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Considerable progress has been made in the development of drug delivery systems for diabetic wounds. However, underlying drawbacks, such as low delivery efficiency and poor tissue permeability, have rarely been addressed. In this study, a multifunctional biohybrid nanorobot platform comprising an artificial unit and several biological components is constructed. The artificial unit is a magnetically driven nanorobot surface modified with antibacterial 2-hydroxypropyltrimethyl ammonium chloride chitosan, which enables the entire platform to move and has excellent tissue penetration capacity. The biological components are two-step engineered extracellular vesicles that are first loaded with mangiferin, a natural polyphenolic compound with antioxidant properties, and then glycoengineered on the surface to enhance cellular uptake efficiency. As expected, the platform is more easily absorbed by endothelial cells and fibroblasts and exhibits outstanding dermal penetration performance and antioxidant properties. Encouraging results are also observed in infected diabetic wound models, showing improved wound re-epithelialization, collagen deposition, angiogenesis, and accelerated wound healing. Collectively, a biohybrid nanorobot platform that possesses the functionalities of both artificial units and biological components serves as an efficient delivery system to promote diabetic wound repair through dual-enhanced cell and tissue penetration and multistep interventions., (© 2024 The Author(s). Advanced Science published by Wiley‐VCH GmbH.)

Optimal tissue recovery and organismal survival are achieved by spatiotemporal tuning of tissue inflammation, contraction and scar formation 1 . Here we identify a multipotent fibroblast progenitor marked by CD201 expression in the fascia, the deepest connective tissue layer of the skin. Using skin injury models in mice, single-cell transcriptomics and genetic lineage tracing, ablation and gene deletion models, we demonstrate that CD201 + progenitors control the pace of wound healing by generating multiple specialized cell types, from proinflammatory fibroblasts to myofibroblasts, in a spatiotemporally tuned sequence. We identified retinoic acid and hypoxia signalling as the entry checkpoints into proinflammatory and myofibroblast states. Modulating CD201 + progenitor differentiation impaired the spatiotemporal appearances of fibroblasts and chronically delayed wound healing. The discovery of proinflammatory and myofibroblast progenitors and their differentiation pathways provide a new roadmap to understand and clinically treat impaired wound healing., (© 2023. The Author(s).)

Injuries to our skin trigger a cascade of spatially- and temporally-synchronized healing processes. During such endogenous wound repair, the role of fibroblasts is multifaceted, ranging from the activation and recruitment of innate immune cells through the synthesis and deposition of scar tissue to the conveyor belt-like transport of fascial connective tissue into wounds. A comprehensive understanding of fibroblast diversity and versatility in the healing machinery may help to decipher wound pathologies whilst laying the foundation for novel treatment modalities. In this review, we portray the diversity of fibroblasts and delineate their unique wound healing functions. In addition, we discuss future directions through a clinical-translational lens., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Knoedler, Broichhausen, Guo, Dai, Knoedler, Kauke-Navarro, Diatta, Pomahac, Machens, Jiang and Rinkevich.)

Deep skin wounds rapidly heal by mobilizing extracellular matrix and cells from the fascia, deep beneath the dermal layer of the skin, to form scars. Despite wounds being an extensively studied area and an unmet clinical need, the biochemistry driving this patch-like repair remains obscure. Lacking also are efficacious therapeutic means to modulate scar formation in vivo. In this study, we identify a central role for p120 in mediating fascia mobilization and wound repair. Injury triggers p120 expression, largely within engrailed-1 lineage-positive fibroblasts of the fascia that exhibit a supracellular organization. Using adeno-associated virus‒mediated gene silencing, we show that p120 establishes the supracellular organization of fascia engrailed-1 lineage-positive fibroblasts, without which fascia mobilization is impaired. Gene silencing of p120 in fascia fibroblasts disentangles their supracellular organization, reducing the transfer of fascial cells and extracellular matrix into wounds and augmenting wound healing. Our findings place p120 as essential for fascia mobilization, opening, to our knowledge, a previously unreported therapeutic avenue for targeted intervention in the treatment of a variety of skin scar conditions., (Copyright © 2022 The Authors. Published by Elsevier Inc. All rights reserved.)

Wound healing disorders are a societal, clinical, and healthcare burden and understanding and treating them is a major challenge. A particularly important cell type in the wound healing processes is the fibroblast. Fibroblasts are not homogenous; however, there are diverse functional fibroblast subtypes coming from different embryonic origins and residing in dispersed anatomic locations including distinct classes of fibroblasts at various skin depths. In this review, we discuss the implications of fibroblast heterogeneity, with a focus on the fundamental physiological functions of the fibroblast subtypes that govern wound repair and clinical degrees of healing. A better understanding of these diverse functional fibroblast populations will likely lead to novel therapies to enhance wound healing and inhibit excessive scarring., (Copyright © 2023 Cold Spring Harbor Laboratory Press; all rights reserved.)

Strategies for therapeutic lymphangiogenesis are gradually directed toward the use of growth factor preparations. In particular, blood-derived growth factor products, including Hypoxia Preconditioned Serum (HPS) and Platelet-rich Plasma (PRP), are both clinically employed for accelerating tissue repair and have received considerable attention in the field of regenerative medicine research. In this study, a comparative analysis of HPS and PRP was conducted to explore their lymphangiogenic potential. We found higher pro-lymphangiogenic growth factor concentrations of VEGF-C, PDGF-BB, and bFGF in HPS in comparison to normal serum (NS) and PRP. The proliferation and migration of lymphatic endothelial cells (LECs) were promoted considerably with both HPS and PRP, but the strongest effect was achieved with HPS-40% dilution. Tube formation of LECs showed the highest number of tubes, branching points, greater tube length, and cell-covered area with HPS-10%. Finally, the effects were double-validated using an ex vivo lymphatic ring assay, in which the highest number of sprouts and the greatest sprout length were achieved with HPS-10%. Our findings demonstrate the superior lymphangiogenic potential of a new generation blood-derived secretome obtained by hypoxic preconditioning of peripheral blood cells-a method that offers a novel alternative to PRP.

The refractory diabetic wound has remained a worldwide challenge as one of the major health problems. The impaired angiogenesis phase during diabetic wound healing partly contributes to the pathological process. MicroRNA (miRNA) is an essential regulator of gene expression in crucial biological processes and is a promising nucleic acid drug in therapeutic fields of the diabetic wound. However, miRNA therapies have limitations due to lacking an effective delivery system. In the present study, we found a significant reduction of miR-31-5p expression in the full-thickness wounds of diabetic mice compared to normal mice. Further, miR-31-5p has been proven to promote the proliferation, migration, and angiogenesis of endothelial cells. Thus, we conceived the idea of exogenously supplementing miR-31-5p mimics to treat the diabetic wound. We used milk-derived exosomes as a novel system for miR-31-5p delivery and successfully encapsulated miR-31-5p mimics into milk exosomes through electroporation. Then, we proved that the miR-31-5p loaded in exosomes achieved higher cell uptake and was able to resist degradation. Moreover, our miRNA-exosomal formulation demonstrated dramatically improved endothelial cell functions in vitro , together with the promotion of angiogenesis and enhanced diabetic wound healing in vivo . Collectively, our data showed the feasibility of milk exosomes as a scalable, biocompatible, and cost-effective delivery system to enhance the bioavailability and efficacy of miRNAs.

Oxidative damage is a critical cause of diabetic wounds. Exosomes from various stem cells could promote wound repair. Here, we investigated the potential mechanism by which exosomes from adipose-derived stem cells (ADSC-EXOs) promote diabetic wound healing through the modulation of oxidative stress. We found that ADSC-EXOs could promote proliferation, migration, and angiogenesis in keratinocytes, fibroblasts, and endothelial cells. Furthermore, ADSC-EXOs reduced the reactive oxygen species (ROS) levels in these cells and protected them against hypoxic and oxidative stress damage. Finally, the local injection of ADSC-EXOs at wound sites significantly increased collagen deposition and neovascularization while reducing ROS levels and cell death; thus, it led to accelerated diabetic wound closure. The mechanism underlying ADSC-EXO functions involved heat-shock protein 90 (HSP90) expressed on the cell surface; these functions could be inhibited by an anti-HSP90 antibody. Exosomal HSP90 could bind to the low-density lipoprotein receptor-related protein 1 (LRP1) receptor on the recipient cell membrane, leading to activation of the downstream AKT signaling pathway. Knockdown of LRP1 and inhibition of the AKT signaling pathway by LY294002 in fibroblasts was sufficient to impair the beneficial effect of ADSC-EXOs. In summary, ADSC-EXOs significantly accelerated diabetic wound closure through an exosomal HSP90/LRP1/AKT signaling pathway.

Background: Human adipose stem cells (ASCs) have emerged as a promising treatment paradigm for skin wounds. Recent works demonstrate that the therapeutic effect of stem cells is partially mediated by extracellular vesicles, which comprise exosomes and microvesicles. In this study, we investigate the regenerative effects of isolated microvesicles from ASCs and evaluate the mechanisms how ASC microvesicles promote wound healing., Methods: Adipose stem cell-derived microvesicles (ASC-MVs) were isolated by differential ultracentrifugation, stained by PKH26, and characterized by electron microscopy and dynamic light scattering (DLS). We examined ASC-MV effects on proliferation, migration, and angiogenesis of keratinocytes, fibroblasts, and endothelial cells both in vitro and in vivo. Next, we explored the underlying mechanisms by gene expression analysis and the activation levels of AKT and ERK signaling pathways in all three kinds of cells after ASC-MV stimulation. We then assessed the effect of ASC-MVs on collagen deposition, neovascularization, and re-epithelialization in an in vivo skin injury model., Results: ASC-MVs could be readily internalized by human umbilical vein endothelial cells (HUVECs), HaCAT, and fibroblasts and significantly promoted the proliferation, migration, and angiogenesis of these cells both in vitro and in vivo. The gene expression of proliferative markers (cyclin D1, cyclin D2, cyclin A1, cyclin A2) and growth factors (VEGFA, PDGFA, EGF, FGF2) was significantly upregulated after ASC-MV treatment. Importantly, ASC-MVs stimulated the activation of AKT and ERK signaling pathways in those cells. The local injection of ASC-MVs at wound sites significantly increased the re-epithelialization, collagen deposition, and neovascularization and led to accelerated wound closure., Conclusions: Our data suggest that ASC-MVs can stimulate HUVEC, HaCAT, and fibroblast functions. ASC-MV therapy significantly accelerates wound healing, and the benefits of ASC-MVs may due to the involvement of AKT and ERK signaling pathways. This illustrates the therapeutic potential of ASC-MVs which may become a novel treatment paradigm for cutaneous wound healing.

Injured human skin fails to regenerate, resulting in scar formation. Annually, 100 million new skin-scarring incidents, occurring as a result of surgery, disease, burns, or sports-related damage, remain untreated. Here, we review knowledge gained from scar-free experimental animal models that have natural regenerative mechanisms for scar-free skin recovery. We also focus on the unique role of endogenous stem cells and other cellular and molecular factors, including the balance of the transforming growth factor-beta (TGF-β) pathway in the context of human skin regeneration. This new strategy opens a new window in drug development for scar-free skin regeneration treatments in both the clinical and cosmetic practice settings., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

A semi-automated workflow for evaluation of diaphyseal fracture treatment of the femur has been developed and implemented. The aim was to investigate the influence of locking compression plating with diverse fracture-specific screw configurations on interfragmentary movements (IFMs) with the use of finite element (FE) analysis. Computed tomography (CT) data of a 22-year-old non-osteoporotic female were used for patient specific modeling of the inhomogeneous material properties of bone. Hounsfield units (HU) were exported and assigned to elements of a FE mesh and converted to mechanical properties such as the Young's modulus followed by a linear FE analysis performed in a semi-automated fashion. IFM on the near and far cortex was evaluated. A positive correlation between bridging length and IFM was observed. Optimal healing conditions with IFMs between 0.5 mm and 1 mm were found in a constellation with a medium bridging length of 80 mm with three unoccupied screw holes around the fracture gap. Usage of monocortical screws instead of bicortical ones had negligible influence on the evaluated parameters when modeling non-osteoporotic bone. Minimal user input, automation of the procedure and an efficient computation time ensured quick delivery of results which will be essential in a future clinical application.

Delayed wound healing and scar formation are among the most frequent complications after surgical interventions. Although biodegradable surgical sutures present an excellent drug delivery opportunity, their primary function is tissue fixation. Mesenchymal stem cells (MSC) act as trophic mediators and are successful in activating biomaterials. Here biodegradable sutures were filled with adipose-derived mesenchymal stem cells (ASC) to provide a pro-regenerative environment at the injured site. Results showed that after filling, ASCs attach to the suture material, distribute equally throughout the filaments, and remain viable in the suture. Among a broad panel of cytokines, cell-filled sutures constantly release vascular endothelial growth factor to supernatants. Such conditioned media was evaluated in an in vitro wound healing assay and showed a significant decrease in the open wound area compared to controls. After suturing in an ex vivo wound model, cells remained in the suture and maintained their metabolic activity. Furthermore, cell-filled sutures can be cryopreserved without losing their viability. This study presents an innovative approach to equip surgical sutures with pro-regenerative features and allows the treatment and fixation of wounds in one step, therefore representing a promising tool to promote wound healing after injury.

The prevalence of chronic wounds is closely correlated to the aging population and so-called civilizational diseases. Therefore, they are causing morbidity and mortality of millions of patients worldwide, with an unbroken upward trend. As a consequence, chronic wounds induce enormous and rapidly growing costs for our health care systems and society in general. Thus, medically effective and cost-efficient treatment methods are urgently needed. Methods of 'regenerative medicine' might offer innovative scientific solutions, including the use of stem cells, growth factors and new bioactive materials. These tools are experimentally well described but clinically poorly performed. The main reasons for this are both legislative and economic. This review describes state-of-the-art techniques, up-to-date research projects, innovative preclinical and clinical approaches in wound care, and activities to translate these innovative techniques into clinical routine., (Copyright © 2012 S. Karger AG, Basel.)

Background: With the development of cell-based gene transfer techniques, genetically modified human keratinocytes (Kc) and fibroblasts (Fb) have been proven to be a better choice in wound repair., Methods: This study was designed to construct in one step a gene-modified artificial skin by a genetically engineered Kc expressing PDGF-BB and Fb expressing VEGF(165) and bFGF. The wound healing effect in a full-thickness wound model was then observed. Unmodified artificial skin served as control. On the post-operative days 7, 14, and 21, residual wound area was calculated and skin wound tissues were subjected to biopsy for further investigation., Results: Compared with unmodified artificial skin, gene-modified artificial skin resulted in a reduced wound contraction and a well-organized human epidermis and better formed dermis., Conclusions: The results suggest that our two-layer, gene-modified artificial skin improved both vascularization and epidermalization for skin regeneration. This technique could bring about a new approach in the treatment of burns and chronic wounds.

Objective: To evaluate the application value of Optical Coherence Tomography (OCT) in the diagnosis of the depth of burn wound., Methods: Deep partial-thickness scald models of Skh-1 mice were reproduced using self-made steam scald appliance. The scald wounds were scanned with OCT 3 hours, or 3 and 8 days after injury respectively. Scanned wound tissue was harvested for histological examination right after each episode of OCT imaging. Normal skin of mice was scanned and examined with the above-mentioned methods at the same time., Results: Compared with those of the normal skin, collagen in the dermis was denatured after steam scald, and it was imaged as vanishing or reduction in birefringence in OCT detection. The structure change intensity was related to the pathological process of the wounds and consistent with the corresponding histological results., Conclusions: OCT is a noninvasive technique. It can be used to diagnose the depth of burn wound in real time.

It has been shown that Pancreatic Stem Cells (PSCs) share many features with skin stem cells. Yet, their potential role in skin regeneration remains to be elucidated. 5x10(5) PSCs from male Rattus norwegicus were seeded on Matriderm scaffold overnight. Cells survival and proliferation were then tested in vitro showing the survival of the cells and their homogenous distribution in the scaffolds. Afterwards, scaffolds were used to replace bilateral full-thickness skin wounds made on the dorsum of Nu/Nu mice. A control group of nude mice received the Matriderm scaffolds without cells. Two weeks after transplantation, wound areas were harvested and analyzed with respect to epithelialization, vascularization and wound closure. The healing area and regeneration rate were significantly increased in the group used the PSCs-seeded scaffolds (factor of 2.1). Vascularization rate showed a significant increase in the PSCs-seeded scaffolds(factor of 1.5). Morphology and immunohistochemistry showed new skin-like structures positive to epidermal markers in the healing wound bed. PSCs were detected in the regenerated tissues. This study showed that the combined use of PSCs with the Matriderm as a scaffold for dermal regeneration significantly increased the epidermalization, vascularization and healing in full-thickness wounds.

We report on our experience with Integra application in 22 patients treated from February 1996 through April 1999. Nine patients suffered a primary burn injury and 13 patients had secondary unstable scars. Integra is artificial skin that consists of a collagen/glycosaminoglycane matrix and a silicon layer. It was first used by Burke and Yannas in 1980 in the USA. The mechanical and biological features of Integra protect the tissue from fluid loss and bacterial invasion in the acutely burned patient. The integra-induced formation of a neodermis leads to functionally and aesthetically highly acceptable scar formation. We present our clinical experience with Integra in long-term follow-up.