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BMC Complementary Medicine and Therapies

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Open Access 01-12-2023 | Mannitol | Research

A novel therapeutic effect of mannitol-rich extract from the brown seaweed Sargassum ilicifolium using in vitro and in vivo models

Authors: Amal D Premarathna, Rando Tuvikene, MNR Somasiri, MLWP De Silva, Ranjith Adhikari, TH Ranahewa, RRMKK Wijesundara, SK Wijesekera, IPGHU Dissanayake, Phurpa Wangchuk, Vitalijs Rjabovs, Anura P Jayasooriya, RPVJ Rajapakse

Published in: BMC Complementary Medicine and Therapies | Issue 1/2023

Abstract

Background

Wound healing is an active, complex, integrated series of cellular, physiological, and biochemical changes initiated by the stimulus of injury in a tissue. The present study was performed to investigate the potential wound healing abilities of Sargassum ilicifolium crude extracts (CE) that were characterized by ¹H NMR and FTIR Spectrometric measurements.

Materials and methods

Seaweed samples were collected from southern coastal sites of Sri Lanka. To determine the cytotoxicity and proliferation of S. ilicifolium CE were used for the MTT and alamarBlue assays respectively. The scratch and exclusion wound models were used to HaCaT and HDF cells to assess the cell proliferation and migration. RAW 264.7 cells (macrophages) were used to evaluate Nitric Oxide (NO) production and phagocytosis activities. Moreover, Fifteen, 8-week-old, female, New Zealand rabbits were selected and divided into five groups: excision skin wounds (10.40 ± 0.60 mm) were induced in groups I, II, and III. Rabbits in groups I and IV were given S. ilicifolium CE (orally, 100 mg/kg day, two weeks), whereas groups II and V were given equal amounts of distilled water. Wound healing properties were measured and wound tissue samples were collated, formalin-fixed, wax-embedded, stained (Hematoxylin and Eosin; Van Gieson) and examined for the healing process.

Results

Anti-inflammatory and wound healing activities were observed in RAW 264.7, HDF and HaCaT cells treated with S. ilicifolium aqueous extracts when compared to the control groups. S. ilicifolium extracts concentration 8 - 4 μg/μL, (P<0.05) had remarkable the highest proliferative and migratory effects on RAW 264.7, HDF and HaCaT cells when compared with the control. RAW 264.7 cell proliferation and/or migration were higher in S. ilicifolium extracts (4 μg/μL, 232.8 ± 10.07%) compared with the control (100 %). Scratch wound healing were remarkably enhanced in 24 h, 48 h (P<0.05) when treated with S. ilicifolium on HaCaT cells. Rabbits treated with the CE of S. ilicifolium showed a significantly increased wound healing activities (P<0.05) within three days with a close wound area of 57.21 ± 0.77 % compared with control group (26.63 ± 1.09 %). Histopathology, aspartate aminotransferase and alanine aminotransferase levels evidenced no toxic effects on seaweed treated groups. Histopathological results also revealed that the healing process was significantly faster in the rabbit groups which were as treated with CE of S. ilicifolium orally with the evidence of enhanced early granulation tissue (connective tissue and angiogenesis) and significant epithelization compared to the control.

Conclusions

Cell proliferation and migration are significantly faster when treated with S. ilicifolium aqueous extracts. Moreover, there are no toxic effect of S. ilicifolium aqueous extracts on RAW 264.7, HDF and HaCaT cell lines. In this study, it is revealed that S. ilicifolium has potential remedial agent; D-Mannitol for skin wound healing properties that by promote keratinocyte and fibroblast proliferation and migration. These findings show that S. ilicifolium have promising wound healing properties.

Meenakshi S, Raghavan G, Nath V, Ajay Kumar SR, Shanta M. Antimicrobial, wound healing and antioxidant activity of Plagiochasma appendiculatum Lehm. et Lind. J Ethnopharmacol. 2006;107:67–72.CrossRef

hess Thomas C. Clinical Guide: Skin and Wound Care. 6th ed. New York: Lippincott wiliams and wilkins; 1998. p. 8–9.

Guo S, Dipietro LA. Factors affecting wound healing. J Dent Res. 2010;89(3):219–29. https://doi.org/10.1177/0022034509359125.CrossRefPubMedPubMedCentral

Mathieu D, Linke JC, Wattel F. Non-healing wounds. In: Handbook on hyperbaric medicine, Mathieu DE, editor. Dordrecht: Springer, 2006; p. 401–28.

Menke NB, Ward KR, Witten TM, Bonchev DG, Diegelmann RF. Impaired wound healing. Clin Dermatol. 2007;25:19–25.PubMedCrossRef

McCosker L, Tulleners R, Cheng Q, Rohmer S, Pacella T, Graves N, Pacella R. Chronic wounds in Australia: A systematic review of key epidemiological and clinical parameters. Int Wound J. 2019;16(1):84–95. https://doi.org/10.1111/iwj.12996.CrossRefPubMed

Wadinamby JMRWW. Chronic wounds and their management and application in a contemporary Sri Lankan hospital setting. Sri Lankan J Infect Dis. 2013;3(2):2–16.

Misic AM, Gardner SE, Grice EA. The wound microbiome: modern approaches to examining the role of microorganisms in impaired chronic wound healing. Adv Wound Care. 2014;3:502–10.CrossRef

Boateng JS, Matthews KH, Stevens HN, Eccleston GM. Wound healing dressings and drug delivery systems: a review. J Pharm Sci. 2008;97:2892–923.PubMedCrossRef

10.

Suguna L, Singh S, Sivakumar P, Sampath P, Chandrakasan G. Influence of Terminaliachebula on dermal wound healing in rats. Phytother Res. 2002;16(3):227–31.PubMedCrossRef

11.

Sasidharan S, Nilawatyi R, Xavier R, Latha LY, Amala R. Wound healing potential of Elaeis guineensis Jacq leaves in an infected albino rat model. Molecules. 2010;15(5):3186–99.PubMedPubMedCentralCrossRef

12.

Gould L, Abadir P, Brem H, Carter M, et al. Chronic wound repair and healing in older adults: current status and future research. Wound Repair Regen. 2015;63:427–38.

13.

Henshaw FR, Boughton P, Lo L, McLennan SV, Twigg SM. Topically applied connective tissue growth factor/CCN2 improves diabetic preclinical cutaneous wound healing: potential role for CTGF in human diabetic foot ulcer healing. J Diabetes Res. 2015;2015:236238.PubMedPubMedCentralCrossRef

14.

Negut I, Grumezescu V & Grumezescu AM. Treatment Strategies for Infected Wounds. Molecules, 2018;23(9). doi:https://doi.org/10.3390/molecules23092392

15.

Hamdan S, Pastar I, Drakulich S, Dikici E, Tomic-Canic M, Deo S, Daunert S. Nanotechnology-Driven Therapeutic Interventions in Wound Healing: Potential Uses and Applications. ACS Cent Sci. 2017;3(3):163–75. https://doi.org/10.1021/acscentsci.6b00371.CrossRefPubMedPubMedCentral

16.

Coombs GW, Nimmo GR, Pearson JC, Collignon PJ, Bell JM, McLaws ML, Turnidge JD. Australian Group on Antimicrobial Resistance Hospital-onset Staphylococcus aureus Surveillance Programme annual report, 2011. Commun Dis Intell Q Rep. 2013;37(3):E210-218.PubMed

17.

Kumar B, Vijayakumar M, Govindarajan R, Pushpangadan P. Ethnopharmacological approaches to wound healing exploring medicinal plants of India. J Ethnopharmacology. 2007;114(2):103–13.PubMedCrossRef

18.

Balekar N, Katkam NG, Nakpheng T, Jehtae K, Srichana T. Evaluation of the wound healing potential of Wedeliatrilobata (L) leaves. Journal of Ethnopharmacology. 2012;141(3):817–24.PubMedCrossRef

19.

Wang JP, Ruan JI, Cai YI, Luo Q. XuHx, Wu Yx. In vitro and in vivo evaluation of the wound healing properties of Siegesbeckiapubescens. J Ethnopharmacol. 2011;134(3):1033–8.PubMedCrossRef

20.

Sood A, Granick MS, Tomaselli NL. Wound Dressings and Comparative Effectiveness Data. Adv Wound Care. 2014;3(8):511–29. https://doi.org/10.1089/wound.2012.0401.CrossRef

21.

Namvar F, Mahamad R, Baharara J, Balnejah SZ, Fargahi F, Rahman HS. Antioxidant, antiproliferative, and antiangiogenesis effects of polyphenol-rich seaweed (Sargassummuticum). Biomed Res Int. 2013;2013:604787.PubMedPubMedCentralCrossRef

22.

Pangestuti R, Kim SK. Biological activities and health benefit effects of natural pigments derived from marine algae. Journal of Functional Foods. 2011;3:255–66.CrossRef

23.

Wangchuk P, Sastraruji T, Taweechotipatr M. Anti-inflammatory, anti-bacterial and anti-acetylcholinesterase activities of two isoquinoline alkaloids–scoulerine and cheilanthifoline. Journal of Biologically Active Products from Nature. 2016;11(12):1–20. https://doi.org/10.1177/1934578X1601101207.CrossRef

24.

Khan AD, Rastogi V, Lavhale PM, Jain J. Novel Approaches for Herbal Drug Delivery in Wound Healing: A Review. Indian J Pharm Sci. 2022;84(2):247–60.

25.

Moreira DDL, Teixeira SS, Monteiro MHD, De-Oliveira ACA, Paumgartten FJ. Traditional use and safety of herbal medicines. Rev Bras. 2014;24:248–57.

26.

Yende SR, Harle UN, Chaugule BB. Therapeutic potential and health benefits of Sargassum species. Pharmacogn Rev. 2014;8(15):1.PubMedPubMedCentralCrossRef

27.

Abu-Khudir R, Ismail GA, Diab T. Antimicrobial, antioxidant, and anti-tumor activities of Sargassum linearifolium and Cystoseira crinita from Egyptian Mediterranean Coast. Nutr Cancer. 2021;73(5):829–44.PubMedCrossRef

28.

Premarathna AD, Ranahewa TH, Wijesekera SK, Wijesundara RRMKK, Jayasooriya AP, Wijewardana V, Rajapakse RPVJ. Wound healing properties of aqueous extracts of Sargassum illicifolium: An in vitro assay. Wound Medicine. 2019;24(1):1–7.CrossRef

29.

Namvar F, Baharara J, Mahdi AA. Antioxidant and anticancer activities of selected Persian Gulf algae. Indian J Clin Biochem. 2014;29(1):13–20.PubMedCrossRef

30.

Husen SA, Setyawan MF, Syadzha MF, Susilo RJK, Hayaza S, Ansori ANM, Alamsjah MA, Ilmi ZN, Wulandari PAC, Pudjiastuti P, Awang K. A novel therapeutic effects of Sargassum ilicifolium alginate and okra (Abelmoschus esculentus) pods extracts on open wound healing process in diabetic mice. Research Journal of Pharmacy and Technology. 2020;13(6):2764–70.CrossRef

31.

Hira K, Farhat H, Sohail N, Ansari M, Ara J, Ehteshamul-Haque S. Hepatoprotective activity against acetaminophen-induced liver dysfunction and GC-MS profiling of a brown algae Sargassum ilicifolium. Clinical Phytoscience. 2021;7(1):1–11.CrossRef

32.

Premarathna AD, Wijesekera SK, Jayasooriya AP, Waduge RN, Wijesundara RR, Tuvikene R, Harishchandra DL, Ranahewa TH, Perera NA, Wijewardana V, Rajapakse RP. In vitro and in vivo evaluation of the wound healing properties and safety assessment of two seaweeds (Sargassum ilicifolium and Ulva lactuca). Biochemistry and Biophysics Reports. 2021;1(26): 100986.CrossRef

33.

Premarathna AD, Kumara AMCP, Jayasooriya AP, Jayanetti DE, Adhikari RB, Sarvananda L, Amarakoon S. Distribution and Diversity of Seaweed Species in South Coastal Waters in Sri Lanka. J Oceanogr Mar Res. 2020;7:196. https://doi.org/10.35248/2572-3103.19.7.196.CrossRef

34.

O’brien J, Wilson I, Orton T, Pognan F. Investigation of the Alamar Blue (resazurin) fluorescent dye for the assessment of mammalian cell cytotoxicity. Eur j biochem. 2000;267(17):5421–6.PubMedCrossRef

35.

Premarathna AD, Ranahewa TH, Wijesekera SK, Harishchandra DL, Karunathilake KJ, Waduge RN, Wijesundara RR, Jayasooriya AP, Wijewardana V, Rajapakse RPVJ. Preliminary screening of the aqueous extracts of twenty-three different seaweed species in Sri Lanka with In vitro and In vivo assays. Heliyon. 2020;6(6): e03918.PubMedPubMedCentralCrossRef

36.

Chen W, Zhang W, Shen W, Wang K. Effects of the acid polysaccharide fraction isolated from a cultivated Cordyceps sinensis on macrophages in vitro. Cell Immunol. 2010;262(1):69–74.PubMedCrossRef

37.

Douds DA. An acute oral toxcity study in rats with Advantra Z™. Washington, Ave: Nu-traech, Inc.; 1997.

38.

Uhlig C, Krause H, Koch T, Gama de Abreu M, Spieth PM. Anesthesia and Monitoring in Small Laboratory Mammals Used in Anesthesiology, Respiratory and Critical Care Research: A Systematic Review on the Current Reporting in Top-10 Impact Factor Ranked Journals. PLoS ONE 2015;10(8): e0134205. https://doi.org/10.1371/journal.pone.0134205

39.

Gonc alves RV, Mezeencio JMS, Benevides GP, Matta SLP, Neves CA, Sarandy MM, et al. Effect of gal- lium-arsenide laser, gallium-aluminum-arsenide laser and healing ointment on cutaneous wound healing in Wistar rats. Braz J Med Biol Res. 2010;43:350–5.CrossRef

40.

Percival KR, Radi ZA. A Modified Verhoeff-Van Gieson Elastin Histochemical Stain to Enable Pulmonary Arterial Hypertension Model Characterization. European Journal of Histochemistry : EJH. 2016;60(1):2588.PubMedPubMedCentralCrossRef

41.

Kihdze TJ,OCEJ, Mayowa AA, Joseph O, Ghaife TG, et al. Br J Pharmacol Toxicol. 2016;7(1):9–19.CrossRef

42.

Premarathna AD, Tuvikene R, Fernando PH, Adhikari R, Perera MC, Ranahewa TH, Howlader MM, Wangchuk P, Jayasooriya AP, Rajapakse RP. Comparative analysis of proximate compositions, mineral and functional chemical groups of 15 different seaweed species. Sci Rep. 2022;12(1):1–3.CrossRef

43.

Bharti SK, Roy R. Quantitative 1H NMR spectroscopy. TrAC, Trends Anal Chem. 2012;35:5–26.CrossRef

44.

Quiterio E, Soares C, Ferraz R, Delerue-Matos C, Grosso C. Marine Health-Promoting Compounds: Recent Trends for Their Characterization and Human Applications. Foods. 2021;10(12):3100.PubMedPubMedCentralCrossRef

45.

Stumpf TR, Pertile RA, Rambo CR, Porto LM. Enriched glucose and dextrin mannitol-based media modulates fibroblast behavior on bacterial cellulose membranes. Mater Sci Eng, C. 2013;33(8):4739–45.CrossRef

46.

Buranasin P, Mizutani K, Iwasaki K, Pawaputanon Na Mahasarakham C, Kido D, Takeda K, Izumi Y. High glucose-induced oxidative stress impairs proliferation and migration of human gingival fibroblasts. PLoS ONE. 2018;13(8): e0201855.PubMedPubMedCentralCrossRef

47.

Mouritzen MV, Jenssen H. Optimized scratch assay for in vitro testing of cell migration with an automated optical camera. JoVE (Journal of Visualized Experiments). 2018;138: e57691.

48.

Justus CR, Leffler N, Ruiz-Echevarria M, Yang LV. In vitro cell migration and invasion assays. JoVE (Journal of Visualized Experiments). 2014;88: e51046.

49.

Hirayama D, Iida T, Nakase H. The phagocytic function of macrophage-enforcing innate immunity and tissue homeostasis. Int J Mol Sci. 2017;19(1):92.PubMedPubMedCentralCrossRef

50.

Gentleman E, Lay AN, Dickerson DA, Nauman EA, Livesay GA, Dee KC. Mechanical characterization of collagen fibers and scaffolds for tissue engineering. Biomaterials. 2003;24:3805–13.PubMedCrossRef

51.

Smith AN, Willis E, Chan VT, Muffley LA, Isik FF, Gibran NS, Hocking AM. Mesenchymal stem cells induce dermal fibroblast responses to injury. Exp Cell Res. 2010;316(1):48–54.PubMedCrossRef

52.

Sen CK. Wound healing essentials: let there be oxygen. Wound Repair Regen. 2009;17(1):1–18. https://doi.org/10.1111/j.1524-475X.2008.00436.x.CrossRefPubMedPubMedCentral

53.

Wangchuk P. Therapeutic applications of natural products in herbal medicines, biodiscovery programs, and biomedicine. Journal of Biologically Active Products from Nature. 2018;8(1):1–20.CrossRef

54.

Wangchuk P, Loukas A. Techniques and technologies for the biodiscovery of novel molecule drug lead compounds from natural products. Chapter 16, in Natural Products and Drug Discovery. 2018; pp 435-465. https://doi.org/10.1016/B978-0-08-102081-4.00016-2

55.

Balunas MJ, Kinghorn AD. Drug discovery from medicinal plants. Life Sci. 2005;78(5):431–41. https://doi.org/10.1016/j.lfs.2005.09.012.CrossRefPubMed

56.

Chin YW, Balunas MJ, Chai HB, Kinghorn AD. Drug discovery from natural sources. AAPS J. 2006;8(2):E239–53. https://doi.org/10.1007/bf02854894.CrossRefPubMedPubMedCentral

57.

Singer AJ, Clark RA. Cutaneous wound healing. N Eng J Med. 1999;341:738–46.CrossRef

58.

Wu YS, Chen SN. Apoptotic cell: linkage of inflammation and wound healing. Front Pharmacol. 2014;5:1. https://doi.org/10.3389/fphar.2014.00001.CrossRefPubMedPubMedCentral

59.

Stroncek JD, Reichert WM. Overview of Wound Healing in Different Tissue Types. In: Reichert WM, editor. Indwelling Neural Implants: Strategies for Contending with the In Vivo Environment. Boca Raton: CRC Press/Taylor & Francis, 2008;1: p. 3–41.

60.

Ross R. Wound healing. Sci Am. 1969;220:40–50.PubMedCrossRef

61.

Walter LJ, Aalhus JL, Robertson WM, McAllister TA, Gibb DJ, Dugan MER, Aldai N, McKinnon JJ. Evaluation of wheat or corn dried distillers’ grains with solubles on performance and carcass characteristics of feedlot steers. Can J Anim Sci. 2010;90(2):259–69.CrossRef

62.

Trabucchi E, PreisBaruffaldi F, Baratti C, Montorsi W. Topical treatment of experimental skin lesions in rats: macroscopic, microscopic and scanning electronmicroscopic evaluation of the healing process. Int J Tissue React. 1986;8:533–44.PubMed

63.

Lordan S, Paul Ross R, Stanton C. Marine bioactives as functional food ingredients: potential to reduce the incidence of chronic diseases. Mar Drugs. 2011;9(6):1056–100.PubMedPubMedCentralCrossRef

64.

Pomin VH, Mourao PAS. Structure, biology, evolution, and medical importance of sulfated fucans and galactans. Glycobiology. 2008;18(12):1016–27.PubMedCrossRef

65.

Hemmati AA, Mohammadian F. Investigation into the effects of mucilage of quince seeds on wound healing in rabbit. J Herb Spec Med Plants. 2000;7:41–6.CrossRef

66.

Moosavi Z, Meshki M, Hemmati AA, Salehi veisi M, Rafiee M. Evaluation of the healing effect of topical ointment contain quince seed mucilage on human skin wounds (Article in Farsi with English abstract). J Derm Dis. 2006;9:260e263.

67.

Suzuki M, Kurosawa E. Halogenated sesquiterpene phenols and ethers from the red alga Laurencia glandulifera Kutzing. Bull Chem Soc Jpn. 1979;52(11):3349–51.CrossRef

68.

Yu P, Wang Z, Sun X, Chen X, Zeng S, Chen L, Li S. Hydrogen-rich medium protects human skin fibroblasts from high glucose or mannitol induced oxidative damage. Biochem Biophys Res Commun. 2011;409(2):350–5.PubMedCrossRef

69.

Michael B, Yano BL, Sellers RS, Perry R, Morton D, Roome N, Johnson JK, Schafer K. Evaluation of organ weights for rodent and non-rodent toxicity studies: A review of regulatory guidelines and a survey of current practices. Toxicol Pathol. 2007;35(5):742–50.PubMedCrossRef

70.

Sellers RS, Morton D, Michael B, Roome N, Johnson JK, Yano BL, Perry R, Schafer K. Society of Toxicologic Pathology position paper: organ weight recom- mendations for toxicology studies. Toxicol Pathol. 2007;35:751–5.PubMedCrossRef

71.

Recknagel RO. Carbon tetrachloride hepatotoxicity. Pharmacol Rev. 1967;19(2):145–208.PubMed

72.

Achike FI, Kwan CY. Nitric oxide, human diseases and the herbal products that affect the nitric oxide signalling pathway. Clin Exp Pharmacol Physiol. 2003;9:605–15.CrossRef

73.

Eble JA, Niland S. The extracellular matrix of blood vessels. Curr Pharm Des. 2009;15(12):1385–400.PubMedCrossRef

74.

Bondonno CP, Croft KD, Hodgson JM. Dietary nitrate, nitric oxide, and cardiovascular health. Crit Rev Food Sci Nutr. 2016;56(12):2036–52.PubMedCrossRef

75.

Jones V, Harding K. Moist wound healing, in: Krasner DL, Rodeheaver GT, Sibbald RG (Eds.), Chronic Wound Care: a Clinical Source Book for Health Care Professionals, HMP Communications, Wayne, 2001; p. 79–89.

Title: A novel therapeutic effect of mannitol-rich extract from the brown seaweed Sargassum ilicifolium using in vitro and in vivo models
Authors: Amal D Premarathna
Rando Tuvikene
MNR Somasiri
MLWP De Silva
Ranjith Adhikari
TH Ranahewa
RRMKK Wijesundara
SK Wijesekera
IPGHU Dissanayake
Phurpa Wangchuk
Vitalijs Rjabovs
Anura P Jayasooriya
RPVJ Rajapakse
Publication date: 01-12-2023
Publisher: BioMed Central
Keyword: Mannitol
Published in: BMC Complementary Medicine and Therapies / Issue 1/2023
Electronic ISSN: 2662-7671
DOI: https://doi.org/10.1186/s12906-023-03840-0

Keynote webinar | Spotlight on medication adherence

Springer Medicine

A novel therapeutic effect of mannitol-rich extract from the brown seaweed Sargassum ilicifolium using in vitro and in vivo models

Abstract

Background

Materials and methods

Results

Conclusions

Keynote webinar | Spotlight on medication adherence

Springer Medicine

Abstract

Background

Materials and methods

Results

Conclusions

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