Heparin-Encapsulated Metered-Dose Topical “Nano-Spray Gel” Liposomal Formulation Ensures Rapid On-Site Management of Frostbite Injury by Inflammatory Cytokines Scavenging

Abstract

The critical time window between the incidence of frostbite injury and the initiation of treatment in remote snowbound areas is a determining factor for an effective therapeutic response. It is an emergency condition and challenging to treat due to the poor vascularity of affected body parts, and it requires immediate action. In addition to cold trauma-induced tissue damage, the inflammatory mediators majorly contribute to pathologic aggravations. We have designed and evaluated a topical “nano-spray gel (NSG)” formulation, which is based on a combination of liposomal heparin sodium (Hp) and ibuprofen (Ibu) for rapid relief of frostbite injury in extremely low temperatures. The scientific literature suggests that heparin is associated with rapid endothelial cell repair, normalizing blood circulation in capillaries, and has a potential role in wound healing. Hp-containing liposomes were prepared by the extruder method, which suitably formulated an ibuprofen-containing gel to obtain a nano-Spray formulation (HLp-Ibu-NSG) applicable for topical delivery. A single spray puff of the formulation delivers ∼154 mg of the gel, which corresponds to ∼205 U of heparin. In this study, heparin liposomes exhibited significant healing of wound in vitro (scratch assay, fibroblast cells) and in vivo (wound healing in Sprague Dawley rats) at a low dose. In the rat model of frostbite injury, the HLp-Ibu-NSG formulation demonstrated significant reduction in the wound area (up to ∼96%) and improvement of histopathology in 14 days as compared to the control groups. No edema and erythema were detected post-treatment of HLp-Ibu-NSG in the affected area. The underlying mechanism was delineated as a modulation of the inflammatory cytokine (IL-6, TNF-α, IL-10, IL-4) mediators at the wound site and blood circulation to foster frostbite healing. Future clinical studies on the nano-spray gel are required to evaluate its efficacy for the treatment of frostbite symptoms. The instant on-site application of this formulation might be helpful in saving extremities of soldiers, mountaineers, and pilgrims having frostbite.

Modulation of metal-azolate frameworks for the tunable release of encapsulated glycosaminoglycans

“…61,62 An initial rapid release of HP followed by a more sustained delivery is most suitable for the treatment of urgent clinical situations, such as vascular surgery, frostbite, dialysis, etc. 37,43,63 Thus, the development of HP delivery systems with fast responsive rate have attracted signicant attention. [64][65][66] The here prepared HP@MOF composites show release proles that are relevant for urgent medical treatments.…”

“…[64][65][66] The here prepared HP@MOF composites show release proles that are relevant for urgent medical treatments. 37,43,63 To test possible alteration in the biotherapeutic properties of HP due the encapsulation and recovery processes, we used a chromogenic anti-IIa assay to evaluate the anticoagulant activity of heparin before and aer being encapsulated within the three different MOFs (see ESI † for details). The collected data reveals that the HP released from ZIF-8 retains z98% of its initial activity, whereas the HP released from ZIF-90 and MAF-7 retains z95% and z97% activity, respectively (Fig.…”

“…35,36 This method of administration is not compatible with all disease treatments such as wound healing and anti-inammatory applications that require efficient local administration. 37 As a consequence, novel carriers with customisable delivery properties for the administration of GAGs are desirable.…”

Methods of Liposomes Preparation: Formation and Control Factors of Versatile Nanocarriers for Biomedical and Nanomedicine Application

“…Fluorescent confocal micrographs showed that fibroblasts exposed to the HLp (green fluorescence) for 3 h accumulated liposomes (in significant numbers) inside the cytoplasm of the cells (Figure 18B). No significant difference in cellular accumulation was observed between HLp and BLp [181]. HLp exhibited significant healing of a wound in vitro (scratch assay, fibroblast cells) and in vivo (wound healing in Sprague-Dawley rats) at a low dose.…”

Design and Optimization of a Novel-Herbosomal Loaded PEG-Poloxamer Topical Formulation For the Treatment of Cold Injuries: A Quality By Design Approach

“…In skin wound condition a precise there wound appears after exposure to validated wound model, but in case of cold-injury the diameter of affected area is directly proportional to the cytokine response and individual's body pathophysiological response, thus it becomes necessary to carry out morphometric analysis as a routine basis to assess the changes in the received treatment groups and control groups. This analysis was carried out by the method described by Vaghasiya et al, (2019) where routine measurement of the frostbitten area was done with the help of Vernier caliper until complete healing of the cold-injured area. Percentage area (contracted wound) is calculated by the formula given below:…”