells (MSCs) have the potential to directly differentiate into osteogenic cells and efficiently regenerate bone tissue. Adipose-derived stem cells (ASCs) have the potential to differentiate into an osteogenic lineage, too. In addition, ASCs can be readily harvested in large numbers with low donor-site morbidity. Meanwhile, recent reports have demonstrated that platelet-rich plasma (PRP) contains a variety of growth factors and may be a powerful biological autologous cocktail of growth factors for tissue engineering.We have shown that ASC/PRP admixture had dramatic effects on bone regeneration in a rat calvarial defect model, not only through the osteogenic potential of ASCs, but also through the release of cytokines by platelets in PRP, which, in turn, support ASCs.In this chapter, we introduce the bone regeneration using a combination of ASCs and PRP in a rat calvarial defect model.