Collagen Matrices Research Articles

Combining Bone Collagen Material with hUC-MSCs for Applicationto Spina Bifida in a Rabbit Model.

Spina bifida is one of the neural tube defects, with a high incidence in human birth defects, which seriously affects the health and quality of life of patients. In the treatment of bone defects, the source of autologous bone is limited and will cause secondary damage to the patient. At the same time, since the bone tissue in animals needs to play a variety of biological functions, its complex structure cannot be replaced by a single material. The combination of mechanical materials and biological materials has become a common choice. Human umbilical cord mesenchymal stem cells (hUC-MSCs) have the advantages of easy access, rapid proliferation, low immunogenicity, and no ethical issues. It is often used in the clinical research of tissue regeneration and repair. Therefore, in this study, we established a spina bifida model using Japanese white rabbits. This model was used to screen the best regenerative repair products for congenital spina bifida, and to evaluate the safety of regenerative repair products. The results showed that the combination of hUC-MSCs with collagen material had better regeneration effect than collagen material alone, and had no negative impact on the health of animals. This study provides a new idea for the clinical treatment of spina bifida, and also helps to speed up the research progress of regenerative repair products.

Evaluation of wound healing activity of GelMA/PCLMA fibrous composites in diabetic model rats

New strategies are necessary to develop bandages to poor wound healing from diabetic patients. Here, we produced different groups of multifunctional bandages using electrospinning and surface functionalization, then we evaluated the skin tissue regeneration using an in vivo diabetes model. The bandages displayed good tissue integration, evidenced by fibroblast adhesion and mature collagen (type I) formation. The group have developed these biomaterials and it have been using for different biomedical applications.

Collagen and Silk Fibroin as Promising Candidates for Constructing Catalysts.

A catalyst determines the mechanism of an organic chemical reaction, thus enabling the commercially viable formation of desired material products. Biopolymers offer new opportunities for the construction of catalysts by virtue of their biocompatibility, environmental benignity, and sustainability, as well as their low cost. Biopolymers are especially useful as carriers and precursors in catalysis application. The employment of biocompatible and biosustainable collagen and silk fibroin materials will revolutionize state-of-the-art electronic devices and systems that currently rely on conventional technologies. In this review, we first consider the ordered hierarchical structure, origin, and processing methods of collagen and silk fibroin. Then, the unique advantages and applicability of collagen and silk fibroin for constructing catalysts are summarized. Moreover, a summary of the state-of-the-art design, fabrication, and application of collagen- and silk fibroin-based catalysts, as well as the application of collagen- and silk-based catalysts, is presented by focusing on their roles as carriers and precursors, respectively. Finally, challenges and prospects are assessed for the construction and development of collagen and silk fibroin-based catalysts.

Driving role of head and neck cancer cell secretome on the invasion of stromal fibroblasts: Mechanistic insights by phosphoproteomics

BackgroundCancer-associated fibroblasts (CAFs) are major players in tumor-stroma communication, and participate in several cancer hallmarks to drive tumor progression and metastatic dissemination. This study investigates the driving effects of tumor-secreted factors on CAF biology, with the ultimate goal of identifying effective therapeutic targets/strategies for head and neck squamous cell carcinomas (HNSCC). MethodsFunctionally, conditioned media (CM) from different HNSCC-derived cell lines and normal keratinocytes (Kc) were tested on the growth and invasion of populations of primary CAFs and normal fibroblasts (NFs) using 3D invasion assays in collagen matrices. The changes in MMPs expression were evaluated by RT-qPCR and kinase enrichment was analyzed using mass spectrometry phosphoproteomics. ResultsOur results consistently demonstrate that HNSCC-secreted factors (but not Kc CM) specifically and robustly promoted pro-invasive properties in both CAFs and NFs, thereby reflecting the plasticity of fibroblast subtypes. Concomitantly, HNSCC-secreted factors massively increased metalloproteinases levels in CAFs and NFs. By contrast, HNSCC CM and Kc CM exhibited comparable growth-promoting effects on stromal fibroblasts. Mechanistically, phosphoproteomic analysis predominantly revealed phosphorylation changes in fibroblasts upon treatment with HNSCC CM, and various promising kinases were identified: MKK7, MKK4, ASK1, RAF1, BRAF, ARAF, COT, PDK1, RSK2 and AKT1. Interestingly, pharmacologic inhibition of RAF1/BRAF using sorafenib emerged as the most effective drug to block tumor-promoted fibroblast invasion without affecting fibroblast viability ConclusionsOur findings demonstrate that HNSCC-secreted factors specifically fine tune the invasive potential of stromal fibroblasts, thereby generating tumor-driven pro-invasive niches, which in turn to ultimately facilitate cancer cell dissemination. Furthermore, the RAF/BRAF inhibitor sorafenib was identified as a promising candidate to effectively target the onset of pro-invasive clusters of stromal fibroblasts in the HNSCC microenvironment.

Triclocarban triggers osteoarthritis via DNMT1-mediated epigenetic modification and suppression of COL2A in cartilage tissues

Triclocarban (TCC) is a widely used environmental endocrine-disrupting chemical (EDC). Articular injury of EDCs has been reported; however, whether and how TCCs damage the joint have not yet been determined. Herein, we revealed that exposure to TCC caused osteoarthritis (OA) within the zebrafish anal fin. Mechanistically, TCC stimulates the expression of DNMT1 and initiates DNA hypermethylation of the type II collagen coding gene, which further suppresses the expression of type II collagen and other extracellular matrices. This further results in decreased cartilage tissue and narrowing of the intraarticular space, which is typical of the pathogenesis of OA. The regulation of OA occurrence by TCC is conserved between zebrafish cartilage tissue and human chondrocytes. Our findings clarified the hazard and potential mechanisms of TCC towards articular health and highlighted DNMT1 as a potential therapeutic target for OA caused by TCC.

Full Skin Equivalent Models for Simulation of Burn Wound Healing, Exploring Skin Regeneration and Cytokine Response.

Healing of burn injury is a complex process that often leads to the development of functional and aesthetic complications. To study skin regeneration in more detail, organotypic skin models, such as full skin equivalents (FSEs) generated from dermal matrices, can be used. Here, FSEs were generated using de-epidermalized dermis (DED) and collagen matrices MatriDerm® and Mucomaix®. Our aim was to validate the MatriDerm- and Mucomaix-based FSEs for the use as in vitro models of wound healing. Therefore, we first characterized the FSEs in terms of skin development and cell proliferation. Proper dermal and epidermal morphogenesis was established in all FSEs and was comparable to ex vivo human skin models. Extension of culture time improved the organization of the epidermal layers and the basement membrane in MatriDerm-based FSE but resulted in rapid degradation of the Mucomaix-based FSE. After applying a standardized burn injury to the models, re-epithelization occurred in the DED- and MatriDerm-based FSEs at 2 weeks after injury, similar to ex vivo human skin. High levels of pro-inflammatory cytokines were present in the culture media of all models, but no significant differences were observed between models. We anticipate that these animal-free in vitro models can facilitate research on skin regeneration and can be used to test therapeutic interventions in a preclinical setting to improve wound healing.

Approaches to Determine Nuclear Shape in Cells During Migration Through Collagen Matrices.

Fibrillar collagen is an abundant extracellular matrix (ECM) component of interstitial tissues which supports the structure of many organs, including the skin and breast. Many different physiological processes, but also pathological processes such as metastatic cancer invasion, involve interstitial cell migration. Often, cell movement takes place through small ECM gaps and pores and depends upon the ability of the cell and its stiff nucleus to deform. Such nuclear deformation during cell migration may impact nuclear integrity, such as of chromatin or the nuclear envelope, and therefore the morphometric analysis of nuclear shapes can provide valuable insight into a broad variety of biological processes. Here, we describe a protocol on how to generate a cell-collagen model in vitro and how to use confocal microscopy for the static and dynamic visualization of labeled nuclei in single migratory cells. We developed, and here provide, two scripts that (Fidler, Nat Rev Cancer 3(6):453-458, 2003) enable the semi-automated and fast quantification of static single nuclear shape descriptors, such as aspect ratio or circularity, and the nuclear irregularity index that forms a combination of four distinct shape descriptors, as well as (Frantz et al., J Cell Sci 123 (Pt 24):4195-4200, 2010) a quantification of their changes over time. Finally, we provide quantitative measurements on nuclear shapes from cells that migrated through collagen either in the presence or the absence of an inhibitor of collagen degradation, showing the distinctive power of this approach. This pipeline can also be applied to cell migration studied in different assays, ranging from 3D microfluidics to migration in the living organism.

Obtaining a filter material used in gas and air purification

Modern technologies allow obtaining small diameter fibers used for fine filtration. Combined layer filter materials are made from such fibers. Nanofibers from 20 to 200 nm are actively used as components of filter materials. They are made of synthetic (polyamides, polyesters, aromatic polyamides, polyacrylates), biological (proteins, collagen) materials and activated carbon. According to their effect, they are similar to reverse osmosis membranes (they retain large anions, Ca, Mg cations, heavy metal ions, large organic compounds) and at the same time allow obtaining membranes with high permeability for small sodium ions. It is known that one of the main problems of environmental protection is the reduction of atmospheric pollution. Every year, a large amount of harmful compounds - oxides, various metal compounds, as well as sulfur dioxide and nitrogen oxides, volatile organic compounds, stable organic compounds and acids are released into the atmosphere. The results of the study of the influence of the composite composition and the method of obtaining a filter material on its filtration properties and main indicators are presented.

Invadopodia Methods: Detection of Invadopodia Formation and Activity in Cancer Cells Using Reconstituted 2D and 3D Collagen-Based Matrices.

Tumor dissemination involves cancer cell migration through the extracellular matrix (ECM). ECM is mainly composed of collagen fibers that oppose cell invasion. To overcome hindrance in the matrix, cancer cells deploy a protease-dependent program in order to remodel the matrix fibers. Matrix remodeling requires the formation of actin-based matrix/plasma membrane contact sites called invadopodia, responsible for collagen cleavage through the accumulation and activity of the transmembrane type-I matrix metalloproteinase (MT1-MMP). In this article, we describe experimental procedures designed to assay for invadopodia formation and for invadopodia activity using 2D and 3D models based on gelatin (denatured collagen) and fibrillar type-I collagen matrices.

Culturing and Imaging Glioma Stem Cells in 3D Collagen Matrices.

Methods to maintain human glioma stem cells as neurosphere cultures and image their dynamic behavior in 3D collagen matrices are described. Additional approaches to monitor glioma stem cell differentiation into mesenchymal-type cells, along with example data are included. Together, these approaches enable glioma stem cell differentiation to be controlled while maintaining the cells in culture, as well as allowing cell dynamics to be captured and analyzed. These methods should be helpful for those seeking to understand the molecular mechanisms driving the invasion of glioma cells through three-dimensional environments. © 2023 Wiley Periodicals LLC. Basic Protocol 1: Culturing human glioma stem cells as neurospheres Basic Protocol 2: Inducing GSC adherence and monitoring their differentiation into mesenchymal cells Support Protocol 1: Preparing fibronectin-coated dishes for cell microscopy Basic Protocol 3: Embedding GSCs in a 3D collagen matrix to study their invasive behavior Support Protocol 2: Phase-contrast imaging with a tiled matrix to study cell migration in a 3D gel.

<i>In Vitro</i> Evaluation of Two Tissue Substitutes for Gingival Augmentation

Three-dimensional collagen matrices of porcine origin are being used as substitutes for soft tissue grafts in periodontal plastic surgery in search of aesthetic and natural results. This in vitro study aimed to compare Fibro-Gide® (GeistlichBiomaterials) and Mucoderm® (BotissBiomaterials) matrices during the initial phase of soft tissue formation. For this purpose, samples of 5 × 5 mm were obtained, and then human fibroblasts were plated on them. After 24, 48 and 72 h, cell viability was assessed using an MTT assay, and the secretion of type I collagen, MMP-2, TIMP-1 and TIMP-2 was analyzed by ELISA immunoassay. The control group (C) consisted of cells plated on polystyrene without the matrices. The morphology of the surfaces was also examined using scanning electron microscopy (SEM), as was the average roughness (Ra) of the samples by a profilometer. Topographic analysis revealed that roughness was significantly higher on Mucoderm® than on Fibro-Gide® (p 0.05). The synthesis of type I collagen, MMP-2 and TIMP-1 were significantly higher from cells plated on Fibro-Gide® than on Mucoderm®, in all time points (p ® than on Mucoderm® (p ® induced an increase in type I collagen, MMP-2 and TIMP-1 and TIMP-2.

Extracellular Matrix Glycation and Crosslinking in Mammary Tumor Progression.

Breast cancer progression is accompanied by profound extracellular matrix (ECM) remodeling. A greater abundance of aligned fibrillar collagen is characteristic of invasive and aggressive breast cancers and has been associated with elevated activity of collagen crosslinking enzymes, such as lysyl oxidase (LOX) and lysyl hydroxylases (LH) and the formation of more mature collagen matrix crosslinks. Aligned collagen fibers can facilitate metastatic dissemination of tumor cells, and LOX inhibitors have been used to inhibit tumor progression and metastasis in experimental models. Thus, a better understanding of how matrix crosslinking alters tumor cell phenotypes, and behaviors would improve our ability to effectively treat aggressive metastatic breast cancer. Herein described is an experimental approach to glycate and crosslink a collagen-I/basement membrane extract ECM to study the impact of ECM crosslinking on mammary tumor progression in vivo. Moreover, glycation of collagen by sugars to form advanced glycation end products naturally occurs during aging, extending the potential relevance of this approach to research on mechanisms of aging involved in disease progression.

Electrospinning of collagen: enzymatic and spectroscopic analyses reveal solvent-independent disruption of the triple-helical structure.

Electrospinning has become a well-established method for creating nanofibrous meshes for tissue-engineering applications. The incorporation of natural extracellular components, such as electrospun pure collagen nanofibers, has proven to be particularly challenging, as electrospun collagen nanofibers do not constitute native collagen fibers anymore. In this study, we show that this detrimental effect is not only limited to fluorinated solvents, as previously thought. Rat tail collagen was dissolved in acetic acid and ethanol and electrospun at various temperatures. Electrospun collagen mats were analyzed using circular dichroism, enzymatic digestion, SDS-PAGE, western blotting, and Raman spectroscopy and compared to heat-denaturated and electrospun collagen in HFIP. Our data suggest that even non-fluorinated electrospinning solvents, such as acid-based solvents, do not yield structurally intact fibers. Interestingly, neither epithelial cells nor fibroblasts displayed a different cellular response to electrospun collagen compared to collagen-coated polyurethane scaffolds in F-actin staining and metabolic analysis using fluorescent lifetime imaging. The disruption of the structural integrity of collagen might therefore be underestimated based on the cell-material interactions alone. These observations expose the larger than anticipated vulnerability of collagen in the electrospinning process. Based on these findings, the influence of the electrospinning process on the distinct biochemical properties of collagen should always be considered, when ECM-mimicking fibrous constructs are manufactured.

Relative survival and cell adhesion of multipotent mesenchymal cells from primary teeth pulp on the surface membranes and sponges produced from collagen

To study the behavior of SHED cell culture on different types of materials for the regeneration of periodontal tissues with different porosity. Porous collagen material Fibro-Gide (Geitstlich Pharma AG, Switzerland), designed to increase the volume of the gum and the barrier collagen membrane Bio-Gide (Geitstlich Pharma AG, Switzerland) were studied in vitro on SHED cultures. As a control sample, a Spongostan sponge made of gelatin (Johnson & Johnson Medical, UK) with the most pronounced porosity and wettability was used. Acute cytotoxicity was determined using a screening method for assessing the number of living cells in a sample (MTT test). SHED cells were sown on the materials to study the attachment of cells to materials and their migration inside the samples. Before seeding, the cells were stained with vital fluorescent dye PKH26 (red fluorescent cell linker kit, Sigma, Germany) for further visualization. Using the MTT test it was shown that they do not have cytotoxic effects. At the same time by the 8th day of the experiment in the presence of Fibro-Gide and Bio-Gide the cells showed an increase in proliferative activity by 19% and 12%, respectively compared with the control group. The cells attached and spread out on the surface of the materials and migrated into the thickness of porous Fibro-Gide and Spongostan. The in vitro study showed that the most favorable material for SHED cell culture is the collagen material Fibro-Gide with sufficient porosity, elasticity and hydrophilicity. SHED cells attach to the collagen matrix and easily penetrate into the sample, filling the entire internal space, while the proliferative capacity of the cell culture increases.

A novel integrated experimental and computational approach to unravel fibroblast motility in response to chemical gradients in 3D collagen matrices.

Fibroblasts play an essential role in tissue repair and regeneration as they migrate to wounded areas to secrete and remodel the extracellular matrix. Fibroblasts recognize chemical substances such as growth factors, which enhance their motility towards the wounded tissues through chemotaxis. Although several studies have characterized single-cell fibroblast motility before, the migration patterns of fibroblasts in response to external factors have not been fully explored in 3D environments. We present a study that combines experimental and computational efforts to characterize the effect of chemical stimuli on the invasion of 3D collagen matrices by fibroblasts. Experimentally, we used microfluidic devices to create chemical gradients using collagen matrices of distinct densities. We evaluated how cell migration patterns were affected by the presence of growth factors and the mechanical properties of the matrix. Based on these results, we present a discrete-based computational model to simulate cell motility, which we calibrated through the quantitative comparison of experimental and computational data via Bayesian optimization. By combining these approaches, we predict that fibroblasts respond to both the presence of chemical factors and their spatial location. Furthermore, our results show that the presence of these chemical gradients could be reproduced by our computational model through increases in the magnitude of cell-generated forces and enhanced cell directionality. Although these model predictions require further experimental validation, we propose that our framework can be applied as a tool that takes advantage of experimental data to guide the calibration of models and predict which mechanisms at the cellular level may justify the experimental findings. Consequently, these new insights may also guide the design of new experiments, tailored to validate the variables of interest identified by the model.

Comparative morphological assessment of crosslinked and non-crosslinked xenograft biointegration quality

Relevance. Autografts are nowadays the gold standard for soft tissue augmentation and gingival recession coverage. However, as this method has a number of significant disadvantages, xenografts of various origins are more frequently used in dental practice. The study aimed to conduct a comparative morphological assessment of crosslinked and non-crosslinked xenogenous scaffold biointegration quality at different healing times.Materials and Methods. The study used 108 Wistar rats (54 males, six months old, fat mass 462 ± 20; 54 females, six months old, fat mass 414 ± 7) and randomly divided them into three groups. Group 1 had a xenogenous collagen matrix (Mucograft) placed in the subperiosteal tunnel in the mandibular vestibule area; Group 2 had an acellular dermal collagen matrix (Mucoderm) implanted in the subperiosteal tunnel, group 3 had D-ribose crosslinked xenogenous scaffold (Ossix Volumax) placed in the subperiosteal tunnel.Results. The studied xenogenous materials showed different integration and biodegradation extents. By postoperative day 7, Group 1 (Mucograft) showed a more pronounced inflammation. Non-crosslinked collagen materials had almost completely resorbed by the 30th day of observation, while a crosslinked collagen matrix was detected as homogenous masses. By the 90th day, all groups demonstrated the formation of new connective tissue with different functional characteristics; only Group 3 histologically revealed microfragments of the graft.Conclusion. The study shows that the xenogenous crosslinked and non-crosslinked collagen membranes may promote an increase in soft tissue thickness to a different extent. However, it is essential to improve the available materials to reach an outcome comparable to the autologous soft tissue graft treatment result.

3D Porous Collagen Matrices-A Reservoir for In Vitro Simultaneous Release of Tannic Acid and Chlorhexidine.

The treatment of wounds occurring accidentally or as a result of chronic diseases most frequently requires the use of appropriate dressings, mainly to ensure tissue regeneration/healing, at the same time as treating or preventing potential bacterial infections or superinfections. Collagen type I-based scaffolds in tandem with adequate antimicrobials can successfully fulfill these requirements. In this work, starting from the corresponding hydrogels, we prepared a series of freeze-dried atelocollagen type I-based matrices loaded with tannic acid (TA) and chlorhexidine digluconate (CHDG) as active agents with a broad spectrum of antimicrobial activity and also as crosslinkers for the collagen network. The primary aim of this study was to design an original and reliable algorithm to in vitro monitor and kinetically analyze the simultaneous release of TA and CHDG from the porous matrices into an aqueous solution of phosphate-buffered saline (PBS, pH 7.4, 37 °C) containing micellar carriers of a cationic surfactant (hexadecyltrimethylammonium bromide, HTAB) as a release environment that roughly mimics human extracellular fluids in living tissues. Around this central idea, a comprehensive investigation of the lyophilized matrices (morpho-structural characterization through FT-IR spectroscopy, scanning electron microscopy, swelling behavior, resistance against the collagenolytic action of collagenase type I) was carried out. The kinetic treatment of the release data displayed a preponderance of non-Fickian-Case II diffusion behavior, which led to a general anomalous transport mechanism for both TA and CHDG, irrespective of their concentrations. This is equivalent to saying that the release regime is not governed only by the gradient concentration of the releasing components inside and outside the matrix (like in ideal Fickian diffusion), but also, to a large extent, by the relaxation phenomena of the collagen network (determined, in turn, by its crosslinking degree induced by TA and CHDG) and the dynamic capacity of the HTAB micelles to solubilize the two antimicrobials. By controlling the degree of physical crosslinking of collagen with a proper content of TA and CHDG loaded in the matrix, a tunable, sustainable release profile can be obtained.

Hyaluronic acid and chitosan-based injectable and self-healing hydrogel with inherent antibacterial and antioxidant bioactivities

Injectable dressings offering intrinsic antibacterial activity and antioxidation are required, because persistent bacterial infection and oxidative damage are common issues associated with chronic non-healing wounds. Herein, a versatile QCS-l-TA-l-OHA hydrogel is fabricated using ubiquitous nature derived biological macromolecules, quaternized chitosan (QCS), tannic acid (TA) and oxidant hyaluronic acid (OHA), driving by multiple dynamic interactions under physiological conditions. The hydrogel can serve as multiple functional wound dress with injectable and self-healing capabilities coupling with attractive antibacterial activity and antioxidation. The biocompatible and self-biodegradable hydrogel augments free radical scavenging capacity, antibacterial, angiogenesis, mature granulation tissue and collagen disposition, enabling fast wound healing of rat skin full-thickness incision model. This study highlights a wound dressing platform of hyaluronic acid and chitosan based injectable and self-healing hydrogel with inherent antibacterial and antioxidant bioactivities, thereby avoiding the use of antibiotics or antioxidant agents.

Multiplexed Detection of Molecular Interactions with DNA Origami Engineered Cells in 3D Collagen Matrices.

The interactions of cells with signaling molecules present in their local microenvironment maintain cell proliferation, differentiation, and spatial organization and mediate progression of diseases such as metabolic disorders and cancer. Real-time monitoring of the interactions between cells and their extracellular ligands in a three-dimensional (3D) microenvironment can inform detection and understanding of cell processes and the development of effective therapeutic agents. DNA origami technology allows for the design and fabrication of biocompatible and 3D functional nanodevices via molecular self-assembly for various applications including molecular sensing. Here, we report a robust method to monitor live cell interactions with molecules in their surrounding environment in a 3D tissue model using a microfluidic device. We used a DNA origami cell sensing platform (CSP) to detect two specific nucleic acid sequences on the membrane of B cells and dendritic cells. We further demonstrated real-time detection of biomolecules with the DNA sensing platform on the surface of dendritic cells in a 3D microfluidic tissue model. Our results establish the integration of live cells with membranes engineered with DNA nanodevices into microfluidic chips as a highly capable biosensor approach to investigate subcellular interactions in physiologically relevant 3D environments under controlled biomolecular transport.

In Vitro and Pre-Clinical Evaluation of Locally Isolated Phages, vB_Pae_SMP1 and vB_Pae_SMP5, Formulated as Hydrogels against Carbapenem-Resistant Pseudomonas aeruginosa.

The inadequate therapeutic opportunities associated with carbapenem-resistant Pseudomonas aeruginosa (CRPA) clinical isolates impose a search for innovative strategies. Therefore, our study aimed to characterize and evaluate two locally isolated phages formulated in a hydrogel, both in vitro and in vivo, against CRPA clinical isolates. The two phages were characterized by genomic, microscopic, phenotypic characterization, genomic analysis, in vitro and in vivo analysis in a Pseudomonas aeruginosa-infected skin thermal injury rat model. The two siphoviruses belong to class Caudovirectes and were named vB_Pae_SMP1 and vB_Pae_SMP5. Each phage had an icosahedral head of 60 ± 5 nm and a flexible, non-contractile tail of 170 ± 5 nm long, while vB_Pae_SMP5 had an additional base plate containing a 35 nm fiber observed at the end of the tail. The hydrogel was prepared by mixing 5% w/v carboxymethylcellulose (CMC) into the CRPA propagated phage lysate containing phage titer 108 PFU/mL, pH of 7.7, and a spreadability coefficient of 25. The groups were treated with either Phage vB_Pae_SMP1, vB_Pae_SMP5, or a two-phage cocktail hydrogel cellular subepidermal granulation tissues with abundant records of fibroblastic activity and mixed inflammatory cell infiltrates and showed 17.2%, 25.8%, and 22.2% records of dermal mature collagen fibers, respectively. In conclusion, phage vB_Pae_SMP1 or vB_Pae_SMP5, or the two-phage cocktails formulated as hydrogels, were able to manage the infection of CRPA in burn wounds, and promoted healing at the injury site, as evidenced by the histopathological examination, as well as a decrease in animal mortality rate. Therefore, these phage formulae can be considered promising for clinical investigation in humans for the management of CRPA-associated skin infections.