Healing Mechanisms Research Articles

Unraveling the Degradation and Air-Induced Healing Mechanisms of Halide Perovskites under Proton Irradiation

Unraveling the Degradation and Air-Induced Healing Mechanisms of Halide Perovskites under Proton Irradiation

Wound Healing, Cell Viability and Antimicrobial Potency of Mucus from Pangasianodon hypophthalmus

Acute and chronic wounds are the major cause of death according to World Health Organization (WHO), in which, antimicrobial resistance is considered to be a major plight. In this regard, our study is aimed at developing an antimicrobial agent using the mucus of Pangasianodon hypophthalmus against the clinically resistant microbial pathogens and to evaluate the cell cytotoxicity and cell viability followed by an in vitro wound healing analysis. The evaluation of antimicrobial activity was performed through well diffusion method and micro dilution method. The cell cytotoxicity and cell viability were assessed using MTT assay. The cell migration and in vitro wound healing was performed using scratch assay. The acidic extracts of mucus showed antimicrobial activity against the eight different selected bacterial strains while the organic extract showed against seven bacterial strains. L929 showed a cell viability of 102.96% at a concentration of 75 µg/mL and did not show cell toxicity effect up to the concentration of 300 µg/mL. In the in vitro wound healing analysis, the cell migration rate was 99.27% in the treated cells while, the untreated showed only 94.68%. The current research work clearly shows that the mucus of P. hypophthalmus possesses antimicrobial activity and wound healing potency. Furthermore, gene expression analysis and in vivo trials have to be performed for a thorough understanding of the actual cellular mechanism of wound healing.

Prognostic bone fracture healing simulations in an ovine tibia model validated with in vivo sensors.

Bone fracture healing is a complex physiological process influenced by biomechanical and biomolecular factors. Mechanical stability is crucial for successful healing, and disruptions can lead to delayed healing or nonunion. Bone commonly heals itself through secondary fracture healing, which is governed by the mechanical strain at the fracture site. To investigate these phenomena, a validated methodology for capturing the mechanoregulatory process in specimen-specific models of fracture healing could provide insight into the healing process. This study implemented a prognostic healing simulation framework to predict healing trajectories based on mechanical stimuli. Sixteen sheep were subjected to a 3 mm transverse tibial mid-shaft osteotomy, stabilized with a custom plate, and equipped with displacement transducer sensors to measure interfragmentary motion over 8 weeks. Computed tomography scans were used to create specimen-specific bone geometries for finite element analysis. Virtual mechanical testing was performed iteratively to calculate strains in the callus region, which guided tissue differentiation and consequently, healing. The predicted healing outcomes were compared to continuous in vivo sensor data, providing a unique validation data set. Healing times derived from the in vivo sensor and in silico sensor showed no significant differences, suggesting the potential for these predictive models to inform clinical assessments and improve nonunion risk evaluations. This study represents a crucial step towards establishing trustworthy computational models of bone healing and translating these to the preclinical and clinical setting, enhancing our understanding of fracture healing mechanisms. Clinical significance: Prognostic bone fracture healing simulation could assist in non-union diagnosis and prediction.

Atomically Resolved Surface Reconstruction of WO3 (002).

The rearrangement of surface atoms in oxide nanocrystals, namely surface reconstruction, plays an important role in improving the physical and chemical properties of metal oxides. However, structural information pertaining to reconstructed surfaces is scarce due to the challenges associated with directly imaging surface and sub-surface atoms under reconstruction conditions. Herein, the reconstruction of the nanocrystalline tungsten trioxide (002) surface is directly investigated via scanning transmission electron microscope (STEM). The results reveal that the atoms on the reconstructed WO3 (002) surface are rearranged into a (1 × 2) structure, and the structural model is determined by density functional theory (DFT) calculation. In addition, after surface reconstruction, the Fermi level shifted toward the conduction band compared to the initial surface, achieving an effect similar to n-type doping. Surprisingly, analogous atomic rearrangements are also observed in cracks, indicating that sub-nanometer fractures in tungsten trioxide can be remedied through surface reconstruction, thus proposing an unconventional mechanism for crack healing. Furthermore, DFT calculations are used to analyze the models and electronic properties of the reconstruction structures. These findings provide insights into the surface reconstruction of WO3 (002) and the healing of nanoscale cracks in tungsten trioxide.

MYB168 and WRKY20 transcription factors synergistically regulate lignin monomer synthesis during potato tuber wound healing.

Lignin is a critical component of the closing layer of the potato (Solanum tuberosum L.) tuber during healing; however, the molecular mechanism of its formation remains poorly understood. To elucidate the molecular mechanism of tuber healing, we screened the genes encoding transcription factors that regulate lignin synthesis(StMYB24/49/105/144/168, StWRKY19/20/22/23/34) and the key genes involved in lignin monomer synthesis (PHENYLALANINE AMMONIA LYASE 5 (StPAL5) and CINNAMYL ALCOHOL DEHYDROGENASE 14 (StCAD14)) for induced expression after wounding using transcriptome data. DLR, Y1H, EMSA, and ChIP-qPCR assays revealed that StMYB168 could bind directly to the StPAL5 and StCAD14 promoters to activate their expression and that StWRKY20 enhanced this regulation with a synergistic effect. Y2H, BiFC, and Co-IP assays showed that StMYB168 interacted with StWRKY20 to form a MYB-WRKY complex. Furthermore, transient overexpression of StMYB168 and StWRKY20 in Nicotiana benthamiana leaves upregulated the expression of NbPAL and NbCAD10 and promoted lignin accumulation in the leaves. In addition, overexpression of StWRKY20 and StMYB168 together resulted in higher expression levels of NbPAL and NbCAD10 and higher levels of lignin monomer and total lignin. In contrast, silencing of StMYB168 and StWRKY20 in potato significantly reduced the lignin content of wounded tubers. In conclusion, StMYB168 and StWRKY20 are important regulators of lignin biosynthesis in potato tubers during healing and can positively regulate lignin biosynthesis by forming a complex. The elucidation of this regulatory module provides information on the regulatory mechanism of lignin monomer synthesis in wounded tubers at the transcriptional level.

New insights into the role of cellular senescence and chronic wounds

Chronic or non-healing wounds, such as diabetic foot ulcers (DFUs), venous leg ulcers (VLUs), pressure ulcers (PUs) and wounds in the elderly etc., impose significant biological, social, and financial burdens on patients and their families. Despite ongoing efforts, effective treatments for these wounds remain elusive, costing the United States over US$25 billion annually. The wound healing process is notably slower in the elderly, partly due to cellular senescence, which plays a complex role in wound repair. High glucose levels, reactive oxygen species, and persistent inflammation are key factors that induce cellular senescence, contributing to chronic wound failure. This suggests that cellular senescence may not only drive age-related phenotypes and pathology but also be a key mediator of the decreased capacity for trauma repair. This review analyzes four aspects: characteristics of cellular senescence; cytotoxic stressors and related signaling pathways; the relationship between cellular senescence and typical chronic non-healing wounds; and current and future treatment strategies. In theory, anti-aging therapy may influence the process of chronic wound healing. However, the underlying molecular mechanism is not well understood. This review summarizes the relationship between cellular senescence and chronic wound healing to contribute to a better understanding of the mechanisms of chronic wound healing.

Building a protective shield: The role of wound healing in reducing postharvest decay and preserving quality of citrus fruit1

Postharvest citrus fruit is susceptible to pathogenic infestation and quality reduction through wounds, leading to tremendous commercial losses. Herein, wound healing of citrus fruit was obviously at 25°C for five days to form a barrier effective against the development of infectious diseases and water dissipation. Combined with the results of transcriptional and metabolic levels, wound healing activated the expression of CsKCS4, CsKCS11, CsCYP704B1, CsFAH1, CsGPAT3 and CsGPAT9 genes in suberin biosynthesis pathway, and CsPMEI7, CsCesA-D3, CsXTH2, CsXTH6, CsXTH22, CsXTH23, CsXTH24, CsC4H and CsCAD genes in cell wall metabolism pathway, leading to the accumulation of suberin monomers and cell wall components. The results of microscopic observations proved wound healing promoted suberin deposition and cell wall strengthening. Meanwhile, wound healing required the provision of energy and precursor substances by carbohydrate metabolism and amino acid metabolism. We provide new insights into the regulatory mechanism of wound healing on improving disease resistance and maintaining the quality of citrus fruit.

Pressure‐Induced Capacity Recovery and Performance Enhancements in LTO/NMC‐LCO Batteries

AbstractLithium titanate oxide (LTO) batteries are a promising technology, particularly suitable for high‐power applications, owing to their inherent cyclic stability, fast charging capability, and superior safety. However, substantial gas generation and accelerated aging driven by the cathode remain substantial challenges. This study explores the mitigation of these aging mechanisms through the application of external mechanical compression. Continuous pressure of 0.3 MPa applied to pristine cells during cycling reduces capacity loss by 42% compared to unpressurized cells cycled under identical operating conditions. Applying short‐term pressure to aged cells leads to immediate capacity recovery, reclaiming up to 57% of the lost capacity. Subsequent cycling of these aged cells under continuous pressure demonstrates improved capacity retention. In contrast, intermittently applied transient pressure causes notable capacity fluctuations. This study reveals insights into aging and healing mechanisms influenced by external pressure, benefiting both first‐ and second‐life battery applications. Understanding these mechanisms is vital for enhancing performance and lifetime in battery packs, while the findings also highlight promising opportunities for capacity recovery in reused batteries.

Combined Analysis of Second- and Third-Generation Transcriptome Sequencing for Gene Characteristics and Identification of Key Splicing Variants in Wound Healing of Ganxi Goat Skin.

Ganxi goat is a local breed of goat unique to Jiangxi Province, China, named for its primary distribution in the Ganxi region. Ganxi goats are primarily raised through grazing, showing good adaptation to the local humid and hot climate, strong disease resistance, and occupying an important position in the local livestock industry. The skin, as the main barrier of the body, plays an indispensable role in resisting the invasion of external pathogenic factors and has received increasing attention in the medical and scientific fields. In this study, Ganxi goat skin was used as the research subject. Full-length transcriptome sequencing of Ganxi goat skin was performed using PacBio third-generation sequencing technology to supplement and improve the annotation information of the Ganxi goat genome. A combined analysis of second- and third-generation transcriptome sequencing was used to analyze the splicing variant events of hub genes (CDC20, MMP2, TIMP1, and EDN1) and the expression changes in each splicing variant in skin samples on day 0 and day 5 after surgical wounding. The regulatory role of related hub gene splicing variants in wound healing was analyzed. A total of 926,667 full-length non-chimeric sequences were obtained, optimizing the annotation information of 3794 genomic gene loci and identifying 2834 new genes, 256 new LncRNAs, 12,283 alternative splicing events, 549 genes with polyadenylation, and 112 fusion genes. Three splicing variant forms were identified in both the CDC20 and EDN1 genes, seven in MMP2, and two in TIMP1. The expression levels of most splicing variants showed significant changes in the skin samples on days 0 and 5 after wounding, potentially participating in the regulation of wound healing. This study provides fundamental data for the annotation of the goat genome and offers a reference for studying the regulatory mechanisms of wound healing.

Transient Apical Breakdown: Incidence, Pathogenesis, and Healing.

Transient apical breakdown (TAB) is a phenomenon that indicates temporary apical periodontal destruction and root resorption after tooth luxation injuries, followed by the healing process of the dental pulp. Andreasen in 1986, reported that TAB was seen in approximately 4.2% of all luxation injuries. However, there have been no reports thereafter on the types and frequency of the luxation traumatic injuries in which TAB occurs. Therefore, this retrospective study was aimed to assess the incidence and pathogenesis of dental trauma-induced TAB and to suggest a possible mechanism of subsequent healing based on a series of cases. Data analysis included mature teeth (n = 56) of 49 patients aged 9-30 years who presented in a private dental office over a period of 10 years (2012-2022) to investigate the incidence and healing sequala of TAB. TAB was observed in 43.8% of subluxation, 62.5% of extrusive luxation, and 75% of lateral luxation injuries. The average age of patients who developed TAB was 14.5 years, ranging from 9 to 28 years old. TAB can be expected in many cases of luxation injuries with minimal dislocation. Therefore, mild injuries (subluxation, extrusion, and lateral luxation), may exhibit spontaneous healing, recovery of dark discoloration of the crown, disappearance of a periapical radiolucent lesion and return to normal response to EPT as long as 12 months after the traumatic injury. Thus, a decision to perform endodontic treatment in these cases might be postponed until clear evidence for an infection exists.

L. plantarum UBLP-40 Versus the Combined Formula of L. rhamnosus UBLP-58 and B. longum UBBL-64 in Excisional Wound Healing: A Cellular Perspective

Introduction: The utilization of probiotics in enhancing the active healing of skin wounds represents a burgeoning trend in contemporary medicine. Previous research has extensively explored wound healing mechanisms involving the strains of Lactiplantibacillus plantarum, Lacticaseibacillus rhamnosus, and Bifidobacterium longum. This study seeks to compare and interpret cellular findings derived from immunohistochemical and pathological applications. Methods: Three groups (the control, Lactiplantibacillus plantarum (RO1) group, and Lacticaseibacillus rhamnosus and Bifidobacterium longum (PRO2) group) underwent histological analysis, and microscopic cell counting were employed, offering insights into dynamic changes among neutrophils, lymphocytes, plasmacytes, mast cells, fibroblasts, and newly formed vessels across distinct treatment groups and temporal intervals. Results: The neutrophil count was found to be elevated in PRO2 on day 2, while the same group resulted in the highest decline on day 15. The number of fibroblasts peaked on day 4 for the PRO1 group, compared to the other two groups, which peaked on day 8. The lymphocyte count was the highest in the control group, while they peaked on day 4 in PRO2. The mast cells and plasmacytes were variable and sparse among all groups and time frames. Neovascularization was promoted by PRO1 and PRO2 groups on day 4 and remained high on day 8 for PRO2. Conclusions: Probiotic strains can be beneficial to the human population and in assisting skin wound healing, each strain working differently and more effectively in different healing phases. Thus, a combined formula containing different probiotics to modulate various healing phases is desirable.

SUMO: A new perspective to decipher fibrosis.

Fibrosis is characterized by excessive extracellular matrix (ECM) deposition resulting from dysregulated wound healing and connective tissue repair mechanisms. Excessive accumulation of ECM leads to fibrous tissue formation, impairing organ function and driving the progression of various fibrotic diseases. Recently, the role of small ubiquitin-like modifiers (SUMO) in fibrotic diseases has attracted significant attention. SUMO-mediated SUMOylation, a highly conserved posttranslational modification, participates in a variety of biological processes, including nuclear-cytosolic transport, cell cycle progression, DNA damage repair, and cellular metabolism. Conversely, SUMO-specific proteases cleave the isopeptide bond of SUMO conjugates, thereby regulating the deSUMOylation process. Mounting evidence indicates that SUMOylation and deSUMOylation regulate the functions of several proteins, such as Smad3, NF-κB, and promyelocytic leukemia protein, which are implicated in fibrotic diseases like liver fibrosis, myocardial fibrosis, and pulmonary fibrosis. This review summarizes the role of SUMO in fibrosis-related pathways and explores its pathological relevance in various fibrotic diseases. All evidence suggest that the SUMO pathway is important targets for the development of treatments for fibrotic diseases.

In Vivo tissue healing mechanism at the intestinal anastomosis site following high-frequency electric welding

In Vivo tissue healing mechanism at the intestinal anastomosis site following high-frequency electric welding

Synergistic effect of zinc oxide-cinnamic acid nanoparticles for wound healing management: in vitro and zebrafish model studies

Wound infections resulting from pathogen infiltration pose a significant challenge in healthcare settings and everyday life. When the skin barrier is compromised due to injuries, surgeries, or chronic conditions, pathogens such as bacteria, fungi, and viruses can enter the body, leading to infections. These infections can range from mild to severe, causing discomfort, delayed healing, and, in some cases, life-threatening complications. Zinc oxide (ZnO) nanoparticles (NPs) have been widely recognized for their antimicrobial and wound healing properties, while cinnamic acid is known for its antioxidant and anti-inflammatory activities. Based on these properties, the combination of ZnO NPs with cinnamic acid (CA) was hypothesized to have enhanced efficacy in addressing wound infections and promoting healing. This study aimed to synthesize and evaluate the potential of ZnO-CN NPs as a multifunctional agent for wound treatment. ZnO-CN NPs were synthesized and characterized using key techniques to confirm their structure and composition. The antioxidant and anti-inflammatory potential of ZnO-CN NPs was evaluated through standard in vitro assays, demonstrating strong free radical scavenging and inhibition of protein denaturation. The antimicrobial activity of the nanoparticles was tested against common wound pathogens, revealing effective inhibition at a minimal concentration. A zebrafish wound healing model was employed to assess both the safety and therapeutic efficacy of the nanoparticles, showing no toxicity at tested concentrations and facilitating faster wound closure. Additionally, pro-inflammatory cytokine gene expression was analyzed to understand the role of ZnO-CN NPs in wound healing mechanisms. In conclusion, ZnO-CN NPs demonstrate potent antioxidant, anti-inflammatory, and antimicrobial properties, making them promising candidates for wound treatment. Given their multifunctional properties and non-toxicity at tested concentrations, ZnO-CN NPs hold significant potential as a therapeutic agent for clinical wound management, warranting further investigation in human models.

Bio-Sprayed/Threaded Microalgae Remain Viable and Indistinguishable from Controls.

Microalgae are increasingly playing a significant role in many areas of research and development. Recent studies have demonstrated their ability to aid wound healing by their ability to generate oxygen, aiding the healing process. Bearing this in mind, the capability to spray/spin deposit microalgae in suspension (solution) or compartmentalize living microalgae within architectures such as fibers/scaffolds and beads, would have significance as healing mechanisms for addressing a wide range of wounds. Reconstructing microalgae-bearing architectures as either scaffolds or beads could be generated via electric field (bio-electrospraying and cell electrospinning) and non-electric field (aerodynamically assisted bio-jetting/threading) driven technologies. However, before studying the biomechanical properties of the generated living architectures, the microalgae exposed to these techniques must be interrogated from a molecular level upward first, to establish these techniques, have no negative effects brought on the processed microalgae. Therefore these studies, demonstrate the ability of both these jetting and threading technologies to directly handle living microalgae, in suspension or within a polymeric suspension, safely, and form algae-bearing architectures such as beads and fibers/scaffolds.

Dual-feedback healing mechanism redefining anti-oxidation coatings in fiber-reinforced composites

Dual-feedback healing mechanism redefining anti-oxidation coatings in fiber-reinforced composites

Management of Gingivitis: Contemporary Approaches and Recent Therapeutic Advancements

Background: Gingivitis, commonly known as gum disease, refers to several types of inflammatory diseases that impact the connective tissues that surround the teeth. Gingivitis causes swelling, redness, and bleeding of the gums in its early stages. Objective: This article aims to describe the standard gingivitis medication. It emphasizes recent advancements in the initial therapy, treatment, and healing mechanisms of gingivitis for achievement in the clinical testing of medicines that promise to enable disease modification in patients. Also, it aims to review recent advancements and emerging therapeutic developments in the management of gingivitis, including gene-based therapies, nanotherapies, anti-cytokine therapies, stem cell-based therapies, and probiotic therapies. Methods: The information for the review articles was acquired by using Google Scholar and PubMed as search engines, as well as a number of publishers, including Springer Nature, Ben-tham Science, Taylor & Francis, Elsevier, and Frontier. Result and Discussion: Gingivitis is a gum disease and scaling root planning (SRP) is now the most common kind of periodontitis therapy available. It has the potential to deliver significant therapeutic success, but it can also have substantial problems that reduce the quality of life of a patient. Stem cell therapies, gingivitis genetic engineering, nuclear-based medicines, and other advances have given people hope that a wide range of illnesses, especially genetic disorders, can be cured. Conclusion: The current gingivitis therapies are successful and continually evolving, with sev-eral drugs currently in clinical trials. These innovative medicines, when combined, may alter gingivitis treatment in the next few years. Finally, gingivitis therapy requires professional dental care and patient education on oral hygiene. Nonetheless, further research and clinical studies are necessary to validate the efficacy, safety, and long-term benefits of these novel treatment modalities.

Silk Fibroin Nanofibers: Advancements in Bioactive Dressings through Electrospinning Technology for Diabetic Wound Healing

Background: Non-healing diabetic wounds represent a significant clinical challenge globally, necessitating innovative approaches in drug delivery to enhance wound healing. Understanding the pathogenesis of these wounds is crucial for developing effective treatments. Bioactive dressings and polymeric nanofibers have emerged as promising modalities, with silk biomaterials gaining attention for their unique properties in diabetic wound healing. Purpose of Review: The purpose of this review is to examine the challenges and innovations in treating non-healing diabetic wounds, emphasizing the global burden and the need for effective solutions. This review explores the complex mechanisms of wound healing in diabetes and evaluates the therapeutic potential of bioactive dressings and polymeric nanofibers. Special focus is given to the application of silk biomaterials, particularly silk fibroin, for wound healing, detailing their properties, mechanisms, and clinical translation. This review also describes various nanofiber fabrication methods, especially electrospinning technology, and presents existing evidence on the effectiveness of electrospun silk fibroin formulations. Recent Findings: Recent advancements highlight the potential of silk biomaterials in diabetic wound healing, owing to their biocompatibility, mechanical strength, and controlled drug release properties. Electrospun silk fibroin-based formulations have shown promising results in preclinical and clinical studies, demonstrating accelerated wound closure and tissue regeneration. Summary: Non-healing diabetic wounds present a significant healthcare burden globally, necessitating innovative therapeutic strategies. Bioactive dressings and polymeric nanofibers, particularly silk-based formulations fabricated through electrospinning, offer promising avenues for enhancing diabetic wound healing. Further research is warranted to optimize formulation parameters and validate efficacy in larger clinical trials.

Ethyl butyrate inhibits caudal fin regeneration in adult zebrafish by disrupting extracellular matrix remodeling

Wound healing and tissue regeneration are influenced by a variety of factors. Adverse lifestyle habits, such as excessive alcohol consumption, delay wound healing and increase the risk of secondary infections. Ethyl butyrate is a common food additive widely used to enhance the aroma of alcoholic beverages. This additive is generally considered harmless to human health in both industrial and domestic settings. However, the ecotoxicity and its effects on wound healing have not been elucidated. In this study, we used zebrafish as the experimental animal, and the caudal fins were amputated to explore the effects of ethyl butyrate on wound healing and tissue regeneration. The effect of ethyl butyrate on blastema and bone regeneration and its impact on the transcriptional levels of regeneration-related genes and inflammation-related genes were evaluated. RNA-seq was conducted to determine the differentially expressed genes (DEGs) between the treatment and the control groups. KEGG and GO analysis was conducted to explore the functions of DEGs. Significantly enriched GO terms and KEGG pathways were identified to explore the molecular mechanism underlying the inhibition of zebrafish caudal fin regeneration by ethyl butyrate. The results demonstrated that ethyl butyrate significantly inhibited the regeneration of zebrafish caudal fins, including blastema and bone regeneration. Ethyl butyrate exposure significantly downregulated the expression of genes associated with bone and blastema regeneration and inflammation response. KEGG and GO functional analyses revealed that the DEGs were associated with significant enrichment of extracellular matrix-receptor interactions. Ethyl butyrate treatment downregulated the expression of most extracellular matrix-related genes. These findings indicate that ethyl butyrate potentially modulates pathways associated with the structure, adhesion, modification, and degradation of the extracellular matrix, thereby disrupting extracellular matrix remodeling, inhibiting wound inflammation, impairing blastema and bone regeneration and ultimately hindering caudal fin regeneration. In summary, the findings demonstrate that ethyl butyrate disrupts extracellular matrix remodeling and inhibits the regeneration of zebrafish caudal fins. These results provide valuable insights into the rational use of ethyl butyrate and further investigation of wound healing mechanisms.

Experimental and simulation-based engineering of calcium alginate self-healing asphalt capsules

An emerging technology to mitigate the cracking of asphalt pavements is the use of self-healing capsules embedded in asphalt mixtures. In this study, self-healing capsules were fabricated by encapsulating asphalt rejuvenators with calcium alginate shells. While past studies have used a “brute force” design process for such capsules, the design and formulation of these can be optimized through careful consideration of chemical interactions and crack healing mechanisms. In this study, experiments and molecular dynamics (MD) simulation were used to engineer capsule–rejuvenator formulations to mitigate premature failure of capsules and improve self-healing efficiency. To explore the thermodynamic process, the inter-diffusion coefficient and blending degree between materials in the capsule system were evaluated at different temperatures and capsule designs to determine the ratio of capsules which survive mixing and compaction through molecular scale studies. MD provided an understanding of healing behavior by simulating and quantifying the fracture–healing–fracture process of the binder–capsule system. The experiments verified the MD model by quantifying oil release percentage from micro-extraction and recovery and fine aggregate matrix (FAM) healing efficiency. The research revealed that the interaction between asphalt binder, rejuvenator, and capsules highly depended on their chemical compositions and that the dose of capsule content influences the penetration degree, and structural failure process. Polymeric capsules experienced significant premature failure and content release at high temperatures (160 °C) compared to intermediate temperatures (25 °C) due to enhanced thermal movement. To optimize capsule performance, rejuvenators C and E, which had higher viscosities, required 30%–40% calcium alginate, while rejuvenator A with lower viscosity needed 40%–60% calcium alginate. Rejuvenators with more asphaltenes were more sensitive to the capsule shell protection effect, and strengthened healing efficiency, while rejuvenators with more aromatics resulted in better wetting and diffusion processes during healing. The healing index, derived from FAM experimental healing test and validated by simulations, indicated that approximately 40% shell material effectively prevented capsule breakage and resulted in a 50% reduction in healing capacity. This research therefore established a framework for engineering a combination of capsules for use at pavement scale based on an understanding of chemical interactions, mechanical properties, and experimental verification.