Local delivery of platelet-derived factors mitigates ischemia and preserves ovarian function through angiogenic modulation: A personalized regenerative strategy for fertility preservation.

As the most prominent and ideal modality in female fertility preservation, ovarian tissue cryopreservation, and transplantation often confront the challenge of ischemic damage and follicular loss from avascular transplantation. To surmount this impediment, we engineered a novel platelet-derived factors-encapsulated fibrin hydrogel (PFH), a paradigmatic biomaterial. PFH encapsulates autologous platelet-derived factors, utilizing the physiological blood coagulation cascade for precise local delivery of bioactive molecules. In our study, PFH markedly bolstered the success of avascular ovarian tissue transplantation. Notably, the quantity and quality of follicles were preserved with improved neovascularization, accompanied by decreased DNA damage, increased ovulation, and superior embryonic development rates under a Low-concentration Platelet-rich plasma-derived factors encapsulated fibrin hydrogel (L-PFH) regimen. At a stabilized point of tissue engraftment, gene expression analysis mirrored normal ovarian tissue profiles, underscoring the effectiveness of L-PFH in mitigating the initial ischemic insult. This autologous blood-derived biomaterial, inspired by nature, capitalizes on the blood coagulation cascade, and combines biodegradability, biocompatibility, safety, and cost-effectiveness. The adjustable properties of this biomaterial, even in injectable form, extend its potential applications into the broader realm of personalized regenerative medicine. PFH emerges as a promising strategy to counter ischemic damage in tissue transplantation, signifying a broader therapeutic prospect. (197 words).

Exploring the Interfacial Dynamics of Unilamellar and Multilamellar Cationic Liposomes on SiO2 and Their Interactions with Membrane-Active Peptide.

Understanding the interplay between lipid assemblies and solid supports is crucial for advancing model membrane systems and biomedical applications. This study investigates the interfacial behaviors of unilamellar and multilamellar cationic liposomes on silicon dioxide and their interactions with a membrane-active AH peptide. Using QCM-D monitoring, unilamellar liposomes were found to rapidly form SLBs through one-step adsorption kinetics, whereas multilamellar liposomes exhibited slower adsorption. Further addition of liposomes caused fusogenic interactions with SLBs, where multilamellar liposomes formed more rigid lipid membranes. Upon AH peptide exposure, unilamellar-based lipid membranes showed higher susceptibility to structural transformations, achieving complete SLB formation, while multilamellar-based lipid membranes displayed reduced sensitivity and retained residual viscoelastic components, indicative of incomplete SLB formation. These findings underscore the significant influence of liposome lamellarity on their interfacial dynamics and peptide interactions, crucial for designing effective lipid-based delivery and sensing systems.

Injectable Biomimetic Hydrogel Constructs for Cell-Based Menopausal Hormone Therapy with Reduced Breast Cancer Potential.

Hormone replacement therapy (HRT) has been a primary method in menopausal women and patients with ablated ovaries, but safety has been a concern. Cell-based HRT has emerged as an alternative approach without side effects causing pharmaceutical HRT via 3-dimensionally engineered constructs layering ovarian hormone-producing cells. In this study, we applied micro-sized ovarian cell-laden hydrogel beads as an approach to cell-based HRT using a minimally invasive method in the menopausal rat model. Here, we constructed GC/TC-laden microbeads (GTBs; GC, granulosa cell; TC, theca cell) that allow crosstalk between endocrine cells, encapsulating multiple beads for the figuration of the original ovary. We assessed the ovarian hormone production function of GTB through in vitro culture for 90 days. We applied it to a menopausal rat model and confirmed that GTB-injected rats restored their endocrine function, leading to the regeneration of the thinned endometrium and the maintenance of regular estrous cycles in some individuals. Additionally, it was observed to alleviate menopausal symptoms, including body weight gain and osteoporosis. Notably, the GTB-injected rats did not show mammary gland hyperplasia observed in the pharmaceutical HRT groups and exhibited fewer p53- and KI67-positive and an increase in phosphatase and tensin homolog-positive mammary gland epithelial cells compared to pharmaceutical hormone-treated rats. These results suggest that GTB-based HRT could present a lower risk of breast cancer compared to conventional pharmaceutical-HRT use. Our study highlights the potential of cell-based HRT using an injectable artificial ovary, offering a safer alternative for women requiring HRT.

Elucidating Structural Configuration of Lipid Assemblies for mRNA Delivery Systems.

The development of mRNA delivery systems utilizing lipid-based assemblies holds immense potential for precise control of gene expression and targeted therapeutic interventions. Despite advancements in lipid-based gene delivery systems, a critical knowledge gap remains in understanding how the biophysical characteristics of lipid assemblies and mRNA complexes influence these systems. Herein, we investigate the biophysical properties of cationic liposomes and their role in shaping mRNA lipoplexes by comparing various fabrication methods. Notably, an innovative fabrication technique called the liposome under cryo-assembly (LUCA) cycle, involving a precisely controlled freeze-thaw-vortex process, produces distinctive onion-like concentric multilamellar structures in cationic DOTAP/DOPE liposomes, in contrast to a conventional extrusion method that yields unilamellar liposomes. The inclusion of short-chain DHPC lipids further modulates the structure of cationic liposomes, transforming them from multilamellar to unilamellar structures during the LUCA cycle. Furthermore, the biophysical and biological evaluations of mRNA lipoplexes unveil that the optimal N/P charge ratio in the lipoplex can vary depending on the structure of initial cationic liposomes. Cryo-EM structural analysis demonstrates that multilamellar cationic liposomes induce two distinct interlamellar spacings in cationic lipoplexes, emphasizing the significant impact of the liposome structures on the final structure of mRNA lipoplexes. Taken together, our results provide an intriguing insight into the relationship between lipid assembly structures and the biophysical characteristics of the resulting lipoplexes. These relationships may open the door for advancing lipid-based mRNA delivery systems through more streamlined manufacturing processes.

Multifunctional Material Building Blocks from Plant Pollen.

With its multifaceted nature, plant pollen serves not only as a key element in the reproductive cycle of seed plants but also as an influential contributor to environmental, human health, safety, and climate-related concerns. Pollen functions as a carrier of nutrients and organisms and holds a pivotal role in sustaining pollinator populations. Moreover, it is vital in ensuring the safety and quality of our food supply while presenting potential therapeutic applications. Pollen, often referred to as the diamond of the organic world due to its distinctive physical structures and properties, has been underappreciated from a material science and engineering standpoint. We propose adopting a more interdisciplinary and comprehensive approach to its study. Recent groundbreaking research has focused on the development of pollen-based building blocks that transform practically indestructible plant pollen into microgel, paper, and sponge, thereby unveiling numerous potential applications. In this review, we highlight the transformative potential of plant pollen as it is converted into a variety of building blocks, thereby unlocking myriad prospective applications through eco-friendly processing.

Development and Characterization of Low Temperature Wafer-Level Vacuum Packaging Using Cu-Sn Bonding and Nanomultilayer Getter.

Most microsensors are composed of devices and covers. Due to the complicated structure of the cover and various other requirements, it difficult to use wafer-level packaging with such microsensors. In particular, for monolithic microsensors combined with read-out ICs, the available process margins are further reduced due to the thermal and mechanical effects applied to IC wafers during the packaging process. This research proposes a low-temperature, wafer-level vacuum packaging technology based on Cu-Sn bonding and nano-multilayer getter materials for use with microbolometers. In Cu-Sn bonding, the Cu/Cu3Sn/Cu microstructure required to ensure reliability can be obtained by optimizing the bonding temperature, pressure, and time. The Zr-Ti-Ru based nanomultilayer getter coating inside the cap wafer with high step height has been improved by self-aligned shadow masking. The device pad, composed of bonded wafer, was opened by wafer grinding, and the thermoelectrical properties were evaluated at the wafer-level. The bonding strength and vacuum level were characterized by a shear test and thermoelectrical test using microbolometer test pixels. The vacuum level of the packaged samples showed very narrow distribution near 50 mTorr. This wafer-level packaging platform could be very useful for sensor development whereby high reliability and excellent mechanical/optical performance are both required. Due to its reliability and the low material cost and bonding temperature, this wafer-based packaging approach is suitable for commercial applications.

Induction of ferroptosis using functionalized iron-based nanoparticles for anti-cancer therapy.

Ferroptosis, a cell death pathway that is induced in response to iron, has recently attracted remarkable attention given its emerging therapeutic potential in cancer cells. The need for a promising modality to improve chemotherapy's efficacy through this pathway has been urgent in recent years, and this non-apoptotic cell death pathway accumulates reactive oxygen species (ROS) and is subsequently involved in lipid peroxidation. Here, we report cancer-targeting nanoparticles that possess highly efficient cancer-targeting ability and minimal systemic toxicity, thereby leading to ferroptosis. To overcome the limit of actual clinical application, which is the ultimate goal due to safety issues, we designed safe nanoparticles that can be applied clinically. Nanoparticles containing ferroptosis-dependent iron and FDA-approved hyaluronic acid (FHA NPs) are fabricated by controlling physicochemical properties, and the FHA NPs specifically induce ROS production and lipid peroxidation in cancer cells without affecting normal cells. The excellent in vivo anti-tumor therapeutic effect of FHA NPs was confirmed in the A549 tumor-bearing mice model, indicating that the induction of FHA NP-mediated cell death via the ferroptosis pathway could serve as a powerful platform in anticancer therapy. We believe that this newly proposed FHA NP-induced ferroptosis strategy is a promising system that offers the potential for efficient cancer treatment and provides insight into the safe design of nanomedicines for clinical applications.

Ovarian Tissue-Based Hormone Replacement Therapy Recovers Menopause-Related Signs in Mice.

PURPOSE: In women, menopause manifests with a variety of symptoms related to sex-hormone deficiency. Supplementing steroid hormones with pharmacological drugs has been widely practiced. However, considering the possible complications associated with artificial hormone therapy, studies have been conducted to find an alternative to pharmacological hormone replacement therapy. Accordingly, this study aimed to evaluate the efficacy of tissue-based hormone replacement therapy (tHRT) for treating post-menopausal signs and symptoms. MATERIALS AND METHODS: CD-1 mice were ovariectomized, and the ovaries were cryopreserved. Following artificial induction of post-menopausal osteoporosis, cryopreserved ovaries were subcutaneously autografted, and indexes related to bone health were monitored for 12 weeks. Bone mineral density (BMD), bone mineral contents (BMC), total bone volume (BV), and body fat mass were measured by dual energy X-ray absorptiometry. Uterine atrophy was assessed histologically, and bone microstructures were imaged by micro-computed tomography analysis. RESULTS: Regardless of the number of grafted ovaries, the BMC, BMD, and BV values of mice that underwent ovary transplantation were better than those that did not undergo transplantation. The uteruses in these mice were thicker and heavier after auto-transplantation. Furthermore, the bone microstructure recovered after tHRT. CONCLUSION: Recovery of menopause-related bone loss and uterine atrophy was achieved through tHRT. Ovarian tissue cryopreservation and transplantation may be applicable not only in patients wanting to preserve fertility but also in sex hormone-deficient post-menopausal women.

Promotion of angiogenesis toward transplanted ovaries using nitric oxide releasing nanoparticles in fibrin hydrogel.

Transplantation of ovary is one method of facilitating fertility preservation to increase the quality of life of cancer survivors. Immediately after transplantation, ovaries are under ischemic conditions owing to a lack of vascular anastomosis between the graft and host tissues. The transplanted ovaries can suffer damage because of lack of oxygen and nutrients, resulting in necrosis and dysfunction. In the technique proposed in this paper, the ovary is encapsulated with nitric oxide-releasing nanoparticles (NO-NPs) in fibrin hydrogels, which form a carrying matrix to prevent ischemic damage and accelerate angiogenesis. The low concentration of NO released from mPEG-PLGA nanoparticles elicits blood vessel formation, which allows transplanted ovaries in the subcutis to recover from the ischemic period. In experiments with mice, the NO-NPs/fibrin hydrogel improved the total number and quality of ovarian follicles after transplantation. The intra-ovarian vascular density was 4.78 folds higher for the NO-NPs/fibrin hydrogel groups compared to that for the nontreated groups. Finally,in vitrofertilization revealed a successful blastocyst formation rate for NO-NPs/fibrin hydrogel coated ovaries. Thus, NO-NPs/fibrin hydrogels can provide an appropriate milieu to promote angiogenesis and be considered as adjuvant surgery materials for fertility preservation.

Enhanced NO-induced angiogenesis via NO/H2S co-delivery from self-assembled nanoparticles.

Nitric oxide (NO) and hydrogen sulfide (H2S) have been the focus of research as therapeutic agents because of their biological functions. The controlled release of NO and H2S can enhance NO-induced angiogenesis by H2S inhibiting PDE5A. Polymeric carriers have been researched to deliver gasotransmitters and used as therapeutic agents because of their important ability to help control the concentration of NO and H2S. Here, NO/H2S-releasing nanoparticles were self-assembled from carboxyl-functionalized mPEG-PLGH-thiobenzamide [(methoxy poly (ethylene glycol-b-lactic-co-glycolic-co-hydroxymethyl propionic acid)-thiobenzamide)], PTA copolymer and encapsulated diethylenetriamine NONOate (DETA NONOate). The PTA copolymers were characterized by FT-IR and 1H NMR, and the PTA-NO nanoparticles (PTA-NO-NPs) were confirmed to have core-shell structures with a size of about 140 nm. The PTA-NO-NPs were demonstrated to be biocompatible with viabilities above 100% in various cell types, with a sustained NO and H2S releasing behavior over 72 h. Co-releasing NO and H2S accelerated tube formation by HUVECs compared to the only NO- or H2S-releasing groups in vitro. Also, PTA-NO-NPs performed enhanced angiogenesis compared to the control groups with statistically significant differences ex vivo. These results indicate the feasibility of medical applications through NO and H2S crosstalk.

The new biocompatible material for mouse ovarian follicle development in three-dimensional in vitro culture systems.

To establish a protocol of optimized three-dimensional (3D) culture of ovarian follicles, various biomaterials have been investigated with regard to their properties and functions on in vitro follicle growth. The present study aims to compare the new biomaterial, extracellular matrix-derived soft hydrogel (ES-hydrogel) and alginate, and evaluate the effects of biomaterials on further in vitro 3D culture growth of ovarian follicle and oocyte maturation. The isolated follicles from mouse ovaries were randomly divided into two-dimensional (2D) culture, alginate and ES-hydrogel, and just seeded on culture wells (2D culture) or encapsulated with alginate or ES-hydrogel (3D culture). Culture media from each group were collected on days 4, 8 and 10 or 11 for 17ß-oestradiol (E2) and progesterone (P4) measurement. On day 10 of in vitro culture, follicular survival and pseudo-antrum formation rate were examined, and oocyte maturation was induced by adding human chorionic gonadotropin and epidermal growth factor. After 17 h, ovulated mature oocytes collected and analyzed for oocyte diameter, normal spindle and chromosome alignment configuration, reactive oxygen species (ROS) level, and mitochondrial membrane potential level. To compare mechanical properties of two biomaterials, storage modulus was measured with the advanced rheometric expansion system. Our results showed that follicles cultured in ES-hydrogel, were significantly superior to those cultured 2D or alginate in the pseudo-antrum formation rate, cumulus-oocyte complexes (COCs) rate, MII oocyte rate, normal spindle rate, and E2 production. The ES-hydrogel and alginate groups were not significantly different in follicle survival rate, oocyte diameter, P4 production, ROS, and mitochondrial membrane potential levels. The storage modulus of ES-hydrogel was lower than that of alginate, suggesting that the improved follicular physiology and oocyte maturation in the ES-hydrogel group was due to better hormone exchange through a less stiff encapsulating material. This study shows that 3D culture system using ES-hydrogel effectively improve the outcome of in vitro ovarian follicle culture, supporting follicle morphology and growth and enhancing oocyte maturation. It means one of the most important factors for 3D culture of ovarian follicle was the selection of appropriate and effective biomaterial that can preserve the structure and morphology of ovarian follicle and facilitate nutrition and hormone exchange.

Inducing angiogenesis with the controlled release of nitric oxide from biodegradable and biocompatible copolymeric nanoparticles.

PURPOSE: Nitric oxide (NO) can be clinically applied at low concentrations to regulate angiogenesis. However, studies using small molecule NO donors (N-diazeniumdiolate, S-nitrosothiol, etc) have yet to meet clinical requirements due to the short half-life and initial burst-release profile of NO donors. In this study, we report the feasibility of methoxy poly(ethylene glycol)-b-poly(lactic-co-glycolic acid) (mPEG-PLGA) nanoparticles (NPs) as NO-releasing polymers (NO-NPs) for inducing angiogenesis. MATERIALS AND METHODS: The mPEG-PLGA copolymers were synthesized by typical ring-opening polymerization of lactide, glycolide and mPEG as macroinitiators. Double emulsion methods were used to prepare mPEG-PLGA NPs incorporating hydrophilic NONOate (dieth-ylenetriamine NONOate). RESULTS: This liposomal NP encapsulates hydrophilic diethylenetriamine NONOate (70%±4%) more effectively than other previously reported materials. The application of NO-NPs at different ratios resulted in varying NO-release profiles with no significant cytotoxicity in various cell types: normal cells (fibroblasts, human umbilical vein endothelial cells and epithelial cells) and cancer cells (C6, A549 and MCF-7). The angiogenic potential of NO-NPs was confirmed in vitro by tube formation and ex vivo through an aorta ring assay. Tubular formation increased 189.8% in NO-NP-treated groups compared with that in the control group. Rat aorta exhibited robust sprouting angiogenesis in response to NO-NPs, indicating that NO was produced by polymeric NPs in a sustained manner. CONCLUSION: These findings provide initial results for an angiogenesis-related drug development platform by a straightforward method with biocompatible polymers.

Use of gasotransmitters for the controlled release of polymer-based nitric oxide carriers in medical applications.

Nitric Oxide (NO) is a small molecule gasotransmitter synthesized by nitric oxide synthase in almost all types of mammalian cells. NO is synthesized by NO synthase by conversion of l-arginine to l-citrulline in the human body. NO then stimulates soluble guanylate cyclase, from which various physiological functions are mediated in a concentration-dependent manner. High concentrations of NO induce apoptosis or antibacterial responses whereas low NO circulation leads to angiogenesis. The bidirectional effect of NO has attracted considerable attention, and efforts to deliver NO in a controlled manner, especially through polymeric carriers, has been the topic of much research. This naturally produced signaling molecule has stood out as a potentially more potent therapeutic agent compared to exogenously synthesized drugs. In this review, we will focus on past efforts of using the controlled release of NO via polymer-based materials to derive specific therapeutic results. We have also added studies and our future suggestions on co-delivery methods with other gasotransmitters as a step towards developing multifunctional carriers.

Development of Poly(HEMA-Am) Polymer Hydrogel Filler for Soft Tissue Reconstruction by Facile Polymerization.

The number of breast reconstruction surgeries has been increasing due to the increase in mastectomies. Surgical implants (the standard polydimethylsiloxane (PDMS) implants) are widely used to reconstruct breast tissues, however, it can cause problems such as adverse immune reactions, fibrosis, rupture, and additional surgery. Hence, polymeric fillers have recently garnered increasing attention as strong alternatives for breast reconstruction materials. Polymeric fillers offer noninvasive methods of reconstruction, thereby reducing the possible adverse effects and simplifying the treatment. In this study, we synthesized a 2-hydroxylethylmethacrylate (HEMA) and acrylamide (Am) copolymer (Poly(HEMA-Am)) by redox polymerization to be used as a biocompatible filler material for breast reconstruction. The synthesized hydrogel swelled in phosphate buffered saline (PBS) shows an average modulus of 50 Pa, which is a characteristic similar to that of the standard dermal acrylamide filler. To investigate the biocompatibility and cytotoxicity of the Poly(HEMA-Am) hydrogel, we evaluated an in vitro cytotoxicity assay on human fibroblasts (hFBs) and human adipose-derived stem cells (hADSCs) with the hydrogel eluate, and confirmed a cell viability of over 80% of the cell viability with the Poly(HEMA-Am) hydrogel. These results suggest our polymeric hydrogel is a promising filler material in soft tissue augmentation including breast reconstruction.