Biochemically wound healing process is promoted by active agents which compose aqueous and alcohol herbal extracts and those components act synergistically. We propose in this study four methods of soft aqueous extraction (A, B, C, D) of Ageratum conyzoides Linn. leaves, a well-known west african plant for its wound healing properties. Our main objective was to know which of the extraction methods would allow us to obtain specifically tannins and flavonoids. We also investigated which types of flavonoids extracted could inhibit serine proteases like thrombin or serine proteases present in bacterial and then being bactericide. Ageratum conyzoides Linn. samples were collected in the beginning of the raining period in May 2019 in two regions of Togo. After aqueous extractions, TLC and HPLC-ESI+-QTOF analysis were done. Chromatographic fingerprints obtained were quite similar for the four extracts investigated. However, additional spots with different colors appeared for the B extract. Flavonoids were revealed by the numerous colors of spots observed on the chromatograms. Superposing HPLC-ESI+-QTOF chromatographic profiles of the four extracts, we deduced that our proposed extraction methods lead to same chemical compounds but B extract contained higher concentrations of later substances. Identified molecules belong to terpernoids and flavonoids classes.Keywords: Ageratum Conyzoides Linn., Aqueous extractions, Biomolecules, Wound healing.

Based on mechanisms of fibrin clot polymerization and dissolution, it is possible to modulate fibrin formation and removal. Ageratum conyzoides Linn. (Asteraceae) is an annual herb with a long history of traditional medicine. There is high variability in the secondary metabolites of this plant which include flavonoids, and these molecules belong to a class of serine proteases inhibitors. Several plant enzymes belonging to the classes of serine proteases were observed to be active on the cascade of coagulation pathways. The aim of this study was to observe if even Ageratum conyzoides Linn. aqueous leaves extract contained proteases which could structurally modify the fibrin clot formation. To prepare plant extracts, dry leaves of the plant were extracted with distilled water. Fibrin gels were prepared by mixtures containing fibrinogen and thrombin with or without extract. Fibrin networks were disrupted by a denaturation buffer. Samples were deposited in 8% polyacrylamide gel and Coomassie blue was used to reveal migration. Our extract contained phytochemicals class flavonoids which are thrombin inhibitors. But our results support the evidence that the same extract contained plant serine proteases, specifically a fibrinogenase which hydrolyzed fibrinogen but not like thrombin.Keywords: Fibrin/Fibrinogen, structural modification, Ageratum conyzoides Linn., phytoproteases.

Objective: It is of primary importance to develop wound healing sealant that prevent bacteria contamination and growth. We propose to formulate poor platelets plasma material supplemented with a bactericidal plant extract, Ageratum conyzoides Linn. (Asteraceae). Aqueous extract of this plant is used as a bactericide.Methodology: Platelet-poor plasma (PPP) containing less than 10,000 platelets per μL were used for all experimentations. Physiological serum (NaCl 0.9%) had served to prepare calcium chloride (CaCl ) solutions at 2 2M, 4M, 6M, 8M and 10M. Clauss fibrinogen assays were done to determine fibrinogen concentrations in each plasma pocket. Clotting times were measured following the addition of the appropriate calcium chloride concentration to the plasma. Plant extract at a concentration of 250 mg/mL in physiological serum was also added to the plasma and followed by the clotting times measured.Results: Clauss fibrinogen assays reflected a global satisfactory fibrinogen concentration. Overall profile of the evolution of clotting time vs calcium concentrations adopts a curve form and the shortest clotting times are those obtained with 4M calcium concentration. However, the presence of extract deeply disturbed clotting process, as clotting times were extremely elongated.Conclusion: Our objective was difficult to achieve because of the thrombin inhibitory side effects of the extract which were added to antithrombin III inhibitory effects in presence of excess calcium. Keywords: Wound healing, fibrin sealant, Ageratum conyzoides Linn., biomaterial, inhibitors. French title: Formation d'un Biomatériau de Fibrine par Apport Exogène de Calcium et Supplémentation par un Extrait Bactéricide de la planteAgeratum conyzoides Linn. : Equilibre entre un Activateur Biochimique et des Inhibiteurs Phytochimiques de la Thrombine Objectif: Il est d'une importance capitale de développer des pansements cicatrisants qui empêchent la contamination et la prolifération bactérienne. Nous nous proposons de formuler un biomatériau cicatrisant de fibrine à base de plasma pauvres en plaquettes et supplémenté d'un extrait aqueux de Ageratum conyzoides Linn.Méthodes: Du plasma pauvre en plaquettes contenant moins de 10 000 plaquettes par μL a été utilisé pour toutes les expérimentations. Le sérum physiologique (NaCl 0,9%) a servi à la préparation des différentes solutions de calcium aux concentrations de 2M, 4M, 6M, 8M et 10M. Le dosage du fibrinogène par les tests de Clauss a été effectué pour chaque poche de plasma. Les temps de gel ont été déterminés suite à l'addition du calcium à la concentration adéquate. L'extrait de plante à la concentration de 250 mg/mL dans du sérum physiologique a également été additionné au plasma suivi de la détermination des temps de gel.Résultats: Les tests de Clauss mènent à des taux de fibrinogène moyens satisfaisants. Le profil global de l'évolution des temp de gel en fonction de la concentration en calcium adopte une forme hyperbolique et le temps le plus court est obtenu à 4M de calcium. La présence de l'extrait perturbe profondément la formation du gel puisque les temps de gel de ces derniers sont extrêmement rallongés.Conclusion: Notre objectif a été difficile à atteindre à cause des effets secondaires inhibiteurs de l'extrait qui se sont ajoutés à ceux de  l'antithrombin III en présence d'un excès de calcium. Mots clé: Cicatrisation ; Pansement fibrine ; Ageratum conyzoides Linn. ; Activateur; Inhibiteurs.