Injuries represent a major global public health concern, accounting for approximately 10 % of all deaths worldwide, disability and a substantial share of disability-adjusted life years (DALYs). According to recent reports, the economic burden of trauma and wound related morbidity remains enormous, with road traffic accidents, occupational hazards, burns, and falls among the leading causes [[1], [2], [3]]. These injuries often result in acute and chronic wounds that challenge healthcare systems globally. Therefore, the development of cost-effective and biologically robust strategies for wound healing has become a priority in regenerative medicine. Wound healing is a complex, multi-stage process in which alterations at any stage can significantly impact overall recovery [4]. It typically progresses through four overlapping but distinct phases: 1. Hemostasis (coagulation): Occurs within minutes after injury. Vasoconstriction reduces blood flow, while platelets aggregate to form a clot and release clotting factors, resulting in a stable fibrin mesh. 2. Inflammatory phase: Begins shortly after injury and generally lasts 3–7 days. This stage is characterized by an immune response to prevent infection and remove cellular debris. Neutrophils and macrophages migrate to the wound site to clear pathogens and necrotic tissue. While essential, prolonged inflammation can hinder healing and result in chronic wounds. Conversely, shortening this phase excessively may also be detrimental; therefore, regulating its duration is critical in wound care management. 3. Proliferative phase:

Initiates around days 2–3 and may last up to two weeks. Fibroblasts produce collagen, forming the structural framework of new tissue [5]. Angiogenesis ensures adequate oxygen and nutrient delivery, while epithelial cells migrate to cover the wound surface [6]. Granulation tissue comprising fibroblasts, collagen, and new capillaries fills the wound bed. This phase is crucial for effective wound healing management, as most cell migration occurs during this period [7]. 4. Maturation (remodeling) phase: Commences approximately three weeks after injury and may persist for months or years. During this stage, collagen fibers are reorganized, tensile strength increases, and scar tissue matures. Given these mechanisms, the use of agents that modulate both the inflammatory and proliferative phases can significantly improve wound healing outcomes [8,9]. One such agent is Portulaca oleracea L. (purslane), a plant with documented nutritional, medicinal, and antimicrobial properties [10]. Purslane extract promotes wound healing by increasing collagen synthesis and angiogenesis, while reducing inflammation through the inhibition of prostaglandin and thromboxane production from arachidonic acid [11]. Another widely used material in regenerative medicine is the human amniotic membrane (hAM). First applied clinically in the early 20th century, [12] hAM has been employed for treating burns, chronic ulcers, ocular surface disorders, and surgical wound dehiscence [13,14]. Its extracellular matrix components elastin, collagen types I, III, IV, V, and VI, and hyaluronic acid create a biocompatible scaffold for epithelial cell growth [15,16,14,17]. Furthermore, the absence of HLA-A, -B, and -D antigens reduces immunogenicity, and its intrinsic anti-inflammatory properties support tissue regeneration [18]. Platelet-rich plasma (PRP), derived from autologous blood, is another potent wound healing agent. Platelets adhere to injured endothelium, activating coagulation and releasing alpha granules containing over 300 bioactive molecules, including growth factors, cytokines, and extracellular matrix regulators [19]. PRP stimulates angiogenesis, fibroblast proliferation, extracellular matrix remodeling, and mesenchymal stem cell differentiation [20,21].

While each of these agents purslane, hAM, and PRP has been individually shown to enhance wound healing, no study to date has evaluated their combined effect. The present study aimed to develop topical formulations containing these three components and investigate their synergistic potential in promoting wound repair. Additionally, a novel ratio-based approach was introduced to assess phase-specific healing dynamics.