Effect of administration of Lupeol and isolated Lupeol molecule from plant extracts on skin wound healing: A systematic review of In-vitro and In-vivo models

Journal of Medicinal Plants Studies 2024; 12(4): 15-20 ISSN (E): 2320-3862 ISSN (P): 2394-0530 https://www.plantsjournal.com JMPS 2024; 12(4): 15-20 © 2024 JMPS Received: 06-04-2024 Accepted: 10-05-2024 Dhanush KR Department of Pharmacognosy, Faculty of Pharmacy, M.S. Ramaiah University of Applied Sciences, Bengaluru, Karnataka, India Ashoka babu VL Associate Professor & Head, Department of Pharmacognosy, Faculty of Pharmacy, M.S. Ramaiah University of Applied Sciences, Bengaluru, Karnataka, India Effect of administration of Lupeol and isolated Lupeol molecule from plant extracts on skin wound healing: A systematic review of In-vitro and In-vivo models Dhanush KR and Ashoka babu VL Abstract This review set out to methodically assess the body of research on the effectiveness of lupeol in wound healing. We looked through the MEDLINE, SCIDIRECT, and SPRINGER databases for original research that was published till December 2023. Seven reviewers assessed the title, abstract, and whole manuscript for every research. Out of the 635 studies that we found, only 04 underwent additional evaluation on the exclusion criteria, clinical trials were applied and 03 non animal experiment model (Invitro) were included. Lupeol based formulation were more effective for wound recovery, isolation of lupeol from different plants like Betula pendula brich, Derris scandens, Bowdichia virgilioides, Bergia ammannioides etc. The lupeol induced a reduction in time closure, and effective was reported in both invito & In-vivo wound models included diabetic wound. In addition, our study indicates that lupeol appear to promote wound healing; however, Taken together, these findings demonstrate that lupeol are a class of molecules with significant promise that leads for the development of new drugs to treat skin injury. Lupeol have been shown to induce cell migration, cell proliferation, collagen deposition, antiinflammatory, anti-angiogenesis, anti-oxidation & Cytotoxicity effect. Keywords: Lupeol, wound healing, In-vitro, In-vivo Corresponding Author: Dhanush KR Department of Pharmacognosy, Faculty of Pharmacy, M.S. Ramaiah University of Applied Sciences, Bengaluru, Karnataka, India Introduction Skin, the largest organ, protects physiological systems and prevents external invasion. Trauma can disrupt skin integrity, leading to infection, bleeding, and delayed wound healing. Research aims to develop novel dressings. Lupeol, a pentacyclic triterpene found in vegetables and medicinal plants, has strong therapeutic potential due to its anti-inflammatory and wound healing effects Lupeol's biomedical application in wound healing is limited by its poor solubility in aqueous media. Chitosan (CS), a linear polysaccharide, offers biocompatibility, biodegradability, and hemostatic effectiveness. CS nanoparticles can encapsulate alcohol-soluble compounds like curcumin, accelerate platelet aggregation, and display strong blood absorption. CS and its derivatives also increase antibacterial activity, with Ag+-loaded CS nanocomposite showing strong bactericidal activity. A novel temperature-sensitive, self-assembled sericin hydrogel loaded with Ag+-modified CS nanoparticles and lupeol was developed. The hydrogel was characterized and evaluated for antibacterial activities, lupeol-releasing properties, degradation kinetics, hemolysis ratio, and wound healing efficiency. The findings suggest it has potential as a multifunctional therapeutic platform. Medical advisors prioritize wound care to prevent infections, delay healing, and disfigure scars. Topical antibiotics are commonly used to facilitate healing. Genus Bergia, a waterwort family, includes 15 species, including B. ammannioides Henye native to Egypt. These plants are important medicinal plants in India, traditionally used for wound healing and sore treatment. The annual shrub, 8-35 cm tall, has pinkish stems and branches, small, white flowers. Bowdichia virgilioides, also known as "sucupira-preta," is a medicinal plant found in the Brazilian Cerrado region. Its bark and seeds are used in infusions to treat diseases like arthritis, diabetes, bronchitis, and skin wounds. The plant's bark and roots contain alkaloids, terpenoids, volatile constituents, flavonoids, and anthocyanins. Diabetes mellitus is a chronic metabolic disease affecting 171 million people globally, with a ~ 15 ~ Journal of Medicinal Plants Studies https://www.plantsjournal.com Methods Source Three main databases were searched electronically in a methodical manner for peer-reviewed English articles: Medline (PubMed), SCIDIRECT, and SPRINGER (up to December 2023). By using keyword “Lupeol + Wound healing”. projected 366 million by 2030. Its most common complication is altered skin wound healing, leading to complications like Diabetic Foot Ulcers (DFUs). This disease causes major morbidity due to clinical and socioeconomic issues. Research shows that hyperglycemia during diabetes delays wound healing, causing complications like DFUs. The study investigates the efficacy of lupeol gel in enhancing wound healing in streptozotocin-induced hyperglycemic rats. It found that lupeol gel promoted cutaneous wound closure by inducing granulation tissue formation, inhibiting macrophage infiltration, and increasing re-epithelialization. The mechanisms underlying these effects remain unknown. The study aims to understand the mechanisms behind these effects. Data Extraction Duplicate results were eliminated after importing the database results into Microsoft Excel. In order to evaluate titles and abstracts against the In-vitro and In-vivo criteria, they were screened. Primary research studies looking at the application of plant-source isolated lupeol for wound healing were included. Preclinical, in vivo, and In-vitro model investigations were included in the study designs; the results of the search and screening procedures are shown in Figure 2. Results After duplicates were eliminated, 631 articles remained out of the total 635 items found. After screening papers for titles and abstracts, 589 publications were eliminated. Out of the 42 full text publications that were assessed, 35 did not match the critical exclusion criteria. Eventually, the systematic review contained seven publications. Every study that was incorporated was released by December 2024. There were three in vitro, four preclinical, and three in vivo model studies among the study designs. The mean difference between the treatment group and the control group was the primary outcome variable that was reported. Fig 1: Structure of lupeol Fig 2: Decision trial of included studies ~ 16 ~ Journal of Medicinal Plants Studies https://www.plantsjournal.com Table 1: Description of the main characteristics of the studies with fractions obtained from plant extracts. Author year and country Animal Total n wound type Wenhui et al.,(2023) [26], China Sprague dawley female rats, age 8weeks 60 full thickness, (2cm diameter punch) excisional wound on the dorsal area administration Dosage Topical once daily (0.5g gel) Source Formulation Standard drug gel 6. 1. 2. 3. 4. Treatment group interventionand size (n) Citosan nanoparticles (n=12) Citosan-Ag-nanoparticles (n=12), Citosan-loaded-nanoparticles(n=12), citosanAg-loaded nanoparticles(n=12) 5% w/w ethanolic residue ointment 10% w/w ethanolic residue ointment. 5% w/w n-hexane residue ointment. 10% w/w n-hexane residue ointment. 5% w/w ethyl acetae residue ointment. 10% w/w ethyl acetae residue ointment. 5% w/w nbutanol residue ointment. 10% w/w n-butanol residue ointment. ointment base only was applied and this group as the vehicle controle (n=06) standard drug Dermazine 0.1% w/w lupeol cream(n=8) 0.2% w/w lupeol cream (n=8) 0.4% w/w lupeol cream(n=8) treated with lanette cream (vehicle). 1. 2. 3. Treated with 0.2% w/w lupeol cream. (n=8) Treated with lanette cream (vehicle). Treated with insulin based cream 0.5 U/g 1. 2. 3. 1. 2. sprague dawley male Shahira et al., rats(130-150g) and 66 (2015) [27], Egypt adult swiss albino mice (20-50g) Circular wound of 1.5 cm2 area was produced in the dorsal interscapular region of each rat by excising the full thickness skin topical isolated from Bergia ammannioides ? 3. 4. ointment 5. 40 full thickness, (2cm diameter punch) excisional wound on the dorsal area Topical ? isolated from bowdichia virgilioides (stem bark) Fernando pereira beserra et al. male wistat rats (250g) 32 (2019) [29], Brazil full thickness, (2cm diameter punch) excisional wound on the dorsal area topical ? isolated from bowdichia virgilioides (stem bark) Fernando pereira male wistat rats (180beserra et al. 220g) (2020) [29], Brazil Cream Cream Control group and size (n) 1. No treatment (n=12) 1. No treatment (n=06) 1. Treated with collagenase 1.2 U/g (n=8) 1. Sham group without diabetes, wounds, or treated(n=8) Table 2: Preclinical In-vivo animal model studies outcome measurements and result summary Author year and country Wenhui et al., (2023) [26], China Shahira et al., (2015) [27], Egypt Fernando pereira beserra et al. (2020) [28], Brazil Fernando pereira beserra et al. (2019) [29], Brazil Outcomes measures 1. 2. Histopathological appearance Wound closure analysis 1. 2. 3. 4. Estimation of total collagen, Wound healing activity Oxidation Inflammation 1. 2. 3. 4. Macroscopic appearance Wound closure percentage Histopathological appearance Immunohistochemistry straining for NF-Kb, Ki67,EGF,& VEGF Inflammation Macroscopic appearance Wound closure percentage Histopathological Immunohistochemistry Inflammation Acute dermal irritation 5. 1. 2. 3. 4. 5. 6. 2. 3. 4. finding outcomes lupeol-loaded groups (CS-Ag-L-NPs) gel showed increase re-epithelialization, reducing inflammation and enhancing collagen fiber deposition compared to other formulation (P<0.01;****P,0.0001) On day 21, wounds of CS-L-NPs and CS-Ag-L-NPs gel groups were essentially healed (P< 0.01; P< 0.05) The application of ointments containing EtOH, HxFr, and EtFr significantly increased collagen content in granulation tissue by the 6th and 10th days, while BuFr showed no significant activity. (P,0.01) HxFr 10% ointment showed the most pronounced activity compared to other fraction formulation at p<0.01 EtFr showed the strongest antioxidant activity against DPPH compared to other formulation Showed strongest anti-inflammatory property 1. 2. 3. 4. 5. There was no significant change in edema and hemorrhage parameters Showed a strong wound-healing effect of lupeol-based cream after 7& 14 days (p<0.05) Increased blood vessels, proliferation & tissue –remodeling phase Increased collagen treatment, immunolabeling area compared to lanette group Causesd proinflammation 1. 2. Lupeol-treated group showed only scar of the injured region but lesions still presented little clot and granulation Lupeol – based cream notable to decreased wound size, but insulin-treated group showed a significant increase in wound contraction on 13 & 15 day Decreased inflammation & increased proliferation of fibroblasts, Lupeol significantly increased the collagen III- immunolabeled area in the central region of the lesion as compared to lanette group 1. 2. 1. 3. 4. ~ 17 ~ Journal of Medicinal Plants Studies https://www.plantsjournal.com 7. 8. Angiogenesis Oxidative stress 5. Lupeol reduces the inflammation 6. Not show any adverse reactions as compared to control 7. Formation of new blood cells 8. Minimized the oxidative stress and improved the antioxidant property CS-chitosan; Ag-silver ion; L-lupeol; NPs- nanoparticle; EtOH- ethanolic extract; HxFr- n-Hexane fraction; BuFr – n-butanol fraction; EtFr- ethyl acetate fraction Table 3: Preclinical In-vitro animal model studies outcome measurements and result summary Author year and country Source Drug Source Method 1. Fernando pereira beserra et Human neonatal foreskins al. (2018) [30], Brazil Isolated from Bowdichia virgilioides kunth (stem bark) 1. 2. 3. 4. Pathom Somwong et al. (2022) [7], Thailand Magdalena anna malinowska et al. (2021) [4] , Poland Derris scandens stem ethanolic extract (0.0588 & 0.3472% w/w lupeol Human skin fibroblast cell content in extract) Human epidermal cells Extract of brich bark contain lupeol Cell proliferation assay Cytotoxicity assay In-vitro wound healing (scratch) Assay Collagen gel contraction assay 1. 2. Cytotoxicity assay In-vitro wound healing (scratch) Assay 1. 2. 3. 4. 5. 6. Cell proliferation assay In-vitro wound healing (scratch) Assay Antioxidant activity Cytotoxicity Cell morphology & cytoskeleton ~ 18 ~ 2. 3. 4. Outcome Lupeol reduced cell proliferation of both keratinocytes and fibroblasts It did not affect keratinocyte viability but showed cytotoxicity to fibroblasts at high concentration(20µg/ mL) Increased the wound closure rate at 83% compared to control(p<0.001) Lupeol significantly increased the contractile effect on collagen gels capered to control (p<0.01 1. 2. Extract had no cytotoxic effect. Ethanolic extract was effective for wound closure in a scratch assay. 1. Cell proliferation is more in 24h(133, 143& 131%) but no significant effect on the proliferation after 48h Increases the wound closure in scratch assay, in dose dependent manner. Lupeol esters (29%) exhibit better antioxidation activity compared to lupeol (1.40%). No effect were observed Accelerate the wound healing process. 2. 3. 4. 5. Journal of Medicinal Plants Studies https://www.plantsjournal.com Fig 3: Summary of the studies describing the plant species, families, used parts of each species Drukała J. The effect of the new lupeol derivatives on human skin cells as potential agents in the treatment of wound healing. Biomolecules. 2021;11(6):774. 5. Pereira Beserra F, Xue M, Maia GLDA, Leite Rozza A, Helena Pellizzon C, Jackson CJ, et al. Lupeol, a pentacyclic triterpene, promotes migration, wound closure, and contractile effect in vitro: Possible involvement of PI3K/Akt and p38/ERK/MAPK pathways. Molecules. 2018;23(11):2819. 6. Pereira Beserra F, Sergio Gushiken LF, Vieira AJ, Augusto Bérgamo D, Luísa Bérgamo P, Oliveira de Souza M, et al. From inflammation to cutaneous repair: Topical application of lupeol improves skin wound healing in rats by modulating the cytokine levels, NF-κB, Ki-67, growth factor expression, and distribution of collagen fibers. International Journal of Molecular Sciences. 2020;21(14):4952. 7. Somwong P, Kamkaen N. Wound-healing activity and quantification of bioactive compounds from Derris scandens extract. Journal of Advanced Pharmaceutical Technology & Research. 2022;13(1):38-43. 8. Singh H, Ali SS, Khan NA, Mishra A, Mishra AK. Wound healing potential of Cleome viscosa Linn. seeds extract and isolation of active constituent. South African Journal of Botany. 2017;112:460-465. 9. Patel S, Srivastava S, Singh MR, Singh D. Preparation and optimization of chitosan-gelatin films for sustained delivery of lupeol for wound healing. International Journal of Biological Macromolecules. 2018;107:18881897. 10. Harish BG, Krishna V, Kumar HS, Ahamed BK, Sharath R, Swamy HK, et al. Wound healing activity and docking of glycogen-synthase-kinase-3-β-protein with isolated triterpenoid lupeol in rats. Phytomedicine. 2008;15(9):763-767. 11. Herrera-Calderón O, Calero-Armijos LL, Cardona-GW, Conclusions According to available data, lupeol and individual lupeol molecules derived from plant extracts promote healing in both in vitro and in vivo models. Which were evaluated using a range of dosages, may hasten the healing of wounds and raise the success rate of healing in both normal and diabetic patients. The primary impacts of formulations containing lupeol appear to be linked to the promotion of cell migration, proliferation, and collagen deposition, as well as antiinflammatory, anti-angiogenesis, and anti-oxidation and cytotoxic actions during tissue healing. When combined, these factors accelerate the healing process and increase the biomechanical resistance of newly created tissue. 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